Chemical Safety Report

CHEMICAL SAFETY REPORT

Substance Name: diammonium tetrachlorozincate(2-)

EC Number: 238-687-6

CAS Number: 14639-97-5

Registrant's Identity: EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Table of Contents

Part A...... 1 1. SUMMARY OF RISK MANAGEMENT MEASURES...... 1 2. DECLARATION THAT RISK MANAGEMENT MEASURES ARE IMPLEMENTED...... 1 3. DECLARATION THAT RISK MANAGEMENT MEASURES ARE COMMUNICATED...... 1 Part B...... 2 1. IDENTITY OF THE SUBSTANCE AND PHYSICAL AND CHEMICAL PROPERTIES...... 2 1.1. Name and other identifiers of the substance...... 2 1.2. Composition of the substance...... 3 1.3. Physico-chemical properties...... 3 2. MANUFACTURE AND USES...... 6 2.1. Manufacture...... 6 2.2. Identified uses...... 8 2.3. Uses advised against...... 48 3. CLASSIFICATION AND LABELLING...... 48 3.1. Classification and labelling according to CLP / GHS...... 48 3.2. Classification and labelling according to DSD / DPD...... 50 3.2.1. Classification and labelling in Annex I of Directive 67/548/EEC...... 50 3.2.2. Self classification(s)...... 50 3.2.3. Other classification(s)...... 52 4. ENVIRONMENTAL FATE PROPERTIES...... 52 4.1. Degradation...... 53 4.1.1. Abiotic degradation...... 53 4.1.1.1. Hydrolysis...... 53 4.1.1.2. Phototransformation/photolysis...... 53 4.1.1.2.1. Phototransformation in air...... 53 4.1.1.2.2. Phototransformation in water...... 53 4.1.1.2.3. Phototransformation in soil...... 54 4.1.2. Biodegradation...... 54 4.1.2.1. Biodegradation in water...... 54 4.1.2.1.1. Estimated data...... 54 4.1.2.1.2. Screening tests...... 54 4.1.2.1.3. Simulation tests (water and sediments)...... 54 4.1.2.1.4. Summary and discussion of biodegradation in water and sediment...... 55 4.1.2.2. Biodegradation in soil...... 55 4.1.3. Summary and discussion of degradation...... 55 4.2. Environmental distribution...... 55 4.2.1. Adsorption/desorption...... 57 4.2.2. Volatilisation...... 58 4.2.3. Distribution modelling...... 58 4.2.4. Summary and discussion of environmental distribution...... 58 4.3. Bioaccumulation...... 58 4.3.1. Aquatic bioaccumulation...... 58 4.3.2. Terrestrial bioaccumulation...... 60 4.3.3. Summary and discussion of bioaccumulation...... 61 4.4. Secondary poisoning...... 62 4.5. Natural background...... 62 4.6. Additional information on environmental fate and distribution...... 63 5. HUMAN HEALTH HAZARD ASSESSMENT...... 65 5.1. Toxicokinetics...... 66 5.1.1. Non-human information...... 66 5.1.2. Human information...... 72 5.1.3. Summary and discussion of toxicokinetics...... 78 5.2. Acute toxicity...... 79 5.2.1. Non-human information...... 79 5.2.1.1. Acute toxicity: oral...... 79

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 2 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

5.2.1.2. Acute toxicity: inhalation...... 82 5.2.1.3. Acute toxicity: dermal...... 83 5.2.1.4. Acute toxicity: other routes...... 83 5.2.2. Human information...... 83 5.2.3. Summary and discussion of acute toxicity...... 85 5.3. Irritation...... 86 5.3.1. Skin...... 86 5.3.1.1. Non-human information...... 86 5.3.1.2. Human information...... 88 5.3.2. Eye...... 88 5.3.2.1. Non-human information...... 88 5.3.2.2. Human information...... 89 5.3.3. Respiratory tract...... 89 5.3.3.1. Non-human information...... 89 5.3.3.2. Human information...... 90 5.3.4. Summary and discussion of irritation...... 90 5.4. Corrosivity...... 90 5.4.1. Non-human information...... 90 5.4.2. Human information...... 90 5.4.3. Summary and discussion of corrosion...... 90 5.5. Sensitisation...... 90 5.5.1. Skin...... 90 5.5.1.1. Non-human information...... 90 5.5.1.2. Human information...... 92 5.5.2. Respiratory system...... 92 5.5.2.1. Non-human information...... 92 5.5.2.2. Human information...... 92 5.5.3. Summary and discussion of sensitisation...... 92 5.6. Repeated dose toxicity...... 92 5.6.1. Non-human information...... 92 5.6.1.1. Repeated dose toxicity: oral...... 93 Table 35. Overview of experimental studies on repeated dose toxicity after oral administration for ammonium chloride...... 95 5.6.1.2. Repeated dose toxicity: inhalation...... 96 5.6.1.3. Repeated dose toxicity: dermal...... 97 5.6.1.4. Repeated dose toxicity: other routes...... 98 5.6.2. Human information...... 98 5.6.3. Summary and discussion of repeated dose toxicity...... 101 5.7. Mutagenicity...... 102 5.7.1. Non-human information...... 102 5.7.1.1. In vitro data...... 102 5.7.1.2. In vivo data...... 106 5.7.2. Human information...... 109 5.7.3. Summary and discussion of mutagenicity...... 109 5.8. Carcinogenicity...... 110 5.8.1. Non-human information...... 110 5.8.1.1. Carcinogenicity: oral...... 110 5.8.1.2. Carcinogenicity: inhalation...... 110 5.8.1.3. Carcinogenicity: dermal...... 110 5.8.1.4. Carcinogenicity: other routes...... 110 5.8.2. Human information...... 110 5.8.3. Summary and discussion of carcinogenicity...... 112 5.9. Toxicity for reproduction...... 112 5.9.1. Effects on fertility...... 112 5.9.1.1. Non-human information...... 112 5.9.1.2. Human information...... 115 5.9.2. Developmental toxicity...... 115 5.9.2.1. Non-human information...... 115 5.9.2.2. Human information...... 117 5.9.3. Summary and discussion of reproductive toxicity...... 118

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 3 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

5.10. Other effects...... 119 5.10.1. Non-human information...... 119 5.10.1.1. Neurotoxicity...... 119 5.10.1.2. Immunotoxicity...... 119 5.10.1.3. Specific investigations: other studies...... 120 5.10.2. Human information...... 120 5.10.3. Summary and discussion of specific investigations...... 121 5.11. Derivation of DNEL(s) / DMEL(s)...... 121 5.11.1. Overview of typical dose descriptors for all endpoints...... 122 5.11.2. Correction of dose descriptors if needed (for example route-to-route extrapolation), application of assessment factors and derivation of the endpoint specific DN(M)EL...... 127 5.11.3. Selection of the critical DNEL(s) for critical health effects...... 132 6. HUMAN HEALTH HAZARD ASSESSMENT OF PHYSICO-CHEMICAL PROPERTIES...... 132 6.1. Explosivity...... 132 6.2. Flammability...... 133 6.3. Oxidising potential...... 133 7. ENVIRONMENTAL HAZARD ASSESSMENT...... 134 7.1. Aquatic compartment (including sediment)...... 135 7.1.1. Toxicity test results...... 135 7.1.1.1. Fish...... 152 7.1.1.1.1. Short-term toxicity to fish...... 152 7.1.1.1.2. Long-term toxicity to fish...... 154 7.1.1.2. Aquatic invertebrates...... 161 7.1.1.2.1. Short-term toxicity to aquatic invertebrates...... 161 7.1.1.2.2. Long-term toxicity to aquatic invertebrates...... 172 7.1.1.3. Algae and aquatic plants...... 191 7.1.1.4. Sediment organisms...... 201 7.1.1.5. Other aquatic organisms...... 211 7.1.2. Calculation of Predicted No Effect Concentration (PNEC)...... 213 7.1.2.1. PNECwater: freshwater...... 213 7.1.2.2. PNECwater: marine...... 220 7.1.2.3. PNEC sediment...... 223 7.2. Terrestrial compartment...... 235 7.2.1. Toxicity test results...... 235 7.2.1.1. Toxicity to soil macro-organisms...... 247 7.2.1.2. Toxicity to terrestrial plants...... 255 7.2.1.3. Toxicity to soil micro-organisms...... 261 7.2.1.4. Toxicity to other terrestrial organisms...... 273 7.2.2. Calculation of Predicted No Effect Concentration (PNEC soil)...... 273 7.3. Atmospheric compartment...... 277 7.4. Microbiological activity in sewage treatment systems...... 277 7.4.1. Toxicity to aquatic micro-organisms...... 278 7.4.2. PNEC for sewage treatment plant...... 280 7.5. Non compartment specific effects relevant for the food chain (secondary poisoning)...... 280 7.5.1. Toxicity to birds...... 280 7.5.2. Toxicity to mammals...... 281 7.5.3. Calculation of PNECoral (secondary poisoning)...... 281 7.6. Conclusion on the environmental hazard assessment and on classification and labelling...... 281 7.6.1. Classification under Annex I dangerous substances directive 67/548/EEC...... 281 7.6.2. Classification under 2nd Adaptation to Technical Progress (ATP) to the CLP Regulation (2nd ATP CLP)...... 281 8. PBT AND VPVB ASSESSMENT...... 282 8.1. Assessment of PBT/vPvB Properties...... 282 8.1.1. Summary and overall conclusions on PBT or vPvB properties...... 282 9. EXPOSURE ASSESSMENT (with local risk characterisation)...... 283 9.1. Local scenarios...... 284 9.1.1. GES Zn(NH4)Clx-0: Industrial use of primary or secondary zinc bearing material in the manufacture of Zn(NH4)Clx in several process steps, collection of the substance produced and packaging...... 286 9.1.2. GES Zn(NH4)Clx -1: Industrial use of Zn(NH4)Clx in the formulation of preparations by mixing

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 4 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

thoroughly, dry or in a solvent, the starting materials with potentially pressing, pelletising, sintering, possibly followed by packing...... 293 9.1.3. GES Zn(NH4)Clx -2: industrial use of ammonium zinc chloride or Zn(NH4)Clx -formulations in the manufacturing of other inorganic or organic zinc substances in a solvent-based matrix with potentially filtering and packaging...... 302 9.1.4. GES Zn(NH4)Clx -3: Industrial and professional use of Zn(NH4)Clx as active laboratory reagent in aqueous or organic media, for analysis or synthesis...... 309 9.1.5. GES Zn(NH4)Clx -4 : Industrial use of Zn(NH4)Clx or Zn(NH4)Clx -formulations as component for the manufacture of solid blends and matrices for further downstream use...... 316 9.1.6. GES Zn(NH4)Clx -5: Industrial use of Zn(NH4)Clx or Zn(NH4)Clx -formulations as component for the manufacture of dispersions, pastes or other viscous or polymerized matrices...... 327 9.1.7. GES Zn(NH4)Clx - 6 : Industrial and professional use of solid substrates containing less than 25%w/w of Zn(NH4)Clx...... 338 9.1.8. GES Zn(NH4)Clx -7 : Industrial and professional use of dispersions, pastes and polymerised substrates containing less than 25%w/w of Zn(NH4)Clx...... 346 9.1.9. GES Zn(NH4)C1x-8: Generic wide dispersive use of Zn...... 354 9.2. Consumer exposure...... 356 9.3. Indirect exposure of humans via the environment...... 358 9.4. Regional exposure concentrations...... 359 9.4.1. Modelling approach: diffuse source analysis...... 359 9.4.1.1. Overview national emission data...... 359 9.4.1.2. Continental releases and PEC calculations...... 361 9.4.2. Measured regional data in the environment...... 364 9.4.3. Comparison of measured and calculated regional zinc concentrations...... 373 10. RISK CHARACTERISATION...... 375 10.1. Local scenarios...... 375 10.1.1. Human health...... 375 10.1.1.1. Workers...... 375 10.1.1.2. Consumers...... 375 10.1.1.3. Indirect exposure of humans via the environment...... 375 10.1.2. Environment...... 375 10.2. Overall exposure (combined for all relevant emission/release sources)...... 375 10.2.1. Human health (combined for all exposure routes)...... 376 10.2.1.1. Consumers...... 376 10.2.1.2. Indirect exposure of humans via the environment...... 377 10.2.2. Environment (combined for all emission sources)...... 377 10.2.2.1. Risk characterisation based on modelled exposure...... 378 10.2.2.2. Risk characterisation based on measured data...... 378 REFERENCES...... 382 ANNEX 1: Exposure scenario building and environmental release estimation for the waste life stage of the manufacture and the use of zinc and zinc compounds...... 412 ANNEX 2: Evaluation of risks due to the presence of Zinc in European sewage treatment plants...... 414

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 5 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

List of Tables

Table 1. Substance identity...... 2

Table 2. Constituents...... 3

Table 3. Impurities...... 3

Table 4. Overview of physico-chemical properties...... 3

Table 5. Overview of quantities (in tonnes/year)...... 6

Table 6. Waste types, amounts and waste treatment processes for zinc and zinc compounds from manufacturing ...... 7

Table 7. Uses by workers in industrial settings...... 9

Table 8. Uses by professional workers...... 31

Table 9. Uses by consumers...... 42

Table 10. Waste types, amounts and waste treatment processes for zinc from identified uses...... 47

Table 11. Waste types, amounts and treatment of waste from service life sated subsequent to the identified uses for zinc from identified uses...... 47

Table 12. Classification according to Directive 67/548/EEC criteria...... 51

Table 13. Overview of screening tests for biodegradation in water for ammonium chloride...... 54

Table 14. (taken from the RA zinc, ECB 2008): Possible chemical forms (speciation) of dissolved zinc in seawater (Cleven et al., 1993)...... 56

Table 15. Overview of studies on aquatic bioaccumulation for zinc chloride...... 59

Table 16. Overview of studies on terrestrial bioaccumulation...... 60

Table 17. Water solubility values of the eleven zinc compounds covered in this CSR...... 65

Table 18. Grouping based on water solubility...... 66

Table 19. Dermal absorption of Zn (% of dose) through pig skin in vitro within 72 hours...... 67

Table 20. Overview of experimental studies on absorption, metabolism, distribution and elimination...... 70

Table 21. Deposition fractions for oral breathers and for oronasal augmenters, using a polydisperse particle distribution (MMAD 15.2 m, GSD 4.0)...... 74

Table 22. Assumptions used for estimating the inhalation absorption...... 75

Table 23. Percentage estimations for inhalation absorption of soluble, slightly soluble and insoluble zinc compounds...... 76

Table 24. Elimination data obtained following thirty humans dosed with 18 to 900 moles of 65Zn...... 78

Table 25. Overview of experimental studies on acute toxicity after oral administration according to decreasing water solubility of zinc compounds...... 80

Table 26. Re-calculation of oral LD50 rat values...... 81

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 6 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Table 27. Overview of experimental studies on acute toxicity after oral administration or ammonium chloride.81

Table 28. Overview of experimental studies on acute toxicity after inhalation exposure according to decreasing water solubility of zinc compounds...... 82

Table 29. Overview of experimental studies on acute toxicity after dermal exposure...... 83

Table 30. Overview of experimental studies on skin irritation according to decreasing water solubility of zinc compounds...... 86

Table 31. Overview of experimental studies on skin irritation for ammonium chloride...... 87

Table 32. Overview of experimental studies on eye irritation according to decreasing water solubility of zinc compounds...... 88

Table 33. Overview of experimental studies on skin sensitisation according to decreasing water solubility of zinc compounds...... 91

Table 34. Overview of experimental studies on repeated dose toxicity after oral administration...... 93

Table 35. Overview of experimental studies on repeated dose toxicity after oral administration for ammonium chloride...... 95

Table 36. Overview of experimental studies on repeated dose toxicity after inhalation...... 96

Table 37. Overview of experimental in vitro genotoxicity studies according to decreasing water solubility.....102

Table 38. Overview of experimental in vitro genotoxicity studies for ammonium chloride...... 105

Table 39. Overview of experimental in vivo genotoxicity studies according to decreasing water solubility...... 106

Table 40. Overview of experimental in vivo genotoxicity studies for ammonium chloride...... 108

Table 41. Overview of experimental studies on fertility...... 113

Table 42. Overview of experimental studies on developmental toxicity...... 115

Table 43. Overview of experimental studies on developmental toxicity for ammonium chloride...... 117

Table 44. Overview of experimental studies on immunotoxicity...... 119

Table 45. OELs for zinc chloride...... 121

Table 46. OELs for zinc oxide...... 121

Table 49. Summary of absorption rates through different routes of exposure...... 127

Table 50. Assessment factors (AF) for zinc compounds...... 128

Table 51. Corrected dose descriptor(s) per endpoint and endpoint-specific DNELs for workers...... 130

Table 52. Corrected dose descriptor(s) per endpoint and endpoint-specific DNELs for consumers...... 130

Table 53. Overview of information on oxidising potential...... 133

Table 54. Acute aquatic toxicity of zinc by species as a function of pH and hardness...... 136

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 7 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Table 55. Lowest acute aquatic toxicity data observed for zinc...... 137

Table 57. Summary statistics for SSD on chronic NOEC values for zinc in freshwater (N= 23)...... 143

Table 58. Geomean species NOECs of the marine zinc effects database presented by taxonomic group, species name and family name...... 148

Table 59. Summary statistics for the SSD on chronic NOEC values for zinc in saltwater (n=39)...... 149

Table 60. Results of field experiments made on phytoplankton communities coming from various natural sea waters...... 151

Table 61. Overview of short-term effects on fish...... 152

Table 62. Overview of long-term effects on fish...... 155

Table 63. Overview of short-term effects on aquatic invertebrates...... 161

Table 64. Overview of long-term effects on aquatic invertebrates...... 172

Table 65. Overview of effects on algae and aquatic plants...... 191

Table 66. Summary of chronic values that are used as input values for the SSD for deriving the 5th percentile values as a basis for the PNECadd, sediment. New values added after the closure of the RAR database are indicated in bold (see above)...... 202

Table 67. Overview of long-term effects on sediment organisms...... 204

Table 68. Overview of effects on other aquatic organisms: communities...... 211

Table 69. Comparison of NOECs normalised to realistic worst case conditions and the measured NOECs from the generic SSD...... 215

Table 70. Summary of Mesocosm studies reported in the risk assessment...... 216

Table 71. PNEC aquatic...... 223

Table 72. Summary statistics for the SSD on chronic NOEC values for zinc in freshwater sediment...... 224

Table 73. SEM-Zn and AVS in marine sediments...... 234

Table 74. PNEC sediment...... 235

Table 76. Generic species/process mean values...... 238

Table 77. HC5 and HC5-50 (with 5% and 95% confidence interval) for the terrestrial environment based on lab- field corrected and normalised NOEC/EC10 values and a log-normal distribution...... 243

Table 78. Normalised and lab-field corrected species/process mean NOEC/EC10 values for the 8 soil scenarios...... 245

Table 79. Overview of effects on soil macro-organisms...... 247

Table 80. Overview of effects on terrestrial plants...... 256

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 8 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Table 81. Overview of effects on soil micro-organisms...... 261

Table 82. Soil characteristics of the selected toxicity studies...... 274

Table 83. PNEC soil...... 277

Table 84. Summary of study results for STP-PNEC...... 278

Table 85. Overview of effects on micro-organisms...... 278

Table 86. PNEC sewage treatment plant...... 280

Table 87. PNEC oral...... 281

Table 88. Generic exposure scenarios for diammonium tetrachlorozincate...... 284

Table 89. Identified uses for Zn(NH4)Clx and corresponding Generic Exposure Scenario (GES)...... 285

Table 90. GES Zn(NH4)Clx-0...... 286

Table 91. Environmental risk characterisation for the manufacture of Zn(NH4)Clx, crossread fromZnCl2...... 291

Table 92. Exposure assessment for the industrial manufacture Zn(NH4)Clx, crossread fromZnCl2...... 291

Table 93. Occupational exposure data and risk characterisation for manufacture of zinc compounds...... 292

Table 94. GES Zn(NH4)Clx -1...... 293

Table 95. Environmental risk characterisation for the Industrial use of ZnCl2 (crossread to Zn(NH4)Clx) as component for the manufacture of preparations for further downstream use...... 297

Table 96. Exposure assessment for the industrial use of ZnCl2 (crossread to Zn(NH4)Clx) for the manufacture of wet or dry preparations, based on recently reported exposure data...... 298

Table 97. Environmental risk characterisation for the industrial use of Zn compounds (e.g. Zn(NH4)Clx) as component for the manufacture of preparations for further downstream use...... 298

Table 98. Occupational exposure data and risk characterisation for the Industrial formulation of dry or wet preparations/mixtures by mixing thoroughly Zn(NH4)Clx or other zinc compound with the other starting materials, with possible pressing, pelletising, sintering and packaging of the preparations/mixtures...... 301

Table 99. Occupational exposure data and risk characterisation for the scenario “the industrial use of ZnO for the manufacture of ZnCl2” (ZnCl2 GES-0)...... 301

Table 100. GES Zn(NH4)Clx -2...... 302

Table 101. Environmental risk characterisation for the scenario “industrial use of ZnCl2 for the manufacture of other zinc compounds” (crossread for Zn(NH4)Clx)...... 306

Table 102. Exposure assessment for the industrial use of ZnCl2 for the manufacture of other zinc compounds, based on recently reported exposure data (crossread for Zn(NH4)Clx)...... 307

Table 103. Workplace exposure data and risk characterisation for the scenario “industrial use of ZnCl2 for the manufacture of other zinc compounds” (crossread to Zn(NH4)Clx)...... 308

Table 104. GES- Zn(NH4)Clx-3...... 309

Table 105. Environmental release factors for the manufacture of different zinc compounds, to be used for industrial laboratories using zinc compounds...... 314

Table 106. Exposure assessment and risk characterisation for the industrial and professional use of Zn(NH4)Clx in laboratory...... 314

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 9 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Table 107. Occupational exposure data and risk characterisation for the Industrial and professional use of Zn(NH4)Clx and other zinc compounds in the laboratory...... 315

Table 108.GES Zn(NH4)Clx -4...... 316

Table 110. Exposure assessment for the industrial use of Zn(NH4)Clx as component for the manufacture of solid blends and matrices for further downstream use (crossread from other zinc compounds)...... 322

Table 111. Environmental risk characterisation for the Industrial use of Zn compounds as component for the manufacture of solid blends and matrices for further downstream use...... 322

Table 112. Occupational exposure data for the Industrial use of Zn(NH4)Clx and other Zn compounds as component for the manufacture of solid blends and matrices for further downstream use...... 324

Table 113. Additional occupational data and risk characterisations also relevant for the industrial use of

Zn(NH4)Clx and other Zn compounds as component for the manufacture of solid blends and matrices for further downstream use...... 326

Table 114. Occupational exposure data and risk characterisation for the scenario “ZnCl2 manufacture” (crossread to Zn(NH4)Clx)...... 326

Table 115. GES Zn(NH4)Clx -5...... 327

Table 116. Environmental risk characterisation for the Industrial use of Zn(NH4)Clx as component for the manufacture of liquid blends and matrices for further downstream use...... 331

Table 117. Exposure assessment for the industrial use of ZnCl2 as component for the manufacture of liquid blends and matrices for further downstream use (crossread to Zn(NH4)Clx)...... 332

Table 118. Environmental risk characterisation for the Industrial use of Zn compounds as component for the manufacture of liquid blends and matrices for further downstream use...... 332

Table 119. Exposure assessment for the industrial use of Zn compounds as component for the manufacture of liquid blends and matrices for further downstream use...... 333

Table 121. Occupational exposure data for the Industrial use of formulations containing ZnO and/or other zinc compounds as component for the manufacture of mixtures for further downstream use...... 335

Table 122. Occupational exposure data also relevant for the industrial formulation of wet preparations/mixtures by mixing thoroughly zinc compounds with other materials...... 337

Table 123. Occupational exposure data and risk characterisation for the scenario “ZnCl2 manufacture”...... 337

Table 125. GES Zn(NH4)Clx -6...... 338

Table 126. Environmental risk characterisation for the Industrial and professional use of solid substrates containing less than 25% w/w of Zn(NH4)Clx...... 343

Table 127: Occupational exposure data for the Industrial and professional use of solid substrates containing less than 25%w/w of Zn(NH4)Clx...... 344

Table 128. GES Zn(NH4)Clx -7...... 346

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 10 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Table 129. Environmental risk characterisation for the Industrial and professional use of liquid substrates containing less than 25% w/w of Zn(NH4)Clx...... 351

Table 130. Occupational exposure data for the Industrial and professional use of solid substrates containing less than 25%w/w of Zn(NH4)Clx...... 352

Table 131. GES Zn(NH4)C1x 8: Generic wide dispersive use of Zn...... 354

Table 132. Input parameters for EUSES...... 355

Table 133. Zinc emissions to water, soil and air in the Netherlands (data from the RA from 1999, with some updates) (in t/y)...... 359

Table 134. Comparison of total emission rates (tonnes/year) for The Netherlands, Germany, Belgium and Sweden...... 360

Table 135. Conversion of the NL emission data to EU...... 362

Table 136. Input data and results of the regional exposure assessment (all data refer to NL-region)...... 363

Table 137. Assessment of the recent monitored data on zinc, reported by 20 of 27 EU member states, according to the methodology as applied in the EU RAR on Zinc (ECB, 2008)...... 366

Table 138. Monitored total and added zinc concentrations (µg Zn/l) in EU member states...... 366

Table 139. Monitored dissolved zinc concentrations (µg Zn/l) in coastal waters and open sea of the Netherlands (2007-2009)...... 367

Table 140. Monitored dissolved zinc concentrations (µg Zn/l) in coastal waters of Belgium (1995-1998)...... 368

Table 141. Monitored sediment concentrations (mg Zn/kg dry wt) in the Netherlands (2000, 2003 and 2006).368

Table 142. Monitored sediment concentrations (mg Zn/kg dry wt) in Belgium...... 369

Table 143. Monitored total zinc concentrations (mg Zn/kg dwt.) in arable soils in Europe...... 369

Table 144. Monitored total zinc concentrations (mg Zn/kg dwt.) in arable soils in Europe...... 370

Table 145. Former and recent zinc sewage sludge concentrations in various EU countries...... 372

Table 146. Measured zinc concentrations in air (EU RA, for references, see ECB 2008)...... 373

Table 147. Regional concentrations in the environment...... 373

Table 148. Consumer exposure estimates (Table 4.1.3.3 of the RA)...... 376

Table 149. Internal exposure levels via water and air at local scale (taken over from Table 4.1.3.4.A of the RA ZnCl2 (ECB 2008))...... 377

Table 150. Modelled PECadd values and risk characterisation for zinc in the regional analysis...... 378

Table 151. Monitored PEC add values and risk characterisation for the EU freshwater...... 378

Table 152. Monitored PEC add values and risk characterisation for EU marine waters...... 379

Table 153. Monitored PEC add values and risk characterisation for EU sediment (freshwater and marine)...... 379

Table 154. Monitored PECvalues and risk characterisation for EU agricultural soils (arable land and grassland) ...... 380

List of Figures

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 11 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Figure 1. Base case total zinc removal from the water column using EUSES model parameters. The initial total zinc concentration in the water column (C0) is 413 μg/L. The horizontal dashed line represents C/C0 = 0.3 or 70% removal of zinc (from Mutch Associates 2010b)...... 64

Figure 2. Lognormal distribution curve fitting to the freshwater chronic toxicity data for zinc (ETX graphics)...... 144

Figure 3. Cumulative distribution of the 39 species mean NOEC values from Zn toxicity tests in the marine organisms database. Observed data and normal distribution curve fitted on the data...... 150

Figure 4. Overview of endpoints with statistical significant differences between treated and untreated (2.7µg Zn/l) mesocosms. Effect class “A”: no effects, “B”: slight effects, “C” pronounced temporal effects, “D”: pronounced durable effects. A positive or negative difference from the control is indicated with “+” or “-“, respectively (taken from Foekema et al 2012)...... 152

Figure 5. NOEC of algae (µg Zn/L) modelled with the Biotic ligand Model (BLM;Heijerick et al. (2005) for different pH and Ca combinations with DOC of 1 mg C/L...... 218

Figure 6. Species diversity in the marine environment (from ECETOC 2001). The stars highlight taxonomic groups represented in the zinc marine database...... 222

Figure 7. Lognormal distribution curve fitting to the freshwater sediment chronic toxicity data for zinc (ETX graphics)...... 225

Figure 8. Covariance between AVSZn and SEMZn in European sediments (taken from Vangheluwe et al 2003)...... 229

Figure 9. The species sensitivity distribution based on all individual EC10 and NOEC values selected for PNEC derivation...... 240

Figure 10. The species sensitivity distribution based on all individual EC10 and NOEC values with information on soil properties allowing correction for bioavailability among soils...... 240

Figure 11. The species sensitivity distribution based on all species/process mean values...... 241

Figure 12. The species sensitivity distribution based on species/process mean values for data with information on soil properties allowing correction for bioavailability among soils...... 241

Figure 13. Flow chart for the implementation of bioavailability factors into PNEC derivation for Zn...... 243

Figure 14. The species sensitivity distributions for the 8 soil scenarios as fitted by the log-normal distribution...... 244

Figure 15. Zinc concentrations (classes) in sludge from communal waste water treatment plants in the Netherlands in 1981 and 1997 (after CBS, 1999)...... 370

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 12 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Part A 1. SUMMARY OF RISK MANAGEMENT MEASURES The risk management of zinc focuses on the 2 relevant routes of exposure of workers, i.e. by inhalation and dermal exposure. Exposure to zinc containing dust and fumes by inhalation is controlled by the general application of local exhaust ventilation at the workplace, in specific cases complemented by personal protection measures. Inhalation exposure is further prevented by enclosures of systems. Dermal exposure is prevented by the general use of specialised protective clothing, including the wearing of specialised working gloves.

The risk management for environment includes on-site waste water treatment techniques are (if applicable) e.g.: chemical precipitation, sedimentation, filtration, the containment of liquid volumes in sumps to collect/prevent accidental spillage, and the control of air emissions by use of bag-house filters and/or other air emission abatement devices. Under section 9, risk management measures and operational conditions are described in more detail.

2. DECLARATION THAT RISK MANAGEMENT MEASURES ARE IMPLEMENTED

“I, …, declare hereby that risk management measures as described in this CSR are implemented.”

3. DECLARATION THAT RISK MANAGEMENT MEASURES ARE COMMUNICATED

“I, …, declare hereby that risk management measures as described in this CSR are communicated to downstream users.”

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 1 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Part B

General remark:

The chemical safety report of ammonium zinc chloride and diammonium tetrachlorozincate refers to the chemicals safety report of zinc chloride developed by the International Zinc Association, Brussels, Belgium.

Reason for this reference is that the substance hydrolyses in ammonium, zinc and chloride. All data of the endpoints show, that the zinc ion is driving the hazards for environment and human.

1. IDENTITY OF THE SUBSTANCE AND PHYSICAL AND CHEMICAL PROPERTIES

1.1. Name and other identifiers of the substance

The substance diammonium tetrachlorozincate(2-) is a mono constituent substance (origin: inorganic) having the following characteristics and physical–chemical properties (see the IUCLID dataset for further details).

The following public name is used: Ammonium zinc chloride.

Table 1. Substance identity

EC number: 238-687-6 EC name: diammonium tetrachlorozincate(2-) CAS number (EC inventory): 14639-97-5 IUPAC name: diammonium tetrachlorozincate(2-) Molecular formula: Cl4Zn.2H4N Molecular weight range: 243.2979

Structural formula:

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 2 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

1.2. Composition of the substance

Name: diammonium tetrachlorozincate(2-)

Degree of purity: >= 95 - <= 100 % (w/w)

Table 2. Constituents

Constituent Typical concentration Concentration range Remarks >= 95 - <= 100 % (w/w) diammonium tetrachlorozincate(2-)

Table 3. Impurities

Impurity Typical concentration Concentration range Remarks water >= 0 - <= 5 % (w/w)

1.3. Physico-chemical properties

Table 4. Overview of physico-chemical properties

Property Results Value used for CSA / Discussion Physical state at The physical state of the substance is solid Value used for CSA: solid 20°C and 1013 hPa powder, its clour is white, it has a weak ammonia like odour Melting / freezing In air, the substance starts melting at 297°C. Value used for CSA: 279 °C at 1013 hPa point Relative density The density of the substance is 1.92 g/cm3. Value used for CSA: 1.92 at 20°C

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 3 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Water solubility The solubility of Zn in the substance is 291 Value used for CSA: 291 g/L at 20 °C g/L (very soluble). Oxidising properties the substance has no oxidising properties. Value used for CSA: Oxidising: no Granulometry The D50 of the substance is 594 µm, the D80 is 765µm..

Data waiving

Information requirement: Boiling point

Reason: study scientifically unjustified

Justification: Not relevant; the sample decomposes before boiling

Information requirement: Vapour pressure

Reason: other justification

Justification: endpoint is not relevant; the sample is salt and has negligible vapour pressure at 25 °C. The vapour pressure from the minor impurities shown by the analysis is irrelevant at 25 °C.

Information requirement: Vapour pressure

Reason: other justification

Justification: The study does not need to be conducted if the melting point is above 300°C (Column 2 of Annex VII REACH regulation)

Information requirement: Surface tension

Reason: other justification

Justification: endpoint is not relevant for solid powder

Information requirement: Partition coefficient n-octanol/water (log value)

Reason: other justification

Justification: Not applicable to metal compounds; The study does not need to be conducted if the substance is inorganic (column 2 of Annex VII of the REACH regulation)

Information requirement: Flash point

Reason: other justification

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 4 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Justification: not applicable. The study does not need to be conducted if the substance is inorganic (Column 2 of Annex VII of REACH regulation)

Information requirement: Flammability

Reason: other justification

Justification: Based on the thermogravimetric and differential scanning calorimetric measurements and mineral composition the substance has no flammability, explosiveness or auto-inflammability properties.

Information requirement: Explosive properties

Reason: other justification

Information requirement: Self-ignition temperature

Reason: other justification

Information requirement: Stability in organic solvents and identity of relevant degradation products

Reason: other justification

Justification: Stability in organic solvents and identity of relevant degadation products is not an applicable endpoint for inorganic substances according to column 2 of Annex IX of the REACH Regulation.

Information requirement: Dissociation constant

Reason: other justification

Justification: The dissociation constant relating to the acidity constant, pKa, as required by the IUCLID database and REACH Guidance document, is not relevant for the substance.

Information requirement: Viscosity

Reason: study scientifically unjustified

Justification: viscosity is not relevant endpoint for powder

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 5 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Discussion of physico-chemical properties

For generating an updated, consistent and well-referenced database on the physico-chemical properties of the substance, a typical sample from the lead registrant was analysed for all parameters relevant for REACH at the Outotec Oy laboratories, Pori, Finland.

Outotec Research Oy has a certified Quality system ISO 9001:2000, Environmental system ISO14001 and Occupational Health and Safety system ISO18001. Laboratory accreditation according to ISO/IEC 17025 covers gas and emission measurements and metal analyses.

Inspecta Sertifiointi Oy evaluates the management systems and FINAS (Finnish Accreditation Service) evaluates the accredited methods. These audits are carried out annually. A couple of internal audits are also done every year, for instance laboratory functions are audited annually. Besides audits, Outotec Research Oy takes part in interlaboratory comparisons concerning metal analytics and emission measurements.

In 2008, Outotec Research Oy took part in the Finnish Excellence Quality Awards and received the prestigious award, "Recognised for Excellence". Outotec Research Oy achieved a score of over 500 points, which entitles the winner to use the five-star Recognised for Excellence, R4E emblem.

By this approach, a consistent, high quality and complete dataset on physicochemical properties of the substance has been established, using state-of-the-art anaylitical techniques. This updated information is encoded in the IUCLID V format.

2. MANUFACTURE AND USES

Quantities

Table 5. Overview of quantities (in tonnes/year)

Year Total tonnage Own use Used for article Used as intermediate Used for under strictly controlled research conditions purposes 2009 Manufactured: 0.0 0.0 Imported in article: 0.0 Transported: 0.0 0.0

Used in production of Imported: 0.0 article: 0.0 2008 Manufactured: 0.0 0.0 Imported in article: 0.0 Transported: 0.0 0.0

Used in production of Imported: 0.0 article: 0.0 2007 Manufactured: 0.0 0.0 Imported in article: 0.0 Transported: 0.0 0.0

Used in production of Imported: 0.0 article: 0.0

2.1. Manufacture

Manufacturing process

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 6 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

 Reception of zinc-bearing materials, if applicable, and transfer to the reaction tank (chloride and ammonia media)

 Reception of the Intermediate Ammonium zinc chloride solution in the reaction tank, if applicable

 Sequential addition of reagents for purification steps and filtration on press filter, when needed. Ventilation is adapted.

 Concentration by water evaporation, under exhaust hood.

 Pouring on a cooling belt

 Discharge and packaging of produced Zn(NH4)Clx crystals. Workers have to place and adjust the bag or drum under the discharge pipe and to set the process in motion. Filled bags or drums are subsequently closed and carried to the storage area.

 Exposure to dust can occur during packing of the powder. Solutions are packed in intermediate bulk containers (ca. 1 m3 capacity); solids are packed in bags or drums.

 Maintenance activities

WASTE (cfr Annex 1, Arche, 2012) :

Table 6. Waste types, amounts and waste treatment processes for zinc and zinc compounds from manufacturing

Waste Type of Suitable Amount Compositio Waste Informatio from waste waste (t/y) n treatmen n source code t process/ recycling Manufact Sludge from 06 05 02* Range: 0 – Range: 250 – Internal or In house ure on-site 19 02 05* 22,000 t/y. 300,0000 mg external questionnaire WWTP Median: Zn/kg dw. landfilling 2011 2,500 t/y Median: 8,700 Recycling mg Zn/kg dw internally Incineration Recycled in other applications General dust 10 05 03* Range: 0 – Range: 700 – Recycled 10 05 05* 36 t/y. 800,000 mg internally or 10 10 09* Median: 11 Zn/kg dw. externally 10 10 11* t/y Median: (e.g. ZnSO4 12 01 03 400,000 mg production) 12 01 04 Zn/kg dw. Slags 10 05 01 Range: 0 – Range: 12,000 Internal or 10 10 03 135,000 – 450,000 mg external t/y. Zn/kg dw. landfilling. Median: Median: 49,500 Recycled in 49,500 t/y mg Zn/kg dw. another application (road constructio n, profiling, covering layer, zinc recyclers) Sludges from 11 02 02* Range: 0 – Range: 27,000 Internal or

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 7 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

zinc 230,000 – 100,000 mg external hydrometallur t/y. Zn/kg dw. landfilling gy (Jarosite, Median: Median: 30,000 goethite, …) 101,500 t/y mg Zn/kg dw. Cadmium 10 05 06* Range: 0 – Range: 30,000 Recycled cake/sponge 11 01 09* 1,000 t/y. – 200,000 mg internally Median: Zn/kg dw. Internal or 670 t/y Median: external 160,000 mg landfilling Zn/kg dw. Cement Not Range: 0 – Range: 50,000 Recycled in Copper considere 4,000 t/y. – 125,000 mg copper d as waste Median: Zn/kg dw. production 100% 1,450 t/y Median: 79,000 recycled mg Zn/kg dw. Hg residue/ 10 05 06* Range: 0 – Range: 100 – Landfilled in sludge/ 06 04 04* 100 t/y. 9000 mg Zn/kg special calomel Median: 20 dw. concrete t/y Median: 550 bunkers, mg Zn/kg dw. eventually stabilization prior to landfilling. Anodic 11 02 07* Range: 0 – Range: 550 – Internal or sludge/ cell 4857 t/y. 300,000 mg external mud/ Mn Median: Zn/kg dw. landfilling sludge 3,000 t/y Median: 5,000 Recycled in mg Zn/kg dw. other applications Pb sludge/ Range: 0 – Range: 25 – Recycled in leach 40,000 t/y. 200,000 mg lead or lead Median: Zn/kg dw. alloy 24,767 t/y Median: 30,000 production mg Zn/kg dw. Casting/smelt 10 05 11 Range: 0 – Range: 745 – Recycled ing residues: 10 05 03* 4,000 t/y. 950,000 mg internally or Zn skimming, 11 02 03 Median: Zn/kg dw. externally drosses and 2,400 t/y Median: Recycled in ashes 825,000 mg another Zn/kg dw. application Other wastes: 06 03 13* Range: 0 – Range: 592 – Recycled slimes, 06 03 14 150,000 600,000 mg internally sludges, leach 06 03 15* t/y. Zn/kg dw. Recycled in residues, solid 06 04 05* Median: Median: 50,000 other wastes lead 08 01 11* 190 t/y mg Zn/kg dw. applications silver anode, 10 05 99 Internal or precipitates, 10 10 03 external salts, 10 10 05* landfilling/ adsorbents, 10 10 07* mine filling packaging 12 01 12* Incineration materials, 15 02 02* spoilt 15 01 10* products, 16 11 02 soaps, 16 11 03* refractories … 16 11 04 16 11 06

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 8 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

2.2. Identified uses

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 9 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Table 7. Uses by workers in industrial settings

Confidential IU number Identified Use Substance Use descriptors (IU) name supplied to that use 1 diammonium as such Process category (PROC): tetrachlorozincat (substance itself) PROC 2: Use in closed, continuous process with occasional controlled exposure e production and PROC 3: Use in closed batch process (synthesis or formulation) refining PROC 5: Mixing or blending in batch processes for formulation of preparations and articles (multistage and/or significant contact) PROC 8b: Transfer of substance or preparation (charging/discharging) from/to vessels/large containers at dedicated facilities PROC 9: Transfer of substance or preparation into small containers (dedicated filling line, including weighing) PROC 26: Handling of solid inorganic substances at ambient temperature

Market sector by type of chemical product: PC 20: Products such as ph-regulators, flocculants, precipitants, neutralisation agents PC 21: Laboratory chemicals PC 12: Fertilisers PC 14: Metal surface treatment products, including galvanic and electroplating products PC 19: Intermediate

Environmental release category (ERC): ERC 1: Manufacture of substances

Sector of end use (SU): SU 8: Manufacture of bulk, large scale chemicals (including petroleum products) SU 9: Manufacture of fine chemicals

Subsequent service life relevant for that use?: yes 5 Production of as such Process category (PROC): inorganic zinc (substance itself) PROC 2: Use in closed, continuous process with occasional controlled exposure compounds PROC 3: Use in closed batch process (synthesis or formulation)

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 10 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

PROC 8b: Transfer of substance or preparation (charging/discharging) from/to vessels/large containers at dedicated facilities PROC 9: Transfer of substance or preparation into small containers (dedicated filling line, including weighing) PROC 15: Use as laboratory reagent

Market sector by type of chemical product: PC 19: Intermediate PC 20: Products such as ph-regulators, flocculants, precipitants, neutralisation agents PC 21: Laboratory chemicals

Environmental release category (ERC): ERC 1: Manufacture of substances ERC 2: Formulation of preparations ERC 6a: Industrial use resulting in manufacture of another substance (use of intermediates)

Sector of end use (SU): SU 8: Manufacture of bulk, large scale chemicals (including petroleum products) SU 9: Manufacture of fine chemicals SU 10: Formulation [mixing] of preparations and/or re-packaging (excluding alloys)

Subsequent service life relevant for that use?: yes 6 Electroplating as such Process category (PROC): (substance itself) PROC 3: Use in closed batch process (synthesis or formulation) PROC 8b: Transfer of substance or preparation (charging/discharging) from/to vessels/large containers at dedicated facilities PROC 21: Low energy manipulation of substances bound in materials and/or articles

Market sector by type of chemical product: PC 7: Base metals and alloys PC 14: Metal surface treatment products, including galvanic and electroplating products

Environmental release category (ERC): ERC 2: Formulation of preparations ERC 5: Industrial use resulting in inclusion into or onto a matrix

Sector of end use (SU):

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 11 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

SU 15: Manufacture of fabricated metal products, except machinery and equipment SU 17: General manufacturing, e.g. machinery, equipment, vehicles, other transport equipment SU 0: Other: NACE C25.6.1: Treatment and coating of metals

Subsequent service life relevant for that use?: yes

Article category related to subsequent service life (AC): AC 2: Machinery, mechanical appliances, electrical/electronic articles AC 7: Metal articles 7 Production of as such Process category (PROC): Zn(NH4)Clx- (substance itself) PROC 2: Use in closed, continuous process with occasional controlled exposure based fluxing PROC 3: Use in closed batch process (synthesis or formulation) in a mixture agents PROC 5: Mixing or blending in batch processes for formulation of preparations and articles (multistage and/or significant contact) PROC 8b: Transfer of substance or preparation (charging/discharging) from/to vessels/large containers at dedicated facilities PROC 9: Transfer of substance or preparation into small containers (dedicated filling line, including weighing) PROC 15: Use as laboratory reagent

Market sector by type of chemical product: PC 19: Intermediate PC 20: Products such as ph-regulators, flocculants, precipitants, neutralisation agents PC 21: Laboratory chemicals

Environmental release category (ERC): ERC 1: Manufacture of substances ERC 2: Formulation of preparations ERC 5: Industrial use resulting in inclusion into or onto a matrix ERC 6a: Industrial use resulting in manufacture of another substance (use of intermediates)

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 12 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Subsequent service life relevant for that use?: yes 8 steel surface as such Process category (PROC): treatment prior (substance itself) PROC 2: Use in closed, continuous process with occasional controlled exposure to hot-dip PROC 4: Use in batch and other process (synthesis) where opportunity for exposure arises galvanizing PROC 8b: Transfer of substance or preparation (charging/discharging) from/to vessels/large containers at dedicated facilities PROC 13: Treatment of articles by dipping and pouring PROC 25: Other hot work operations with metals

Market sector by type of chemical product: PC 14: Metal surface treatment products, including galvanic and electroplating products PC 38: Welding and soldering products (with flux coatings or flux cores.), flux products

Environmental release category (ERC): ERC 5: Industrial use resulting in inclusion into or onto a matrix ERC 8a: Wide dispersive indoor use of processing aids in open systems ERC 10a: Wide dispersive outdoor use of long-life articles and materials with low release ERC 11a: Wide dispersive indoor use of long-life articles and materials with low release

Sector of end use (SU): SU 8: Manufacture of bulk, large scale chemicals (including petroleum products) SU 14: Manufacture of basic metals, including alloys SU 15: Manufacture of fabricated metal products, except machinery and equipment SU 18: Manufacture of furniture SU 19: Building and construction work SU 0: Other: Nace C23.9.9: Manufacture of other non-metallic mineral products n.e.c.

Subsequent service life relevant for that use?: yes

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 13 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

reagent (substance itself) PROC 1: Use in closed process, no likelihood of exposure PROC 2: Use in closed, continuous process with occasional controlled exposure PROC 3: Use in closed batch process (synthesis or formulation) PROC 4: Use in batch and other process (synthesis) where opportunity for exposure arises PROC 5: Mixing or blending in batch processes for formulation of preparations and articles (multistage and/or significant contact) PROC 8b: Transfer of substance or preparation (charging/discharging) from/to vessels/large containers at dedicated facilities PROC 9: Transfer of substance or preparation into small containers (dedicated filling line, including weighing) PROC 15: Use as laboratory reagent

Market sector by type of chemical product: PC 19: Intermediate PC 21: Laboratory chemicals PC 28: Perfumes, fragrances PC 39: Cosmetics, personal care products

Environmental release category (ERC): ERC 1: Manufacture of substances ERC 2: Formulation of preparations ERC 4: Industrial use of processing aids in processes and products, not becoming part of articles ERC 6a: Industrial use resulting in manufacture of another substance (use of intermediates) ERC 6b: Industrial use of reactive processing aids ERC 8a: Wide dispersive indoor use of processing aids in open systems ERC 8d: Wide dispersive outdoor use of processing aids in open systems

Sector of end use (SU): SU 10: Formulation [mixing] of preparations and/or re-packaging (excluding alloys) SU 24: Scientific research and development

Subsequent service life relevant for that use?: yes 11 Production of as such Process category (PROC): organic zinc (substance itself) PROC 1: Use in closed process, no likelihood of exposure compounds PROC 2: Use in closed, continuous process with occasional controlled exposure in a mixture PROC 3: Use in closed batch process (synthesis or formulation) PROC 4: Use in batch and other process (synthesis) where opportunity for exposure arises

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 14 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Market sector by type of chemical product: PC 19: Intermediate PC 20: Products such as ph-regulators, flocculants, precipitants, neutralisation agents PC 21: Laboratory chemicals PC 24: Lubricants, greases, release products PC 29: Pharmaceuticals PC 39: Cosmetics, personal care products

Environmental release category (ERC): ERC 1: Manufacture of substances ERC 2: Formulation of preparations ERC 6a: Industrial use resulting in manufacture of another substance (use of intermediates)

Sector of end use (SU): SU 9: Manufacture of fine chemicals SU 10: Formulation [mixing] of preparations and/or re-packaging (excluding alloys) SU 8: Manufacture of bulk, large scale chemicals (including petroleum products)

Subsequent service life relevant for that use?: yes 12 Production of as such Process category (PROC): coatings, paints, (substance itself) PROC 1: Use in closed process, no likelihood of exposure inks, enamels, PROC 2: Use in closed, continuous process with occasional controlled exposure in a mixture varnishes PROC 3: Use in closed batch process (synthesis or formulation) PROC 4: Use in batch and other process (synthesis) where opportunity for exposure arises PROC 5: Mixing or blending in batch processes for formulation of preparations and articles (multistage and/or significant contact) PROC 8b: Transfer of substance or preparation (charging/discharging) from/to vessels/large containers at dedicated facilities PROC 9: Transfer of substance or preparation into small containers (dedicated filling line, including weighing)

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 15 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Market sector by type of chemical product: PC 1: Adhesives, sealants PC 9a: Coatings and paints, thinners, paint removes PC 9b: Fillers, putties, plasters, modelling clay PC 9c: Finger paints PC 14: Metal surface treatment products, including galvanic and electroplating products PC 15: Non-metal-surface treatment products PC 18: Ink and toners PC 26: Paper and board dye, finishing and impregnation products: including bleaches and other processing aids PC 32: Polymer preparations and compounds

Environmental release category (ERC): ERC 1: Manufacture of substances ERC 2: Formulation of preparations ERC 3: Formulation in materials ERC 4: Industrial use of processing aids in processes and products, not becoming part of articles ERC 5: Industrial use resulting in inclusion into or onto a matrix ERC 7: Industrial use of substances in closed systems

Sector of end use (SU): SU 5: Manufacture of textiles, leather, fur SU 8: Manufacture of bulk, large scale chemicals (including petroleum products) SU 9: Manufacture of fine chemicals SU 10: Formulation [mixing] of preparations and/or re-packaging (excluding alloys) SU 11: Manufacture of rubber products SU 12: Manufacture of plastics products, including compounding and conversion SU 13: Manufacture of other non-metallic mineral products, e.g. plasters, cement SU 14: Manufacture of basic metals, including alloys

Subsequent service life relevant for that use?: yes 13 Component for as such Process category (PROC): paper coating or (substance itself) PROC 3: Use in closed batch process (synthesis or formulation) treatment for PROC 4: Use in batch and other process (synthesis) where opportunity for exposure arises in a mixture paper products PROC 5: Mixing or blending in batch processes for formulation of preparations and articles (multistage and/or significant contact)

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 16 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

PROC 6: Calendering operations PROC 8b: Transfer of substance or preparation (charging/discharging) from/to vessels/large containers at dedicated facilities PROC 9: Transfer of substance or preparation into small containers (dedicated filling line, including weighing) PROC 13: Treatment of articles by dipping and pouring

Market sector by type of chemical product: PC 9a: Coatings and paints, thinners, paint removes PC 15: Non-metal-surface treatment products PC 18: Ink and toners PC 21: Laboratory chemicals PC 35: Washing and cleaning products (including solvent based products) PC 20: Products such as ph-regulators, flocculants, precipitants, neutralisation agents PC 23: Leather tanning, dye, finishing, impregnation and care products PC 34: Textile dyes, finishing and impregnating products; including bleaches and other processing aids

Environmental release category (ERC): ERC 2: Formulation of preparations ERC 6b: Industrial use of reactive processing aids

Sector of end use (SU): SU 6b: Manufacture of pulp, paper and paper products SU 7: Printing and reproduction of recorded media SU 8: Manufacture of bulk, large scale chemicals (including petroleum products) SU 9: Manufacture of fine chemicals SU 10: Formulation [mixing] of preparations and/or re-packaging (excluding alloys)

Subsequent service life relevant for that use?: yes 14 Use of in a mixture Process category (PROC): Zn(NH4)Clx- PROC 4: Use in batch and other process (synthesis) where opportunity for exposure arises containing paper PROC 5: Mixing or blending in batch processes for formulation of preparations and articles coatings (multistage and/or significant contact) PROC 6: Calendering operations PROC 8b: Transfer of substance or preparation (charging/discharging) from/to vessels/large containers at dedicated facilities

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 17 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

PROC 9: Transfer of substance or preparation into small containers (dedicated filling line, including weighing) PROC 10: Roller application or brushing PROC 13: Treatment of articles by dipping and pouring PROC 19: Hand-mixing with intimate contact and only PPE available.

Market sector by type of chemical product: PC 1: Adhesives, sealants PC 9a: Coatings and paints, thinners, paint removes PC 9b: Fillers, putties, plasters, modelling clay PC 9c: Finger paints PC 15: Non-metal-surface treatment products PC 18: Ink and toners PC 14: Metal surface treatment products, including galvanic and electroplating products

Environmental release category (ERC): ERC 8a: Wide dispersive indoor use of processing aids in open systems ERC 8d: Wide dispersive outdoor use of processing aids in open systems ERC 10a: Wide dispersive outdoor use of long-life articles and materials with low release ERC 10b: Wide dispersive outdoor use of long-life articles and materials with high or intended release (including abrasive processing)

Sector of end use (SU): SU 6b: Manufacture of pulp, paper and paper products SU 10: Formulation [mixing] of preparations and/or re-packaging (excluding alloys)

Subsequent service life relevant for that use?: yes

Article category related to subsequent service life (AC): AC 01: Other (non intended to be released): coatings in art and creative items 15 Textile and as such Process category (PROC): leather coating (substance itself) PROC 3: Use in closed batch process (synthesis or formulation) treatment PROC 4: Use in batch and other process (synthesis) where opportunity for exposure arises in a mixture PROC 5: Mixing or blending in batch processes for formulation of preparations and articles (multistage and/or significant contact) PROC 6: Calendering operations PROC 8b: Transfer of substance or preparation (charging/discharging) from/to vessels/large

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 18 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Market sector by type of chemical product: PC 9a: Coatings and paints, thinners, paint removes PC 15: Non-metal-surface treatment products PC 19: Intermediate PC 20: Products such as ph-regulators, flocculants, precipitants, neutralisation agents PC 21: Laboratory chemicals PC 23: Leather tanning, dye, finishing, impregnation and care products PC 34: Textile dyes, finishing and impregnating products; including bleaches and other processing aids PC 35: Washing and cleaning products (including solvent based products)

Environmental release category (ERC): ERC 2: Formulation of preparations ERC 6b: Industrial use of reactive processing aids

Subsequent service life relevant for that use?: yes

Article category related to subsequent service life (AC): AC 6: Leather articles AC 5: Fabrics, textiles and apparel 16 Use of in a mixture Process category (PROC): Zn(NH4)Clx- PROC 4: Use in batch and other process (synthesis) where opportunity for exposure arises containing PROC 5: Mixing or blending in batch processes for formulation of preparations and articles coatings for (multistage and/or significant contact) textile and PROC 6: Calendering operations leather PROC 19: Hand-mixing with intimate contact and only PPE available.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 19 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Market sector by type of chemical product: PC 15: Non-metal-surface treatment products PC 23: Leather tanning, dye, finishing, impregnation and care products PC 34: Textile dyes, finishing and impregnating products; including bleaches and other processing aids

Environmental release category (ERC): ERC 8a: Wide dispersive indoor use of processing aids in open systems ERC 8d: Wide dispersive outdoor use of processing aids in open systems ERC 10a: Wide dispersive outdoor use of long-life articles and materials with low release ERC 11a: Wide dispersive indoor use of long-life articles and materials with low release

Sector of end use (SU): SU 5: Manufacture of textiles, leather, fur SU 10: Formulation [mixing] of preparations and/or re-packaging (excluding alloys)

Subsequent service life relevant for that use?: yes

Article category related to subsequent service life (AC): AC 5: Fabrics, textiles and apparel AC 6: Leather articles 17 Batteries /fuel as such Process category (PROC): cells (substance itself) PROC 3: Use in closed batch process (synthesis or formulation) PROC 5: Mixing or blending in batch processes for formulation of preparations and articles in a mixture (multistage and/or significant contact) PROC 14: Production of preparations or articles by tabletting, compression, extrusion, pelletisation PROC 13: Treatment of articles by dipping and pouring

Market sector by type of chemical product: PC 14: Metal surface treatment products, including galvanic and electroplating products

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 20 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

PC 19: Intermediate PC 20: Products such as ph-regulators, flocculants, precipitants, neutralisation agents PC 21: Laboratory chemicals

Environmental release category (ERC): ERC 2: Formulation of preparations ERC 5: Industrial use resulting in inclusion into or onto a matrix

Sector of end use (SU): SU 16: Manufacture of computer, electronic and optical products, electrical equipment SU 0: Other: Nace C27.2: Manufacture of batteries and accumulators

Subsequent service life relevant for that use?: yes

Article category related to subsequent service life (AC): AC 3: Electrical batteries and accumulators 18 Component for as such Process category (PROC): production of (substance itself) PROC 10: Roller application or brushing rubber, resins PROC 3: Use in closed batch process (synthesis or formulation) in a mixture and related PROC 5: Mixing or blending in batch processes for formulation of preparations and articles preparations (multistage and/or significant contact) PROC 8b: Transfer of substance or preparation (charging/discharging) from/to vessels/large containers at dedicated facilities PROC 9: Transfer of substance or preparation into small containers (dedicated filling line, including weighing) PROC 13: Treatment of articles by dipping and pouring PROC 14: Production of preparations or articles by tabletting, compression, extrusion, pelletisation PROC 23: Open processing and transfer operations with minerals/metals at elevated temperature PROC 24: High (mechanical) energy work-up of substances bound in materials and/or articles

Market sector by type of chemical product: PC 9a: Coatings and paints, thinners, paint removes PC 9b: Fillers, putties, plasters, modelling clay PC 9c: Finger paints PC 18: Ink and toners PC 19: Intermediate PC 20: Products such as ph-regulators, flocculants, precipitants, neutralisation agents

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 21 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

PC 24: Lubricants, greases, release products PC 32: Polymer preparations and compounds PC 33: Semiconductors

Environmental release category (ERC): ERC 2: Formulation of preparations ERC 3: Formulation in materials ERC 4: Industrial use of processing aids in processes and products, not becoming part of articles ERC 5: Industrial use resulting in inclusion into or onto a matrix ERC 6d: Industrial use of process regulators for polymerisation processes in production of resins, rubbers, polymers ERC 10a: Wide dispersive outdoor use of long-life articles and materials with low release ERC 11a: Wide dispersive indoor use of long-life articles and materials with low release

Sector of end use (SU): SU 10: Formulation [mixing] of preparations and/or re-packaging (excluding alloys) SU 11: Manufacture of rubber products

Subsequent service life relevant for that use?: yes

Article category related to subsequent service life (AC): AC 10: Rubber articles 19 Production of as such Process category (PROC): polymer- (substance itself) PROC 2: Use in closed, continuous process with occasional controlled exposure matrices, plastics PROC 3: Use in closed batch process (synthesis or formulation) in a mixture and related PROC 5: Mixing or blending in batch processes for formulation of preparations and articles preparations (multistage and/or significant contact) PROC 6: Calendering operations PROC 8b: Transfer of substance or preparation (charging/discharging) from/to vessels/large containers at dedicated facilities PROC 9: Transfer of substance or preparation into small containers (dedicated filling line, including weighing) PROC 10: Roller application or brushing PROC 13: Treatment of articles by dipping and pouring PROC 14: Production of preparations or articles by tabletting, compression, extrusion, pelletisation PROC 21: Low energy manipulation of substances bound in materials and/or articles PROC 24: High (mechanical) energy work-up of substances bound in materials and/or articles

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 22 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Market sector by type of chemical product: PC 19: Intermediate PC 20: Products such as ph-regulators, flocculants, precipitants, neutralisation agents PC 32: Polymer preparations and compounds PC 33: Semiconductors

Environmental release category (ERC): ERC 1: Manufacture of substances ERC 3: Formulation in materials ERC 5: Industrial use resulting in inclusion into or onto a matrix ERC 6a: Industrial use resulting in manufacture of another substance (use of intermediates)

Sector of end use (SU): SU 10: Formulation [mixing] of preparations and/or re-packaging (excluding alloys) SU 12: Manufacture of plastics products, including compounding and conversion

Subsequent service life relevant for that use?: yes

Article category related to subsequent service life (AC): AC 1: Vehicles AC 2: Machinery, mechanical appliances, electrical/electronic articles AC 3: Electrical batteries and accumulators AC 13: Plastic articles 20 Additive / as such Process category (PROC): component for (substance itself) PROC 3: Use in closed batch process (synthesis or formulation) the production of PROC 5: Mixing or blending in batch processes for formulation of preparations and articles in a mixture Sealants / (multistage and/or significant contact) Adhesives / PROC 8b: Transfer of substance or preparation (charging/discharging) from/to vessels/large Mastics containers at dedicated facilities PROC 9: Transfer of substance or preparation into small containers (dedicated filling line, including weighing) PROC 10: Roller application or brushing PROC 11: Non industrial spraying PROC 13: Treatment of articles by dipping and pouring PROC 14: Production of preparations or articles by tabletting, compression, extrusion, pelletisation PROC 20: Heat and pressure transfer fluids in dispersive, professional use but closed systems

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 23 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

PROC 21: Low energy manipulation of substances bound in materials and/or articles PROC 24: High (mechanical) energy work-up of substances bound in materials and/or articles

Market sector by type of chemical product: PC 1: Adhesives, sealants PC 9a: Coatings and paints, thinners, paint removes PC 14: Metal surface treatment products, including galvanic and electroplating products PC 19: Intermediate PC 20: Products such as ph-regulators, flocculants, precipitants, neutralisation agents PC 24: Lubricants, greases, release products PC 32: Polymer preparations and compounds

Environmental release category (ERC): ERC 1: Manufacture of substances ERC 2: Formulation of preparations ERC 3: Formulation in materials ERC 5: Industrial use resulting in inclusion into or onto a matrix ERC 6a: Industrial use resulting in manufacture of another substance (use of intermediates) ERC 6d: Industrial use of process regulators for polymerisation processes in production of resins, rubbers, polymers ERC 8b: Wide dispersive indoor use of reactive substances in open systems ERC 8c: Wide dispersive indoor use resulting in inclusion into or onto a matrix ERC 10a: Wide dispersive outdoor use of long-life articles and materials with low release ERC 10b: Wide dispersive outdoor use of long-life articles and materials with high or intended release (including abrasive processing) ERC 11a: Wide dispersive indoor use of long-life articles and materials with low release

Sector of end use (SU): SU 8: Manufacture of bulk, large scale chemicals (including petroleum products) SU 10: Formulation [mixing] of preparations and/or re-packaging (excluding alloys)

Subsequent service life relevant for that use?: yes

Article category related to subsequent service life (AC): AC 1: Vehicles AC 2: Machinery, mechanical appliances, electrical/electronic articles AC 7: Metal articles

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 24 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

AC 11: Wood articles 22 Additive / as such Process category (PROC): component for (substance itself) PROC 3: Use in closed batch process (synthesis or formulation) the production of PROC 4: Use in batch and other process (synthesis) where opportunity for exposure arises in a mixture Lubricants / PROC 5: Mixing or blending in batch processes for formulation of preparations and articles Grease / Metal (multistage and/or significant contact) working fluids PROC 8b: Transfer of substance or preparation (charging/discharging) from/to vessels/large containers at dedicated facilities PROC 9: Transfer of substance or preparation into small containers (dedicated filling line, including weighing) PROC 10: Roller application or brushing PROC 13: Treatment of articles by dipping and pouring

Market sector by type of chemical product: PC 14: Metal surface treatment products, including galvanic and electroplating products PC 24: Lubricants, greases, release products PC 25: Metal working fluids PC 32: Polymer preparations and compounds

Environmental release category (ERC): ERC 1: Manufacture of substances ERC 2: Formulation of preparations ERC 3: Formulation in materials ERC 5: Industrial use resulting in inclusion into or onto a matrix ERC 6a: Industrial use resulting in manufacture of another substance (use of intermediates) ERC 6d: Industrial use of process regulators for polymerisation processes in production of resins, rubbers, polymers ERC 8b: Wide dispersive indoor use of reactive substances in open systems ERC 9a: Wide dispersive indoor use of substances in closed systems ERC 9b: Wide dispersive outdoor use of substances in closed systems ERC 11a: Wide dispersive indoor use of long-life articles and materials with low release

Sector of end use (SU): SU 10: Formulation [mixing] of preparations and/or re-packaging (excluding alloys) SU 18: Manufacture of furniture

Subsequent service life relevant for that use?: yes

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 25 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Article category related to subsequent service life (AC): AC 1: Vehicles AC 2: Machinery, mechanical appliances, electrical/electronic articles AC 7: Metal articles 24 Additive / as such Process category (PROC): component for (substance itself) PROC 3: Use in closed batch process (synthesis or formulation) the production of PROC 4: Use in batch and other process (synthesis) where opportunity for exposure arises in a mixture Polishes / wax PROC 5: Mixing or blending in batch processes for formulation of preparations and articles blends (multistage and/or significant contact) PROC 7: Industrial spraying PROC 8b: Transfer of substance or preparation (charging/discharging) from/to vessels/large containers at dedicated facilities PROC 9: Transfer of substance or preparation into small containers (dedicated filling line, including weighing) PROC 10: Roller application or brushing PROC 11: Non industrial spraying PROC 13: Treatment of articles by dipping and pouring PROC 19: Hand-mixing with intimate contact and only PPE available.

Market sector by type of chemical product: PC 9c: Finger paints PC 9a: Coatings and paints, thinners, paint removes PC 9b: Fillers, putties, plasters, modelling clay PC 14: Metal surface treatment products, including galvanic and electroplating products PC 25: Metal working fluids PC 31: Polishes and wax blends

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 26 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

ERC 8b: Wide dispersive indoor use of reactive substances in open systems

Sector of end use (SU): SU 9: Manufacture of fine chemicals SU 10: Formulation [mixing] of preparations and/or re-packaging (excluding alloys) SU 18: Manufacture of furniture

Subsequent service life relevant for that use?: yes

Article category related to subsequent service life (AC): AC 1: Vehicles AC 2: Machinery, mechanical appliances, electrical/electronic articles AC 7: Metal articles 26 Use of in a mixture Process category (PROC): Zn(NH4)Clx- PROC 1: Use in closed process, no likelihood of exposure containing PROC 2: Use in closed, continuous process with occasional controlled exposure catalysts PROC 3: Use in closed batch process (synthesis or formulation) PROC 5: Mixing or blending in batch processes for formulation of preparations and articles (multistage and/or significant contact) PROC 8b: Transfer of substance or preparation (charging/discharging) from/to vessels/large containers at dedicated facilities PROC 9: Transfer of substance or preparation into small containers (dedicated filling line, including weighing) PROC 14: Production of preparations or articles by tabletting, compression, extrusion, pelletisation

Market sector by type of chemical product: PC 2: Adsorbents PC 9a: Coatings and paints, thinners, paint removes PC 9b: Fillers, putties, plasters, modelling clay PC 9c: Finger paints PC 19: Intermediate PC 20: Products such as ph-regulators, flocculants, precipitants, neutralisation agents PC 40: Extraction agents

Environmental release category (ERC): ERC 1: Manufacture of substances ERC 4: Industrial use of processing aids in processes and products, not becoming part of articles

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 27 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

ERC 5: Industrial use resulting in inclusion into or onto a matrix ERC 6a: Industrial use resulting in manufacture of another substance (use of intermediates) ERC 6b: Industrial use of reactive processing aids

Subsequent service life relevant for that use?: yes 27 Additive as such Process category (PROC): component for (substance itself) PROC 3: Use in closed batch process (synthesis or formulation) production of de- PROC 5: Mixing or blending in batch processes for formulation of preparations and articles in a mixture icing products (multistage and/or significant contact) PROC 8b: Transfer of substance or preparation (charging/discharging) from/to vessels/large containers at dedicated facilities PROC 9: Transfer of substance or preparation into small containers (dedicated filling line, including weighing)

Market sector by type of chemical product: PC 4: Anti-freeze and de-icing products PC 20: Products such as ph-regulators, flocculants, precipitants, neutralisation agents PC 35: Washing and cleaning products (including solvent based products)

Environmental release category (ERC): ERC 2: Formulation of preparations ERC 5: Industrial use resulting in inclusion into or onto a matrix ERC 8c: Wide dispersive indoor use resulting in inclusion into or onto a matrix ERC 8f: Wide dispersive outdoor use resulting in inclusion into or onto a matrix

Sector of end use (SU): SU 8: Manufacture of bulk, large scale chemicals (including petroleum products) SU 9: Manufacture of fine chemicals

Subsequent service life relevant for that use?: yes 31 Additive for the as such Process category (PROC):

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 28 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

formulation of (substance itself) PROC 5: Mixing or blending in batch processes for formulation of preparations and articles biocidal products (multistage and/or significant contact) in a mixture PROC 8b: Transfer of substance or preparation (charging/discharging) from/to vessels/large containers at dedicated facilities PROC 9: Transfer of substance or preparation into small containers (dedicated filling line, including weighing)

Market sector by type of chemical product: PC 8: Biocidal products (e.g. disinfectants, pest control) PC 37: Water treatment chemicals

Environmental release category (ERC): ERC 2: Formulation of preparations

Sector of end use (SU): SU 9: Manufacture of fine chemicals SU 10: Formulation [mixing] of preparations and/or re-packaging (excluding alloys)

Subsequent service life relevant for that use?: yes 32 Additive for the as such Process category (PROC): formulation of (substance itself) PROC 5: Mixing or blending in batch processes for formulation of preparations and articles cleaning (multistage and/or significant contact) in a mixture products PROC 8b: Transfer of substance or preparation (charging/discharging) from/to vessels/large containers at dedicated facilities PROC 9: Transfer of substance or preparation into small containers (dedicated filling line, including weighing)

Market sector by type of chemical product: PC 8: Biocidal products (e.g. disinfectants, pest control) PC 35: Washing and cleaning products (including solvent based products) PC 37: Water treatment chemicals

Environmental release category (ERC): ERC 2: Formulation of preparations ERC 8a: Wide dispersive indoor use of processing aids in open systems ERC 8b: Wide dispersive indoor use of reactive substances in open systems

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 29 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Sector of end use (SU): SU 9: Manufacture of fine chemicals SU 10: Formulation [mixing] of preparations and/or re-packaging (excluding alloys)

Subsequent service life relevant for that use?: yes 29 Additive for the as such Process category (PROC): formulation of (substance itself) PROC 1: Use in closed process, no likelihood of exposure fertilizers PROC 2: Use in closed, continuous process with occasional controlled exposure in a mixture PROC 3: Use in closed batch process (synthesis or formulation) PROC 4: Use in batch and other process (synthesis) where opportunity for exposure arises PROC 5: Mixing or blending in batch processes for formulation of preparations and articles (multistage and/or significant contact) PROC 8b: Transfer of substance or preparation (charging/discharging) from/to vessels/large containers at dedicated facilities PROC 9: Transfer of substance or preparation into small containers (dedicated filling line, including weighing) PROC 13: Treatment of articles by dipping and pouring

Market sector by type of chemical product: PC 9b: Fillers, putties, plasters, modelling clay PC 12: Fertilisers PC 20: Products such as ph-regulators, flocculants, precipitants, neutralisation agents PC 21: Laboratory chemicals

Environmental release category (ERC): ERC 2: Formulation of preparations ERC 3: Formulation in materials ERC 10a: Wide dispersive outdoor use of long-life articles and materials with low release ERC 10b: Wide dispersive outdoor use of long-life articles and materials with high or intended release (including abrasive processing) ERC 5: Industrial use resulting in inclusion into or onto a matrix

Sector of end use (SU): SU 1: Agriculture, forestry and fishing SU 8: Manufacture of bulk, large scale chemicals (including petroleum products) SU 10: Formulation [mixing] of preparations and/or re-packaging (excluding alloys)

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 30 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Subsequent service life relevant for that use?: yes 34 Additive in the as such Process category (PROC): formulation of (substance itself) PROC 1: Use in closed process, no likelihood of exposure cosmetics PROC 2: Use in closed, continuous process with occasional controlled exposure in a mixture PROC 3: Use in closed batch process (synthesis or formulation) PROC 5: Mixing or blending in batch processes for formulation of preparations and articles (multistage and/or significant contact) PROC 8b: Transfer of substance or preparation (charging/discharging) from/to vessels/large containers at dedicated facilities PROC 9: Transfer of substance or preparation into small containers (dedicated filling line, including weighing) PROC 13: Treatment of articles by dipping and pouring PROC 14: Production of preparations or articles by tabletting, compression, extrusion, pelletisation PROC 15: Use as laboratory reagent

Market sector by type of chemical product: PC 28: Perfumes, fragrances PC 35: Washing and cleaning products (including solvent based products) PC 39: Cosmetics, personal care products

Environmental release category (ERC): ERC 2: Formulation of preparations ERC 5: Industrial use resulting in inclusion into or onto a matrix

Sector of end use (SU): SU 10: Formulation [mixing] of preparations and/or re-packaging (excluding alloys)

Subsequent service life relevant for that use?: yes 36 Additive in the as such Process category (PROC): formulation of (substance itself) PROC 1: Use in closed process, no likelihood of exposure pharma / PROC 2: Use in closed, continuous process with occasional controlled exposure in a mixture veterinary PROC 3: Use in closed batch process (synthesis or formulation) products PROC 5: Mixing or blending in batch processes for formulation of preparations and articles (multistage and/or significant contact) PROC 8b: Transfer of substance or preparation (charging/discharging) from/to vessels/large

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 31 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

containers at dedicated facilities PROC 9: Transfer of substance or preparation into small containers (dedicated filling line, including weighing) PROC 13: Treatment of articles by dipping and pouring PROC 14: Production of preparations or articles by tabletting, compression, extrusion, pelletisation PROC 15: Use as laboratory reagent

Market sector by type of chemical product: PC 20: Products such as ph-regulators, flocculants, precipitants, neutralisation agents PC 21: Laboratory chemicals PC 29: Pharmaceuticals

Environmental release category (ERC): ERC 2: Formulation of preparations ERC 5: Industrial use resulting in inclusion into or onto a matrix ERC 8a: Wide dispersive indoor use of processing aids in open systems ERC 8d: Wide dispersive outdoor use of processing aids in open systems

Sector of end use (SU): SU 9: Manufacture of fine chemicals SU 10: Formulation [mixing] of preparations and/or re-packaging (excluding alloys) SU 20: Health services SU 0: Other: Nace C21.1: Manufacture of basic pharmaceutical products

Subsequent service life relevant for that use?: yes

Table 8. Uses by professional workers

Confidential IU number Identified Use Substance Use descriptors (IU) name supplied to that use 9 Use of as such Process category (PROC): Zn(NH4)Clx- (substance itself) PROC 2: Use in closed, continuous process with occasional controlled exposure based fluxing PROC 4: Use in batch and other process (synthesis) where opportunity for exposure arises agents before PROC 8b: Transfer of substance or preparation (charging/discharging) from/to vessels/large welding/solderin containers at dedicated facilities

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 32 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

g processes PROC 13: Treatment of articles by dipping and pouring PROC 25: Other hot work operations with metals

Market sector by type of chemical product: PC 7: Base metals and alloys PC 25: Metal working fluids PC 38: Welding and soldering products (with flux coatings or flux cores.), flux products

Environmental release category (ERC): ERC 3: Formulation in materials ERC 5: Industrial use resulting in inclusion into or onto a matrix ERC 8a: Wide dispersive indoor use of processing aids in open systems ERC 8d: Wide dispersive outdoor use of processing aids in open systems ERC 10a: Wide dispersive outdoor use of long-life articles and materials with low release ERC 10b: Wide dispersive outdoor use of long-life articles and materials with high or intended release (including abrasive processing)

Sector of end use (SU): SU 18: Manufacture of furniture SU 0: Other:Nace C23.9.9: Manufacture of other non-metallic mineral products n.e.c. SU 14: Manufacture of basic metals, including alloys SU 15: Manufacture of fabricated metal products, except machinery and equipment SU 19: Building and construction work

Subsequent service life relevant for that use?: yes

Article category related to subsequent service life (AC):

AC 1: Vehicles AC 2: Machinery, mechanical appliances, electrical/electronic articles AC 3: Electrical batteries and accumulators AC 7: Metal articles 10 Laboratory as such Process category (PROC): reagent (substance itself) PROC 3: Use in closed batch process (synthesis or formulation) PROC 4: Use in batch and other process (synthesis) where opportunity for exposure arises PROC 5: Mixing or blending in batch processes for formulation of preparations and articles (multistage and/or significant contact) PROC 1: Use in closed process, no likelihood of exposure

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 33 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

PROC 2: Use in closed, continuous process with occasional controlled exposure PROC 8b: Transfer of substance or preparation (charging/discharging) from/to vessels/large containers at dedicated facilities PROC 9: Transfer of substance or preparation into small containers (dedicated filling line, including weighing) PROC 15: Use as laboratory reagent

Market sector by type of chemical product: PC 19: Intermediate PC 21: Laboratory chemicals PC 28: Perfumes, fragrances PC 39: Cosmetics, personal care products

Sector of end use (SU):

SU 10: Formulation [mixing] of preparations and/or re-packaging (excluding alloys) SU 24: Scientific research and development

Subsequent service life relevant for that use?: yes 14 Use of in a mixture Process category (PROC): Zn(NH4)Clx- PROC 4: Use in batch and other process (synthesis) where opportunity for exposure arises paper coatings PROC 5: Mixing or blending in batch processes for formulation of preparations and articles (multistage and/or significant contact) PROC 6: Calendering operations PROC 8b: Transfer of substance or preparation (charging/discharging) from/to vessels/large containers at dedicated facilities PROC 9: Transfer of substance or preparation into small containers (dedicated filling line, including weighing)

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 34 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

PROC 10: Roller application or brushing PROC 13: Treatment of articles by dipping and pouring PROC 19: Hand-mixing with intimate contact and only PPE available.

Market sector by type of chemical product: PC 1: Adhesives, sealants PC 9a: Coatings and paints, thinners, paint removes PC 9b: Fillers, putties, plasters, modelling clay PC 9c: Finger paints PC 15: Non-metal-surface treatment products PC 18: Ink and toners PC 14: Metal surface treatment products, including galvanic and electroplating products

Environmental release category (ERC): ERC 8a: Wide dispersive indoor use of processing aids in open systems ERC 8d: Wide dispersive outdoor use of processing aids in open systems ERC 10a: Wide dispersive outdoor use of long-life articles and materials with low release ERC 11a: Wide dispersive indoor use of long-life articles and materials with low release

Sector of end use (SU): SU 6b: Manufacture of pulp, paper and paper products SU 10: Formulation [mixing] of preparations and/or re-packaging (excluding alloys)

Subsequent service life relevant for that use?: yes

Article category related to subsequent service life (AC):

AC 01: Other (non intended to be released):coating for art, creative items 16 Use of in a mixture Process category (PROC): Zn(NH4)Clx- PROC 4: Use in batch and other process (synthesis) where opportunity for exposure arises formulations in PROC 5: Mixing or blending in batch processes for formulation of preparations and articles textile and (multistage and/or significant contact) leather PROC 6: Calendering operations coating/treatmen PROC 8b: Transfer of substance or preparation (charging/discharging) from/to vessels/large t containers at dedicated facilities PROC 9: Transfer of substance or preparation into small containers (dedicated filling line, including weighing) PROC 13: Treatment of articles by dipping and pouring

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 35 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

PROC 10: Roller application or brushing PROC 19: Hand-mixing with intimate contact and only PPE available.

Market sector by type of chemical product: PC 15: Non-metal-surface treatment products PC 34: Textile dyes, finishing and impregnating products; including bleaches and other processing aids PC 23: Leather tanning, dye, finishing, impregnation and care products

Environmental release category (ERC): ERC 11a: Wide dispersive indoor use of long-life articles and materials with low release ERC 8a: Wide dispersive indoor use of processing aids in open systems ERC 8d: Wide dispersive outdoor use of processing aids in open systems ERC 10a: Wide dispersive outdoor use of long-life articles and materials with low release

Sector of end use (SU):

SU 5: Manufacture of textiles, leather, fur SU 10: Formulation [mixing] of preparations and/or re-packaging (excluding alloys)

Subsequent service life relevant for that use?: yes

Article category related to subsequent service life (AC):

AC 6: Leather articles AC 5: Fabrics, textiles and apparel 21 Use of as such Process category (PROC): Zn(NH4)Clx- (substance itself) PROC 7: Industrial spraying containing PROC 8a: Transfer of substance or preparation (charging/discharging) from/to vessels/large Sealants / containers at non-dedicated facilities Adhesives / PROC 8b: Transfer of substance or preparation (charging/discharging) from/to vessels/large Mastics containers at dedicated facilities PROC 9: Transfer of substance or preparation into small containers (dedicated filling line, including weighing) PROC 10: Roller application or brushing PROC 11: Non industrial spraying PROC 13: Treatment of articles by dipping and pouring PROC 14: Production of preparations or articles by tabletting, compression, extrusion, pelletisation

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 36 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

PROC 17: Lubrication at high energy conditions and in partly open process PROC 19: Hand-mixing with intimate contact and only PPE available. PROC 21: Low energy manipulation of substances bound in materials and/or articles

Market sector by type of chemical product: PC 1: Adhesives, sealants PC 9a: Coatings and paints, thinners, paint removes PC 9b: Fillers, putties, plasters, modelling clay PC 9c: Finger paints PC 14: Metal surface treatment products, including galvanic and electroplating products PC 19: Intermediate PC 20: Products such as ph-regulators, flocculants, precipitants, neutralisation agents PC 24: Lubricants, greases, release products PC 32: Polymer preparations and compounds

Environmental release category (ERC): ERC 8a: Wide dispersive indoor use of processing aids in open systems ERC 8c: Wide dispersive indoor use resulting in inclusion into or onto a matrix ERC 8d: Wide dispersive outdoor use of processing aids in open systems ERC 8f: Wide dispersive outdoor use resulting in inclusion into or onto a matrix

Sector of end use (SU):

SU 5: Manufacture of textiles, leather, fur SU 6a: Manufacture of wood and wood products SU 6b: Manufacture of pulp, paper and paper products SU 11: Manufacture of rubber products SU 12: Manufacture of plastics products, including compounding and conversion SU 13: Manufacture of other non-metallic mineral products, e.g. plasters, cement SU 15: Manufacture of fabricated metal products, except machinery and equipment SU 19: Building and construction work

Subsequent service life relevant for that use?: yes

Article category related to subsequent service life (AC):

AC 1: Vehicles AC 2: Machinery, mechanical appliances, electrical/electronic articles

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 37 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

AC 7: Metal articles AC 11: Wood articles 23 Use of in a mixture Process category (PROC): Zn(NH4)Clx- PROC 7: Industrial spraying containing PROC 8a: Transfer of substance or preparation (charging/discharging) from/to vessels/large Lubricants / containers at non-dedicated facilities Grease / Metal PROC 8b: Transfer of substance or preparation (charging/discharging) from/to vessels/large working fluids containers at dedicated facilities PROC 9: Transfer of substance or preparation into small containers (dedicated filling line, including weighing) PROC 10: Roller application or brushing PROC 11: Non industrial spraying PROC 13: Treatment of articles by dipping and pouring PROC 14: Production of preparations or articles by tabletting, compression, extrusion, pelletisation PROC 17: Lubrication at high energy conditions and in partly open process PROC 19: Hand-mixing with intimate contact and only PPE available. PROC 21: Low energy manipulation of substances bound in materials and/or articles

Sector of end use (SU):

SU 17: General manufacturing, e.g. machinery, equipment, vehicles, other transport equipment SU 18: Manufacture of furniture

Subsequent service life relevant for that use?: yes

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 38 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Article category related to subsequent service life (AC):

AC 1: Vehicles AC 2: Machinery, mechanical appliances, electrical/electronic articles AC 7: Metal articles AC 11: Wood articles 25 Use of in a mixture Process category (PROC): Zn(NH4)Clx- PROC 7: Industrial spraying containing PROC 8b: Transfer of substance or preparation (charging/discharging) from/to vessels/large Polishes / wax containers at dedicated facilities blends PROC 9: Transfer of substance or preparation into small containers (dedicated filling line, including weighing) PROC 10: Roller application or brushing PROC 11: Non industrial spraying PROC 13: Treatment of articles by dipping and pouring PROC 14: Production of preparations or articles by tabletting, compression, extrusion, pelletisation PROC 19: Hand-mixing with intimate contact and only PPE available. PROC 21: Low energy manipulation of substances bound in materials and/or articles PROC 8a: Transfer of substance or preparation (charging/discharging) from/to vessels/large containers at non-dedicated facilities

Market sector by type of chemical product: PC 9a: Coatings and paints, thinners, paint removes PC 9b: Fillers, putties, plasters, modelling clay PC 9c: Finger paints PC 14: Metal surface treatment products, including galvanic and electroplating products PC 25: Metal working fluids PC 31: Polishes and wax blends

Sector of end use (SU):

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 39 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

SU 9: Manufacture of fine chemicals SU 18: Manufacture of furniture

Subsequent service life relevant for that use?: yes

Article category related to subsequent service life (AC):

AC 1: Vehicles AC 2: Machinery, mechanical appliances, electrical/electronic articles AC 7: Metal articles AC 11: Wood articles 28 Use of in a mixture Process category (PROC): Zn(NH4)Clx- PROC 7: Industrial spraying containing de- PROC 8a: Transfer of substance or preparation (charging/discharging) from/to vessels/large icing products containers at non-dedicated facilities PROC 8b: Transfer of substance or preparation (charging/discharging) from/to vessels/large containers at dedicated facilities PROC 9: Transfer of substance or preparation into small containers (dedicated filling line, including weighing) PROC 10: Roller application or brushing PROC 11: Non industrial spraying PROC 13: Treatment of articles by dipping and pouring PROC 14: Production of preparations or articles by tabletting, compression, extrusion, pelletisation PROC 19: Hand-mixing with intimate contact and only PPE available. PROC 21: Low energy manipulation of substances bound in materials and/or articles

Environmental release category (ERC): ERC 8a: Wide dispersive indoor use of processing aids in open systems ERC 10a: Wide dispersive outdoor use of long-life articles and materials with low release

Sector of end use (SU):

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 40 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

SU 9: Manufacture of fine chemicals SU 18: Manufacture of furniture

Subsequent service life relevant for that use?: yes 30 Use of in a mixture Process category (PROC): Zn(NH4)Clx- PROC 2: Use in closed, continuous process with occasional controlled exposure containing PROC 7: Industrial spraying fertilizer's PROC 8a: Transfer of substance or preparation (charging/discharging) from/to vessels/large formulations containers at non-dedicated facilities PROC 8b: Transfer of substance or preparation (charging/discharging) from/to vessels/large containers at dedicated facilities PROC 9: Transfer of substance or preparation into small containers (dedicated filling line, including weighing) PROC 10: Roller application or brushing PROC 11: Non industrial spraying PROC 13: Treatment of articles by dipping and pouring PROC 19: Hand-mixing with intimate contact and only PPE available. PROC 26: Handling of solid inorganic substances at ambient temperature

Market sector by type of chemical product: PC 9b: Fillers, putties, plasters, modelling clay PC 12: Fertilisers PC 20: Products such as ph-regulators, flocculants, precipitants, neutralisation agents

Environmental release category (ERC): ERC 8a: Wide dispersive indoor use of processing aids in open systems ERC 8b: Wide dispersive indoor use of reactive substances in open systems ERC 8d: Wide dispersive outdoor use of processing aids in open systems ERC 8e: Wide dispersive outdoor use of reactive substances in open systems ERC 9b: Wide dispersive outdoor use of substances in closed systems ERC 10b: Wide dispersive outdoor use of long-life articles and materials with high or intended release (including abrasive processing)

Sector of end use (SU):

SU 1: Agriculture, forestry and fishing SU 9: Manufacture of fine chemicals

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 41 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Subsequent service life relevant for that use?: yes 33 Use of in a mixture Process category (PROC): Zn(NH4)Clx- PROC 8a: Transfer of substance or preparation (charging/discharging) from/to vessels/large containing containers at non-dedicated facilities cleaning PROC 8b: Transfer of substance or preparation (charging/discharging) from/to vessels/large products containers at dedicated facilities PROC 9: Transfer of substance or preparation into small containers (dedicated filling line, including weighing) PROC 10: Roller application or brushing PROC 11: Non industrial spraying PROC 13: Treatment of articles by dipping and pouring

Environmental release category (ERC): ERC 8a: Wide dispersive indoor use of processing aids in open systems

Sector of end use (SU):

SU 9: Manufacture of fine chemicals

Subsequent service life relevant for that use?: yes 35 Use of cosmetics in a mixture Process category (PROC): PROC 8b: Transfer of substance or preparation (charging/discharging) from/to vessels/large containers at dedicated facilities PROC 8a: Transfer of substance or preparation (charging/discharging) from/to vessels/large containers at non-dedicated facilities PROC 9: Transfer of substance or preparation into small containers (dedicated filling line, including weighing) PROC 10: Roller application or brushing PROC 11: Non industrial spraying

Market sector by type of chemical product:

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 42 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

PC 28: Perfumes, fragrances PC 35: Washing and cleaning products (including solvent based products) PC 39: Cosmetics, personal care products

Environmental release category (ERC): ERC 8a: Wide dispersive indoor use of processing aids in open systems

Sector of end use (SU):

SU 9: Manufacture of fine chemicals

Subsequent service life relevant for that use?: yes 37 Use of in a mixture Process category (PROC): Pharma/veterinar PROC 8b: Transfer of substance or preparation (charging/discharging) from/to vessels/large y products containers at dedicated facilities PROC 9: Transfer of substance or preparation into small containers (dedicated filling line, including weighing) PROC 10: Roller application or brushing PROC 11: Non industrial spraying

Market sector by type of chemical product: PC 20: Products such as ph-regulators, flocculants, precipitants, neutralisation agents PC 21: Laboratory chemicals PC 29: Pharmaceuticals

Environmental release category (ERC): ERC 8a: Wide dispersive indoor use of processing aids in open systems

Sector of end use (SU):

SU 20: Health services

Subsequent service life relevant for that use?: yes

Table 9. Uses by consumers

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 43 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Confidential IU number Identified Use Use descriptors (IU) name 9 Use of Chemical product category (PC): Zn(NH4)Clx- PC 7: Base metals and alloys based fluxing PC 25: Metal working fluids agents before PC 38: Welding and soldering products (with flux coatings or flux cores.), flux products welding/solderin g processes Environmental release category (ERC): ERC 3: Formulation in materials ERC 5: Industrial use resulting in inclusion into or onto a matrix ERC 8a: Wide dispersive indoor use of processing aids in open systems ERC 8d: Wide dispersive outdoor use of processing aids in open systems ERC 10a: Wide dispersive outdoor use of long-life articles and materials with low release ERC 11a: Wide dispersive indoor use of long-life articles and materials with low release

Subsequent service life relevant for that use?: yes

Article category related to subsequent service life (AC): AC 1: Vehicles AC 2: Machinery, mechanical appliances, electrical/electronic articles AC 3: Electrical batteries and accumulators AC 7: Metal articles 21 Use of Chemical product category (PC): Zn(NH4)Clx- PC 1: Adhesives, sealants containing PC 9a: Coatings and paints, thinners, paint removes Sealants/ PC 9b: Fillers, putties, plasters, modelling clay Adhesives/ PC 9c: Finger paints Mastics PC 14: Metal surface treatment products, including galvanic and electroplating products PC 19: Intermediate PC 20: Products such as ph-regulators, flocculants, precipitants, neutralisation agents PC 24: Lubricants, greases, release products PC 32: Polymer preparations and compounds

Environmental release category (ERC): ERC 8a: Wide dispersive indoor use of processing aids in open systems

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 44 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

ERC 8c: Wide dispersive indoor use resulting in inclusion into or onto a matrix ERC 8d: Wide dispersive outdoor use of processing aids in open systems ERC 8f: Wide dispersive outdoor use resulting in inclusion into or onto a matrix

Subsequent service life relevant for that use?: yes

Article category related to subsequent service life (AC): AC 1: Vehicles AC 2: Machinery, mechanical appliances, electrical/electronic articles AC 7: Metal articles AC 11: Wood articles 23 Use of Chemical product category (PC): Zn(NH4)Clx- PC 14: Metal surface treatment products, including galvanic and electroplating products containing PC 24: Lubricants, greases, release products Lubricants / PC 25: Metal working fluids Grease / Metal PC 32: Polymer preparations and compounds working fluids Environmental release category (ERC): ERC 8a: Wide dispersive indoor use of processing aids in open systems ERC 8c: Wide dispersive indoor use resulting in inclusion into or onto a matrix ERC 8d: Wide dispersive outdoor use of processing aids in open systems ERC 8f: Wide dispersive outdoor use resulting in inclusion into or onto a matrix

Subsequent service life relevant for that use?: yes

Article category related to subsequent service life (AC): AC 1: Vehicles AC 2: Machinery, mechanical appliances, electrical/electronic articles AC 7: Metal articles AC 11: Wood articles 25 Use of Chemical product category (PC): Zn(NH4)Clx- PC 9a: Coatings and paints, thinners, paint removes containing PC 9b: Fillers, putties, plasters, modelling clay Polishes / wax PC 9c: Finger paints blends PC 14: Metal surface treatment products, including galvanic and electroplating products PC 25: Metal working fluids

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 45 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

PC 31: Polishes and wax blends

Subsequent service life relevant for that use?: yes

Environmental release category (ERC): ERC 8a: Wide dispersive indoor use of processing aids in open systems ERC 10a: Wide dispersive outdoor use of long-life articles and materials with low release

Subsequent service life relevant for that use?: yes 30 Use of Chemical product category (PC): Zn(NH4)Clx- PC 9b: Fillers, putties, plasters, modelling clay containing PC 12: Fertilisers fertilizer's PC 20: Products such as ph-regulators, flocculants, precipitants, neutralisation agents formulations Environmental release category (ERC): ERC 8a: Wide dispersive indoor use of processing aids in open systems ERC 8b: Wide dispersive indoor use of reactive substances in open systems ERC 8d: Wide dispersive outdoor use of processing aids in open systems ERC 8e: Wide dispersive outdoor use of reactive substances in open systems ERC 9b: Wide dispersive outdoor use of substances in closed systems

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 46 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

ERC 10b: Wide dispersive outdoor use of long-life articles and materials with high or intended release (including abrasive processing)

Subsequent service life relevant for that use?: yes 33 Use of Chemical product category (PC): Zn(NH4)Clx- PC 8: Biocidal products (e.g. disinfectants, pest control) containing PC 35: Washing and cleaning products (including solvent based products) cleaning PC 39: Cosmetics, personal care products products Environmental release category (ERC): ERC 8a: Wide dispersive indoor use of processing aids in open systems

Subsequent service life relevant for that use?: yes 35 Use of cosmetics Chemical product category (PC): PC 28: Perfumes, fragrances PC 35: Washing and cleaning products (including solvent based products) PC 36: Water softeners

Environmental release category (ERC): ERC 8a: Wide dispersive indoor use of processing aids in open systems

Subsequent service life relevant for that use?: yes 37 Use of Pharma / Chemical product category (PC): veterinary PC 20: Products such as ph-regulators, flocculants, precipitants, neutralisation agents products PC 21: Laboratory chemicals PC 29: Pharmaceuticals

Environmental release category (ERC): ERC 8a: Wide dispersive indoor use of processing aids in open systems

Subsequent service life relevant for that use?: yes

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 47 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

WASTE (cfr Annex 1, Arche, 2012) :

Table 10. Waste types, amounts and waste treatment processes for zinc from identified uses

Waste Type of Suitabl Amount Comp Waste Information from waste e (t/y) ositio treatmen source waste n t code process/ recycling Downstream Scraps, 02 01 10* 0.7% from Treated as Reck 2008 use cuttings, dusts, 10 10 10 the hazardous solvents, 12 01 03* Zndownstre waste Waste stream solutions, 15 01 04* am use profiles EC sludges, 16 01 04* tonnage 2010 contaminated 16 01 06* ends up in material, off- 16 01 18* hazardous Waste report specification 16 06 02* waste ARCHE 2011 batches, … 16 08 02* 16 08 03* 17 04 07* 17 04 09* 17 09 04* 19 10 02* 19 12 03* …

Table 11. Waste types, amounts and treatment of waste from service life sated subsequent to the identified uses for zinc from identified uses

Waste Type of Suitabl Amount Compositio Waste Informatio from waste e (t/y) n treatment n source waste process/ code recycling Municipal Solid 20 01 34 179,430 Average Municipal waste EUROSTAT waste and municipal 20 01 40 ktonnes concentration: landfill 2009 EoL waste: 20 03 01 dry 71 mg Ni/kg Municipal waste Paper/card 20 03 07 weight dw incineration Waste report -board, Recycling ARCHE 2011 Metal, Glass, Plastics, Textile, Organic matter, Other

Most common technical function of substance (what it does): Flux agents for casting Laboratory chemicals Plating agents and metal surface treating agents component in batteries

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 48 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Process regulators, other than polymerisation or vulcanisation processes Intermediates Anti-freezing agents Complexing agents

2.3. Uses advised against

None 3. CLASSIFICATION AND LABELLING 3.1. Classification and labelling according to CLP / GHS

Name: diammonium tetrachlorozincate(2-)

Implementation: EU

Classification

The substance is classified as follows:

←for physical-chemical properties:

Explosives: Reason for no classification: conclusive but not sufficient for classification

Flammable gases: Reason for no classification: conclusive but not sufficient for classification

Flammable aerosols: Reason for no classification: conclusive but not sufficient for classification

Oxidising gases: Reason for no classification: conclusive but not sufficient for classification

Gases under Reason for no classification: conclusive but not sufficient for classification pressure:

Flammable liquids: Reason for no classification: conclusive but not sufficient for classification

Flammable solids: Reason for no classification: conclusive but not sufficient for classification

Self-reacting Reason for no classification: conclusive but not sufficient for classification substances and mixtures:

Pyrophoric liquids: Reason for no classification: conclusive but not sufficient for classification

Pyrophoric solids: Reason for no classification: conclusive but not sufficient for classification

Self-heating Reason for no classification: conclusive but not sufficient for classification substances and mixtures:

Substances and Reason for no classification: conclusive but not sufficient for classification mixtures which in contact with water

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 49 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 emits flammable gases:

Oxidising liquids: Reason for no classification: conclusive but not sufficient for classification

Oxidising solids: Reason for no classification: conclusive but not sufficient for classification

Organic peroxides: Reason for no classification: conclusive but not sufficient for classification

Corrosive to metals: Reason for no classification: conclusive but not sufficient for classification

←for health hazards:

Acute toxicity - oral: Acute Tox. 4 (Hazard statement: H302: Harmful if swallowed.)

Acute toxicity - Reason for no classification: conclusive but not sufficient for classification dermal:

Acute toxicity - Reason for no classification: conclusive but not sufficient for classification inhalation:

Skin Skin Mild Irrit. 3 (Hazard statement H316: Causes mild skin irritation). corrosion/irritation:

Serious damage/eye Eye Irrit. 2B (Hazard statement: H320: Causes eye irritation.) irritation:

Respiration Reason for no classification: conclusive but not sufficient for classification sensitization:

Skin sensitation: Reason for no classification: conclusive but not sufficient for classification

Aspiration hazard: Reason for no classification: conclusive but not sufficient for classification

Reproductive Reason for no classification: conclusive but not sufficient for classification Toxicity:

Reproductive Reason for no classification: conclusive but not sufficient for classification Toxicity: Effects on or via lactation:

Germ cell Reason for no classification: conclusive but not sufficient for classification mutagenicity:

Carcinogenicity: Reason for no classification: conclusive but not sufficient for classification

Specific target organ Reason for no classification: conclusive but not sufficient for classification toxicity - single:

Specific target organ Reason for no classification: conclusive but not sufficient for classification toxicity - repeated:

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 50 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

←for environmental hazards:

Hazards to the Aquatic Acute 1 (Hazard statement: H400: Very toxic to aquatic life.) aquatic environment (acute/short- term):

M-Factor acute: 1

Hazards to the Aquatic Chronic 2 (Hazard statement: H411: Toxic to aquatic life with long lasting aquatic environment effects.) (long-term):

Hazardous to the Reason for no classification: data lacking atmospheric environment:

Labelling

Signal word: Warning

Hazard pictogram:

GHS07: exclamation mark

GHS09: environment

Hazard statements:

H302: Harmful if swallowed. H315: Causes skin irritation. H400: Very toxic to aquatic life. H411: Toxic to aquatic life with long lasting effects.

Precautionary statements:

P202: Do not handle until all safety precautions have been read and understood. P270: Do no eat, drink or smoke when using this product. P273: Avoid release to the environment. P391: Collect spillage. P281: Use personal protective equipment as required. P405: Store locked up. P308+P313: IF exposed or concerned: Get medical advice/attention. P501: Dispose of contents/container to...

3.2. Classification and labelling according to DSD / DPD

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 51 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

3.2.1. Classification and labelling in Annex I of Directive 67/548/EEC

3.2.2. Self classification(s)

Chemical name: diammonium tetrachlorozincate(2-)

Table 12. Classification according to Directive 67/548/EEC criteria

Endpoints Classification Reason for no Justification for classification (non) classification can be found in section Explosiveness conclusive but not 6.1 sufficient for classification Oxidising properties conclusive but not 6.3 sufficient for classification Flammability conclusive but not 6.2 sufficient for classification Thermal stability conclusive but not sufficient for classification Acute toxicity Xn; R22 Harmful; Harmful if 5.2 swallowed. Acute toxicity- irreversible conclusive but not 5.2 damage after single sufficient for exposure classification Repeated dose toxicity conclusive but not 5.6 sufficient for classification Irritation / Corrosion Xi; R38 Irritating to skin. 5.3.4 and 5.4.3 Sensitisation conclusive but not 5.5.3 sufficient for classification Carcinogenicity conclusive but not 5.8.3 sufficient for classification Mutagenicity - Genetic conclusive but not 5.7.3 Toxicity sufficient for classification Toxicity to reproduction- conclusive but not 5.9.3 fertility sufficient for classification Toxicity to reproduction- conclusive but not 5.9.3 development sufficient for classification Toxicity to reproduction - conclusive but not 5.9.3 breastfed babies sufficient for classification Environment N; R50/53 Dangerous for the 7.6

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 52 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

environment; Very toxic to aquatic organisms, may cause long-term adverse effects in the aquatic environment.

Labelling

Indication of danger:

Xn - harmful N - dangerous for the environment Xi- irritant

R-phrases:

R22 - harmful if swallowed R50/53 Very toxic to aquatic organisms, may cause long-term adverse effects in the aquatic environment R38- irritating to skin

S-phrases:

S46 - if swallowed, seek medical advice immediately and show this container or label S60 - this material and its container must be disposed of as hazardous waste S61 - avoid release to the environment. refer to special instructions/safety data sheets 3.2.3. Other classification(s)

4. ENVIRONMENTAL FATE PROPERTIES

General introduction to chapters 4, 5, 6 and 7.

Under Regulation 793/93/CEE, an extensive risk assessment on Zinc and 5 zinc compounds (ZnO, ZnCl2, ZnSO4, Zn orthophosphate and zinc distearate) has been recently prepared by the Dutch authorities for the EU. The risk assessment report (RAR) on these 6 zinc substances has been recently published (ECB 2008). Diammonium tetrachlorozincate was not assessed specifically in the EU RA, but the analysis was for a greater part generically related to the hazard profile of the Zn++ ion. Therefore the analysis of the RA is considered also relevant for diammonium tetrachlorozincate. Since these RARs were the result of intensive discussions between all stakeholders, and were approved by experts from all the member states; since they provide a recent review of the available evidence on zinc and zinc compounds (the file was closed in September 2006), they will be used as the main reference for this chemical safety report. In this chemical safety report, the information, data and conclusions of the RAR will be summarised, focusing on the principles applied, the assumptions made and the conclusions. Where available and relevant, new information and data will be included and discussed.

General remarks on the chapter on environmental fate properties.

Zinc is a natural element, which is essential for all living organisms. It occurs in the metallic state, or as zinc compound, with one valency state (Zn++). All environmental concentration data are expressed as “Zn”, while toxicity is caused by the Zn++ ion. For this reason, the sections on human toxicity and ecotoxicity are applicable

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 53 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 to all zinc compounds, from which zinc ions are released into the environment. Some zinc compounds have however very low solubility and will therefore not release zinc ions; this strongly decreases their potential (eco-)toxicity. As a consequence, distinction is being made between zinc compounds, as a function of their solubility (see chapters 5 and 7). For checking the potential of metal substances to release ions in the environment, a specific test, the transformation/dissolution (T/D) test is used. For metallic zinc and some of the zinc compounds, this test has been performed. If applicable, the results of such T/D test are discussed in section 4.6. (data in IUCLID section 5.6.).

The issue of degradation (section 4.1.) is not applicable to inorganic compounds. However, the speciation of zinc in the environment compartments is relevant and is discussed under section 4.2.

When zinc ions are formed in the environment, they will further interact with the environmental matrix and biota. As such, the concentration of zinc ions that is available to organisms, the bioavailable fraction, will depend on processes like dissolution, absorption, precipitation, complexation, inclusion into (soil) matrix, etc. These processes are defining the fate of zinc in the environment and, ultimately, its ecotoxic potential. This has been recognised e.g. in the guidance to the CLP regulation 1272/2008 (metals annex): “Environmental transformation of one species of a metal to another species of the same does not constitute degradation as applied to organic compounds and may increase or decrease the availability and bioavailability of the toxic species. However as a result of naturally occurring geochemical processes metal ions can partition from the water column. Data on water column residence time, the processes involved at the water – sediment interface (i. e. deposition and re-mobilisation) are fairly extensive, but have not been integrated into a meaningful database. Nevertheless, using the principles and assumptions discussed above in Section IV.1, it maybe possible to incorporate this approach into classification.“

In the water, the bioavailability of zinc through interaction with components of the water and biota has been studied in detail in the zinc RA (ECB 2008). This has resulted in an approach for quantifying zinc bioavailability into risk assessment. The ultimate fate of zinc in water (in the water column) is assessed via the “unit world model”, that can quantify the “removal from the water column” of the zinc species. As such, it is shown that zinc (ions) brought into water will be rapidly removed from the water column (>70% removal within 28days). This phenomenon is described in section 4.6. (data in IUCLID 5.6), and is considered for classification.

In sediment, zinc binds to the sulphide fraction to form insoluble ZnS. As such, zinc is not bioavailable anymore to organisms. This has been discussed in the EU RA (ECB 2008), and has resulted in an approach for quantifying zinc bioavailability into risk assessment. Based on experimental data, a default conservative bioavailability factor of 0.5 was proposed in the RA. This approach can be refined when the relevant data on sulphide and Zn in sediment are available. Due to the insolubility of the ZnS (K=9.2 x 10-25) zinc will be sequestered in the (anaerobioc) sediments, and the re-mobilisation of zinc ions into the water column will be prevented. This is also quantified in the unit world model, see section 4.6.

In soil, short-term interaction of zinc ions upon spiking, and long term interactions (“ageing”) have been extensively discussed in the zinc RA (ECB 2008). This has resulted in an approach for quantifying zinc bioavailability into risk assessment. Based on experimental data, a general ageing factor of 3 was derived in the RA; according to soil type, the bio-availability of zinc can be further determined, when the relevant data on e.g. pH, CEC are available.

4.1. Degradation 4.1.1. Abiotic degradation

4.1.1.1. Hydrolysis

Data waiving

Reason: study scientifically unjustified

Justification: Waived: According to Annex IX of REACH Regulation, information on hydrolysis is not required for inorganics

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 54 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

The following 4.1.-related endpoints are not relevant for inorganics:

4.1.1.2. Phototransformation/photolysis

4.1.1.2.1. Phototransformation in air

4.1.1.2.2. Phototransformation in water

4.1.1.2.3. Phototransformation in soil

4.1.2. Biodegradation

4.1.2.1. Biodegradation in water

4.1.2.1.1. Estimated data

4.1.2.1.2. Screening tests

Table 13. Overview of screening tests for biodegradation in water for ammonium chloride Method Results Remarks Reference not mentioned nitrification 4 (not assignable) Cutler, D.W, (1933)

% Degradation of test supporting study substance: Test material (EC (no data) name): ammonium chloride no data Nitrification 4 (not assignable) Painter, H.A., (1970) supporting study

Test material (EC name): ammonium chloride Nitrification 4 (not assignable) Fisher, T., et al (1956) supporting study

Test material (EC name): ammonium chloride

The information mentioned above does not provide reliable results. In general, it can be stated that ammonium chloride is an inorganic substance, for which a general waiver is applied for this endpouint, see below.

Data waiving for diammonium tetrachlorozincate

Reason: other justification Justification: Biodegradation is not applicable to metals/inorganic substances, study does not need to be conducted if substance is inorganic (Annex VII of REACH regulation).

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 55 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

4.1.2.1.3. Simulation tests (water and sediments)

Data waiving

Reason: study scientifically unjustified

Justification: Biodegradation is not applicable to metals/inorganic substances, study does not need to be conducted if substance is inorganic (Annex VII of REACH regulation).

4.1.2.1.4. Summary and discussion of biodegradation in water and sediment

Discussion (screening testing)

Discussion (simulation testing)

4.1.2.2. Biodegradation in soil

Data waiving

Reason: other justification

Justification: Biodegradation is not applicable to metals/inorganic substances, study does not need to be conducted if substance is inorganic (Annex VII of REACH regulation).

4.1.3. Summary and discussion of degradation

Abiotic degradation

Waived: Inorganic substance: According to Annex IX of REACH Regulation, information on hydrolysis is not required for inorganics.

Also the other endpoints under 4.1. e. g; phototransformation in air, water and soil, are not applicable to the substance

Biotic degradation

Biodegradation is not applicable to metals/inorganic substances. Tests are not to be conducted if the substance is inorganic (Column 2 of Annex VII of REACH regulation)

For water, information is available on the removal of metals from the water column (given under 5.6.). The removal from the water column was modelled referring to the EUSES model parameters and different conditions of pH. Zinc is removed by > 70% under the reference conditions for the EU regional waters (EUSES) (see section 5.6.: "removal from the water column").

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 56 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

4.2. Environmental distribution

The environmental fate and release of zinc and zinc compounds has been discussed extensively in the RAR (ECB 2008). Environmental distribution in water Zinc in freshwater or seawater can occur in both suspended and dissolved forms and is partitioned over a number of chemical species. Depending on the concentration of suspended matter, about 25-40% of the zinc entering the surface water is in dissolved form, the remaining part is bound to the suspended matter. For toxicity, only the fraction not bound is important. Dissolved forms of zinc in freshwater are e.g.: hydrated zinc ions, zinc ions complexed by inorganic or organic ligands (humic and fulvic acids), zinc oxy ions and zinc adsorbed to solid matter (RAR 2008).

Possible chemical forms of zinc in seawater are presented in the table below. In this table the variation in the percentages of total zinc can for instance be explained by analytical differences or by the different ion strengths of the examined seawaters.

Table 14. (taken from the RA zinc, ECB 2008): Possible chemical forms (speciation) of dissolved zinc in seawater (Cleven et al., 1993). Percentage of total zinc Zn species Reference 1 Reference 2 Reference 3 Reference 4 Zn2+ 17 16.1 12.5 5.7 2-n ZnCln (n:1-4) 11.4 63.7 79 17.8 + ZnOH , ZN(OH)2 62.2 2.3 0.6 71.8

ZnCO3 6 3.3 1.6 2.4 + ZnHCO 3 0.7 0.3 - 0.2 ZnOHCl - 12.5 - -

ZnSO4 4 1.9 1.6 2.2

The speciation of zinc in the aquatic compartment is of high complexity and depends highly on abiotic factors, such as pH, (dissolved) organic matter content, redox potential, etc. It is assumed that speciation is very relevant for the migration of zinc through sediment, for the distribution of zinc among its truly dissolved and non- dissolved forms, and for the uptake of zinc by some aquatic and sediment organisms. The relationship between physicochemical factors driving the speciation of zinc in water, and the bioavailability, and consequently, the toxicity of Zinc has been experimentally elucidated and has been quantified in the biotic ligand model (BLM) (see further).

Environmental distribution in soil; adsorption/desorption of zinc in soil Speciation of zinc in soil

The speciation of zinc in soils has been extensively reviewed in the EU risk assessment on zinc (ECB 2008). The following is being summarised from the risk assessment: in soils, zinc interacts with various reactive soil surfaces. The most important in this respect are soil organic matter, amorphous soil oxides (Al, Fe, Mn) and clay minerals. The major process by which metals are bound to these surfaces is adsorption. Other processes including precipitation of carbonate type minerals can occur but are, in non- and moderately polluted soils, unlikely to control the solubility of metals in soils. An exception to this is the formation of sulphide minerals that are formed, in the presence of sulphate under reducing conditions. Zinc in soil is distributed between the following fractions (ECB 2008): 1. Dissolved in pore water (which includes many species) 2. Exchangeable, bound to soil particles

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 57 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

3. Exchangeable, bound to organic ligands (of which a small part in the dissolved fraction and the major part in the solid fraction) 4. Present in secondary clay minerals and metal oxides/hydroxides 5. Present in primary minerals

So, zinc is present in the soil in various forms, that have varying degree of extractability. The soil pH is an important parameter that affects the speciation and the distribution of the zinc species over the soil and the solution. Zinc tends to be more sorbed and complexed at higher pH (pH > 7) than at lower pH. Below pH 7, the amount of zinc in solution was reported to be inversely related to soil pH (Janssen et al., 1997). The pH of the soil not only determines the degree of complexation and adsorption of zinc, but also the solubility of the various zinc minerals. The solubility of zinc in soil decreases with increasing pH (Cleven et al., 1993). After addition of a metal to a soil, often a slow decrease in the soil solution concentration, or the available fraction as determined in an extraction solution (e.g. by CaCl2) decreases as a result of (presumably) slow diffusion processes of metals into the matrix of the reactive surfaces. It is this process, or sum of as of yet poorly defined slow processes, that can be defined as ‘ageing’ (ECB 2008). The challenge is to develop models that scale from the theoretical and laboratory level to the field scale. Following an extensive discussion in the risk assessment process, an integrative research program has been conducted aiming to reveal the relevant information required for using bioavailability corrections within the framework of the terrestrial risk assessment. The various relationships between on the one hand abiotic soil parameters and on the other hand the toxicity of zinc to plants, invertebrates and microbial endpoints were used to develop “soil sensitivity” functions i.e. relationships that express the potential toxicity of zinc in various soil types as a function of soil characteristics. Long-term distribution of zinc in soil was also recognised as an important process that affects the distribution of zinc and bioavailability in soil and toxicity towards soil species. Based on recent studies and a recent evaluation of older studies, the ‘ageing’ phenomenon was also quantitatively taken into account in the EU RAR (ECB 2008).

4.2.1. Adsorption/desorption

Data waiving

Reason: study scientifically unjustified

Justification: For metals, adsorption/desorption translates in the distribution of the metals between the different fractions of the environmental compartment, e. g. the water (dissolved fraction, fraction bound to suspended matter), soil (fraction bound or complexed to the soil particles, fraction in the soil pore water...). This distribution between the different compartments is translated in the partition coefficients between these different fractions. information on partition coeficients is given under section 5.6. of IUCLID.

Discussion

For metals, adsorption/desorption translates in the distribution of the metals between the different fractions of the environmental compartment, e. g. the water (dissolved fraction, fraction bound to suspended matter), soil (fraction bound or complexed to the soil particles, fraction in the soil pore water...). This distribution between the different compartments is translated in the partition coefficients between these different fractions. Study records on partition coeficients are given under section 5.6. of IUCLID.

Partition coefficients for zinc in freshwater has been reviewed in the RAR (ECB 2008). Based on the extensive experimental evidence, a partition coefficient for the distribution between solid particulate matter and water (Kpsusp) of 5.04 (log value) has been defined for EU waters and used throughout the RAR.

The Kp for the distribution between sediment and water (Kpsed) was estimated in the RAR from that for particulate matter, as follows: Kpsed= Kpsusp/ 1.5, based on the average difference in concentrations of zinc and other metals in both media. For zinc this results in a Kpsedof 73,000 l/kg. (ECB 2008)

The marine Kd was derived based on data from several marine waters. the geomean value for zinc in seawater is 6010 l/kg

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 58 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

For soil, a solids-water partitioning coefficient of 158.5 l/kg (log value 2.2) was determined experimentally on 11 American soils. This value was used in the RA Zinc (ECB 2008).

The following information is taken into account for any environmental exposure assessment:

Kp for solid particulate matter and water (Kpsusp): 110000 l/kg (log value: 5.04) (ECB 2008) Kp for water and sediment (Kpsed); 73000l/kg (log value:4.86) (ECB 2008) Kd for marine waters is 6010 l/kg (log value: 3.78) Kd for solids-water in soil is 158.5 l/kg (log value: 2.2)

4.2.2. Volatilisation

Not relevant 4.2.3. Distribution modelling

4.2.4. Summary and discussion of environmental distribution

For metals, the transport and distribution over the different environmental compartments e. g. the water (dissolved fraction, fraction bound to suspended matter), soil (fraction bound or complexed to the soil particles, fraction in the soil pore water...) is described and quantified by the metal partition coefficients between these different fractions. Information on these partition coefficients is given under section 5.6. of IUCLID.

Partition coefficients for zinc in freshwater have been reviewed in the RAR (ECB 2008). Based on this experimental evidence, a partition coefficient for the distribution between solid particulate matter and water (Kpsusp) of 5.04 (log value) has been defined for EU waters and was used throughout the RAR.

These partition coefficients have been used since then in other legislative processes in the EU (e. g. the water framework directive) and will also be used for REACH.

For the marine water, a partition coefficient water/suspended matter of 6010 l/kg has been derived.

For soil, a solids-water partitioning coefficient of 158.5 l/kg (log value 2.2) was determined experimentally on 11 American soils. This value was used in the RA Zinc.

4.3. Bioaccumulation

Due to homeostatic control mechanisms, bioaccumulation is not relevant to essential elements in general and to zinc in particular. In experimental work, high BCF factors are observed generally at the lowest zinc exposure levels, due to the fact that organisms will concentrate zinc to satisfy internal physiological needs for the essential element. For the same reason of homeostasis, the BCF will strongly decrease when exposure concentrations increase. This results in a general negative relationship between BCF and exposure (McGeer et al 2003).

On bioaccumulation, the EU risk assessemnt report (ECB 2008) concludes that“it is concluded that secondary poisoning is considered to be not relevant in the effect assessment of zinc. Major decision points for this conclusion are the following. The accumulation of zinc, an essential element, is regulated in animals of several

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 59 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 taxonomic groups, for example in molluscs, crustaceans, fish and mammals. In mammals, one of the two target species for secondary poisoning, both the absorption of zinc from the diet and the excretion of zinc, are regulated. This allows mammals, within certain limits, to maintain their total body zinc level (whole body homeostasis) and to maintain physiologically required levels of zinc in their various tissues, both at low and high dietary zinc intakes. The results of field studies, in which relatively small differences were found in the zinc levels of small mammals from control and polluted sites, are in accordance with the homeostatic mechanism. These data indicate that the bioaccumulation potential of zinc in both herbivorous and carnivorous mammals will be low.

4.3.1. Aquatic bioaccumulation

The studies on aquatic bioaccumulation are summarised in the following table:

Table 15. Overview of studies on aquatic bioaccumulation for zinc chloride

Method Results Remarks Reference Palaemon elegans (crustaceae) BCF: 28960 (whole body d.w.) 2 (reliable with Rainbow PS and (steady state) restrictions) White SL. (1989) aqueous (saltwater) BCF: 2558 (whole body d.w.) key study semi-static (steady state) read-across based on Total uptake duration: 28 d BCF: 843 (whole body d.w.) grouping of (steady state) substances (category Total depuration duration: 1 min approach) BCF: 277 (whole body d.w.) The decapod Palaemon elegans (steady state) Test material was exposed to sublethal (IUPAC name): zinc concentrations of zinc over 28 BCF: 123 (whole body d.w.) chloride (See days time period; total zinc (steady state) endpoint summary accumulation was measured. for justification of BCF: 38 (whole body d.w.) read-across) (steady state) Echinogammarus pirloti BCF: 60960 (whole body d.w.) 2 (reliable with Rainbow PS and (steady state) restrictions) White SL. (1989) aqueous (saltwater) BCF: 5658 (whole body d.w.) key study semi-static (steady state) read-across based on Total uptake duration: 21 d BCF: 2024 (whole body d.w.) grouping of (steady state) substances (category Total depuration duration: 1 min approach) BCF: 819 (whole body d.w.) The amphipod Echinogammarus (steady state) Test material pirloti was exposed to sublethal (IUPAC name): zinc concentrations of zinc over 21 BCF: 328 (whole body d.w.) chloride (See days time period; total zinc (steady state) endpoint summary accumulation was measured. for justification of BCF: 116 (whole body d.w.) read-across) (steady state) various wildlife species BAF: 4060 (whole body d.w.) 2 (reliable with Pascoe GA, (steady state) (aquatic restrictions) Blanchet RJ and feed and aqueous (freshwater) invertebrates: control water) Linder G (1996) key study field study BAF: 3483 (whole body w.w.) (steady state) (aquatic read-across based on Type of sediment: natural invertebrates, contaminated water) grouping of sediment

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 60 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 food chain analysis in a BAF: 2600 (whole body d.w.) substances (category contaminated natural (steady state) (snails, control approach) environment water) Test material BAF: 779 (whole body d.w.) (IUPAC name): zinc (steady state) (snails, (See endpoint contaminated water) summary for justification of read- BAF: 177 (whole body w.w.) across) (steady state) (fish, contaminated water)

Data waiving for ammonium chloride

Reason: other justification

Justification: In accordance with Column 2 of REACH Annex IX, the bioaccumulation in aquatic species (required in section 9.3.2) does not need to be conducted as the substance has a low potential for bioaccumulation. Simple inorganic salts with high aqueous solubility will exist in a dissociated form in an aqueous solution. Such a substance has a low potential for bioaccumulation.

4.3.2. Terrestrial bioaccumulation

The results of terrestrial bioaccumulation studies are summarised in the following table:

Table 16. Overview of studies on terrestrial bioaccumulation

Method Results Remarks Reference various wildlife species BCF: 3.3 (whole body d.w.) 2 (reliable with Pascoe GA, (steady state) (grasshoppers, restrictions) Blanchet RJ and food chain analysis in a control soil) Linder G (1996) contaminated natural key study environment BCF: 0.14 (whole body d.w.) (steady state) (grasshoppers, read-across based on contaminated soil) grouping of substances (category BCF: 1.84 (whole body d.w.) approach) (steady state) (earthworms, control soil) Test material (IUPAC name): zinc BCF: 0.54 (whole body d.w.) (See endpoint (steady state) (earthworms, summary for contaminated soil) justification of read- across) BCF: 0.015 (whole body d.w.) (steady state) (small mammals, contaminated soil)

BCF: 0.27 (whole body d.w.) (steady state) (above ground fodder, control soil)

BCF: 0.11 (whole body d.w.) (steady state) (above ground

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 61 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

fodder, contaminated soil)

BCF: 0.66 (whole body d.w.) (steady state) (below ground fodder, control soil)

BCF: 0.21 (whole body d.w.) (steady state) (below ground fodder, contaminated soil)

BCF: 0.73 (whole body d.w.) (steady state) (above ground grasses, control soil)

BCF: 0.079 (whole body d.w.) (steady state) (above ground grasses, contaminated soil)

BCF: 1.5 (whole body d.w.) (steady state) (below ground grasses, control soil)

BCF: 0.45 (whole body d.w.) (steady state) (below ground grasses, contaminated soil)

4.3.3. Summary and discussion of bioaccumulation

Aquatic bioaccumulation

Ammonium:

Simple inorganic salts with high aqueous solubility will exist in a dissociated form in aqueous solution. Such a substance has a low potential for bioaccumulation.

Zinc:

Bioaccumulation is not considered relevant for essential elements because of the general presence of homeostatic control mechanisms.

McGeer et al (2003) recently extensively the reviewed evidence on bioconcentration and bioaccumulation of zinc as a function of exposure concentration in a number of taxonomic groups (algae, molluscs, arthropods, annelids, salmonid fish, cyprinid fish, and other fish). The data clearly illustrated that internal zinc content is well regulated. All eight species taxonomic groups investigated exhibited very slight increases in whole body concentration over a dramatic increase in exposure concentration. In fact, most species did not show significant increases in zinc accumulation when exposure levels increased, even when exposure concentrations reached those that would be predicted to cause chronic effects. This suggests that adverse effects related to Zn exposure are independent of whole body accumulation. Due to the general lack of increased whole body and tissue concentrations at higher exposure levels, the zinc BCF data showed an inverse relationship to exposure concentrations. In all cases, the relationship of BCF to exposure was significant and negative. The slopes of the BCF/BAF – exposure relations were: algae: -1.0, insects: -0.79, arthropods: -0.73, molluscs: -0.83, salmonids: -0.92, Centrarchids: -0.80, Killifish: -0.84, other fish: -0.87. Overall, species mean slope was -0.85 +/- 0.03 (McGeer et al 2003).

The physiological basis for the inverse relationship of BCF to zinc exposure concentration arises from Zn uptake and control mechanisms. At low environmental zinc levels, organisms are able to sequester and retain Zn in tissues for essential functions. When Zn exposure is more elevated, aquatic organisms are able to control uptake. There is clear evidence that many species actively regulate their body Zn concentrations, including crustaceae, oligochaetes, mussels, gastropods, fish, amphipods, chironomids by different mechanisms (McGeer

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 62 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 et al 2003).

The following information is taken into account for any hazard / risk / bioaccumulation assessment:

Zinc is an essential element which is actively regulated by organisms, so bioconcentration/bioaccumulation is not considered relevant.

Terrestrial bioaccumulation

Bioaccumulation is not considered relevant for essential elements because of the general presence of homeostatic control mechanisms. the data from a field food chain transfer study indicate that bioconcentration of zinc is indeed very low. It is in all cases also lower in contaminated soil, as compared to control soil.

The following information is taken into account for any hazard / risk / bioaccumulation assessment:

Zinc is an essential element which is actively regulated by organisms, so bioconcentration/bioaccumulation is not considered relevant.

4.4. Secondary poisoning

Based on the available information, there is no indication of a bioaccumulation potential and, hence, secondary poisoning is not considered relevant (see CSR chapter 7.5.3 "Calculation of PNECoral (secondary poisoning) ".

Justification for no PNEC oral derivation: Zinc is an essential element that is actively regulted within the body of all organisms. Due to the general lack of increased whole body and tissue concentrations at higher exposure levels, the zinc BCF data show generally an inverse relationship to exposure concentrations (McGeer et al 2003). The physiological basis for the inverse relationship of BCF to zinc exposure concentration arises from Zn uptake and control mechanisms. At low environmental zinc levels, organisms are able to sequester and retain Zn in tissues for essential functions. When Zn exposure is higher, aquatic organisms are able to control uptake. There is clear evidence that many species actively regulate their body Zn concentrations, including crustaceans, oligochaetes, mussels, gastropods, fish, amphipods, chironomids by different mechanisms (McGeer et al 2003). The bioaccumulation potential in mammals is also considered low. Based on this, the EU risk assessment concludes that secondary poisoning is considered to be not relevant in the effect assessment for zinc.

4.5. Natural background

4.5.1. Natural background in surface waters Data on natural background of zinc in surface waters in the EU were discussed in the RAR (ECB 2008). Based on values reported for a number of EU countries, incl. NL, D, F, SF and others, two values were selected for total background concentration, to be used in the risk characterisations: -3µg Zn/l which is the lower limit of the range reported by the member states in the risk assessment process, -12µg Zn/l which is the geometric mean value of an extensive EU database on Zn concentrations in lowland brooks in unpolluted areas in N-Europe, (10P and 90P in this review are 4 and 35 µg total Zn/l resp. - Zuurdeeg 1999). -Using a Kpsusp of 110,000 l/kg (RA; ECB 2008) and a default particulate matter content of 15 mg/l, these values are equivalent to 4.44 µg dissolved Zn/l, and 1.11 µg dissolved Zn /l, respectively.

Due to geochemical differences, the natural background concentrations in surface waters will differ throughout Europe. It is thus not appropriate to set one value for the whole of the EU. Recently, an extensive database has become available on a.o. zinc background values in surface waters, covering the whole of the EU (FOREGS 2005). The 10P and 90P values for dissolved zinc following from this analysis performed also on non-polluted head waters (cfr Zuurdeeg 1999) are 0.7 and 18 µg dissolved zinc /l, resp. (http://www.gtk.fi/publ/foregsatlas/maps/Water/w_icpms_zn_edit.pdf); the median value of 2.7 µg dissolved

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 63 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Zn/l corresponds well with the one used in the RAR (ECB 2008).

The FOREGS study thus confirms the data reported in the EU RAR and the Zuurdeeg (1999) analysis. It confirms that the value of 12µg total Zn/l – 4.44 µg dissolved Zn/l can be seen as the median value for zinc background in European freshwaters.

Regarding the sediment, the RAR concluded, based on reported data from a number EU countries, incl NL, D, F, N, SF, SW, that the range of the background data for sediment was more or less in the same order of magnitude (range 70-175 mg/kg dwt). Based on the data from several EU-regions the value of 140 mg/kg dwt was set for use as a natural background for correcting the EU sediment monitoring data.

It was noted that, if available monitoring data can unequivocally be linked with a particular natural background value in an area, preference should be given to that specific background value (ECB 2008). For comparison, FOREGS (2005) reports for zinc in sediments 10P and 90P values of 18 and 270 ppm, resp., demonstrating the big variation, due to local geology. The median value is 65 ppm. Because of the great variability, more localised assessments are preferred.

4.5.2. Natural background in the terrestrial compartment The natural zinc concentrations in soils are highly variable and dependent on soil type and soil properties. The EU RAR concluded from the available soil data for a number of EU countries that there is a large variation in the natural zinc background concentrations. This variation is related to the native soil material and the present soil characteristics like humus and lutum. The RAR observed a clear relationship between natural background levels and various soil parameters, but noted that a quantification of the exact natural background level for a specific EU soil type is at present still an extremely difficult and complex issue. The RAR mentioned zinc backgrounds usually in the range 50-100 mg Zn/kg DW, which is confirmed by data from the more recent FOREGS database, reporting 10P, 50P and 90P zinc concentrations of 8, 52, and 140 mg/kg DW, respectively in unpolluted soils throughout Europe. Important to note is that, due to the high variability on the data, the RAR concluded that it was only possible to use monitored soil data for risk characterisation when it was possible to make a correction with the natural zinc background concentration(s) typical for that soil type.

4.6. Additional information on environmental fate and distribution

Transformation dissolution For diammonium tetrachlorozincate(2-), no transformation/dissolution tests have been done, given its high solubility.

Removal from the water column The removal of zinc ions from the water column was assessed by using the “Unit World Model” (UWM), a screening level model used to explicitly assess the effects of chemical speciation on metal partitioning, transport and bioavailability in the water column and underlying sediments. Specific processes considered in the UWM include 1) dissolved and particulate phase transport between the overlying water and sediment, 2) metal binding to inorganic ligands, dissolved and particulate carbon (using WHAM V), and iron hydroxides in the water column, 3) metal precipitation, 4) dissolution kinetics for metal powders, massives, etc., and 5) average-annual cycling of organic matter and sulfide production in the lake (Mutch Associates 2010b).

The numerical engine for the model calculations is the Tableau Input Coupled Kinetics Equilibrium Transport (TICKET) model (Farley et al., 2008). As parameters for the calculation, reference was made to the conditions as prescribed by ECHA (2009) for use with the European Union System for the Evaluation of Substances (EUSES) to carry out rapid and efficient assessments of the general risks posed by chemical substances.Zinc removal was evaluated at different water chemistries relative to the REACH Implementation Project (RIP) 3.6 definition of rapid removal for soluble metals of greater than 70% removal in a 28-day period. The model was used to calculate the removal of zinc ions for a standard EUSES lake (“base case”). Additional sensitivity analysis was applied to check on the removal with varying conditions, e.g. pH, zinc concentration (the initial zinc concentrations in the water column were specified based upon reference zinc toxicity levels at different pH), settling velocity. Finaly, some “real world” experimental evidence is discussed.

The removal of zinc in the base case is described in figure below.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 64 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

1.2

0.8 0 C

/ 0.6 C

0.4

0.2

0 0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 Time (days)

TICKET-UWM 30% Remaining

Zinc removal from the water column is rapid: 70% zinc removal is achieved within 3 days of dosing. Based on the suspended solids concentration of 15 mg/L and the log KD of 5.04, approximately 62% of the zinc in the water column is associated with settling particles. After 28 days, the total zinc concentration in the water column is 1.74 µg/L.

The following sensitivity analysis was done (Mutch Associates 2010b):  When decreasing settling velocity from the EUSES value of 2.5 m/d to 0.24 m/d, the zinc removal rate is decreased, but the rapid removal benchmark is still met.  Varying the hydraulic residence time between 300 years and 40 days (EUSES value) has a minimal effect on zinc removal. The rapid removal benchmark is met within 3 days.

 The output from the TICKET-UWM indicates that the WHAM V-computed log KD varies over the course of the 28-day simulation between 4.32 and 4.55. This variation is associated with decreasing water column zinc concentrations (partitioning in WHAM V not necessarily linear). This range of log

KD values is lower than the zinc risk assessment document value of 5.04. As a result of the generally lower log KD (and associated lower fPart values), zinc is removed from the water column at a slower rate but the rapid removal benchmark is still achieved (in approximately 5.5 days).  When the standard EUSES lake parameters are applied, but pH is varied between 6 and 8, the removal rate is basically similar under all 3 conditions, and zinc is removed > 70% within 28 days (Mutch Associates 2010a).

Experimental evidence on the removal of zinc from is available from the work of Hart et al, 1992. The concentration of zinc added to large (40m3) in situ enclosures of natural water, containing natural phytoplankton community, was followed over >40 days. In this system, the total zinc fraction, the particulate zinc fraction and the ion-exchangeable zinc fraction all decreased steadily over the study period. 70% removal of zinc in the 3 fractions was observed after 11-12 days. A mass balance indicated that most of the zinc ended up in the sediments (84%), about 6% remained in the water column and about 10% was associated with wall epiphytes (Hart et al 1992).

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 65 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Further experimental evidence for the removal of zinc from the water column is found in a recent freshwater microcosm study (Rand et al 2012). In this study, a significant decrease of dissolved zinc in the microcosm water column was monitored over a period of 4 days. Extrapolation of the data indicated that zinc would be removed >> 70% within 28 days.

Re-mobilisation of zinc from the sediment is prevented by binding of zinc to the sulphide fraction to form insoluble ZnS. Due to the insolubility of the ZnS (K=9.2 x 10-25) zinc will be effectively sequestered in the (anaerobioc) sediments.

In the systems described above, zinc was added as soluble salt that dissolves instantaneously. This is in contrast to e.g. metal powders or powders from insoluble zinc compounds which may dissolve at slower rates, may be only sparingly soluble, and, depending on particle size and density, may be subject to rapid settling. Use of a soluble zinc salt in the TICKET-UWM simulations, therefore, represents a worst-case scenario for metal release and persistence in the water column.

In conclusion, model calculations supported by experimental field evidence show that zinc is rapidly removed from the water column, at a rate of > 70% within 28 days. This result has implications for the classification for aquatic toxicity.

5. HUMAN HEALTH HAZARD ASSESSMENT General considerations This chemical safety assessment and chemical safety report focuses on zinc metal and ten zinc compounds (i.e., zinc oxide-ZnO; zinc hydroxide-Zn(OH)2; zinc phosphate-Zn3(PO4)2; zinc carbonate-ZnCO3; zinc sulphide-ZnS; zinc sulphate-ZnSO4; zinc chloride–ZnCl2; diammonium tetrachlorozincate– ZnCl2/2NH4Cl; triamonium pentachlorozincate- ZnCl2/3NH4Cl; zinc bis(dihydrogen phosphate)-Zn(H2PO4)2). The zinc compounds have been grouped into three categories on the basis of their water solubility as described in Table below:

Table 17. Water solubility values of the eleven zinc compounds covered in this CSR Zinc compound Water solubility Water solubility in mg/L2 Ranking of solubility in mg/L1 6 6 ZnCl2 4.3 x10 0.851x10 Soluble 6 6 ZnSO4 0.22x10 0.210x10 Soluble 4 6 Zn(H2PO4)2 >1x10 >1 x10 Soluble 4 6 ZnCl2/2NH4Cl >1x10 0.291 x10 Soluble 4 6 ZnCl2/3NH4Cl >1x10 0.155 x10 Soluble

Zn(OH)2 1.6 648 Slightly soluble ZnO 1.6 2.9 Slightly soluble

Zn 3(PO4)2 0.1 2.7 Slightly soluble

ZnCO3 <0.2 1.3 Slightly soluble Zn metal <0.1 0.1 Slightly soluble ZnS <10 0.00045x10-3 Insoluble 1Values are taken from ESIS database http://ecb.jrc.ec.europa.eu/esis/; ATSDR 2005; EU RARs (EU RAR, 2004a-f) 2Values are taken from section 4 of the IUCLID files on the respective substances. Data by Outotec Research Oy, Pori, Finland.

Collectively this group of zinc compounds is considered “data rich” with a multitude of information available in the public domain regarding the effects of zinc compounds on human health. The wealth of available toxicological data has been carefully reviewed and scrutinised by the Rapporteur in the framework of the discussions on the EU Risk Assessment Reports (RAR) developed according to EU Regulation 793/93/EEC (EU RAR, 2004a-f). The Rapporteur’s analysis of the available toxicological data was extensively discussed by the experts from

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 66 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Member States and other stakeholders during the meetings of the “Technical committee on new and existing substances” (TCNES), during which the relevant data sets were approved. Therefore, the data used in the RARs (EU RAR, 2004a-f) will be the main data source for this chapter. Decisions on data quality and relevancy approved by TCNES will be used as in the EU risk assessment process. Consequently, the current analysis will focus on the data considered useful for deriving the MOS in the RARs as such (EU RAR, 2004a-f). Also, the data considered not useful in the EU risk assessment process will also not be used in the current analysis. Given the substantial amount of data, only pertinent data in the IUCLID5 files have been included in this CSR. Data which were considered not sufficient during the EU risk assessment process have been summarised in this CSR, with reference to the author mentioned in the EU RARs for completeness. In addition, the dataset from the EU RARs have been complemented with relevant and reliable information that became available between 2005- 2009 (i.e., after the EU RARs were finalised in 2004). The additional data have also been reported and summarised in IUCLID5 as well as in the following subsequent sections. Assumptions Zinc exists in different chemical forms and the bioavailability of these forms depends on various physicochemical parameters of which water solubility is the main determining factor. It is accepted that the actual bioavailable concentration of the zinc cation in both animals and in humans is an important determinant of toxicity, and although there is information available on the various zinc compounds, adequate information is lacking on how to quantitatively determine or estimate the bioavailable fraction of all the different zinc compounds in either laboratory animals or humans (Windholz et al., 1983). Since water solubility is the main determinant of bioavailability, zinc compounds with similar solubility characteristics have been grouped in Table below and, where necessary, the local or systemic toxicity have been read-across within the same group of zinc compounds.

Table 18. Grouping based on water solubility Zinc compound Ranking of solubility Solubility-based grouping

ZnCl2 Soluble ZnCl2ZnSO4

ZnSO4 Soluble Zn(H3PO4)2

Zn(H3PO4)2 Soluble

ZnCl2/2NH4Cl Soluble ZnCl2/2NH4Cl

ZnCl2/3NH4Cl Soluble ZnCl2/3NH4Cl ZnO Slightly soluble ZnO

Zn(OH)2 Slightly soluble Zn(OH)2

Zn3(PO4)2 Slightly soluble Zn3(PO4)2

ZnCO3 Slightly soluble ZnCO3 Zn metal Slightly soluble Zn metal ZnS Insoluble ZnS As such, this section in the CSR makes an integrated case on the zinc compounds mentioned above and is relevant for all of them. For reasons of consistency, it was decided not to develop partial cases on separate zinc substances.

5.1. Toxicokinetics 5.1.1. Non-human information

Absorption In vitro dermal penetration studies

2 The dermal absorption of zinc 2-pyrrolidone 5-carboxylate, ZnO and ZnSO4 (16 mg formulation/cm ; 0.02 – 5.62% zinc) in different formulations (3 emulsions and 2 ointments) using human abdominal skin was investigated. The receptor medium was 0.9% NaCl. After application for 72 hours, the skin was washed and stripped twice. The percutaneous absorption was determined as a percentage of the applied dose found in receptor medium and cutaneous bioavailability. It never exceeded 2%. The percentages for the absorption of

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 67 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

zinc from ointments containing ZnO and ZnSO4 were 0.36% and 0.34%, respectively. The percutaneous absorption of zinc from the emulsion containing zinc 2-pyrrolidone 5-carboxylate was 1.60% of the applied dose. Furthermore the experiment showed a vehicle effect on absorption (Pirot et al., 1996a).

2 The dermal absorption of ZnSO4 and ZnCl2 (in 20 mg formulation/cm ) in petrolatum and hydrophilic gels using human breast or abdominal skin was also investigated. The receptor medium used was isotonic saline. After application for 72 hours, the skin was washed and the epidermis was removed from the dermis. The result showed that the absorption was low (i.e., ≤ 2%) regardless of the choice of vehicle (Pirot et al., 1996b). The utility of the data generated by Pirot et al., 1996a, 1996b, is limited due to the absence of membrane integrity measurements. Moreover, it is not clear whether the skin was occluded. The cutaneous bioavailability might be underestimated in the first study due to double stripping and in the second study, absorption is based on zinc in fresh dermis and receptor fluid, the fresh epidermis is not included. An industry in vitro percutaneous absorption testing programme on two representative zinc compounds (ZnO and ZnSO4) was conducted by (Grötsch, 1999). In this programme, a solution of ZnSO4 monohydrate and a suspension of ZnO, each at concentration of 40 mg/mL in water, were tested for cutaneous penetration and absorption through pig skin in vitro. Skin preparations measuring 1 mm in thickness with stratum corneum, stratum germinativum and blood-vessel-containing parts of the dermis were obtained from pigs using a modified dermatome. In two independent experiments for each compound seven skin preparations were mounted in Teflon flow-through diffusion chambers which were continuously rinsed with physiological receptor fluid (0.9% NaCl in aqua bidest with antibiotics). Following an integrity check using the marker substance caffeine, each of the test formulations were applied to six skins at a dose of 1 mg/cm2 for 8 hours without occlusion, and subsequently washed off with a neutral shampoo. After 0, 2, 4, 6, 8, 16, 24, 40, 48, 64 and 72 hours, the cutaneous permeation was determined by quantifying zinc with atomic absorption spectroscopic analysis (detection limit: 10 ng/mL) in the receptor fluid. The experiment was stopped at 72 hours. Zinc was analysed in the skin preparations and the rinsing fluids. In addition, blanks were measured in an unloaded control chamber. Results are summarized in Table below.

Table 19. Dermal absorption of Zn (% of dose) through pig skin in vitro within 72 hours

a a Soluble ZnSO4 Slightly soluble ZnO Receptor fluid 0.3 % 0.03 % Horny layer 1.3 % 12.3 % Residual skin 0 % 2.6 % Potentially absorbed dose 1.6% 14.9% a Corrected for background levels of zinc in receptor fluid and skin.

Total recoveries of applied zinc in both experiments ranged from 82.0 to 109.6%. The results of the analysis of the receptor fluid used and of the blank chambers without topical application of zinc compounds indicated that both the receptor fluid and porcine skin contain an intrinsic level of zinc. The amounts of zinc detected in receptor fluid and different layers of the skin were therefore corrected for background levels. The authors concluded that dermal penetration of zinc was below 1% based on the cumulative amount recovered from the receptor fluid at 72 hours. However, the amount retained in the skin should be regarded as being absorbed because it may become available at a later stage. Hence, the rapporteur concluded that the dermal absorption of zinc from a solution of ZnSO4 monohydrate and a suspension of ZnO in this in vitro system may amount to 1.6% and 14.9%, respectively (Grötsch, 1999). Animal studies Oral Zinc acetate was added to the diet of Sprague-Dawley rats (9/group) to reach zinc concentrations of 58 (no zinc acetate added; normal zinc concentration in “control” feed), 117, 175, 293, 410 or 664 mg/kg via the feed, corresponding to ca. 3, 6, 9, 14.5, 20.5 or 33 mg Zn/kg bw. After 28 days, the unfasted animals were dosed with 65 1.2 Ci of ZnCl2 (ca. 0.15 ng). Whole-body radioactivity was determined at various time points up to 11 days post-dosing using a whole-body gamma counter. In the group which received the non-supplemented diet (i.e., 58 mg Zn/kg feed) ca. 20% of the administered radioactivity was retained at 24 h post-dosing which gradually decreased to about 9% at day 11. The amount of radioactivity retained at 24 h post-dosing declined with increasing dietary zinc levels to about 13% for the group with the highest dietary zinc. In this group after 11 days only ca. 2.3% of the administered radioactivity was left. The data indicated that low dietary zinc intake results in increased zinc retention and that at higher dietary zinc levels absorption of zinc is reduced (Furchner and Richmond, 1962).

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 68 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

After a pre-exposure period of 7 days, male Wistar rats, kept on a semi-synthetic diet, were dosed orally with 86 65 - 130 g Zn as ZnCl2 (n=15), ZnCO3 (n=15) or Zn5(OH)8Cl2H2O (n=20) added to a test meal. It was assumed that during the first 5 days post-dosing non-absorbed zinc was excreted via the faeces. Absorption of labelled zinc was calculated from in vivo whole-body gamma counting results over the period 5-14 days post-dosing.

The uptake was calculated to be 40, 45 or 48 % for Zn5(OH)8Cl2H2O, ZnCl2 and ZnCO3, respectively (Galvez- Morros et al., 1992). Inhalation The rate or percentage of absorption of zinc following inhalation exposure is not available, but there are several studies investigating the zinc retention in the lung. Male and female rats were exposed to 15 mg ZnO dust/m3 (particle size < 1 m) for 4 hours/day, 5 days/week during 1 day or for 2, 4 or 8 weeks. Animals were killed 24 hours after the last exposure and the zinc content of the lungs, liver, kidneys, tibia and femur was measured. After 1 day of exposure the total zinc content of the lung in males and females were approximately 46 and 49 g, respectively. In the lung, liver, kidney and bone only minimal differences in tissue zinc content was seen during the experiment. As tissue zinc levels in non-treated animals were not studied, it is not clear whether tissue zinc comes from the experimental or from dietary exposure. However, as the pulmonary zinc level did not rise throughout the study it can be assumed that pulmonary deposition is very low and/or that pulmonary clearance of zinc particles is very high (Pistorius et al., 1976). In another experiment, following exposure to 4.3 mg (rat), 6.0 mg (rabbit), 11.3 mg (guinea pig) mg ZnO (aerosol)/m3 (aerosol mass median diameter was 0.17 m) for 2-3 hours, the pulmonary retention in rats, rabbits and guinea pigs was determined to be 11.5%, 4.7% and 19.8%, respectively (Gordon et al., 1992). The lung clearance rate of zinc aerosols was determined in male Wistar rats (8/group) 0, 2, 4, 8 and 24 hours after exposure to ZnO aerosol at a concentration of 12.8 mg/m3 (mean aerodynamic diameter of 1 m) for 17 hours. The ZnO aerosol was created by pyrolysis of a micronized zinc acetate aerosol at 500 o C. Eight animals were kept in clean air and served as controls. The lungs and trachea of the animals were removed and their zinc content was determined by flame photometry. In comparison with the controls, the lungs of exposed rats were increased in weight (presumably because of oedema), of which the increase was significant at 8 hours and even more pronounced at 24 hours. The zinc content in the trachea was not uniform but was above control values except after 24 hours. The zinc content in the lungs decreased mono-exponentially and was 7% of the initial burden after 24 hours. According to the short half-life of 6.3 hours found in this study for the pulmonary zinc content, a fast dissolution of the particles must occur, as the alveolar clearance of an inert Fe 2O3 aerosol occurred with a half-life of about 34 hours. It was not clear whether the clearance of Zn particles from the lungs was affected by the pathological condition of the lungs (Oberdörster et al., 1980). Intratracheal instillation In a time course experiment male Wistar rats (3/group) received a single intratracheal instillation of 0.4 ml ZnO suspension (i.e., ZnO particles < 2 m; particles appeared to form aggregates of 10-20 particles) at a dose of 100 g Zn/rat and the rats were killed 1, 2, 3, 5, 7, 14 and 21 days after administration. In a dose-response experiment 0.4 ml ZnO suspension (ZnO particles < 2 m, but they appeared to form aggregates of 10-20 particles) was instilled in the lungs of male Wistar rats (3/group) at doses of 20, 50, 100, 200, 500 and 1000 g Zn/rat. The rats were killed after 2 days. Control animals were included in the experiments. A significant increase in lung wet weight 1 day after instillation remained throughout the study. Only a limited portion of zinc could be retrieved in the bronchoalveolar lavage fluid (BALF). No measurable amount of exogenous zinc was observed after 5 days. The half-life of ZnO instilled in the lung was calculated to be 14 hours. In the dose- response experiment, the lung wet weight increased with increasing dose of ZnO, two days after instillation. The results indicated that the rat lung was able to clear ZnO particles up to a dose of 50 g Zn/rat at least within two days. No measurable accumulation of zinc was observed in the liver and kidneys even at a dose of 1000 g Zn/rat (Hirano et al., 1989). Dermal

65 The percutaneous uptake of ZnCl2 by the dorsal skin of the guinea pig was estimated by monitoring the 65 decline of radioactivity emitted by ZnCl2 in at least 10 trials for each concentration tested ranging from 0.8 to 4.87 M ZnCl2 (pH 1.8-6.1). It appeared that the loss of radioactivity after 5 hours was less than 1% except for the trials with the lowest pH where it might have been between 1 and 2%. The study gives too little details to be used for risk assessment as cited in EU 2004, a, b, c, d, e, f (Skog and Wahlberg 1964). Zinc oxide, zinc omadine, zinc sulphate and zinc undecylenate (131 Ci/mole of 65Zn) were topically applied to shaved skin on the back of rabbits. Each application consisted of 2.5 mg zinc compound containing 5 Ci 65Zn.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 69 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Two animals received one application on four skin areas left of the spine, while the four skin areas on the right side received two applications, the second one 24 hours after the first one. The rabbits were killed 6 and 24 hours after the second application. One rabbit served as the control. No significant differences were found in the amount and location of 65Zn in skin treated with 4 different zinc compounds. High concentrations of 65Zn were observed in the cortical and cutical zones of the hair shaft, being the highest in the keratogenous zone. Accumulation of 65Zn in epidermis was very low but heavy in the subdermal muscle layer. No difference in the rates of absorption and concentrations of zinc compounds with different oil/water solubility, pH, and molecular weight were seen. It was therefore suggested that the major mode of 65Zn uptake in skin is by diffusion through the hair follicles due to the heavy localization of 65Zn primarily in the hair shaft and hair follicles. According to the author, this emphasizes that chemical differences in the compounds may not play a very important role in the skin uptake of 65Zn. No data were given on systemic absorption (Kapur et al., 1974).

65 The dermal absorption of Zn from ZnCl2 and ZnO was studied by applying zinc preparations under occlusion on the shaven intact skin on the back of male Sprague-Dawley rats. The zinc absorption, being the ratio between 65Zn-activity in the carcass, liver and gastrointestinal tract, and the 65Zn-activity in carcass, liver, gastrointestinal tract, skin and bandage, was reported to range from 1.6 to 6.1%. It should be noted that the higher percentages (3.6 to 6.1%) were achieved after application of ZnCl2 in acidic solution (pH = 1). Less acidic solutions with ZnCl2 or with ZnO resulted in a dermal absorption of less than 2%. In this study, only the absorption into the body, excluding the skin, was determined. No data were available as to the effect of ZnCl 2 solutions with pH = 1 on dermal integrity (Hallmans and Lidén, 1979).

Topical application of ZnCl2 in an oil vehicle to pregnant Sprague-Dawley rats which were fed a zinc-deficient diet for 24 hours resulted in an increase in plasma concentration of zinc cations to normal or slightly above normal levels. The absorbed fraction was not determined therefore it can be concluded that dermal absorption is possible but no quantification can be given (Keen and Hurley, 1977). The application of ZnO dressings (containing 250 g Zn/cm2) to rats for 48 hours with full-thickness skin excision resulted in a 12% delivery of zinc ions from the dressing to each wound, while application of ZnSO 4 dressings (containing 66 g Zn/cm2) resulted in a 65% delivery of ions to each wound. The data suggest that the application of ZnO resulted in sustained delivery of zinc cations causing constant wound-tissue zinc cation levels due to its slow dissociation rate, while the more water soluble ZnSO4 delivers zinc ions more rapidly to the wound fluid with subsequent rapid transferral into the blood (Agren et al., 1991a). Distribution The highest levels of radioactivity were found in the small intestine followed by the kidney, liver and large 65 2+ intestine six hours after a single oral administration of 0.1 Ci of Zn as ZnCl2 to Wistar rats. Smaller amounts were found in the lungs and spleen. Fourteen days after administration, the highest levels of radioactivity were found in the hair, testicles, liver and large intestines (Kossakowski and Grosicki, 1983). Organs with high zinc concentrations (ranging from 20 to 60 mg/kg fresh weight) are liver, gut, kidney, skin, lung, brain, heart and pancreas as cited in EU RARs (Bentley and Grubb, 1991; He et al., 1991; Llobet et al., 1988). High concentrations of zinc were also detected in the retina and in sperm as cited in EU RARs (EU 2004, a, b, c, d, e, f; Bentley and Grubb, 1991). Metabolism As described in EU RARs, zinc is primarily bound to organic ligands rather than existing free in solution as a cation (Gordon et al., 1981). It is found in diffusible and non-diffusible forms in the blood. About 66% of the diffusible form of zinc in the plasma is freely exchangeable and loosely bound to albumin (Cousins et al., 1985).

A small amount of the non-diffusible form of zinc is tightly bound to 2-macroglobulin in the plasma and is not freely exchangeable with other zinc ligands. Zinc is incorporated into and dissociated from 2-macroglobulin only in the liver (Henkin et al., 1974). Excretion

65 65 After a single oral dose of 86 – 130 g of Zn as ZnCl2, ZnCO3 or Zn5(OH)8Cl2H2O, male rats eliminated Zn from the body with a rate of about 1.7% of the absorbed dose during day 5 to 14 post-dosing as determined from stool, urinary and in vivo whole-body gamma counting results. Male rats who received 25 mg ZnCO 3/kg feed or 65 100 mg Zn5(OH)8Cl2H2O/kg feed for 14 days, the radioactivity from a subcutaneous dose of 37 kBq of ZnCl2 disappeared from the body with a rate of approximately 1% during the period 5 to 14 days post dosing (Galvez- Morros et al., 1992). As described in EU RARs (EU RAR, 2004a-f) within certain limits, mammals can maintain the total body zinc

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 70 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 and the physiologically required levels of zinc in the various tissues, constant, both at low and high dietary zinc intakes. The sites of regulation of zinc metabolism are: absorption of zinc from the gastrointestinal tract, excretion of zinc in urine, exchange of zinc with erythrocytes, release of zinc from tissue, and secretion of zinc into the gastrointestinal tract. Regulation of gastrointestinal absorption and gastrointestinal secretion most likely contributes the most to zinc homeostasis. In spite of the mechanism for whole-body zinc homeostasis, a regular exogenous supply of zinc is necessary to sustain the physiological requirements because of the limited exchange of zinc between tissues ((EU RAR, 2004a-f). It has been hypothesized by Hempe and Cousins (1992) that zinc entering the luminal cells is associated with cysteine-rich intestinal protein (CRIP), a diffusible intracellular zinc carrier, and that a small amount is bound to metallothionein; however, as the luminal zinc concentration increases, the proportion of cytosolic zinc associated with CRIP is decreased and zinc binding to metallothionein is increased. CRIP binds 40% of radiolabelled zinc entering the intestinal cells of rats when zinc concentration is low; but only 14% when the concentration is high. Zinc is initially concentrated in the liver after ingestion, and is subsequently distributed throughout the body. When plasma zinc levels are high, liver metallothionein synthesis is stimulated, which facilitates the retention of zinc by hepatocytes (EU RAR, 2004a-f).

Table 20. Overview of experimental studies on absorption, metabolism, distribution and elimination Method Results Remarks Reference human (not applicable) Toxicokinetic parameters: 2 (reliable with Prasad AS , Beck male/female Cmax: 225.2 ± 45.5 µg/dL (Test No.: #1) restrictions) FWJ & Nowak J (1993) oral: capsule Tmax: 2.5 hr (Test No.: #1) key study AUC: ca. 800 ( for percent change in Exposure regime: plasma zinc) (Test No.: #1) read-across based on Single administration grouping of Metabolites identified: not measured substances (category Doses/conc.: Test approach) material equivalent to Details on metabolites: Not applicable 50 mg elemental zinc Test material Evaluation of results: One of the highest (IUPAC name): zinc Oral zinc tolerance test levels of absorption among all tested sulfate (See endpoint was performed in compounds summary for humans to study the justification of read- absorption of zinc from across) various zinc compounds including zinc sulfate. rat (Wistar) male Metabolites identified: not measured 2 (reliable with Kossakowski S & restrictions) Grosicki A (1983) oral: gavage Details on metabolites: Not applicable key study Exposure regime: Single administration read-across based on grouping of Doses/conc.: 0.1 µCi substances (category (3.7 kBq) of Zn (65) approach)

Distribution of zinc to Test material different organs (IUPAC name): zinc measured for up to 14 dichloride (See d after single oral endpoint summary administration of for justification of radiolabelled zinc read-across) chloride to male Wistar rats. rat (Wistar) male Transfer : not determined 2 (reliable with Galvez-Morros M,

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 71 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

restrictions) Garcia-Martinez O, oral: feed Toxicokinetic parameters: Wright AJA & Not applicable key study Southon S (1992) Exposure regime: Single exposure, rats Metabolites identified: not measured read-across based on were allowed to grouping of consume the test meal Details on metabolites: Not applicable substances (category for 1 h approach)

Doses/conc.: 130 µg Test material Zn as 65Zn-Zinc (IUPAC name): zinc chloride dichloride (See endpoint summary Zinc absorption and for justification of excretion in Wistar rats read-across) from 65Zn-Zinc chloride was measured. rat (Sprague-Dawley) Metabolites identified: not measured 2 (reliable with Furchner JE & male restrictions) Richmond CR Details on metabolites: Not applicable (1962) oral: gavage key study

Exposure regime: Zinc read-across based on chloride: Single grouping of exposure substances (category approach) Doses/conc.: Zinc chloride: 1.2 µC Test material Zinc acetate: 58, 117, (IUPAC name): zinc 175, 293, 410 and 644 dichloride (See ppm Zinc endpoint summary for justification of Effect of dietary zinc read-across) administration as Zinc acetate was evaluated on the absorption of orally administered Zn65 in rats. mouse (LACA mouse) Transfer (not determined): not determined 2 (reliable with He LS, Yan XS & female restrictions) Wu DC (1991) Toxicokinetic parameters: oral (gavage) and Half-life 1st: 0.3, 0.1 and 0.3 at age 20, 70key study intraperitoneal and 100 d respectively read-across based on Exposure regime: Half-life 2nd: 3.2, 3.1, 4.1 and 18.6 at age grouping of Single dose 1, 20, 70 and 100 d respectively substances (category Half-life 3rd: 8.7, 42.2, 24.8 and 96.7 at approach) Doses/conc.: (a) Oral age 1, 20, 70 and 100 d respectively exposure: 8.6 kBq Test material (sucklings) and 19.3 Metabolites identified: not measured (IUPAC name): zinc kBq (adolescent, young dichloride (See adults and adults) (For Details on metabolites: Not applicable endpoint summary details see attached for justification of study report, Table 1, read-across) Pg 908) (b) Intraperitoneal exposure: 19·3 kBq (for all other age groups except

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 72 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 sucklings)

Mice were gavaged with test material solution, 8.6-19.3 kBq per mouse, and the whole-body retention and organ content of test material were measured at different times after administration. The age-dependence of the fractional absorption of test material from the gastrointestinal tract (f1), the endogenous faecal excretion fraction of test material (EFEF), tissue distribution and whole- body retention were determined. rat (Wistar) male Transfer (Not reported): 2 (reliable with Pullen RGL, restrictions) Franklin PA & Hall intraperitoneal Toxicokinetic parameters: GH (1990) Not applicable supporting study Exposure regime: No data Metabolites identified: not measured read-across based on grouping of Doses/conc.: 15 µCi Details on metabolites: Not applicable substances (category 65Zn2+ approach) Evaluation of results: bioaccumulation The tissue uptake of potential cannot be judged based on study Test material 65Zn2+ (as zinc results (IUPAC name): zinc chloride) was dichloride (See determined in adult endpoint summary male Wistar rats after for justification of intraperitoneal read-across) injection of 15 µCi 65Zn2+. Main ADME results: 2 (reliable with Plodikova, P., Ammonium chloride is rapidly absorbed restrictions) Rostislav, C. (2010) and almost complete. supporting study

statement

Test material (Common name): ammonium chloride

5.1.2. Human information Absorption Oral A wide range of absorption (8-80%) is observed in humans (EU RAR, 2004a-f). This is likely due to differences

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 73 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 in eating habits (Hunt et al., 1991; Reinhold et al., 1991; Sandstrom and Sandberg, 1992). Persons with adequate nutritional levels of zinc absorb approximately 20-30% of all ingested zinc. Those who are zinc- deficient absorb greater proportions of administered zinc while persons with excessive zinc intake, gastrointestinal uptake can be less (Babcock et al., 1982; EU RAR, 2004a-f). Zinc absorption in the gastrointestinal tract occurs throughout the entire small intestine with the highest rate in the jejunum and the rate of total absorption appears to be concentration-dependent (Lee et al., 1989) as cited EU RARs (EU RAR, 2004a-f). The zinc absorption process in the intestines includes both passive diffusion and a carrier-mediated process (Tacnet et al., 1990). At low zinc concentrations CRIP is involved in this process. This protein binds zinc entering the intestinal cells from the lumen but this process appears to be saturable. Metallothionein, a metal- binding protein (also rich in cystein), may be involved at higher zinc concentrations (Gunshin et al., 1999; Hempe and Cousins et al., 1992; Struniolo et al., 1991). Zinc cations can induce metallothionein production in intestinal mucosa cells (Richards and Cousins, 1975; EU 2004 a-f).

65 The intestinal absorption following a single oral administration of ZnCl2 to 6 groups of 5 healthy adult volunteers has been determined by comparison of whole body radioactivity counting and faecal excretion data. 65 The individuals fasted overnight prior to dosing. Approximately 55% of the administered ZnCl2 was absorbed at doses of 18, 45 and 90 mol of zinc (i.e., approximately 1.2, 2.9 or 5.8 mg Zn). The absorption was reduced with increasing dose, indicating that zinc absorption is saturable. At test dose levels of 180, 450 and 900 mol (i.e., approximately 11.6, 29 or 58 mg Zn), only 51, 40 and 25% of the 65Zn was absorbed, respectively. Additional studies in 15 human volunteers with various intestinal diseases indicated that absorption of zinc occurred mainly in the proximal parts of the intestine, This study suggests that uptake levels vary maximally by a factor of 2 in healthy persons with intake levels differing by a factor of 10 (Payton et al., 1982). The absorption of orally administered 65Zn was studied in 50 patients with taste and smell dysfunction. The study was conducted in three phases: Prior to the start of the study 10 patients were admitted to a metabolic ward and put on a fixed daily diet containing 8-13 mg Zn. In the first phase, all patients were studied for 21 days 65 after receiving a single oral dose of 3-18 Ci of Zn (i.e., approximately 0.4 to 1.2 ng Zn) as ZnCl2 after an overnight fast. In the second phase, which started after 21 days and continued for 290 to 440 (mean 336) days, all 50 patients received a placebo. To study the effect of additional zinc intake on the elimination of previously sequestered radioactivity, in the third phase of the study 14 patients continued on the placebo while 36 received ZnSO4 (100 mg Zn/day) for 112 to 440 (mean 307) days. Phases two and three were a controlled clinical trial of the effects of zinc on retention of the 65Zn tracer. The results of phase two and three are described under excretion. Total body retention and activity in plasma and red cells were measured for all patients throughout the study. It was estimated that for the ten in-patients ca. 55% of the administered radioactivity was absorbed while for the whole group of 50 patients the absorption was approximately 60 %. From the study description it is not clear whether the radioactivity was administered as pure radioactive ZnCl 2 or whether it was diluted with 65 unlabelled ZnCl2. The authors stated that “patients were given 3 to 18 Ci carrier free Zn”, therefore for the calculation of the dose of 65Zn in terms of nanogram zinc, it has been assumed that all zinc administered was in fact radiolabelled zinc (Aamodt et al., 1982). The absorption of zinc from soluble zinc acetate, zinc sulphate, zinc aminoate, zinc methionine and insoluble zinc oxide were compared in ten human volunteers who were dosed orally with 50 mg zinc in various forms separated by two week intervals. The bioavailability of zinc from the various forms was compared on the basis of plasma zinc levels and area under the plasma curve analysis. Plasma peak levels were observed after about 2.5 h for all forms, but maximal plasma zinc concentration amounted to 221 and 225 g/dL for the acetate and the sulphate form while the peak plasma level for zinc from the oxide was only 159 g/dL. When AUC values for the different zinc forms were compared, it appeared that the bioavailability of insoluble ZnO was about 60% of the bioavailability of the soluble forms. Information on absolute bioavailability was not obtained (Prasad et al., 1993).

A study to measure the absorption half-life of zinc as ZnSO4 was performed. Gelatine capsules containing 45 mg zinc as ZnSO4 was administered once to 10 healthy young men. Serum concentrations were measured frequently during a total investigation time of 8 hours. A mean maximum concentration of 8.2 mol Zn/L serum was found after 2.3 hours (tmax). There was evidence of an enteral recirculation, the first rebound effect appeared after 1.4 hours during the absorption phase before tmax was reached, and exhibited mean reabsorption rates of 70% of the dose given. The subsequent ones (max. of 5) appeared at regular intervals of 1.2 hours with a decrease of the quantity reabsorbed. The absorption half-life of zinc administered as ZnSO 4 was 0.4 hours (Nève et al., 1991). Factors that influence the gastrointestinal absorption of zinc cations include ligands (for example a decreased

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 74 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 zinc absorption may occur by intake of plant proteins, such as soy and phytate), by intake of alcohol, use of EDTA or other trace elements in the diet (EU 2004 a-f). Also the zinc status of the body, the endogenous zinc secretion into the intestinal lumen via epithelial cells, bile and pancreatic secretion, and the intracellular transport have an influence on the zinc absorption in the gastrointestinal tract (Cunnane, 1988); Flanagan et al., 1983). However, the mechanism by which zinc is transferred to or across the mucosal surface of the microvilli is unknown (Cousins, 1989). Inhalation Elevated zinc concentrations in blood and urine of persons occupationally exposed to ZnO fumes suggest that there is some pulmonary absorption, but no quantitative human data are available (Hamdi et al., 1969 and Trevisan et al., 1982 cited in EU RAR, 2004a-f). Data on the particle size distribution of zinc aerosol in three different industry sectors, i.e. the galvanising sector (three plants, 4 samples each), the brass casting sector (two plants, 3 and 4 samples respectively) and the zinc oxide production sector (one plant, 10 samples), has been provided using personal cascade impactors with cut- off diameters of 0.52, 0.93, 1.55, 3.5, 6.0 and 21.3 m, and a final filter diameter of 0.3 m (Groat et al., 1999; EU 2004, a-f). These data served as input for the Multiple Path Particle Deposition Model (MPPDep version V1.11; Freijer et al., 1999) in order to estimate the airway deposition (in head, tracheobronchial and pulmonary region) for workers, by using:  The human – five lobar lung model;  A polydisperse particle distribution (i.e. this distribution contains a wide range of particle sizes), by taking the mean size distribution of the 10 samples for zinc oxide production (MMAD 15.2 m, GSD 4.0). Using this MMAD and GSD for the total polydisperse distribution is preferred above treating the polydisperse particles on individual impactor stages (with given cut-off diameters) as being monodisperse particles, also because the maximum particle size in the MPPDep model (20 m) is lower than the largest size fraction of the cascade impactor (21.3 m).  Both the oral breathing and the oronasal (normal augmenter) mode, but not the nasal breathing mode. The latter is considered to present an underestimate because (1) many people are oronasal or oral breathers, independent of their activities, (2) people with a cold will not normally breath nasally and (3) with heavy ex - ercise, short-term deep oral breathing will occur, resulting in increased deep pulmonary deposition.  The possibility of inhalability adjustment for the oronasal augmenter. Inhalability is defined as that fraction of particles in an aerosol that can enter the nose or mouth upon inhalation. It must be noted that inhalability is different from respirability (which relates to the deposition of particles after making their entrance inside the airways). If “inhalability adjustment” is “off”, the calculations start by assuming that the airflow is in line with the direction of the nasal entrance. However, in reality this will not be the case because the airflow has to make turns to enter the nose. This results in losses that are larger with increasing particle size. Mén- ache et al., (1995) described the relationship between exposure concentration and concentration at the entrance of the airways for laboratory animals and humans as cited in EU RARs (EU RAR, 2004a-f).  A tidal volume and breathing frequency corresponding to the default breathing rate of 10 m 3 for an 8-hr shift (1100 mL and 20 breaths/min, respectively).This breathing rate is more representative for light exercise activities than for more moderate or heavy exercise activities (EPA, 1997), which can be expected to take place in the zinc industry. Therefore, also a non-default tidal volume and breathing frequency corresponding to a breathing rate of 19 m3 for an 8-hr shift have been taken (1700 mL and 23 breaths/min, respectively, based on a breathing volume of 40 L/min for moderate exercise activities (EPA, 1997)). It must be noted that at a minute volume <35.3 L/min for normal augmenters breathing is only through the nose, while at a minute volume 35.3 mL/min there is combined nose and mouth breathing. For oral breathers, breathing is always only through the mouth, independent of the minute volume used. The results of the MPPDep modelling are given in Table below. It must be noted that the MPPDep only models deposition, not clearance and retention.

Table 21. Deposition fractions for oral breathers and for oronasal augmenters, using a polydisperse particle distribution (MMAD 15.2 m, GSD 4.0) Inhalability Tidal Breaths Deposition region Adjustment volume (min-1) Head Tracheo- Pulmonary Total (mL) bronchial Oral Off 1100 20 0.638 0.071 0.139 0.848

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 75 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

1700 23 0.676 0.100 0.101 0.877 Oronasal Off 1100 20 0.927 0.011 0.021 0.960 1700 23 0.804 0.064 0.064 0.932 Oronasal On 1100 20 0.519 0.011 0.021 0.551 1700 23 0.585 0.063 0.064 0.713

From the above table it can be seen that for oral as well as for oronasal breathers the largest part of the deposition takes place in the head region when inhalability adjustment is “off”, irrespective of the breathing rate. When inhalability adjustment is “on” the head region deposition is reduced. However, as stated above, the corrections for inhalability of particles is based on relationships derived by Ménache et al., (1995). For humans this is based on experiments with 4 healthy adult volunteers. From the available data it is not possible to conclude that this correction is valid for all human subjects in all situations (children, elderly, exercise activity, etc). Therefore it is reasonable to estimate the deposition with the inhalability adjustment “off” which leads to a worst case scenario and therefore the inhalability adjustment “on” will not be considered further. The fate and uptake of deposited particles depends on the clearance mechanisms present in the different parts of the airway. In the head region, most material will be cleared rapidly, either by expulsion (not the case for oral breathers) or by translocation to the gastrointestinal tract (half-life 10 min). A small fraction will be subjected to more prolonged retention, which can result in direct local absorption. This is concluded to be almost the same for the tracheobronchial region, where the largest part of the deposited material will be cleared to the pharynx (mainly by mucociliary clearance (half-life 100 min)) followed by clearance to the gastrointestinal tract, and only a small fraction will be retained (ICRP, 1994). Higher uptake rates may be assumed for the pulmonary region than for the head and tracheobronchial region. Once translocated to the gastrointestinal tract, uptake will be in accordance with oral uptake kinetics. Hence, for the part of the material deposited in head and tracheobronchial region that is cleared to the gastrointestinal tract, the oral absorption figures 20% for soluble zinc compounds and 12% for slightly soluble and insoluble zinc compounds can be estimated. However, there are no data available on zinc to estimate the part that is cleared to the gastrointestinal tract and the part that is absorbed locally in the different airway regions. With respect to the latter, there are some data available for radionuclides. After instillation of small volumes (2-3 L for rats, 10 L for hamsters, 0.3 mL for dogs) of solutions or suspensions of radionuclides into each region of the respiratory tract, retention and absorption into blood was measured. For the more soluble chemical forms (a.o. citrate and nitrate) absorption values of 4.8-17.6% in the nasopharynx, 12.5-48% in the tracheobronchial region and up to 100% in the pulmonary region was found. For the slightly soluble chemical forms (i.e. oxide) retention and absorption in the nasopharynx and tracheobronchial region was negligible (ICRP, 1994). There are no data on how the solubility of the different chemical forms of the radionuclides compares to the solubility of the soluble zinc compounds. Although the applicability of the radionuclide figures to the zinc compounds is not quite clear, it is probably a reasonable worst case scenario to take the upper values found (i.e. 20, 50 and 100% in head, trachebronchial and pulmonary region, respectively) for local absorption of the soluble zinc compounds (zinc chloride and zinc sulphate). For the slightly soluble and insoluble zinc compounds (zinc oxide, zinc phosphate and zinc metal) it is probably safe to assume negligible absorption for the head and tracheobronchial region and 100% absorption for the pulmonary region. This is supported by the findings in the study by Oberdörster et al., (1980), where the dissolution half-life of 1 m diameter zinc oxide particles in the deep lung was approximately 6 hrs. Given that the clearance to the gastrointestinal tract occurs within a time frame of minutes (10-100 min in head and tracheobronchial region), there will be no significant dissolution in these areas. Furthermore, most of the particles in these areas will have a diameter >1 m, thus dissolution half-lives for these larger particles will be longer. Based on the above, Table below describes the assumptions used in estimating the absorption by inhalation.

Table 22. Assumptions used for estimating the inhalation absorption Soluble zinc compounds Slightly soluble to insoluble zinc compounds (e.g., ZnCl2, ZnSO4) (e.g., Zn; ZnO; Zn3(PO4)2) Fraction absorbed in 20% head 0% head airway region 50% tracheobronchial 0% tracheobronchial 100% pulmonary 100% pulmonary Fraction cleared to GI 80% head x 20% 100% head x 12% tract, followed by oral 50% tracheobronchial x 20% 100% tracheobronchial x 12% uptake kinetics 0% pulmonary 0% pulmonary

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 76 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

By applying the above assumptions to the deposition fractions (Table 22), the % of inhalatory absorption of the soluble zinc compounds (zinc chloride and zinc sulphate) and slightly soluble to insoluble zinc compounds (zinc oxide, zinc phosphate and zinc metal) can be estimated as described in Table below. Table 23. Percentage estimations for inhalation absorption of soluble, slightly soluble and insoluble zinc compounds Inhalability Tidal Breaths Soluble zinc compounds Slightly soluble to insoluble volume zinc compounds -1 (mL) (min ) (e.g., ZnCl2, ZnSO4) (e.g., Zn; ZnO; Zn3(PO4)2)

Oral off 1100 20 41.1 22.4 1700 23 40.4 19.4 Oronasal off 1100 20 36.1 13.4 1700 23 39.2 16.8 Inhalation absorption for the soluble zinc compounds (zinc chloride and zinc sulphate) is at maximum 40%, while for the slightly soluble and insoluble zinc compounds (zinc oxide, zinc phosphate and zinc metal) inhalation absorption is at maximum 20%. These values are assumed to be a reasonable worst case and are thought to cover existing differences between the different zinc industry sectors with respect to the type of activities (therefore breathing rate) and the particle size distribution. Dermal Zinc has been reported to be absorbed through damaged or burned skin however in the absence of quantitative data it is difficult to assume that zinc can be absorbed through intact skin (EHC, 1996). An increase in serum zinc levels was observed in 8 patients suffering from second and third degree burns, who were treated with adhesive zinc-tape (ca. 7.5 g ZnO/100 g dry weight). The maximum value (up to 28.3 mol/litre) was reached within 3-18 days during treatment. It is noted that the absorption through intact skin could not be assessed (Hallmans, 1977). The systemic absorption from topical application of 40% zinc oxide ointment (with petrolatum) was investigated in 6 healthy subjects in a cross-over study. On two separate days, one week apart the subjects received a topical application of 100 g of the 40% zinc oxide ointment or 60 g of control ointment (100% white petrolatum base) to the chest, upper legs and lower legs (exposed skin area: not specified; occlusion: not specified) for 3 hours. Each subject fasted for 12 hours before treatment started (only water ad libitum). During the study no food or water was consumed. Blood samples were taken after the 12 hr-fast (baseline value), and at 1, 2 and 3 hours after the start of the topical application. Mean serum zinc concentrations at these time points were 107.3, 116.1, 105.3 and 112.6 g/dL for the zinc ointment and 115.2, 103.5, 105.5 and 110.5 for the control ointment, respectively. Normal serum zinc concentrations were considered to be in the range of 68 to 136 g/dL. An increase in serum zinc over the baseline value was observed in 4/6 subjects. In 3 of them, the rise was most pronounced after 1 hr. In 2/6 no increase was observed throughout the treatment. Overall, 1 hour after application, there was a mean serum zinc increase of 8.8 g/dL over the baseline. This represented an 8.2% rise in serum zinc which was not statistically significant (Derry et al., 1993). The systemic absorption was also investigated in patients receiving total parenteral nutrition (TPN) for a minimum of 3 days prior to the start of the experiment. TPN is known to result in zinc deficiency (mean decrease 6.6 g/dL/week), and the longer the period of TPN without zinc supplementation, the greater the decrease in serum zinc concentration. In a controlled, cross-over study (on two separate days, one week apart) 6 patients received a topical application of 15 g of the 40% zinc oxide ointment onto the upper legs (10x15 cm) once daily for 8 consecutive days under occlusion. Blood samples were taken before treatment (baseline value), at 4, 6 and 8 days (just prior to application), and at day 10. The mean baseline level of the patients (88.6 g/dL) differed significantly from the mean baseline level of the healthy subjects. The mean zinc concentration in the 3 patients that completed the study remained relatively stable over the 10 day period (78-93 g/dL) (Derry et al., 1993). It can be concluded that topical applications of 40% zinc oxide ointment did not result in a significant increase in serum zinc concentration in healthy human subjects over a 3-hr period nor in TPN-patients over 10 days. The authors suggested that after topical application, zinc is locally absorbed and stored in the hair follicles where it is relatively unavailable for immediate systemic absorption in subjects with normal serum zinc concentrations. In subjects that are hypozincemic, there is absorption from the storage depot at a rate sufficient to prevent a decline in serum zinc concentration. The authors concluded that the 3-hr sampling time in normal subjects may have

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 77 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 been insufficient to allow for appreciable systemic absorption from the storage depot (Derry et al., 1983). When ZnO-mediated occlusive dressings (25% w/w; 4x5 cm) were applied to the lower arm of 10 healthy volunteers for 48 hours it appeared that the mean release rate of zinc to normal skin was 5 g/cm2/hour. After treatment of 5 other volunteers with the ZnO dressings for 48 hours the zinc content was significantly increased in the epidermis and in the accumulated blister fluid (to model percutaneous absorption, suction blisters were used). It should be noted, however, that the zinc penetration was enhanced during the formation of blisters, indicating that the barrier function was impaired (Agren, 1990). In another study, five human volunteers were exposed to different occlusive ZnO dressings (with hydrocolloid vehicle or gum rosin). After 48 hours, suction blisters on treated skin were raised and zinc concentration in blister fluid was determined. Furthermore the zinc concentration in the stratum corneum (10 successive tape strippings) was determined. The absorbed amount could not be determined from the data presented but it appeared that the vehicle is an important factor for zinc penetration (Agren, 1991b). Distribution After absorption from the gastrointestinal tract, the zinc is bound in plasma primarily to albumin and then transported to the liver and subsequently throughout the body. The normal plasma zinc concentration is ca. 1 mg/L, the total zinc content of the human body (70 kg) is in the range of 1.5-2 g (ATSDR, 2005). Zinc is found in all tissues and tissue fluids and it is a co-factor in over 200 enzyme systems. In humans, the major part of total body zinc is found in muscle and bone, approximately 60% and 30%, respectively (Wastney et al., 1986). Under normal conditions, the highest zinc concentration per tissue weight is found in bone, hair and in the prostate (Cleven et al., 1993). The distribution of zinc in humans appears to be influenced by age. The zinc concentration levels increases in the liver, pancreas and prostate and decreases in the uterus and aorta with age. Levels in kidneys and heart peak at approximately 40-50 years of age and then declines. Levels in the aorta decline after 30 years of age (Schroeder et al., 1967). The tissue uptake of 65Zn (as zinc chloride) was determined in adult male Wistar rats after intraperitoneal injection of 15 Ci 65Zn. The liver displayed the greatest uptake for zinc ions, followed by the kidney, pancreas, spleen, ileum, lung, heart, bone, testis, blood cells, muscle and brain. Additional data on Zn uptake by the brain indicates that the blood-brain barrier is minimally permeable to zinc cations (Pullen et al., 1990). Eight hours following intravenous administration of 65[Zn]-chloride to rabbits, tissue levels were highest in the liver, intestine and kidney with levels being  10%/g in tissue (Lorber et al., 1970). Metabolism Zinc is mostly bound to organic ligands rather than existing free in solution (Gordon et al., 1981). Zinc is found in diffusible and non-diffusible forms in the blood and about 66% of the diffusible form of zinc in the plasma is freely exchangeable and loosely bound to albumin (Cousins, 1985). A small amount of the non-diffusible form of zinc is tightly bound to 2-macroglobulin in the plasma and is not freely exchangeable with other zinc ligands. Zinc is incorporated into and dissociated from 2-macroglobulin only in the liver (Henkin, 1974).

Excretion In humans, the faecal zinc consists of un-absorbed dietary zinc and endogenous zinc from bile, pancreatic juice and other secretions. About 70-80% of the ingested amount of zinc is excreted via faeces (5 to 10 mg/day depending upon the dietary zinc concentration) (Spencer et al., 1976; Venugopal and Lucky, 1978; Reinhold et al., 1991; Wastney et al., 1986). In humans, of the amount of zinc consumed, about 10% is lost through urine (approximately 200 to 600 g zinc/day). The urinary zinc excretion appears to be sensitive to alterations in the zinc status (Babcock et al., 1982; Aamodt et al., 1982). Saliva, hair loss, mother’s milk and sweat appear to be minor routes for zinc excretion. In tropical climates about 2-3 mg Zn/day may be lost in sweat (Venugopal and Lucky, 1978; Rivlin, 1983; Prasad et al., 1963; Rossowka and Nakamoto, 1992; Henkin et al., 1975). In humans with no excessive intake of zinc, the half-life of absorbed radio-labelled zinc ranges from 162 to 500 days. After parenteral administration of 65Zn, half-lives ranged from 100 to 500 days (Elinder, 1986).

65 Sixteen healthy adult human volunteers were given oral administration of 92 mol of Zn (as ZnCl2) to investigate the body retention of zinc at 7 to 10 days after dosing. The results showed that about 10% of the

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 78 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 initially absorbed amount of zinc was excreted during the first 10 days post dosing. Thirty other volunteers were dosed with 18 to 900 moles of 65Zn. Table below shows the elimination data following 10 to 60 days post- dosing. Table 24. Elimination data obtained following thirty humans dosed with 18 to 900 moles of 65Zn Dose group Excretion rate Biological half-life (moles; (mg)) (% of remaining Zn/day ) (days) 18 (1.2) 0.44 157 45 (2.9) 0.62 111 90 (5.8) 0.37 186 180 (11.6) 0.49 141 450 (29) 0.37 186 900 (58) 0.74a 93 a significantly different from the 18 moles group

The excretion rates for the 18 to 450 moles dose groups were not significantly different. The 900 mole dose group showed a significant increase in elimination rate (Payton et al., 1982). The effect on excretion following oral administration of radiolabelled zinc as zinc chloride in 50 patients with taste and smell dysfunction was investigated. The study was conducted in three phases. In the first phase all patients were studied for 21 days after receiving a single oral dose of 3-18 Ci of 65Zn (i.e., approximately 0.4 to 1.2 ng zinc) as ZnCl2 after an overnight fast. In the second phase, which started after 21 days and continued for 290 to 440 (mean 336) days, all 50 patients received placebo. To study the effect of additional zinc intake on elimination of previously sequestered radioactivity, in the third phase of the study 14 patients continued on placebo while 36 received ZnSO4 (100 mg Zn/day) for 112 to 440 (mean 307) days. In the controlled clinical trial of phases two and three, observations were made to see the effects of zinc on retention of the 65Zn tracer. The results from the first phase of the study are described under absorption section. Total body retention and activity in plasma and red cells were measured for all patients throughout the study. About one-third of the absorbed radioactivity was eliminated from the body with a half-life of ca. 19 days, while after about 100 days post dosing the remainder of the absorbed dose was eliminated with a biological half-life of 380 days (i.e. phase 65 two of the study). During the third phase patients receiving ZnSO4 showed an accelerated loss of total body Zn (half-life ca. 230 days) which was significantly different (P>0.001) from half-life values during placebo treatment. Accelerated loss of 65Zn from the thigh was apparent immediately while that from the liver began after a mean delay of 107 days. There was no apparent effect of zinc on loss of mean 65Zn activity from red blood cells (Aamodt et al., 1982). From the study description it is not clear whether the radioactivity was administered as pure radioactive zinc chloride or whether it was diluted with unlabelled zinc chloride. As the authors stated that “patients were given 3 to 18 Ci carrier free 65Zn” for the calculation of the dose of 65Zn in terms of nanogram zinc, it has been assumed that all zinc administered was 65Zn (Aamodt et al. 1982). In ten of the patients from the study described above (Aamodt et al. 1982), the kinetics of 65Zn was studied in more detail by Babcock et al. (1982). These patients received a fixed diet containing 8 – 13 mg Zn per day for 4 to 7 days before and after the single 65Zn dose, followed by 290-440 (mean 336) days of non-restricted diet, followed by an additional intake of 100 mg/day of non-radioactive zinc ion (as ZnSO4) over the next 112-440 days (mean 307). The overall kinetic parameters of these 10 patients did not differ from those of the other patients (Aamodt et al., 1982). The authors further submitted retention-time curve data for whole body, plasma, red blood cells, liver and thigh to a multi-compartment kinetic model. From this model analysis it could be demonstrated that the increase in elimination of Zn during the third phase of the study by Aamodt et al. (1982) can be ascribed entirely to the change in parameters: reduction in absorption in the gastrointestinal tract (5-fold: from 43% absorption at the beginning of the study to 9% during the period in which patients were dosed with ZnSO4) and to an increase in the urinary elimination rate (about 2-fold upon administration of ZnSO4 during phase three of the study). Michaelis-Menten type saturation mechanisms were adequate to explain the observed parameter changes. These changes also accounted for the observed mean plasma zinc mass increase of only 37% above pre-load levels in face of an 11-fold increase in zinc intake (from ca. 10 mg/day to ca. 110 mg/d) (Babcock et al., 1982). From this model analysis it was estimated that the total body Zn contents of these 10 patients at the start of the study was 1.4 g. Babcock et al. (1982) indicated that normally the body contents of zinc is in the range of 2.1 to 2.5 g. This may indicate that the patients studied by Babcock et al. (1982) and possibly by Aamodt et al. (1982) were deficient in total body zinc. 5.1.3. Summary and discussion of toxicokinetics

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 79 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

As described in Section 5 General considerations (assumptions), zinc compounds release, depending on their solubility, zinc cations which determine the biological activity of the respective zinc compounds. Sufficient data is available on the soluble zinc compounds zinc chloride and zinc sulphate and on the slightly soluble zinc compounds ZnO and ZnCO3. Zinc is an essential trace element which is regulated and maintained in the various tissues mainly by the gastrointestinal absorption and secretion during high and low dietary zinc intake and because of the limited exchange of zinc between tissues, a constant supply of zinc is required to sustain the physiological requirements. The zinc absorption process in the intestines includes both passive diffusion and a carrier-mediated process. The absorption can be influenced by several factors such as ligands in the diet and the zinc status. Persons with adequate nutritional levels absorb 20-30% and animals absorb 40-50%. Persons that are zinc deficient absorb more, while persons with excessive zinc intake absorb less. For the soluble zinc compounds, the available information suggests an oral absorption value of 20%. This value can be considered as the lower bound range at adequate nutritional levels. The oral absorption of the slightly soluble zinc oxide has been shown to be 60% of that of the soluble zinc compounds. This corresponds to approximately 12-18%. No oral absorption information is available for the remaining slightly soluble and insoluble zinc compounds (i.e., ZnO, Zn(OH)2, Zn3(PO4)2, ZnCO3, Zn, ZnS). However, considering that these substances have lower water solubility than ZnO, it can be conservatively assumed that the oral absorption of these compounds is ≤ 12%. Animal data suggests that there is pulmonary absorption following inhalation exposure. Half-life values of 14 and 6.3 hours were reported for dissolution of zinc oxide. The absorption of inhaled zinc depends on the particle size and the deposition of these particles therefore data was provided on the particle size distribution of zinc aerosol from three different industry sectors. The particle size distribution data was evaluated by using a multiple path particle deposition (MPPDep) model. This model revealed that for zinc aerosols the largest part of the deposition is in the head region and much less in the tracheobronchial and pulmonary region. Although most of the material deposited in the head and tracheobronchial region is rapidly translocated to the gastrointestinal tract, a part will also be absorbed locally. Based on data for local absorption of radionuclides in the different airway regions, it can be assumed that the local absorption of the soluble zinc compounds will be approximately 20% of the material deposited in the head region, 50% of the material deposited in the tracheobronchial region and 100% of the material deposited in the pulmonary region. For the slightly soluble and insoluble zinc compounds a negligible absorption can be assumed for materials deposited in the head and the tracheobronchial region. 100% of the deposited slightly or insoluble zinc compounds are assumed to be absorbed in the pulmonary tract. The deposited material will be cleared via the lung clearance mechanisms into the gastrointestinal tract where it will follow oral absorption kinetics. Therefore the inhalation absorption for the soluble zinc compounds is a maximum of 40% and for the slightly soluble and insoluble zinc compounds inhalation absorption is at a maximum of 20%. These values can be assumed as a reasonable worst case, because they are considered to cover existing differences between the different zinc industry sectors with respect to the type of exercise activities (and thus breathing rate) and particle size distribution. The available information from in vivo as well as the in vitro studies suggests the dermal absorption of zinc compounds through intact skin to be less than 2%. In vitro studies that estimated dermal absorption values only on the basis of the zinc levels in the receptor medium without taking into account the zinc present in the stratum corneum appear to underestimate absorption values derived from in vivo studies. Such zinc trapped in the skin layers may become systemically available at a later stage. Quantitative data to evaluate the relevance of this skin depot are however lacking. Given the efficient homeostatic mechanisms of mammals to maintain the total body zinc and the physiologically required levels of zinc in the various tissues to be constant, the anticipated slow release of zinc from the skin is not expected to disturb the homeostatic zinc balance of the body. Considering the available information on dermal absorption, the default for dermal absorption of all zinc compounds (solutions or suspensions) is 2%. Based on the physical appearance, for dust exposure to zinc and zinc compounds a 10- fold lower default value of 0.2% is a reasonable assumption. Zinc appears to be distributed to all tissues and tissue fluids and it is a cofactor in over 200 enzyme systems. The excretion of zinc is primarily via faeces, but also via urine, saliva, hair loss, sweat and mothers-milk. 5.2. Acute toxicity 5.2.1. Non-human information

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 80 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

5.2.1.1. Acute toxicity: oral Acute oral toxicity studies are available in rats and mice on zinc compounds across all water solubilities. Acute oral toxicity studies have been conducted on soluble zinc compounds (zinc chloride, zinc sulphate and zinc bis(dihydrogen phosphate)) and on the slightly soluble (zinc oxide, zinc phosphate, zinc metal) and insoluble zinc compound (zinc sulphide). Table 25. Overview of experimental studies on acute toxicity after oral administration according to decreasing water solubility of zinc compounds Test Study Species Endpoint Exposure Result Remarks Reference Substance Type LD50

Zinc chloride Acute Rat LD50 Single dose 1,100 2 (reliable with Domingo J L, oral mg/kg restrictions) Llobet J M, bw key study Paternain J L and Corbella J (1988a)

Zinc chloride Acute Mouse LD50 Single dose 1,260 2 (reliable with Domingo J L, oral mg/kg restrictions) Llobet J M, bw key study Paternain J L and Corbella J (1988b)

Zinc sulphate Acute Rat LD50 50, 100, 920 2 (reliable with Litton oral 1,000 or mg/kg restrictions) bionetics 3,000 bw supporting study (1974) mg/kg bw used in RAR, (EU 2004 e)

Zinc sulphate Acute Mouse LD50 No 926 2 (reliable with Domingo J L, oral information mg/kg restrictions) Llobet J M, available bw key study Paternain J L and Corbella J (1988b)

Zinc sulphate Acute Rat LD50 200- 2000 1,000 2 (reliable with Sanders A oral mg/kg bw – restrictions) (2001b) 2,000 supporting study mg/kg used in RAR, bw (EU 2004 e)

Zinc sulphate Acute Rat LD50 Single dose 1,710 2 (reliable with Domingo J L, oral mg/kg restrictions) Llobet J M, bw key study Paternain J L and Corbella J (1988a)

Zinc sulphate Acute Mouse LD50 No further 1,891 2 (reliable with Courtois Ph, oral information mg/kg restrictions) Guillard O, available bw supporting study Pouyollon M, Piriou A and Warnet J-M, (1978)

Zinc sulphate Acute Rat LD50 No further 2,280 2 (reliable with Lorke D oral information mg/kg restrictions) (1983) available bw supporting study used in RAR, (EU 2004 e)

Zinc sulphate Acute Rat LD50 Single dose; > 2 (reliable with Sanders A oral 2,000 restrictions) (2001a) mg/kg supporting study bw used in RAR, (EU 2004 e)

Zinc sulphate Acute Rat LD50 No further 2,949 2 (reliable with Courtois Ph, oral information mg/kg restrictions) Guillard O,

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 81 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Test Study Species Endpoint Exposure Result Remarks Reference Substance Type LD50 available bw supporting study Pouyollon M, Piriou A and Warnet J-M, (1978)

Zinc Acute Rat LD50 Single dose 300- 1 (reliable Van bis(dihydrogen oral 2000 without Huygevoort phosphate) mg/kg restriction) AHBM bw key study (2007)

Zinc oxide Acute Rat LD50 Single; dose >5,000 2 (reliable with Loser E oral mg/kg restrictions) (1977) bw key study

Zinc oxide Acute Mouse LD50 No further ca. 4 (not Shumskaya oral information 7,950 assignable) NI, available mg/kg supporting study Mel’nikova bw VV, Zhilenko VN and Berezhnova LI (1986)

Zinc oxide Acute Rat LD50 Single dose > 2 (reliable with Löser E oral 15,000 restrictions) (1972) mg/kg supporting study bw used in RAR, (EU 2004 b)

Zinc Acute Rat LD50 Single dose; > 2 (reliable with Klein and phosphate oral 5,000 restrictions) Glaser (1989) mg/kg supporting study bw used in RAR, (EU 2004 d)

Zinc metal Acute Rat LD50 Single dose >2,000 2 (reliable with Prinsen MK oral mg/kg restrictions) (1996) bw key study used in RAR, (EU 2004 d)

Zinc sulfide Acute Rat LD50 No > 4 (not Sachtleben oral information 15,000 assignable) Chemie available mg/kg supporting study GmbH (2000 bw a) The acute oral toxicity studies with zinc sulphate do not specifically state which hydrated form of the zinc sulphate was tested. As this impacts the LD50 value, all available LD50 values were re-calculated to provide an understanding the range of LD50 values of currently marketed zinc sulphate products (i.e., mono-, hexa-, and heptahydrate). Table below summarises the results of this re-calculation

Table 26. Re-calculation of oral LD50 rat values

Reported for LD50 LD50 (mg/kg bw) recalculated for Reference Zinc sulphate (mg/kg bw) Mono Hexa Hepta form Dihydrate 1,710 1,554 2,334 2,490 Domingo et al., (1988) Heptahydrate 2,280 1,423 2,137 2,280 Lorke, (1983) Unspecified 2,949 1,840 1 2,764 1 2,9491 Courtois et al., 2,949 2 4,429 2 4,7252 (1978) Unspecified 920 574 1 862 1 9201 Litton Bionetics, 920 2 1,382 2 1,4742 (1974) Hexahydrate > 2,500 > 1,665 > 2,500 > 2,667 Sanders, (2001a)

Heptahydrate 1,000 < LD50 < 624 < LD50 < 937 < LD50 < 1000 < LD50 < Sanders, (2001b) 2,000 1,248 1,875 2,000 1 Assumes testing of the heptahydrate (worst case) ; 2 Assumes testing of the monohydrate

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 82 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Table 27. Overview of experimental studies on acute toxicity after oral administration or ammonium chloride

Method Results Remarks Reference rat (Wistar) male/female LD50: ca. 1220 mg/kg bw 2 (reliable with n.n. (2003) (female) based on: test mat. restrictions) oral: gavage LD50: 1630 mg/kg bw (male) supporting study equivalent or similar to OECD based on: test mat. Guideline 401 (Acute Oral Toxicity) experimental result LD50: ca. 1410 mg/kg bw (male/female) based on: test Test material (CAS mat. name): ammonium chloride

5.2.1.2. Acute toxicity: inhalation Acute inhalation data is available on zinc chloride, zinc oxide as well as on zinc metal. Table 28. Overview of experimental studies on acute toxicity after inhalation exposure according to decreasing water solubility of zinc compounds

Test Study Specie End- Exposure Result LC50 Remarks Reference substance type s point time (mg/L) (4hrs) (mg/L)

Zinc Acute Rat LC50 10 min < 2 Not 2 (reliable Karlsson chloride inhalation applicable with N, Cassel restrictions) G, key study Fangmark I and Bergman F (1986)

Zinc oxide Acute Mouse LC50 No 2.5 Not known- 4 (not RTECS, inhalation informati assignable) (1991) on supporting available study used in RAR, (EU 2004 b)

Zinc oxide Acute Rat LC50 4 hours >5.7 >5.7 2 (reliable Klimisch inhalation with H-J, restrictions) Hilde- key study brand B used in RAR, and (EU 2004 b) Freisberg KO (1982)

Zinc metal Acute Rat LC50 4 hours >5.41 >5.41 2 (reliable Arts, MHE inhalation with (1996) restrictions) key study used in RAR, (EU 2004 a)

Additional inhalation data Male Syrian hamsters were exposed via inhalation to zinc sulphate aerosols in doses of 1.3 to 34.2 mg /m 3 (1.1- 7.3 mg Zn) for 4 hours. The activity median aerodynamic diameter (AMAD) and geometric standard deviation (GSD) of the aerosols were 0.59 μm and 1.46, respectively. The rate of phagocytosis of insoluble particles by pulmonary macrophages was determined in situ by introduction of insoluble gold colloid in the respiratory tract

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 83 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

under anesthesia. From a dose of 5.2 up to 34.2 mg ZnSO4 /m³ macrophage endocytosis of colloidal gold was significantly reduced 1 h after exposure compared with that in unexposed control animals. After 24 hours the rate of phagocytosis was still depressed, whereas after 48 hours it had returned to normal values. An increase in macrophage cell number was seen at low concentrations followed by depressions in macrophage numbers at high concentrations. No effects were observed at 1.3 mg/m3 (0.2 mg Zn) (Skornik and Brain, 1983). In anesthetized dogs the pulmonary mechanics were not significantly changed after inhalation exposure to 3 3 submicron aerosols of ZnSO4 up to 17.3 mg/m for 7.5 minutes. Also an exposure of 4 hours to 4.1 to 8.8 mg/m ZnSO4 to anesthetized dogs showed no effect on breathing mechanics, hemodynamic, or on arterial blood gases (Sackner et al., 1981). In a lung function test, 23 guinea pigs were exposed by inhalation to 0.9 mg ZnO/m3 (furnace-generated aerosol; 0.05 microns) for 1 hour. A progressive decrease in lung compliance was observed (from 9% below control value at the end of exposure to 16% after one hour post-exposure), but no change in air flow resistance (Amdur et al., 1982). In contrast to these results, no effects on ventilation, lung mechanics, diffusing capacity of carbon monoxide, or most lung volume parameters were observed in another lung function test with 10 guinea pigs exposed for 3 hours to 7.8 mg ZnO/m3 (furnace-generated aerosol; 0.05 microns). However functional residual capacity was significantly decreased (10% below control value) with only minimal changes in other lung volume subdivisions (Lam et al., 1982). The effect of inhaled ZnO was studied in guinea pigs, rats, and rabbits. Animals were exposed to 0, 2.5 or 5 mg ZnO/m3 (furnace-generated aerosol; 0.06 microns) for up to 3 hours and their lungs lavaged at 24 hours thereafter. The lavage lung fluid of both guinea pigs and rats exposed to the highest dose showed significant increases in total cells (guinea pigs 2.5-fold; rats 2-fold), lactate dehydrogenase (guinea pigs 24-fold, rats 9- fold), -glucuronidase (guinea pigs 13-fold; rats 27-fold), and protein content (guinea pigs 3.5-fold and rats 5.6- fold). Guinea pigs exposed to 2.5 mg ZnO/m³ for 3 hours resulted in significant increases in LDH (16-fold), β- glucuronidase (5-fold), and protein (1.4-fold). Exposure of rats to 2.5 mg ZnO/m³ resulted in significant increases in lactate dehydrogenase (4.5-fold), β-glucuronidase (11-fold), and protein (5-fold). Rabbits, exposed to 2.5 or 5 mg ZnO/m3 (furnace-generated aerosol; 0.06 microns) for 2 hours, showed no changes in the biochemical or cellular parameters (Gordon et al., 1992).

5.2.1.3. Acute toxicity: dermal Acute dermal toxicity has been investigated with the soluble zinc sulphate. Table below presents the respective study details and results Table 29. Overview of experimental studies on acute toxicity after dermal exposure Test Study Species Endpoint Exposure Result Study Reliability Reference substance Type (Klimisch Score)

Zinc Acute Rat LD50 Single >2,000 2 (reliable with Van sulphate dermal application; mg/kg restrictions) Huygevoort 24 hours; bw key study AHBM semi- (1999a) occlusive

5.2.1.4. Acute toxicity: other routes

Male Wistar rats (5/group) were given an intratracheal dose of 2.5 mg ZnCl 2/kg bw and sacrificed 3, 14, 28 or 35 days after dosing. Within 3 hours after dosing all rats were subdued and showed respiratory distress. Histology showed alveolitis around the major bronchi, most severe on day 3 after treatment. A change from macrophage to lymphocyte infiltration was seen in the damaged areas at day 14, without evidence of fibrosis. At 28 days, early alveolar thickening with increased interstitial reticulum deposition was observed, and at 35 days these changes had amounted to mature, discrete areas of parenchymal scarring (Brown et al., 1990).

After intratracheal administration of ZnCl2 to male Wistar rats at dose levels of 0, 0.25, 0.5, 1, 2, 4 or 5 mg/kg bw, no histological effects on the lung tissue were seen up to dose level of 0.5 mg ZnCl 2/kg bw. At higher dose levels, a dose-related intra-alveolar oedema was observed (Richards et al., 1989)

Exposure of male Wistar rats to a dose of 2.5 mg ZnCl2/kg bw by instillation caused oedema of the lung and a ten-fold increase in the level of alveolar surface protein within 6 hours of treatment. The pulmonary oedema was most severe between 6 hours and 3 days after exposure, with more than 50% of the rats showing oedema (Richards et al., 1989).

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 84 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

5.2.2. Human information Soluble zinc compounds Oral Oral intake of ‘one tablespoon’ by a 16-month old boy (McKinney et al., 1994, 1995) or ‘about three ounces’ of a zinc chloride solution (soldering flux) by a 24-yr old male (Chobanian, 1981) led to local caustic effects, nausea, vomiting, abdominal pain, hyperamylasemia and lethargy. A 15-year-old girl with no history of dyspepsia ingested zinc sulphate tablets of 220 mg twice daily (440 mg

ZnSO4/day  2.6 mg Zn/kg bw/day) for the treatment of acne. After each capsule the girl experienced epigastric discomfort. After 1 week gastrointestinal haemorrhages accompanied by anemia was observed. No other medicines were used (Moore, 1978). Inhalation Inhalation exposure to concentrations between 0.07 and 0.4 mg/m3 zinc chloride fume for 30 minutes failed to elicit sensory effects. In the same study, an average concentration of 4.8 mg/m 3 over a 30-minute period caused mild, transient irritation of the respiratory tract in bearing manufacture workers (Ferry, 1966; 1974). Exposure to 40 mg/m3 zinc chloride aerosol a metallic taste was detected. Experimental exposure to zinc chloride for 2 minutes resulted in slight nausea and some cough at 80 mg/m 3 in the majority of human subjects, whereas at 120 mg/m3 irritation of the nose, throat and chest were noted (Cullumbine, 1957). Exposure to 4,800 mg/m3 for 30 minutes induced pulmonary effects. No further data available (Lewis, 1992). Accidental exposure to zinc chloride fume resulted in intoxications (Evans, 1945; Hjortsø et al., 1988; Homma et al., 1992; Johnson and Stonehill, 1961; Macaulay and Mant, 1964; Matarese and Matthews, 1986; Milliken et al., 1963; Pare and Sandler, 1954; Schenker et al., 1981), but quantitative data are lacking except for one study (Johnson and Stonehill, 1961), where the concentration was 4,075 mg/m3 (duration of exposure not indicated). After inhalation, shortness of breath, pain in the throat, acute inflammation of the respiratory tract, cyanosis, bronchopneumonia, painful cough with sputum, chest pain and tightness, nausea and vomiting, headache, pulmonary oedema and fibrosis, acute respiratory insufficiency was experienced more or less in increasing order of seriousness. In several cases the symptoms receded one or two hours after exposure, but occasionally aggravated a few hours up to 2 weeks later. In a few cases the high exposure concentration was fatal. Slightly soluble and insoluble zinc compounds Oral A 16-year old boy, who ingested 12 g metallic zinc in 2 days (114 mg/kg bw on the first and 57 mg/kg bw on the second day) in order to hasten healing of a minor laceration, experienced light-headedness, lethargy, staggering gait, and difficulty writing legibly, but no gastrointestinal distress. He showed an increase in serum lipase and amylase, measured eight days after dosing (Murphy, 1970). Inhalation Very specific operations using very high temperatures such as cutting or welding of galvanised steel can give rise to the formation of fumes containing ultrafine particulate zinc oxide (<0.1 micron in diameter) (EU 2004 a). Exposure to these fumes can cause metal fume fever, expressing itself in certain typical symptoms including dry and sore throat, fever, coughing, dyspnoea, pyrexia, muscular pains, headache and metallic taste (Heydon and Kagan, 1990; Gordon et al., 1992; Mueller and Seger, 1985). In addition to these symptoms, gastrointestinal disturbance may be associated with exposure to ultrafine particulate fumes (NIOSH, 1975). A number of studies have measured exposure levels associated with metal fume fever. In a human study, subjects (n=4) were exposed in a single-blind fashion to control furnace gases or ultrafine ZnO particles (5 mg/m3) for 2 hours. All 4 persons exposed to ZnO showed the typical metal fume fever symptoms beginning 4 to 8 hours after exposure and disappearing within 24 hours. The reported symptoms include fever, chills, dry or sore throat, chest tightness, and headache. No changes were observed in pulmonary function immediately after exposure. The specific airway resistance increased with 16% in all subjects exposed to ZnO (Gordon et al., 1992). Therefore, an effect level of 5 mg ZnO/m3 for metal fume fever can be derived. Occupational exposure (6-8 hours) to zinc oxide fume generated during welding operations was investigated. Spirometric lung-function measurements were conducted 5 days before and after the work shift of 11 welders of zinc-coated steel, ten non-welders who were indirectly exposed to welding fumes, and 17 controls. The personnel exposure to zinc was monitored using PAS-6 samplers. The geometric mean concentration for welders was 0.034 mg Zn (as ZnO)/m3, for exposed non-welders 0.019 mg ZnO/m³, and for controls 0.004 mg

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 85 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

ZnO/m³. No changes in lung function parameters were observed at a 5% significance level. No symptoms of metal fume fever were reported (Marquart et al., 1989). In another study, the response of humans after exposure to zinc welding fume was investigated. Fourteen welders were acutely exposed to zinc oxide welding fume over a 15- to 30-minute period. The personal exposure to zinc oxide was monitored and the mean cumulative exposure was 2.3 ± 1.7 g.min/m³ resulting in an exposure of 77-153 mg ZnO/m3. Pulmonary function, airway reactivity, serum zinc levels and blood cell counts were measured. A bronchoalveolar lavage (BAL) was carried out to assess the cellular inflammatory response in the lung. Changes in pulmonary function and measured airway resistance were minimal. Cumulative zinc exposure and polymorphonuclear leukocyte count were positively correlated. A significant dose-dependent increase of immunological activity (i.e. increased polymorphonuclear leukocytes) was found in the BAL fluid 22 hours after exposure (Blanc et al., 1991). Blanc et al. (1993), 26 experimental welding fume exposures in 23 volunteers, with a representative range of welding experience, were carried out. Subjects performed electric arc welding on galvanized mild steel over a 15- to 30-min period. Postexposure BAL was performed at 3, 8, or 22 hours after exposure in 6, 11, and 9 subjects, respectively, and compared with BAL obtained from 17 control subjects. The mean zinc exposures were 1.8, 2.0, and 2.6 gmin/m3 for groups lavaged after 3, 8, and 22 hours, respectively, resulting in an exposure of 20-170 mg zinc/m3 (equal to 25-212 mg ZnO/m3; calculation based on a 30-min exposure to the reported exposure range of 0.6-5.1 g.min/m3). Besides inflammatory cells, BAL fluid supernatant concentrations of several cytokines, i.e. tumor necrosis factor (TNF), interleukin-6 (IL-6), and interleukin-8 (IL-8) increased in time and exposure-dependent fashion after zinc oxide welding fume exposure. In another study, 14 volunteers were studied after inhalation exposure to purified zinc oxide fume and after sham exposure to air. The exposure concentrations ranged from 2.76-37 mg zinc/m3 (3.4-46 mg ZnO/m3) for a period of 15 to 120 minutes (cumulative zinc exposure 165-1110 mg.min/m3). Twenty hours after exposure BAL was performed and analysed for cell contents and cytokines including TNF, IL-8, and interleukin-1 (IL-1). Polymorphonuclear leukocytes were significantly increased in the BAL fluid obtained post-exposure compared to sham. Cumulative zinc exposure correlated positively with changes in BAL supernatant concentrations of both TNF (r2=0.58) and IL-8 (r2=0.44). Cigarette smoking was not associated with differences in BAL TNF or IL-8. The data suggests a threshold for zinc exposure-related increased TNF and IL-8 at approximately 500 mg.min/m3 expressed as zinc (625 mg.min/m3 as ZnO). However, the correlation coefficients between cumulative exposure levels and rise in TNF or IL-8 were low (Kuschner et al., 1995). The data was also analysed for the presence of a concentration-effect relationship, but these correlation coefficients appeared to be even lower. It can be concluded that whether the onset of metal fume fever is governed by the cumulative exposure rather than the exposure concentration cannot be drawn due to the limited amount of data points and the large variability of the data. Hence it is impossible to derive a NOAEL for metal fume fever from this study with reasonable certainty. Therefore, the data from this study is considered not to supersede the study results found by Gordon et al., (1992), from which a 5 mg ZnO/m3 effect level for metal fume fever can be derived. A number of reports have addressed the etiology of metal fume fever as well, e.g. Barceloux et al., (1999), and Kelleher et al., (2000). However, these studies, as well as several case reports (e.g. Vogelmeier et al., 1987; Langham Brown, 1988; Malo et al., 1990; Ameille et al., 1992) do not allow the establishment of a clear NOAEL for metal fume fever. It is clear that metal fume fever is restricted to very specific operations using very high temperatures such as cutting or welding of galvanised steel. It is not related to the production and use of commercial grade zinc oxide. Metal fume fever is exclusively associated with freshly formed ultrafine particulate zinc oxide (<0.1 m). As these ultrafine particles rapidly agglomerate to bigger particles, which are normally encountered at production and processing sites, at these sites there is no indication for metal fume fever. By means of a questionnaire all zinc companies were asked for the incidence of metal fume fever at their site over the past decades of operation. Medical surveillance programs have been carried. Eleven companies (mainly zinc oxide producers) reported no observations of zinc metal fume fever over the last decade or in recent occupational practice (EU RAR, 2004a- f). 5.2.3. Summary and discussion of acute toxicity The acute toxicity of zinc and its compounds depends on the type of zinc compound as well as on the route of application. While the slightly soluble and insoluble zinc compounds (i.e., zinc oxide, zinc hydroxide, zinc phosphate, zinc carbonate, zinc metal and zinc sulphide) are of low acute, dermal and inhalation toxicity not requiring a classification for acute toxicity according to the EC criteria, the soluble forms of zinc displayed a higher level of acute toxicity requiring classification for oral and possibly inhalation exposure.

Soluble zinc chloride is harmful following acute oral exposure (LD50 range 1,100 to 1,260 mg/kg bw) and is

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 86 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

classified as harmful if swallowed (Xn; R22) according EC criteria (Council Directive 67/548/EEC). zinc 3 chloride has also demonstrated acute toxicity via the inhalation route (LC50 ≤ 1,975 mg/m ). However, since the exposure of the animals to the size of the particles is not truly representative of exposure to humans under normal conditions, it is difficult to assess whether or not, zinc chloride is acutely toxic since a four hour LC 50 value could not be derived and a clear dose-response relationship could not be established. Airway irritation has been observed both in animals and in humans, zinc chloride has the potential to be a respiratory tract irritant.

Soluble zinc sulphate (monohydrate, hexahydrate and heptahydrate) has LD50 oral values ranging from 574 to 2,949 mg/kg bw, 862 to 4,429 mg/kg bw and 920 to 4,725 mg/kg bw, respectively for the three forms of zinc sulphate and is classified as harmful if swallowed (Xn; R22) according EC criteria (Council Directive 67/548/EEC). Zinc sulphate is not acutely toxic via the dermal route (LD50 >2,000 mg/kg bw). Effects of inhalation exposure to zinc sulphate were limited to pulmonary effects only. Soluble zinc bis(dihydrogen phosphate) is also harmful following acute oral exposure (LD50 range 300 to 2000 mg/kg bw) and meets the classification criteria for harmful if swallowed (Xn; R22) according to EC criteria (Council Directive 67/548/EEC). While no specific acute toxicity data were identified for diammonium tetrachlorozincate and triamonium pentachlorozincate, it is (due to its similar solubility characteristics) likely to display a toxicity profile similar to that of zinc chloride, zinc sulphate or zinc bis(dihydrogen phosphate).

With LD50 values consistently exceeding 2,000 mg/kg bw, slightly soluble or insoluble zinc compounds such as, zinc oxide (LD50 ranges between 5,000 and 15,000mg/kg bw), zinc phosphate (LD50 is >5,000mg/kg bw), zinc metal (LD50 >2,000mg/kg bw) or zinc sulphide (LD50 is >15,000mg/kg bw) show low level of acute oral toxicity. Moreover, zinc oxide and zinc metal were further shown to be of low acute inhalation toxicity (i.e., LC50 values of > 5.41 and 5.7 mg/L/4hrs). Given the common characteristics shared via their solubility characteristics, the remaining slightly soluble zinc compounds are also considered to be of low acute inhalation toxicity. Of significance for humans from an acute toxicity standpoint is the occurrence of metal fume fever following exposure to ultrafine particles of special grades of zinc oxide in context of very specific operations. According to the response from 11 zinc companies to a questionnaire, there have been no observations of zinc metal fume fever over the last decade and in recent occupational practice. However in light of responsible care and since no studies are available that allow the establishment of a NOAEL for metal fume fever with a reasonable degree of certainty, a LOAEL (5 mg ZnO/m3) for 2 hours (showed the typical metal fume fever symptoms beginning 4 to 8 hours after exposure and disappearing within 24 hours) can be used for metal fume fever based on the study by Gordon et al. (1992).

5.3. Irritation 5.3.1. Skin

5.3.1.1. Non-human information Table 30. Overview of experimental studies on skin irritation according to decreasing water solubility of zinc compounds Test Study Species Endpoint Exposure Result Remarks Reference substance type Zinc Skin Guinea-pig Erythema, Daily; 0.5 Moderately 2 (reliable with Lansdown chloride irritation eschar and mL of 1% irritating restrictions) ABG, oedema solution; key study (1991) formation open-patch; 5 days Zinc Skin Rabbit Erythema, Daily; 0.5 Severely 2 (reliable with Lansdown chloride irritation eschar and mL of 1% irritating restrictions) ABG, oedema solution; key study (1991) formation open-patch; 5 days Zinc Skin Rabbit Erythema, Daily; 0.5 Severely 2 (reliable with Lansdown chloride irritation eschar and mL of 1% irritating restrictions) ABG, oedema solution; key study (1991) formation occluded; 5 days

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 87 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Test Study Species Endpoint Exposure Result Remarks Reference substance type Zinc Skin Mouse Erythema, Daily; 0.5 Severely 2 (reliable with Lansdown chloride irritation eschar and mL of 1% irritating restrictions) ABG, oedema solution; key study (1991) formation open-patch; 5 days Zinc Skin Rabbit Erythema, 0.5g; Not irritating 2 (reliable with Van sulphate irritation eschar and moistened restrictions) Huygevoort oedema substance; key study AHBM formation semi- (1999b) occlusive; 4 hours Zinc Skin Rabbit Erythema, Daily; 0.5 Not irritating 2 (reliable with Lansdown sulphate irritation eschar and mL; open- restrictions) ABG, oedema patch; 5 days key study (1991) formation Zinc Skin Guinea Pig Erythema, Daily; 0.5 Not irritating 2 (reliable with Lansdown sulphate irritation eschar and mL; open- restrictions) ABG, oedema patch; 5 days key study (1991) formation Zinc Skin Mouse Erythema, Daily; 0.5 Not irritating 2 (reliable with Lansdown sulphate irritation eschar and mL; open- restrictions) ABG, oedema patch; 5 days key study (1991) formation Zinc oxide Skin Rabbit Erythema, 500 mg; Not irritating 2 (reliable with Löser, irritation eschar and occlusive; 24 restrictions) (1977) oedema hours key study formation Zinc oxide Skin Rabbit Erythema, Daily; 0.5 Not irritating 2 (reliable with Lansdown irritation eschar and mL; restrictions) ABG, oedema occlusive; 5 key study (1991) formation days Zinc oxide Skin Rabbit Erythema, Daily; 0.5 Not irritating 2 (reliable with Lansdown irritation eschar and mL; open restrictions) ABG, oedema patch; 5 days key study (1991) formation Zinc oxide Skin Guinea-pig Erythema, Daily; 0.5 Not irritating 2 (reliable with Lansdown irritation eschar and mL; open restrictions) ABG, oedema key study (1991) formation Zinc oxide Skin Mouse Erythema, Daily; 0.5 Not irritating 2 (reliable with Lansdown irritation eschar and mL; open restrictions) ABG, oedema key study (1991) formation

Table 31. Overview of experimental studies on skin irritation for ammonium chloride Method Results Remarks Reference rabbit (New Zealand moderately irritating 2 (reliable with Microbiological and White) restrictions) Biochemical Assay Redness/Edema/Eschar: 0 of max. 3 Laboratories (1985) Coverage: occlusive (animal: all) (Time point: 48) (all supporting study (abraded) observed effects were reversible within 48 hours) (see below) experimental result Federal Register, vol.43, no 163-Tuesday, Aug.22, Test material (EC 1978 name): Ammonium

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 88 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Method Results Remarks Reference

chloride rabbit moderately irritating 4 (not assignable) BASF AG (1968)

Coverage: occlusive supporting study

patch test experimental result

Test material (EC name): Ammonium chloride

5.3.1.2. Human information Slightly soluble and insoluble zinc compounds Zinc sulphide was not irritating to human skin (Sachtleben Chemie GmbH, 2000b). No signs of skin irritation were noted when an occlusive 25% zinc oxide patch (2.9 mg Zn/cm 2) was placed on the human skin for 48 hours (Agren, 1990). A patient who was treated with 40% zinc oxide ointment (15 g on 150 cm2) under occlusive dressing displayed a rash and follicular pustules at 24 hours post-treatment. The dermal reaction disappeared 2 days after removal of the ointment and treatment with cool saline compresses, but reappeared after application of 5% zinc oxide. From the study it could not be derived whether the dermal effects were a result of zinc oxide or from other treatment- related stimuli. In 5 other patients who were treated with 40% zinc oxide ointment in a similar way and in 6 volunteers who received 100 g of 40% zinc oxide ointment on chest and legs, no signs of dermal reactions were reported (Derry et al., 1983).

5.3.2. Eye

5.3.2.1. Non-human information The results of experimental studies on eye irritation are summarised in the following table: Table 32. Overview of experimental studies on eye irritation according to decreasing water solubility of zinc compounds Test substance Study Species Endpoint Exposure Result Remarks Reference type Zinc sulphate Occular Rabbit Effects on 98.1 mg Severely 2 (reliable with Van irritation iris, cornea neat product irritating restrictions) Huygevoort and instilled into key study (1999 f) conjunctiv one eye; used in RAR, ae unrinsed (EU 2004 e) Diammonium Occular Rabbit Effects on 98.1 mg Moderately 4 (not E.I.Dupont tetrachlorozincate irritation iris, cornea neat product irritating assignable) de Nemours and instilled into supporting and Co conjuncti- one eye; study (1992) vae rinsed and unrinsed observed Zinc oxide Occular Rabbit Effects on 64 mg Not irritating 1 (reliable Van irritation iris, cornea (0.1mL) without Huygevoort

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 89 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Test substance Study Species Endpoint Exposure Result Remarks Reference type and neat product restriction) AHBM conjuncti- instilled into key study (1999 e) vae one eye; used in RAR, rinsed after (EU 2004 b) 24 hours Zinc oxide Occular Rabbit Effects on 50 mg neat Not irritating 2 (reliable with Thijssen J irritation iris, cornea product restrictions) (1978) and instilled into supporting conjuncti- one eye study vae used in RAR, (EU 2004 b) Zinc oxide Occular Rabbit Effects on 50 mg neat slightly 2(reliable with Löser E irritation iris, cornea product irritating restrictions) (1977) and instilled into supporting conjuncti- one eye study vae used in RAR, (EU 2004 a) Zinc phosphate Occular Rabbit Effects on 100 mg neat Not irritating 1(reliable Mirbeau T, irritation iris, cornea product without Guillaumat and instilled into restriction) PPO and conjuncti- one eye; key study Pelcot C vae unrinsed used in RAR, (1999) (EU 2004 d) Zinc dust Occular Rabbit Effects on 100 mg Minimally 2 (reliable with Van irritation iris, cornea instilled into irritating restrictions) Huygevoort and one eye; key study AHBM conjuncti- Median used in RAR, (1999 c) vae particle (EU 2004 a, b) diameter 4µm; rinsed after 24 hours Zinc powder Occular Rabbit Effects on 100 mg neat Minimally 2 (reliable with Van irritation iris, cornea product irritating restrictions) Huygevoort and instilled into key study AHBM conjuncti- one eye; used in RAR, (1999 d) vae Median (EU 2004 a, b) particle diameter 150µm; unrinsed

5.3.2.2. Human information Soluble zinc compounds Accidental splash of ammonium zinc chloride into three eyes of two patients resulted in corneal oedema and some permanent corneal scarring. Recovery required 6 to 28 weeks. The patient who had also splashes in his nasal passages lost all sense of smell permanently, in spite of medical treatment (Houle and Grant, 1973). Slightly soluble and insoluble zinc compounds Zinc sulphide is not irritating to human eyes (Sachtleben Chemie GmbH, 2000c). 5.3.3. Respiratory tract

5.3.3.1. Non-human information Soluble zinc compounds Rats exposed to zinc chloride in single exposure studies exhibited signs of respiratory distress and oedema (see

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 90 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 acute inhalation toxicity). Slightly soluble and insoluble zinc compounds Zinc oxide did not show any signs of upper airway irritation in acute inhalation studies. Single and repeated inhalation exposure to ultra-fine zinc oxide fumes showed changes in pulmonary function and induction of airway inflammatory responses, however a well-performed acute inhalation toxicity study in rats, did not yield any indication of signs of upper airway irritation from commercial zinc oxide aerosol (particle size: MMAD 4 m ± 2.9 (GSD)) (Klimisch et al.,1982).

5.3.3.2. Human information No information available 5.3.4. Summary and discussion of irritation Slightly soluble zinc oxide, zinc phosphate, zinc metal and insoluble zinc sulphide are not irritating to skin or eyes. The soluble zinc compounds (i.e., zinc chloride, zinc sulphate and diammonium tertachlorozincate), displayed varying degrees of skin and eye irritation ranging from moderate to severely irritating. Based on the available data of the soluble zinc compounds, soluble zinc chloride is classified as corrosive (C;R34) according to EC criteria due to severe skin irritancy seen in animals at concentrations of 1% solution and irreversible damage to eyes caused by zinc chloride after accidental exposure in humans. zinc chloride has also shown signs of respiratory tract irritation in single exposure studies (see acute inhalation toxicity). On the other hand zinc sulphate was not irritating to skin but is a severe eye irritant and has been classified as a severe eye irritant (Xi R41) according to EC criteria. While no pertinent data exists on zinc bis(dihydrogen phosphate) in vitro data with questionable reliability suggests zinc bis(dihydrogen phosphate) is not irritating to eyes. Diammonium tetrachlorozincate appears to be a moderate eye irritant however no classification has been assigned. Ammonium chloride appears to be a moderate skin irritant. While no specific irritation data were identified for triammonium pentachlorozincate, it is (due to its similar solubility characteristics) likely to display a toxicity profile similar to that of the soluble diammonium tetrachlorozincate.

Based on the available information it appears that the slightly soluble zinc oxide and insoluble zinc sulphide are not skin irritants and therefore slightly soluble zinc hydroxide, zinc phosphate, zinc carbonate and zinc metal are also expected to be not irritating to skin. Zinc oxide, zinc phosphate, zinc metal and zinc sulphide are not eye irritants and therefore zinc carbonate and zinc hydroxide are also expected to be not irritating to eyes. None of the slightly soluble or insoluble zinc compounds appear to cause respiratory tract irritation. 5.4. Corrosivity 5.4.1. Non-human information Refer to section 5.3 5.4.2. Human information Refer to section 5.3 5.4.3. Summary and discussion of corrosion As discussed in section 5.3, irritation studies indicate that soluble zinc chloride is corrosive to skin and is classified as such according to Annex I of Directive 67/548/EEC. The remaining soluble zinc compounds are not classified as corrosive. Zinc phosphate and diammonium tetrachlorozincate were moderate to severe eye irritants. The slightly soluble and insoluble zinc compounds (zinc oxide, zinc hydroxide, zinc phosphate, zinc carbonate, zinc metal and zinc sulphide) are not corrosive based on the available irritation data and therefore no classification is required according to Annex I of Directive 67/548/EEC for corrosivity. 5.5. Sensitisation 5.5.1. Skin

5.5.1.1. Non-human information

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 91 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

The results of experimental studies on skin sensitisation are summarised in the following table: Table 33. Overview of experimental studies on skin sensitisation according to decreasing water solubility of zinc compounds Test substance Method Results Remarks Reference Zinc sulphate Mouse local lymph node Negative 2 (reliable with Ikarashi Y, assay restrictions) Tsuchiya T and key study Nakamura A (1992)

Zinc sulphate Guinea pig (Dunkin- Negative 2 (reliable with Van Huygevoort Hartley) female restrictions) (1999 i) Guinea pig maximization supporting study test used in RAR, (EU 2004 e) Zinc oxide Guinea pig maximization Negative 1 (reliable without Van Huygevoort test restriction) AHBM (1999 g) key study used in RAR, (EU 2004 b) Zinc oxide Guinea pig maximization Ambiguous 1 (reliable without Van Huygevoort test restriction) AHBM (1999h1) key study Van Huygevoort AHBM (1999h2)

Zinc sulphate (ZnSO4•7 H2O) was tested in a mouse local lymph node assay (Ikarashi et al., 1992), according to the testing methods developed by Kimber et al., (1989 and 1990). After gentle dermal abrasion, 25 l of a 5% zinc sulphate solution in 20% ethanol was applied for three consecutive days at the dorsal side of both ears of 3 Balb/c mice. On the fourth day the animals were sacrificed and the ear-draining lymph nodes were collected. Lymph node lymphocyte proliferation was determined by tritiated thymidin incorporation. The results were compared to those of vehicle-treated controls. Zinc sulphate did not induce proliferative activity, whereas for potassium bichromate, nickel sulphate and cobalt chloride (known dermal sensitizers) positive results were obtained.

The skin sensitising potential of zinc sulphate (ZnSO4•7 H2O) was also investigated in guinea pigs. A well- performed maximisation test, conducted according to Directive 96/54/EC B.6 and OECD guideline 406, was carried out in female Dunkin Hartley guinea pigs. Based on the results of a preliminary study, in the main study 10 experimental animals were intradermally injected with a 0.1% concentration and epidermally exposed to a 50% concentration. Five control animals were similarly treated, but with vehicle (water) alone. Approximately 24 hours before the epidermal induction exposure all animals were treated with 10% SDS. Two weeks after the epidermal application all animals were challenged with a 50% test substance concentration and the vehicle. A second challenge followed one week after the first. In response to the 50% test substance concentration, in some experimental animals and controls skin reactions of grade 1 were observed 48 hours after the first (5/10 and 2/5, respectively) and the second challenge (4/10 and 2/5, respectively). As the skin reactions were comparable among the experimental and control animals, and as there was poor consistency of the skin reactions among individual experimental animals after the first and second challenge, the observed skin reactions can be considered to be non-specific signs of irritation. Hence, it can be concluded that zinc sulphate did not induce hypersensitivity in experimental animals (Van Huygevoort, 1999i). The skin sensitising potential of zinc oxide (purity 99.69%) was investigated in female Dunkin Hartley guinea pigs in two well-performed maximisation tests, conducted according to Directive 96/54/EC B.6 and OECD guideline 406. Based on the results of a preliminary study, in the main studies experimental animals (10 in each test) were intradermally injected with a 20% concentration and epidermally exposed to a 50% concentration (i.e. the highest practically feasible concentration). Control animals (5 in each test) were similarly treated, but with vehicle (water) alone. Approximately 24 hours before the epidermal induction exposure all animals were treated with 10% SDS. Two weeks after the epidermal application all animals were challenged with a 50% test substance concentration and the vehicle. In the first study, in response to the 50% test substance concentration skin reactions of grade 1 were observed in 4/10 experimental animals 24 hours after the challenge (40% sensitisation rate), while no skin reactions were evident in the controls. In contrast, in the second study no skin reactions were evident in the experimental animals (0% sensitisation rate), while a skin reaction grade 1 was

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 92 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 seen in one control animal. The skin reaction observed in one control animal is probably a sign of non specific irritation (Van Huygevoort, 1999h1, 1999h2). In a third well-performed maximisation test, conducted according to the same guidelines and with the same experimental design, another analytical grade zinc oxide was tested (Zincweiß Pharma A; purity 99.9%). The only difference with the studies described above was the intradermal induction concentration, which was 2% as for Zincweiß Pharma A this was considered the highest concentration that could reproducibly be injected. In this test no skin reactions were evident in both experimental and control animals, hence a 0% sensitisation rate for Zincweiß Pharma A. White staining of the treated skin by the test substance was observed in some animals 24 and 48 hours after challenge (Van Huygevoort, 1999g).

5.5.1.2. Human information Slightly soluble zinc compound In a human patch test performed with 100 selected leg-ulcer patients, 11/100 patients gave an allergic reaction with zinc ointment (60% ZnO and 40% sesame oil). However, 14/81 patients gave a positive response when treated with sesame oil alone. This study does not give any indication for a skin sensitizing potential of zinc oxide in humans (Malten and Kuiper, 1974). The effect of zinc oxide on contact allergy to colophony was investigated. With 14 patients with earlier history of moderate patch test reactions to colophony (a patch test) with 10% ZnO (2.3 mg Zinc/cm²) with and without colophony was performed. No positive response was observed in the 14 patients when only a 10% solution of zinc oxide was used. The addition of zinc oxide to colophony decreased the allergic reaction induced by colophony (Söderberg et al., 1990). 5.5.2. Respiratory system

5.5.2.1. Non-human information While is no particular study addressing respiratory sensitisation in experimental animals, there is no information suggesting zinc compounds to cause such effects animals. Taking into account the complete absence of skin sensitization potential of zinc compounds, respiratory sensitisation is not expected to be of concern for the zinc and zinc compounds considered in this chemical safety report.

5.5.2.2. Human information No reports were identified in the literature that associated zinc metal or zinc compounds with respiratory sensitization in humans. 5.5.3. Summary and discussion of sensitisation Skin sensitisation The data on slightly soluble zinc oxide indicated no skin sensitising potential (negative in animal and human studies) therefore classification for skin sensitisation is not required according to Annex I of Directive 67/548/EEC. Based on the assumption that zinc compounds with similar water solubility characteristics can be read across, it can be concluded that the other slightly soluble and insoluble zinc compounds are also expected to be non-skin sensitisers. The data on soluble zinc sulphate indicates no sensitisation potential and therefore no classification is required according to Annex I of Directive 67/548/EEC. Sensitisation is not expected from soluble zinc chloride, zinc bis(dihydrogen phosphate), diammonium tetrachlorozincate and triammonium pentachlorozincate based on the data for zinc sulphate since the soluble zinc compounds share similar solubility characteristics.

Respiratory sensitisation Considering the absence of evidence of respiratory sensitization responses in, this endpoint is not expected to be of concern for zinc and zinc compounds. 5.6. Repeated dose toxicity The repeated dose toxicity section provides an overview of the available studies for all zinc compounds which are considered key studies. The subsection “Additional supporting studies” comprises studies conducted in non standard laboratory animals, special investigations into specific parameters and which are limitedly reported.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 93 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

5.6.1. Non-human information

5.6.1.1. Repeated dose toxicity: oral The results of experimental studies are summarised in the following table: Table 34. Overview of experimental studies on repeated dose toxicity after oral administration Test substance Species Method Results Remarks Reference Zinc sulphate Mouse 90-Day oral feeding in male/female NOAEL: 2 (reliable with Maita K, Hirano ICR ICR mice; 458 mg ZnSO4/kg restrictions) M, Mitsumori K, bw/day; equalling key study Takashi K and Similar to OECD Guideline 408; 104 mg Zn/kg bw/day Shirasu Y (1981) Dietary doses: 0, 300, 3,000, 30,000 ppm; equivalent to: At LOEL of 30,000ppm blood and biochemical 42.7/46.4, 458/479, 4927/4,878 mg effects noted. ZnSO4/kg bw/day (males/females) Pathological and histopathological changes observed in kidneys, thyroid, GI tract and pancreas Zinc sulphate Rats 90-Day oral feeding in male/female NOAEL: 2 (reliable with Maita K, Hirano Wistar rats; 234 mg ZnSO4/kg restrictions) M, Mitsumori K, bw/day; equalling key study Takashi K and Similar to OECD Guideline 408; 53.5 mg Zn/kg bw/day Shirasu Y (1981) Dietary doses: 0, 300, 3,000, 30,000 ppm; equivalent to: At LOEL of 2,486 mg ZnSO4/kg bw/day blood 23.2/24.5, 234/243, and 2,514/2,486 effects and pancreatic mg/kg bw/day (males/females) damage noted;

Zinc Rats 90-Day oral feeding in male/female NOAEL: 2 (reliable with Edwards K and monoglycerolate Sprague SD rats; 31.52 mg Zn mg/kg restrictions) Buckley P (1995) Dawley bw/day; equalling key study Similar to OECD Guideline 408; 13.3 mg Zn/kg bw/day Dietary doses: 0, 0.05%, 0.2%, 30000 ppm; equivalent to: At the LOEL of 53.8 mg Zn/kg bw/day, rats 31.5/35.8, and 127.5/145.9 mg/kg displayed changes in bw/day (males/females) haematological Exposure: 13 weeks parameters, pancreatic cell necrosis; no effects were seen at LOEL in reproductive organs;

ICR mice (12/sex/group) were given daily doses of 300, 3000 or 30000 mg ZnSO4•7 H2O/kg feed (equivalent to 42.7/46.4, 458/479 and 4927/4878 mg/kg bw for males/females, respectively) during 13 weeks. A control group was included. At the highest dose level 4 males and 1 female were found dead or killed in extremis. Histological findings of these animals revealed impairment of the urinary tract and regressive changes in the exocrine gland of the pancreas. Only the high dose animals showed moderately lower haematocrit (males: from 42% in controls to 29% in high dose animals; females: from 44% in controls to 31% in high dose animals) and haemoglobin concentrations (males and females: 14 to 10 g/dL). The leucocyte counts of high dose males were moderately decreased (lymphocytes 70 to 60%; monocytes 5.3 to 4.9%). Total protein, glucose and cholesterol were reduced and alkaline phosphatase and urea nitrogen were increased in high dose animals. High dose females showed reduced ALAT and increased calcium levels, ASAT was increased in high dose males. Absolute and relative (in parentheses) thyroid weights of males were increased from 3.3 mg (0.007%) in control animals to 4.2 mg (0.0011%) in the highest dose group. Kidney weights of females were also increased from 0.42 g

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 94 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

(0.93%) in controls to 0.53 g (1.62%) at the highest dose. Gross pathology and histopathology showed changes in kidneys, thyroids, pancreas (degeneration/necrosis of acinar cells, clarification of nucleoli), gastrointestinal tract, and spleen. No effects were found on the reproductive organs (i.e. ovaries, testes, accessory sex organs). The NOAEL in this study is 458 and 479 mg ZnSO4•7 H2O/kg bw/day for males and females, respectively (equivalent to approximately 104 mg Zn/kg bw/day) (Maita et al., 1981).

Wistar rats (12/sex/group) were given daily doses of 300, 3000 or 30000 mg ZnSO 4•7 H2O/kg feed (equivalent to 23.2/24.5, 234/243, and 2514/2486 mg/kg bw for males/females, respectively) during 13 weeks. A control group was included. At the highest dose level a moderate reduction in leucocyte counts was seen in both sexes (males: from 7.3 x10³/mm³ in controls to 4.7x10³/mm³ in high dose animals; females: from 4.5x10³/mm³ in controls to 3.3x10³/mm³ in high dose animals). Compared to controls, males also showed slightly decreased haematocrit (42 to 40%), decreased total protein (5.2 to 4.4 g/dL) and cholesterol values (96 to 62 mg/dL). Absolute and relative (in parentheses) liver weights were decreased in the high dose males (from 16.1 g (3.55%) in controls to 11.9 g (3.20%) at the highest dose). Absolute kidney weights were decreased in high dose males (2.29 g vs. 2.93 g in controls). Histopathology showed pancreatic damage (degeneration, necrosis of acinar cells, clarification of centroacinar cells and interstitial fibrosis) in high dose animals. No effects were found on the reproductive organs (i.e. ovaries, testes, accessory sex organs). The NOAEL is 234 and 243 mg ZnSO4•7 H2O/kg bw/day for males and females, respectively (equivalent to approximately 53.5 mg Zn/kg bw) (Maita et al., 1981). Groups of 20 male and 20 female Sprague-Dawley rats were fed zinc monoglycerolate at dietary levels of 0, 0.05 or 0.2% (equal to 0, 31.52 or 127.52 mg/kg bw/day for males and 0, 35.78 or 145.91 mg/kg bw for females, respectively) for a period of 13 weeks in a study performed according to OECD 408. A similar group was fed 1% (equal to 719 and 805 mg/kg bw/day for males and females, respectively) of zinc monoglycerolate up to day 58 of the study when a deterioration in their clinical condition (poor physical health and reduced food intake) necessitated reducing the dietary level to 0.5% (equal to 632 and 759 mg/kg bw/day for males and females, respectively). However, as no improvement was noted, these rats were killed on humane grounds on day 64 of the study. These rats developed hypocupremia manifested as a hypochromic microcytic regenerative type anaemia (low haemoglobin and haematocrit, decreased MCV and MCH, and increased MCHC, red blood cell and reticulocyte count). Enlargement of the mesenteric lymph nodes and slight pitting of the surface of the kidneys were noted. Severe pancreatic degeneration and pathological changes in the spleen, kidneys, incisors, eyes and bones were observed. The testes of all males showed hypoplasia of the seminiferous tubules to a varying degree and in addition the prostate and seminal vesicles showed hypoplasia. In all but one female the uterus was hypoplastic. All other rats survived to the end of the 13 weeks treatment. At a dietary level of 0.2% increases in plasma ALAT, alkaline phosphatase and creatine kinase were observed in males and in plasma creatine kinase in females. Total plasma cholesterol was reduced in both males and females. The changes were statistically significant but small in absolute terms. No changes in haematological parameters were seen at 0.05 and 0.2%. A dose related reduction in the quantity of abdominal fat was noted in male rats at 0.05 and 0.2%. Enlargement of the mesenteric lymph nodes was apparent in 6 out of 20 rats fed 0.2% and in one male fed 0.05%. Microscopic examination showed a reduction in the number of trabeculae in the metaphysis of the tibia of 5 male and 3 female rats fed 0.2%, 4 males and 1 female had a similar reduction in the metaphysis of the femur. Pancreatic cell necrosis was seen in both sexes at 0.2% and a slight, but statistically not significant increase could be noted at 0.05% (3 males and 1 female). This pancreatic cell necrosis was seen also in 1 control male. A reduction in the number of pigmented macrophages in the red pulp of the spleen was observed in both sexes at 0.2% and a marginal reduction was also seen in males at 0.05%. In the animals given 0.05 and 0.2% no effects were found on the reproductive organs. Since the pancreatic cell necrosis, being without statistical significance at 0.05%, was also apparent in 1 control male and because the reduction in pigmented macrophages in the spleen was only marginal at 0.05% without any haematological changes the dose level of 0.05%, is considered as a NOAEL. This dose level is equal to 31.52 or 35.78 mg zinc monoglycerolate/kg bw/day for males and females, respectively, so the NOAEL in this study is 31.52 mg/kg bw/day equalling approximate 13.26 mg Zn/kg bw/day) (Edwards and Buckley, 1995).

Additional supporting studies Oral

A group of 150 C3H mice was given daily doses of 0.5 g ZnSO 4 (unspecified)/l drinking water for 1 year. This exposure equals approximately 100 mg ZnSO4/kg bw/day and 22.6 mg Zn/kg bw in case heptahydrate was used. A 2 months post observation period and a control group were included. At monthly intervals, 5 control and 5 test animals were investigated for plasma zinc, glucose and insulin, and for zinc in skin, liver and spleen. Histology, histochemistry and microscopy was performed on adrenals and pancreas, and on adenohypophysis only microscopy. The animals remained healthy throughout the study. Hypertrophy of the adrenal glands

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 95 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

(cellular enlargement) and hypertrophy and vacuolization of pancreatic islets and fasciculata cells in adrenal cortex from month 3 onwards. Changes indicating hyperactivity in the anterior pituitary were noted, such as increased cell size of all cell types in the pituitary. All the other parameters remained the same during the study. The study was undertaken to further investigate the effects of supplemental zinc on endocrine glands and correlate these effects with any change in body zinc levels produced (Aughey et al., 1977). Minks (3/sex/group) were given diets supplemented with 0, 500, 1000 or 1,500 mg/kg feed zinc sulphate for 144 days. Zinc concentrations in liver, pancreas and kidney increased with increasing zinc content in the diet. No histological lesions were found in these organs (Aulerich et al. 1991). Wistar rats (2 months, 16 males and 14 females) were given 0.12 mg Zn/mL drinking water (equivalent to 12 mg Zn/kg bw/day; 25 mg ZnCl2/kg bw/day) for 4 consecutive weeks. A control group was included. The body weights of exposed males and food and water intakes of both exposed sexes decreased. A statistically significant decrease in Hb level (85% of control value) and erythrocyte count was reported in the peripheral blood of treated rats. An increased leucocyte count, due to increased lymphocyte numbers was noted in treated males. No inhibition of erythropoiesis in the bone marrow was found. No more details were given in this limited study performed to investigate the effect of simultaneous oral administration of zinc and vanadium (Zaporowska and Wasilewski, 1992). Special studies were conducted to examine the morphological and histoenzymatic changes of the brain. Twelve Wistar rats were given daily doses of 100 mg ZnO (i.e., approximately 600 mg ZnO/kg bw/day or 480 mg Zn/kg bw/day) intragastrically for 10 consecutive days. A control group was included. After 10 days the rats were sacrificed and the brains were examined for morphological and histoenzymatic changes. Morphological changes included degenerative changes of neurocytes, accompanied with moderate proliferation of the oligodendroglia and glial proliferation in the white matter. Furthermore, endothelial oedema was observed in the small arterial and capillary walls. Histoenzymatic changes included decreased activities of AcP (acid phosphatase), ATPase (adenosinetriphosphatase), AChE (acetylcholinesterase), and BuTJ (Butyrylthiocholin-esterase). The activities of TPPase (thiamine pyrophophatase) and NsE (non specific esterase) were increased. No details on quantitative aspects of enzymatic changes were given. No change was seen in the alkaline phosphatase. The authors indicated that observed morphological and histoenzymatic changes were unspecific, indistinctive and most likely reversible (Kozik et al., 1980). Examination of the neurosecretory function of the hypothalamus and the hypophysis in these animals showed an increased neurosecretion in cells of the supraoptic and paraventricular nucleus of the hypothalamus along with a declined neurosecretion in the hypophysis and an enhanced release of antidiuretic hormone in the neurohypophysis (Kozik et al., 1981). It is not clear whether these observations represent an adverse effect of zinc on the brain or whether they are secondary to changes somewhere else in the body. Four groups of ferrets (3-5/group) were given 0, 500, 1,500 or 3,000 mg zinc oxide/kg feed (i.e., equivalent to 0, 81.3, 243.8 or 487.5 mg ZnO/kg bw/day). At the highest dose level (i.e., 487.5 mg ZnO/kg bw/day) all animals (3) were killed in extremis within 13 days. Macroscopic examination showed pale mucous membranes, dark coloured fluid in the stomach, blood in the intestines, orange coloured liver and enlarged kidneys showing diffuse necrosis, haemorrhages in the intestine and a severe macrocytic hypochromic anaemia. Histology showed nephrosis and extramedullary haematopoesis in the spleen. At the mid dose level of 243.8 mg ZnO/kg bw/day, the animals (4) were killed on day 7, 14 and 21 (1/2 in extremis) showing poor condition. Macroscopy showed pale livers with fatty infiltration and enlarged kidneys. Histology was comparable with the highest dose group. The heamogram showed macrocytic hypochromic anaemia, increased reticulocytes and leucocytosis.At the lowest dose level (81.3 mg ZnO/kg bw) the animals (3) were killed on day 48, 138 and 191, respectively. No clinical signs of toxicity or pathological changes were seen, apart from an extramedullary heamatopoesis in the spleen (Straube et al., 1980). Ellis et al. (1984) conducted a 14-day and a 49-day feeding study in 3 different breeds of sheep that were receiving feed containing 31 mg Zn/kg feed. The sheep received additional amounts of Zn (from ZnO) at dose levels of 261 and 731 (14 day study) or 731 and 1431 mg Zn/kg feed (49-day study). No effects were seen after 261 mg Zn/kg feed. In all other groups pancreatic lesions were seen. Administration of 240 mg Zinc (as ZnO)/kg bw for 3 times/week during 4 weeks to 42 castrated sheep resulted in an increased incidence of pancreatic lesions (Smith and Embling, 1993).

Table 35. Overview of experimental studies on repeated dose toxicity after oral administration for ammonium chloride

Method Results Remarks Reference

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 96 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 rat (Sprague-Dawley) male NOEL: 684 mg/kg bw/day 2 (reliable with Arnold et al. (1997) subchronic (oral: feed) (nominal) (male) based on: test restrictions) 0 (nominal in diet) mat. 12300ppm in the diet (= 684 mg/kg supporting study bw day) (nominal in diet) Exposure: 70 d (7 days per week7) experimental result Test material (CAS name): ammonium chloride

5.6.1.2. Repeated dose toxicity: inhalation The results of experimental studies are summarised in the following table: Table 36. Overview of experimental studies on repeated dose toxicity after inhalation Test Species Method Results Remarks Reference substance Zinc Rat Subchronic inhalation in Study focused on the 2 (reliable with Wallenborn JG, sulphate Wistar Wistar Kyoto rats evaluation of effects of zinc restrictions) Evansky P, Kyoto sulphate on cardiac changes. supporting study Shannahan JH, Aerosol concentrations No cardiac pathology, but Vallanat B, ‘nose only’ of 10, 30 and cardiac gene array analysis Ledbetter AD, 100 g zinc/m3 μ indicated small changes in (2008) Exposure: 16 weeks (5 gene expression; hrs/day for 3 days/week) No NOAEL identified; Zinc oxide Guinea pigs 5day inhalation in guinea NOAEL: 2.7 mg ZnO/m³ 2 (reliable with Lam HF, Chen Hartley pigs; restrictions) LC, Ainsworth Decreased lung volume: 7 supporting study D, Peoples S and Ultrafine particle (0.05 µm) mg/m³ air (analytical) (male) Amdur MO ‘nose only’ exposure (Other: Pulmonary function (1988) concentrations of 0, 2.7, and measurement) 7 mg ZnO/m3; decrease of Lung volumes and diffusing capacity at peaks Exposure: 5 days (3hrs/day) occurs rapidly and to a greater extent: 25 — 34 mg/m³ air (analytical) (male) (Pulmonary function measurement) Pulmonary damage: 7 mg/m³ air (analytical) (male) (Wet- lung weight/Body weight ratio and Wet-lung weight/Dry-lung weight ratio) Increased pulmonary damage at peak concentrations: 25 — 34 mg/m³ air (analytical) (male) (Wet-lung weight/Body weight ratio and Wet-lung weight/Dry-lung weight ratio) Zinc oxide Guinea pigs 6 day inhalation in guinea Decreased Vital capacity, 2(reliable with Lam HF, Conner Hartley pigs; functional residual capacity, restrictions) MW, Rogers alveolar volume and single supporting study AE, Fitzgerald S Ultrafine particle (0.05 µm) breath diffusing capacity for and Amdur MO ‘nose only’ exposure carbon monoxide and elevated (1985) concentrations of 0 and 5 lung weights due to mg ZnO/m3; inflammation Exposure: 6 days (3hrs/day)

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 97 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Animal examination at 24, 48, and 72 hours post exposure; Zinc oxide Guinea pigs 1, 2, or 3 day inhalation in Marginal LOAEL: 2.3 mg 2 (reliable with Conner MW, Hartley guinea pigs; ZnO/m3; restrictions) Flood WH and supporting study Rogers AE Ultrafine particle (0.05 µm) Morphological damage and (1988) ‘nose only’ exposure increase of inflammation concentrations of 0, 2.3, 5.9 markers and enzymes in and 12.1 mg ZnO/m3; pulmonary lavage fluid at dose levels of 5.9 and 12.1 mg Exposure: 1, 2, or 3 ZnO/m3; minimal changes in consecutive days (3hrs/day); enzyme levels in lavage fluid Microscopical examination but no morphological changes of lung tissue as well as in lung tissue 2.3 mg ZnO/m3; ; examination of pulmonary lavage fluid;

A subchronic inhalation study was conducted to evaluate the toxic effects of zinc sulphate on cardiac changes in male Wistar Kyoto rats. Rats were exposed 3days/week for 5hours/day to zinc sulphate heptahydrate concentrations of 10, 30 and 100 μg zinc/m3 nose only. A control group was exposed to filtered air only. Animals were sacrificed 48 hours after the last exposure. No significant changes were observed in neutrophil or macrophage count, total lavageable cells, or enzyme activity levels (lactate dehydrogenase, n-acetyl β- Dglucosaminidase, γ-glutamyl transferase) in bronchoalveolar lavage fluid, indicating minimal pulmonary effect. In the heart, cytosolic glutathione peroxidase activity decreased, while mitochondrial ferritin levels increased and succinate dehydrogenase activity decreased, suggesting a mitochondria-specific effect. Although no cardiac pathology was seen, cardiac gene array analysis indicated small changes in genes involved in cell signalling, a pattern concordant with known zinc effects. While changes are small in healthy rats, these may be especially relevant in individuals with pre-existing cardiovascular disease (Wallenborn et al., 2008). Male Hartley guinea pigs were exposed to 0, 2.7 or 7 mg ultrafine (0.05 m in diameter) ZnO/m3 3 hours a day for 5 days. Lung function measurements were performed every day after exposure in 5-8 animals. After the last exposure the animals were sacrificed. At the highest exposure level a gradual decrease in total lung capacity (18%) and vital capacity (22%) was seen during the exposure period. At day 4 the carbon monoxide diffusing capacity dropped to below 30% of the control level. Wet-lung weights were increased with 29%, indicating the presence of edema. Exposures up to 2.7 mg ZnO/m3 did not alter any parameters measured (Lam et al., 1988). Male Hartley guinea pigs (73) were exposed (nose-only) 3 hours a day for 6 days to 5 mg ZnO/m 3 (0.05 m in diameter). A group of 53 animals served as control group. Lung function tests (in 38 animals) were performed and the respiratory tract of the animals was morphologically examined 1, 24, 48 and 72 hours after the last exposure. Furthermore epithelial permeability (5 animals at 1 and 24 hours) and DNA synthesis in epithelial cells (5 animals at 24, 48 and 72 hours) were determined. Vital and functional residual capacity, alveolar volume and carbon monoxide diffusing capacity were all decreased and did not return to normal values 72 hours after the last exposure. Lung weights were elevated due to inflammation, still present at 72 hours after last exposure (Lam et al., 1985). Male Hartley guinea pigs were exposed to 0, 2.3, 5.9 or 12.1 mg/m3 of ZnO (as ultrafine particles with an average diameter of 0.05 m) 3 hours a day for 1, 2 or 3 consecutive nose-only exposures. Three animals from each group were examined after each exposure period, were sacrificed and lung tissues were microscopically examined, and the pulmonary lavage fluid was also examined. Exposure to 12.1 mg/m 3 increased the number of nucleated cells in lavage fluid. Exposures to 5.9 and 12.1 mg ZnO/m3 were associated with increased protein, neutrophils, and activities beta-glucuronidase, acid phosphatase, alkaline phosphatase, lactate dehydrogenase, and angiotensin-converting enzyme. The increases were dose dependent and were detectable after the second exposure, and generally increased after the third exposure. Significant morphologic damage characterized by centriacinar inflammation in the lung was seen at 5.9 and 12.1 mg/m3. Minimal changes in neutrophils and activities of lactate dehydrogenase and alkaline phosphatase were seen in the pulmonary fluid at the lowest dose level of 2.3 mg/m3 after 3 exposures but no morphologic changes were observed at this dose level. Based on these results 2.3 mg ZnO/m3 is considered as a marginal LOAEL in this study (Conner et al., 1988).

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 98 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

5.6.1.3. Repeated dose toxicity: dermal Considering that the dermal absorption of zinc compounds is low (see toxicokinetics section 5.1) and the lack of acute dermal effects (see acute toxicity), this endpoint is considered not to be of concern.

5.6.1.4. Repeated dose toxicity: other routes Considering the available information on repeated dose toxicity via oral and inhalation routes of exposure, other routes are considered not pertinent for this chemical safety assessment. 5.6.2. Human information Dietary levels were not measured in all of the studies reported here. According to a Total Diet Study performed by the US Food and Drug Administration (FDA) over the period 1982 to 1986, adult males (25-35 years of age) consumed an average of 16.4 mg Zn/day. Adult females (25-30 years of age) consumed an average of 9.72 mg Zn/day (Pennington, 1989). Oral In a double-blind cross-over trial 47 healthy volunteers (26 females and 21 men) ingested zinc sulphate capsules containing 220 mg zinc sulphate, three times a day with each meal (resulting in a total daily dose of 150 mg Zn which equals approximately 2.1 and 2.5 mg Zn/kg bw /day for males and females, respectively) for six weeks. Plasma zinc and copper levels, plasma cholesterol, plasma low-density-lipoprotein (LDL) and high-density- lipoprotein (HDL) cholesterol, serum ceruloplasmin and erythrocyte superoxide dismutase (ESOD) were determined. In 84% of the women and 18% of the men symptoms were reported which included headaches, nausea, vomiting, loss of appetite and abdominal cramps. The study authors reported that the gastric discomfort went together with lower body weights and taking the capsules with small meals (breakfast or morning tea) or no food. Plasma zinc levels rose significantly in both men and women (36% and 57%, respectively). Plasma copper levels did not change significantly. Total plasma cholesterol and HDL were unchanged in both sexes. In females the LDL cholesterol decreased significantly from 2.38 to 2.17 mmol/L. In females a decrease was also found in serum ceruloplasmin (13% reduction) and in ESOD (20% reduction) (Samman and Roberts, 1987, 1988). Hooper et al. (1980) examined the effect of oral zinc administration on human lipoprotein values. Twelve healthy adult men were given oral doses of 440 mg zinc sulphate/day (equals approximate 2.3 mg Zn/kg bw/day in the form of two zinc sulphate capsules containing 220 mg zinc sulphate (80 mg elemental zinc per capsule resulting in a total daily dose of 160 mg Zn), each capsule to be eaten with a main meal for 35 days. Fasting lipid levels were determined on a weekly basis and continued two weeks after zinc supplementation stopped, with a final determination at 16 weeks after start of the experiment. HDL cholesterol levels were decreased by 25% at the 7th week, but had returned to baseline levels at 16 weeks. Total serum cholesterol, triglyceride and LDL cholesterol levels were not changed. There is a discrepancy between the dosimetric data in the Samman and Roberts study (1987/1988) as compared to the Hooper et al. study (1980). In the first study, a daily dose of 660 mg zinc sulphate was declared to be equivalent to a dose of 150 mg Zn per day, while in the second study a daily dose of 440 mg zinc sulphate was stated to have resulted in a daily dose of 160 mg Zn. This discrepancy can only be explained by assuming that in the Samman and Roberts study zinc sulphate was administered in the form of the heptahydrate, while in the Hooper et al. study the monohydrate has been used. As this is not clearly stated in either of the two studies, the dosimetric data which are presented here are the same as those given in the respective publications. Chandra (1984) examined the effect of zinc on immune response and serum lipoproteins. Zinc sulphate was administered twice daily to 11 adult men for a total (extra) intake of 300 mg elemental zinc/day (i.e., approximately 4.3 mg Zn/kg bw/day). Dietary zinc intake amounted to ca 11 mg/person/day. None of the subjects showed evidence of any untoward side-effects. There was a significant increase in serum zinc levels and reduction in lymphocyte stimulating response to PHA after 4 and 6 weeks of treatment. A slight increase in LDL was observed together with a significant reduced level of HDL cholesterol. In two studies, the side-effects of zinc administration as a medication in the treatment chronic leg ulcers were investigated. In a double-blind trial, 13 humans received 200 mg zinc sulphate (i.e., approximately 135 mg Zn) three times a day for 18 weeks, while 14 humans received a placebo. No signs of nephrotoxicity associated with the zinc treatment were reported, but the study was not sufficiently documented to fully appreciate the relevance of its result (Hallbook and Lanner, 1972). In a study of Greaves and Skillen (1970) no indications for heamatotoxicity, hepatotoxicity or nephrotoxicity, was determined by several clinical biochemical and haematological parameters, were seen in 18 humans after

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 99 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 administration of 220 mg zinc sulphate (i.e., approximately 150 mg Zn) 3 times a day for 16-26 weeks. In a 12-week double blind study Black et al. (1988) administered zinc gluconate tablets to 2 groups of healthy male volunteers for 12 weeks at doses equivalent to 50 or 75 mg zinc/kg bw/day (i.e., approximately 0.71 and 1.1 mg Zn/kg bw/day). A control group received a placebo tablet. No changes in serum cholesterol, triglyceride, and LDL and very-low-density-lipoprotein (VLDL) cholesterol levels were observed. In a 10-week single-blind oral study conducted by Yadrick et al., (1989), 9 healthy female volunteers were given 50 mg Zn/day in form of zinc gluconate (i.e., approximately 0.83 mg Zn/kg bw/day) and 9 other healthy female volunteers were given 50 mg Zn /day plus (as zinc gluconate) 50 mg iron (as iron sulphate monohydrate) in two daily doses via their diet to investigate the effect of zinc supplementation on iron, copper and zinc status. The subjects served as their own controls. In both groups the erythrocyte superoxide dismutase (ESOD) activity was significantly reduced with 47% after 10 weeks. In the zinc supplemented group, significant decreases in haematocrit (by 4%) and serum ferritin levels (with 23%) were seen after 10 weeks, whereas the haemoglobin levels were unchanged. In the zinc iron supplemented group, serum ferritin levels were significantly increased by approximately 25%, whereas the haematocrit and haemoglobin levels were unchanged. The ceruloplasmin concentration, another indicator for copper status besides ESOD, was not altered in both groups, but the serum zinc concentration was significantly increased. The NOAEL in this study was less than 0.83 mg Zn/kg bw/day. A significant decrease of 15% in ESOD activity was reported by Fischer et al., (1984) who administered 50 mg Zn /day in form of zinc gluconate (i.e., approximate 0.71 mg Zn/kg bw/day) divided in two daily doses to 13 healthy young men with assumed mean body weight of 70 kg for 6 weeks in a double-blind study design. The other two indices of copper status, i.e. the ceruloplasmin activity and plasma copper levels were not changed compared to the controls at 2, 4 or 6 weeks, but the serum zinc levels were significantly increased from 2 weeks of supplementation onwards. Serum zinc showed a significant inverse correlation with ESOD activity at 6 weeks. In a controlled metabolic-unit study by Davis et al., (2000), various indicators of zinc status were measured in 25 healthy postmenopausal women (mean age 64.9 years) to evaluate the usefulness of these indicators as a marker for the functional assessment of zinc status in humans. The subjects were kept under close supervision for 200 days, divided into two 90-day dietary periods, each preceded by a 10-day equilibration period. The subjects received a daily diet with a total energy content of 8.4 MJ (or 2000 kcal). In the equilibration periods the subjects received a diet containing 2 mg copper/day and 9 mg zinc/day. For the 90-day dietary periods the subjects were randomly divided into two groups, one group (n=12) was fed a low copper diet (1 mg Cu/day) and the other group (n=13) a high copper diet (3 mg Cu/day). In the first 90-day dietary period both groups received no zinc supplement (low zinc; 3 mg Zn/day), while in the second 90-day dietary period both groups received a zinc supplement of 50 mg per day (high zinc; 53 mg Zn/day). Zinc was supplemented as zinc gluconate and copper as cupric sulphate. Blood samples were taken (after overnight fasting for 12 hours) during each of the equilibration periods and one to twice monthly during the dietary periods, and analysed for various zinc-status indicators. Zinc concentrations in erythrocytes and erythrocyte membranes, plasma and erythrocyte membrane alkaline phosphatase activities, and erythrocyte membrane 5’nucleotidase activity did not change statistically significantly with the different dietary treatments. Zinc supplementation significantly increased plasma zinc concentrations and activities of mononuclear 5’nucleotidase and extracellular superoxide dismutase (P<0.0001). For all three indicators the effect of zinc supplementation was dependent on the copper intake although this was not statistically significant for plasma zinc. In case of mononuclear 5’nucleotidase activity, the difference caused by zinc supplementation was apparent when subjects were fed high dietary copper (92% change) but not when they were fed low dietary copper (5% change). The effects for plasma zinc and for extracellular superoxide dismutase activity were more apparent when subjects were fed low dietary copper (35 vs. 22% and 21 vs. 8% change, respectively). Independent of copper intake, zinc supplementation caused relatively small increases in free thyroxine (7-8%) and triiodothyronine (7-9%) concentrations, platelet zinc concentrations (10- 13%) and bone specific alkaline phosphatase activity (18%) (0.00218.4 mol/L. Decreased activities upon zinc supplementation were found for plasma 5’nucleotidase activity (P<0.0001), thyroid stimulating hormone concentrations (P<0.07) and erythrocyte superoxide dismutase activity (ESOD; not statistically significant). For these three indicators the decrease was somewhat more apparent when fed high dietary copper (28 vs. 29%, 5 vs. 9%, and 3 vs. 5%, respectively). However, for plasma 5’nucleotidase and

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 100 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

ESOD the levels at high dietary copper were higher than at low dietary copper (only at high copper/low zinc the levels were elevated from equilibration values). For thyroid stimulating hormone the levels were depressed from equilibration values at all dietary treatments. Limited data suggested that zinc supplementation in combination with low dietary copper depresses amyloid precursor protein expression in platelets (Davis et al., 2000). In the same dietary experiment as described by Davis et al., (2000) other parameters (i.e. copper-status and iron- status indicators) were investigated to study the effect of moderately excessive and deficient intakes of zinc on copper metabolism and utilization in humans fed low and luxuriant amounts of copper (Milne et al., 2001). For that purpose, urine and faeces were collected during the last 78 days of each 90-day dietary period and copper and zinc were determined (in faeces in 6-day composite samples). Once weekly blood was sampled (after overnight fasting for 12 hours), and blood samples were analysed for various copper-status and iron-status indicators. Women fed low copper were in negative copper balance. Zinc intake (low or high) did not alter this. Women fed high copper were put into negative copper balance by low zinc. Upon transition to high zinc, women fed high copper came into positive copper balance, which apparently was the result of a lower amount of dietary copper lost in the faeces; urinary copper was not affected. The zinc balance reflected dietary zinc intake (more positive with increased zinc intake) and was not significantly affected by copper intake. Copper-status indicators were variably affected by dietary treatment. The concentrations of serum ceruloplasmin (enzymatically determined), HDL and VLDL cholesterol, triglycerides and red blood cell zinc did not change statistically significantly with the different dietary treatments. Independent of zinc intake, plasma copper concentrations were significantly lower on low dietary copper than on high dietary copper (P<0.07). Although plasma copper concentrations were depressed from equilibration values at all dietary treatments, the depression was less for high than for low dietary copper (P<0.03). Independent of copper intake, zinc supplementation caused increases in the concentrations of serum ceruloplasmin (immunochemically determined; 4-8%, P<0.05) and plasma zinc (19-32%, P<0.0001) and in platelet cytochrome c oxidase activity (on a platelet number basis; 19-27%, P<0.0007), and decreases in the concentrations of red blood cell copper (8-16%, P<0.0008) and whole blood glutathione (8-12%, P<0.009) and in the activities of specific ceruloplasmin (defined as the ratio between enzymatic and immunoreactive ceruloplasmin; 8-11%, P<0.0003) and erythrocyte glutathione peroxidase (11-15%, P<0.002). The levels of these indicators were elevated from equilibration values at all dietary treatments, with the exception of serum immunoreactive ceruloplasmin concentration (reduced at all dietary treatments), platelet cytochrome c oxidase activity (reduced at high copper/low zinc), specific ceruloplasmin activity and whole blood glutathione concentration (essentially at equilibration values at low copper/high zinc and high copper/high zinc), and red blood cell copper concentration (essentially at equilibration value at low copper/low zinc and reduced at low copper/high zinc). Zinc supplementation significantly decreased ESOD activity (5-7%, P<0.03) as well as the concentrations of total cholesterol (3-4%, P<0.005) and LDL cholesterol (2-6%, P<0.003), but not by much. The effect on ESOD was dependent on copper intake (P<0.0001): compared to equilibration values, ESOD activity decreased on low copper but increased on high copper. Total cholesterol and LDL cholesterol concentrations were significantly higher on low dietary copper than on high dietary copper (P<0.02 and P<0.03, respectively). This suggests a dependency on copper intake, but it should be noted that women fed low copper had higher equilibration values for both indicators than women fed high copper. The authors stated that measured indicators of iron status (serum iron, haemoglobin, haematocrit and percent transferrin saturation) were unaffected by dietary treatment (no data presented), with the exception of haemoglobin, which was lower on high zinc than on low zinc in both the low and high copper groups. The drop in haemoglobin occurred especially during the last month of zinc supplementation, possibly due to the frequent blood sampling. Data from another two volunteers (one on a low copper diet and one on a high copper diet) were not included, because they were using an adhesive containing extremely high amounts of zinc for their false teeth. In the human studies described above, the effects of high or moderately high dietary zinc on several indicators known to be associated with copper status have been investigated. These indicators included plasma zinc and copper concentrations, cholesterol and lipoprotein cholesterol concentrations, and several enzyme activities (e.g. ESOD and ceruloplasmin). Effects of zinc on the latter are thought to precede changes in plasma and tissue levels of the elements, given the primary role of zinc as a component of different enzymes. In humans supplemented with zinc, plasma zinc concentration was elevated, while plasma copper concentration was not affected. In the earlier studies by Samman and Roberts (1987/1988), Yadrick et al., (1989) and Fischer et al. (1984) reductions in ESOD activity were found upon zinc supplementation. This was thought to be associated with copper deficiency, as was the reduction in ceruloplasmin activity found by Samman and Roberts (1987/1988). In the more recent and more sophisticated studies by Davis et al., (2000) and Milne et al., (2001), however, only very small reductions in ESOD activity were observed that did not correlate with changes in

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 101 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 copper balance. The clinical significance of this ESOD reduction is questionable, because the findings in these studies on more specific copper deprivation signs (i.e., decreased serum ceruloplasmin and platelet cytochrome c oxidase) indicate that sub-optimal intake of zinc was more effective than a moderately high intake of zinc in inducing changes associated with a decreased copper status in postmenopausal women. It might also be that the small decrease in ESOD activity with high zinc intake was not caused by an interference with copper metabolism, but was more reflective of reduced oxidative stress given the serum glutathione and erythrocyte glutathione peroxidase findings. However, one can only conclude from the Grand Forks studies (Davis et al., 2000; Milne et al., 2001) that very subtle changes were induced by the different dietary treatments. From various studies (e.g. Fischer et al., 1990; Barnett and King, 1995; Verhagen et al., 1996 and Puscas et al., 1999), it can be concluded that ESOD activities in healthy human volunteers may show a coefficient of variation of at least 10 to 20%. Although it is impossible to compare the absolute ESOD activities as reported by these authors to those from the Grand Forks studies, due to methodological differences, the relative changes in activities as reported by Davis et al., (2000) and Milne et al., (2001) can be compared to the coefficient of variation of ESOD activity, showing that the changes found in the Grand Forks studies are within the range of natural variation. In addition, Fischer et al., (1990) have demonstrated that in a large group of male and female human volunteers of different ages, ceruloplasmin and serum copper levels were highly correlated, but that no correlation between serum copper concentration and ESOD could be established. ESOD activity was independent of sex, age, pre-post menopausal status, estrogen use (including that in post-menopausal women), smoking or drinking habits, or level of physical exercise. The general function of ESOD, also within red blood cells, is to catalyze the dismutation of superoxide anion radicals to hydrogen peroxide and oxygen, thus preventing damage of cell constituents and structures by this radical intermediate generated during the oxygen transport function. Concentrations of superoxide anion radicals are in the order of 0.01 – 0.001 nmol/l under non-pathological conditions. Hydrogen peroxide, on the other hand, is destroyed by catalase being present in high amounts within erythrocytes resulting in concentrations between 1 and 100 nmol/L. According to our knowledge there are only few measured data available showing a direct relationship between changes of intracellular concentrations of free radicals and tissue damage. Assuming that there is a considerable reduction of the ESOD activity then higher concentration of superoxide radical anions should occur in red blood cells which may lead to destructive effects. Such effects should be detectable, e.g. by changes in haematological parameters (e.g., increased hemolysis, decreased number of erythrocytes, increase in reticulocytes). However, such findings have not been observed in any study. In the Grand Forks studies (Milne et al., 2001) hematocrit, serum iron, and transferrin saturation were unaffected by a dose of 50 mg Zn/day leading to a 3-7% reduction of ESOD activity. Yadrick et al., (1989) reported a 47% decrease of ESOD activity after giving 50 mg Zn/day over 10 weeks However, this decrease of ESOD is accompanied by a small decrease in hematocit value. The subtle changes in clinical-biochemical parameters, as reported in the Grand Forks studies, are hardly indicative for zinc-induced perturbations of the copper homeostasis. These biochemical changes do not lead to detectable deterioration of red blood cell functioning. Therefore, these changes are also of marginal biological significance, if any. Hence, it is concluded that in women supplemented with zinc, a dose of 50 mg Zn/day is the NOAEL.

5.6.3. Summary and discussion of repeated dose toxicity

Ammonium:

The NOEL for NH4Cl was determined to be 684 mg/kg bw day.

The clinical signs form NH4Cl are an increase of bladder weight and an inrease of calcium in urine. The pH of unrine decreased.

Zinc:

The biological activities of zinc compounds are determined by their ability to release zinc under the respective exposure conditions. Hence, information on the effects of systemically available zinc allows the repeated dose toxicity assessment across all those zinc compounds covered in this safety report. Non-human information

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 102 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

The repeated dose toxicity of water soluble zinc sulphate and zinc monoglycerolate has been examined in a total of 3 subchronic oral feeding studies. Due to the different dosing regimens, the lowest NOAEL was determined to be 31.5 mg/kg bw/day of zinc monoglycerolate which equals a total zinc exposure of approximate 13 mg/kg bw/day. The zinc NOAEL derived from the feeding studies with zinc sulphate was determined to be 104 mg Zn/kg bw/day in mice and approximately 53.5 mg/kg bw/day in rats. At higher doses the most important effects in the rats were the development of hypocupremia, and significant changes in the pancreas (i.e., focal acinar degeneration and necrosis) and a decreased number of pigmented macrophages in spleen. No longer term inhalation studies allowing to derive a robust NOEL for the inhalatory exposure of the respective zinc compounds has been identified. In a short term 3-day inhalation study with guinea pigs, a concentration of 2.3 mg ultrafine ZnO/m3 (3 hours/day) resulted in changes in neutrophils and activities of lactate dehydrogenase and alkaline phosphatase in the pulmonary fluid. At higher concentrations increased protein concentration, neutrophils, and enzyme activities in lung lavage fluids were seen, together with significant centriacinar inflammation of the pulmonary tissue. Inhalatory doses of 2.7 mg ultrafine ZnO/m 3 for 5 days 3hours/day did not alter the lung function parameters in guinea pigs, but at 5 and 7 mg ultrafine ZnO/m 3 exposure according to a similar pattern, a gradual decrease in total lung capacity, vital capacity and reduction of the carbon monoxide diffusing capacity was seen in combination with inflammatory changes and edema. The relevance of the findings in studies with ultra-fine zinc oxide fumes is unclear with respect to commercial grade zinc oxide, as the latter is of much larger particle size and can have different toxicological characteristics. Human information Upon supplementing men and women with 150 mg Zn/day (as zinc sulphate capsules), women appeared to be more sensitive than men to the effects of high zinc intake: clinical signs such as headache, nausea and gastric discomfort were more frequent among women and women but not men had decreased activities of serum ceruloplasmin and ESOD. In some earlier oral studies in which humans were supplemented with moderately high amounts of zinc (50 mg Zn/day), a reduction in ESOD activity was also observed and again women appeared to be more sensitive to this effect. Hence, a reduction in ESOD was thought to be a sensitive indicator of copper status. However, in more recent and more sophisticated studies using the same dose level, ESOD was only marginally reduced (without a correlation with changes in copper balance), while findings on more specific copper deprivation signs (decreased serum ceruloplasmin and platelet cytochrome c oxidase) indicated that a sub-optimal intake of zinc was more effective than a moderately high intake of zinc in inducing changes associated with a decreased copper status in postmenopausal women. Given this, and the degree of the observed ESOD reduction in comparison to the natural variability in its activity, the zinc-induced decrease in ESOD activity is considered to have marginal biological significance, if any and also because it may not have been caused by an interference with copper metabolism as deep tissue SOD increases as a function of zinc exposure was observed. Overall, it can be concluded that from studies in which humans were supplemented with zinc (as zinc gluconate), that women are more sensitive to the effects of high zinc intake and that a dose of 50 mg Zn/day is the human NOAEL. This equals a daily exposure of 0.83 mg/kg bw. At the LOAEL of 150 mg Zn/day, clinical signs and indications for disturbance of copper homeostasis have been observed.

5.7. Mutagenicity 5.7.1. Non-human information

5.7.1.1. In vitro data The results of experimental studies are summarised in the following table: Table 37. Overview of experimental in vitro genotoxicity studies according to decreasing water solubility

Test substance Endpoint Species Method Results Remarks Reference Zinc chloride Bacillus Bacillus Bacillus Negative 2 (reliable Kada et al., subtilis subtilis subtilis with (1980) recombination recombination restrictions) assay (DNA assay supporting repair) study

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 103 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Test substance Endpoint Species Method Results Remarks Reference Zinc chloride Bacterial E. coli Other: Ambiguous 4 (not Rossman et al., assay (gene (strain WP2s induction of λ (two-fold assignable) (1984) mutation) ()) prophage increase of supporting (adaptation of λ prophage study McCarroll et induction) al. 1981); conc. 3200 μmol/l; m.a. unknown Zinc chloride Eukaryotic Mouse Unknown: Negative 2 (reliable Amacher and assay (gene lymphoma without m.a. with Paillet (1980) mutation) cells restrictions) key study Zinc chloride Cytogenetic Human Doses: Negative 2 (reliable Someya et al., assay dental pulp Concentration with (2008) (chromosomal cells (D824 in (uM) restrictions) aberrations) cells) 30 100, 300 supporting (met. act.: study with and without) Zinc chloride Cytogenetic Human Other: m.a. Ambiguous 2 (reliable Deknudt and assay lymphocytes unknown; with Deminatti (chromosomal 0, 30 and 300 restrictions) (1978) aberrations) μM (3mM supporting toxic) study Zinc chloride Cytogenetic Human Other: Negative 2 (reliable Deknudt (1982) assay lymphocytes without m.a.; with (chromosomal 0, 20, and 200 restrictions) aberrations) μg/culture supporting (2000 μg study toxic) used in RAR (EU 2004 c) Zinc chloride Cell Syrian Unknown; up Negative 2 (reliable Di Paolo and transformation hamster to 20 μg /ml with Casto (1979) assay embryo cells restrictions) supporting study Zinc sulphate Bacterial test S. Ames test: Negative 2 (reliable Gocke et al., (gene typhimurium with and with (1981) mutation) (5 strains) without m.a. ; restrictions) 5 doses, up to key study 3600 µg/plate Zinc sulphate Bacterial test S. Other: Negative 2 (reliable Marzin and Phi (gene typhimurium without m.a.; with (1985) mutation) (1 strain) up to 3000 restrictions) nM/plate supporting study Zinc sulphate Eukaryotic S. cerevisiae Other: Weakly 2 (reliable Singh, (1983) assay (gene (1 strain) without m.a.; positive with mutation) single (no details restrictions) concentration given) supporting (0.1 mol/L study screening assay

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 104 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Test substance Endpoint Species Method Results Remarks Reference Zinc sulphate Eukaryotic S. cerevisiae Unknown: Negative 2 (reliable Siebert et al., assay (gene (1 strain) m.a. with (1970) mutation) Unknown; restrictions) 1000 and supporting 5000 ppm study Zinc sulphate Cytogenetic Human Unknown: Negative 2 (reliable Litton Bionetics assay embryonic without m.a.; with (1974) lung 0.1, 1.0 and restrictions) cells:WI-38 10 µg/plate supporting study Zinc Bacterial test S. Ames test: Negative 1 (reliable Research bis(dihydrogen (gene typhimurium with and without Institute of phosphate) mutation) (4 strains) without m.a; restriction) Organic and E. coli 50- key study Synthesis inc., (strain WP2 5000µg/plate CETA (2010) uvrA) Zinc oxide Bacterial test S. Ames test; Negative 2 (reliable Crebelli et al., (gene typhimurium 1000 – 5000 with (1985) mutation) (4 strains) μg/plate restrictions) key study Zinc oxide Bacterial test S. Ames test Negative 2 (reliable Litton Bionetics (gene typhimurium with (1976) mutation) (3 strains) restrictions) supporting study Zinc oxide Eukaryotic Mouse Unknown: Positive 2 (reliable Cameron (1991) assay (gene lymphoma with and with mutation) cells without m.a. restrictions) supporting study used in RAR (EU 2004 b) Zinc oxide Cytogenetic Syrian Unknown; Ambiguous 2 (reliable Suzuki (1987) assay (sister hamster m.a. unknown with chromatide embryo cells restrictions) exchange) supporting study Zinc oxide Cytogenetic Human Doses: Positive 2 (reliable Someya et al., assay dental pulp Concentration with (2008) (chromosomal cells (D824 in (uM) restrictions) aberrations) cells) 30 100, 300 supporting (met. act.: study without) Zinc oxide Cytogenetic Syrian Doses: Positive 2 (reliable Hikiba et al., assay hamster Concentration with (2005) (chromosomal embryo in (uM) restrictions) aberrations) cells) 0, 60, 120, supporting 180(met. act.: study without Zinc oxide Unscheduled Syrian Unknown: Positive  2 (reliable Suzuki (1987) DNA hamster without m.a.; 1 μ g/mL with synthesis embryo cells 0.3, 1, 3, 10 restrictions) and 30 µg/mL supporting study

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 105 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Test substance Endpoint Species Method Results Remarks Reference Zinc oxide Cell Syrian Unknown: Positive 1 2 (reliable Suzuki (1987) transformation hamster without m.a.; and 3 with assay embryo cells 0, 1, 3 μg μg/mL restrictions) ZnO/ mL supporting study Zinc Bacterial test S. According to Negative 2 (reliable Jones and Gant, monoglycerolate (gene typhimurium OECD with (1994) mutation) (4 strains) guideline No. restrictions) 471; supporting 50 - 5000 study μg/plate; no toxicity up to 5000 μg/plate Zinc Eukaryotic Mouse According to Positive: 2 (reliable Adams and monoglycerolate assay (gene lymphoma OECD without with Kirkpatrick mutation) cells guideline No. m.a. from restrictions) (1994) 476; 10 μg/mL supporting without m.a. with m.a. study 1-15 μg/mL from 15 (toxic at 15 μg/mL μg/mL) with m.a. 1- 30 μg/mL (toxic at 30 μg/mL) Zinc Cytogenetic Human According to Positive in 2 (reliable Akhurst and monoglycerolate assay lymphocytes OECD the with Kitching (1994) (chromosomal guideline No. presence of restrictions) aberrations) 473; m.a. at 30 supporting cytotoxicity at and 40 study 40 μg/mL (MI μg/mL 51%), con. tested: without m.a. 5 – 20μg/mL, with m.a. 10 – 40 μg/mL

Table 38. Overview of experimental in vitro genotoxicity studies for ammonium chloride

Method Results Remarks Reference bacterial reverse mutation assay Evaluation of results: 1 (reliable without n.n. (2003) (e.g. Ames test) (gene mutation) negative restriction)

S. typhimurium TA 1535, TA Test results: supporting study 1537, TA 98 and TA 100 (met. act.: with and without) not determined for S. experimental result typhimurium TA 1535, TA E. coli WP2 uvr A (met. act.: 1537, TA 98 and TA 100(all Test material (CAS with and without) strains/cell types tested); met. name): ammonium act.: with and without; chloride Doses: 0, 4, 20, 100, 500, 2500, cytotoxicity: no 5000 micro gram/plate not determined for E. coli WP2 uvr A(all strains/cell types OECD Guideline 471 (Bacterial tested); met. act.: with and Reverse Mutation Assay) without; cytotoxicity: no, but tested up to limit concentrations

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 106 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Method Results Remarks Reference chromosomal aberration test in Evaluation of results: 2 (reliable with n.n. (2003) Chinese hamster lung cells positive without activation in restrictions) (CHL/IU) (chromosome due to physical-chemical supporting study aberration) properties of the substance experimental result Chinese hamster lung cells. (met. Test results: act.: without) not determined for CHL/IU Test material (CAS Doses: 0 - 0.4mg/ml cell(all strains/cell types tested);name): ammonium met. act.: without; cytotoxicity: chloride equivalent or similar to yes chromosomal aberration test in Chinese hamster lung cells (CHL/IU) bacterial reverse mutation assay Evaluation of results: 2 (reliable with n.n. (2003) (e.g. Ames test) (gene mutation) negative restrictions) Ishidate et al. S. typhimurium TA 1535, TA Test results: supporting study (1983) 1537, TA 98 and TA 100 (met. not determined for S. typhimuriumexperimental result act.: with and without) TA 1535, TA 1537, TA 98 and Test material Doses: Six different TA 100(all strains/cell types (compound): concentrations. Maximum tested); met. act.: with and unknown inorganic concentration was 10mg/plate without; cytotoxicity: no ammonium compund equivalent or similar to OECD in food Guideline 471 (Bacterial Reverse Mutation Assay)

In vitro genotoxicity assays are only available for soluble and slightly soluble zinc compounds. No data were identified for zinc sulphide. However, like for the other toxicity endpoints, there is common agreement that the ionic form of zinc is responsible for the biological activities of zinc compounds in general. Hence, information on the in vitro genotoxicity of soluble or slightly soluble zinc compounds is considered to be suitable for the assessment of any potential genotoxic activity of zinc metal. The genotoxicity of soluble zinc compounds zinc chloride and zinc sulphate as well as slightly soluble zinc compounds zinc oxide and zinc monoglycerolate in vitro has been extensively studied in various bacterial and mammalian test systems. This included mutagenicity and clastogenicity assays as well as in vitro UDS and cell transformation assays. All investigated zinc compounds were predominantly negative in bacterial and mammalian mutagenicity assays. Conflicting information was, however, found in clastogenicity (i.e., chromosomal aberrations, sister chromatide exchange) and the cell transformation assays where negative as well as positive results were obtained. In case clastogenic effects were observed, these were generally considered to be weak and occurred only at high, often cytotoxic concentrations. While zinc acetate and zinc 2,4-pentanedione were negative, Zinc oxide was positive in the in vitro UDS assay.

5.7.1.2. In vivo data The results of experimental studies are summarised in the following table:

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 107 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Table 39. Overview of experimental in vivo genotoxicity studies according to decreasing water solubility Test substance Endpoint Species Method Results Remarks Reference Zinc chloride Cytogenetic Mouse Other: 0.5% Slightly 2 (reliable Deknudt assay zinc in positive in with (1982) (chromosomal calcium- case of restrictions) aberrations) deficient calcium supporting (0.03% Ca) or deficient diet study standard diet in the (1.1% Ca) for survivors 30 days (0.5% Zn with poor Ca- diet resulted in 50% mortality after 30 days) Zinc chloride Cytogenetic Mouse Other; single Single dose 2 (reliable Gupta et assay i.p. injections study: with al., (1991) (chromosomal of 0, 7.5, 10 or positive; restrictions) aberrations) 15 mg repeated dose supporting ZnCl2/kg bw study: study and repeated Positive i.p. injections every other day of 2 and 3 mg ZnCl2/kg bw for 8, 16 or 24 days. Zinc chloride Drosophila Drosophila Unknown; Negative 2 (reliable Carpenter SLRL test melanogaster 0.247 mg/mL with and Ray adult feeding restrictions) (1969) supporting study Zinc sulphate Cytogenetic Rat Other: 2.75, Negative 2 (reliable Litton assay 27.5 or 275 with Bionetics (chromosomal mg/kg bw by restrictions) (1974) aberrations) gavage once or supporting daily for 5 study consecutive days Zinc sulphate Micronucleus Mouse Other: i.p. Negative 2 (reliable Gocke et 28.8, 57.5 or with al. (1981) 86.3 mg/kg bw restrictions) at 0 and 24 key study hours Zinc sulphate Host- Mouse Other: 2.75, Weakly 2 (reliable Litton Mediated 27.5 or 275 positive with Bionetics Assay mg/kg bw by restrictions) (1974) gavage once or supporting daily for 5 study consecutive days

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 108 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Test substance Endpoint Species Method Results Remarks Reference Zinc sulphate Dominant Rat other: 2.75, Negative 2 (reliable Litton lethal assay 27.5 or 275 with Bionetics mg/kg bw by restrictions) (1974) gavage once or supporting daily for 5 study consecutive days Zinc sulphate Drosophila Drosophila other; 5 mM Negative 2 (reliable Gocke et SLRL test melanogaster (in 5% with al., (1981) saccharose) restrictions) adult feeding supporting method study Zinc oxide Cytogenetic Rat Other: 5 Slight 2 (reliable Voroshilin assay months increases of with et al., (chromosomal inhalation of chromosomal restrictions) (1978) aberrations) 0.1 to 0.5 aberrations supporting mg/m3 were seen; study primarily hyperdiploid cells were seen. Zinc Micronucleus Rat Other: Negative 2 (reliable Windebank monoglycerolate resembling with et al., OECD restrictions) (1995) guideline No. supporting 474; 0.05%, study 0.2%, and 1% in purified diet over a 13 week period

The in vivo genotoxicity of zinc compounds has been studied in various test systems including the micronucleus test, sister chromatide exchange and chromosomal aberration test, dominant lethal mutation assay as well as for sex-linked recessive lethal mutations in drosophila melanogaster. Neither zinc sulphate nor zinc monoglycerolate induced micronuclei in two reliable mouse and rat micronucleus tests. Further, both zinc sulphate and zinc chloride did not increase the incidence of sex-linked recessive lethal mutations in Drosophila melanogaster (Gocke et al., 1981; Carpenter and Ray, 1969). Zinc sulphate was further negative in a dominant lethal assay in rats. Equivocal and sometimes contradicting results were found for the induction of chromosomal aberrations which have been studied in bone marrow cells harvest from animals exposed to zinc compounds zinc chloride, and zinc oxide. Negative findings for chromosome aberrations have been produced after intraperitoneal injection of zinc chloride into mice (Vilkina et al., 1978) or when rats were given zinc sulphate by gavage once or daily for 5 consecutive days (Litton Bionetics, 1974). In contrast, increased aberrations have been reported in rats after inhalation exposure to zinc oxide (Voroshilin et al., 1978), in rats after oral exposure to zinc chloride and in mice after multiple intraperitoneal injections of zinc chloride (Gupta et al., 1991). Moreover, increased chromosomal aberrations were found in calcium-deficient mice when fed zinc (in form of zinc chloride) via the diet (Deknudt, 1982). Table 40. Overview of experimental in vivo genotoxicity studies for ammonium chloride Method Results Remarks Reference micronucleus assay (chromosome Evaluation of results: negative 2 (reliable with Hayashi M et al. aberration) restrictions) (1988) Test results: mouse (T23-48:ddY) male Genotoxicity: negative (male); supporting study toxicity: no effects intraperitoneal experimental result

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 109 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

(1)Single injection: 0, 62.5, 125, Test material (EC 250, 500mg/kg|(2)Four injections name): Ammonium with 24hr intervals: 0, 31.3, 62.5, chloride 125, 250mg/kg no data

5.7.2. Human information The only identified publicly available genotoxicity study in humans related to the identification of chromosomal aberrations in lymphocytes of 24 workers in a zinc smelting plant (Bauchinger et al., 1976). This study was, however, not suitable to draw any conclusions to the association of these effects with zinc exposure, as the workers displayed also increased blood levels of lead and cadmium, and the clastogenic effects were predominantly attributed to cadmium exposure. There were no further reports in the accessible literature on genotoxic effects of zinc compounds in human populations. 5.7.3. Summary and discussion of mutagenicity The genotoxicity of soluble and slightly soluble zinc compounds have been extensively investigated in a wide range of in vitro and in vivo studies. The in vitro investigations included non-mammalian and mammalian test systems covering the endpoints of gene mutation, chromosomal aberrations, sister chromatide exchange, unscheduled DNA synthesis (UDS), as well as cell transformation. Available in vivo genotoxicity assays included the micronucleus test, sister chromatide exchange (SCE) and chromosomal aberration test and the dominant lethal mutation assay in mouse or rat as well as investigations for sex-linked recessive lethal mutation in drosophila melanogaster. The investigated zinc compounds did not increase the mutation frequencies in the majority of bacterial or mammalian cell culture systems. For example, zinc chloride, zinc sulphate, zinc bis(dihydrogen phosphate), zinc oxide or zinc monoglycerolate were consistently negative in the Ames test. While zinc chloride was also negative for gene mutations in the mouse lymphoma assays, there was some evidence that zinc oxide, zinc acetate or zinc monoglycerolate induced in the absence of metabolic activation the formation of mutation colonies. Several reviewers noted, however, that these mutations were observed at cytotoxic concentrations and that the analysis did not distinguish between big and small colonies which could be caused by gene mutation or chromosomal aberrations (Thompson et al., 1989, WHO, 2001; EU RAR, 2004; MAK, 2009). Conflicting information was further found when zinc compounds were examined for their potential to induce chromosomal aberrations or sister chromatide exchange in mammalian cell systems or when evaluated in the cell transformation assay. Positive as well as negative results were obtained in these cell systems with either soluble or slightly soluble zinc compounds. In those studies where chromosomal aberrations or sister chromatide exchange were observed, these were generally considered to be weak and occurred only at high, often cytotoxic concentrations. Moreover, these positive in vitro findings have also to be seen in context of the impact that changes in zinc levels can have on cell system processes that are controlled by a strict metal homeostasis. A change of this metal homeostasis due to increased zinc levels, may lead to a binding of zinc to amino acids like cystein and therefore to an inhibition of certain enzymes. This can lead to interactions with the energy metabolism, signal transmission and apoptotic processes which can lead to the observed clastogenic or aneugenic effects in in vitro systems (EU RAR, 2004; MAK, 2009). In addition to above mentioned in vitro investigations, various soluble and slightly soluble zinc compounds have also been studied in a range of in vivo studies including the micronucleus test, SCE and chromosomal aberration test or dominant lethal mutation assay in mice or rats as well as in the Drosophila Melanogaster SLRL test. The zinc compounds were consistently negative in the micronucleus and in the assay with Drosophila Melanogaster. Zinc sulphate was further negative in a dominant lethal assay in rats. As discussed in section 5.7.1.2, equivocal and sometimes contradictory results were obtained in the in vivo chromosomal aberration assays. These equivocal finding likely a reflection of inter-study differences in routes, levels, and duration of zinc exposure, the nature of lesions scored (gaps compared to more accepted structural alterations) and great variability in the technical rigour of individual studies (WHO, 2001). The German MAK committee reviewed the existing in vivo evidence and concluded that particularly those studies indicating

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 110 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 clastogenic effects involved a lot of methodological uncertainties which do not allow overruling those in vivo studies which did not provide any evidence for chromosomal aberrations in vivo. Moreover, the Dutch rapporteur of EU risk assessment of zinc compounds under the EU existing substance legislation considered the positive in vitro findings for chromosomal aberration and SCE assays to be overruled by the overall weight of evidence of negative in vivo tests for this endpoint (EU RAR, 2004). The overall weight of the evidence from the existing in vitro and in vivo genotoxicity assays suggests that zinc compounds do not have biologically relevant genotoxic activity. This conclusion is in line with those achieved by other regulatory reviews of the genotoxicity of zinc compounds (WHO, 2001; EU RAR, 2004, MAK, 2009). Hence, no classification and labelling for mutagenicity is required for any of those zinc compounds covered in this chemical safety report.

5.8. Carcinogenicity 5.8.1. Non-human information

5.8.1.1. Carcinogenicity: oral A one-year drinking water study was conducted to evaluate the carcinogenic potential of zinc sulphate in Chester Beatty stock mice. The doses of zinc sulphate were 4.4 g/L (1,000 ppm zinc) and 22 g/L (5,000 ppm zinc) in drinking water along with a control group fed a basal diet and normal drinking water. The animals were examined thoroughly once a week throughout the experiment and a more cursorily examination daily when fed. Weighing was done once every 2 weeks. Deaths of animals occurred during the first 8 week of experiment due to an epizootic of ectromelia. The survivors were vaccinated with sheep lymph and animals showing a negative or accelerated response were sacrificed. New group of weanling mice (4 -5 wk old) were added to supplement the control group. All the surviving animals were sacrificed after 1 year of treatment and examined for gross pathology. Histopathological examination was done for suspected neoplastic lesions. Stomachs were examined for tumours and other changes in the forestomach and glandular epithelium. No differences in carcinogenic effects were observed between treatment and control groups under the test conditions. Under the test conditions, the test material was found to be non-carcinogenic in mice (Walters and Roe, 1965).

5.8.1.2. Carcinogenicity: inhalation Presently information is unavailable.

5.8.1.3. Carcinogenicity: dermal Presently information is unavailable.

5.8.1.4. Carcinogenicity: other routes Presently information is unavailable. 5.8.2. Human information Human experience is predominantly available from the use of zinc compounds as dietary supplements. Isolated epidemiological studies examined also the association between occupational exposures to zinc and carcinogenicity. The following presents some of the key studies in this context: A population based case-control study was conducted to determine the association of dietary zinc level and brain tumour development. The study was conducted between 2001 and 2004 in the UK and comprised adults aged 18–69 years. Dietary information was collected from 637 cases diagnosed with a glioma or meningioma, and 876 controls. Data were obtained from a self-completed food frequency questionnaire (FFQ). Multivariate logistic regression analysis was conducted, adjusting for socio-demographic factors, season of questionnaire return, multivitamin supplementation and energy intake. Although a weak protective effect was observed for the third quartile of intake (normal compared with low intake) in the meningioma group, this was limited to the specific brain tumour subtype and quartile, and was not significant after also adjusting for intake of other elements. Overall there was no significant effect of zinc intake. In conclusion, no association or dose–response relationship was observed between increased vs. low zinc intake and risk of glioma or meningioma (Dimitropoulou et al., 2008).

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 111 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

A case-control study was conducted for analysing the association of prostatic cancer with the intake of particular nutrients, namely fat, vitamins A and C and zinc. A total of 452 cases of prostatic cancer, identified through the population based Hawaii Tumor Registry during the period 1977-1983, and 899 age-matched population controls were interviewed on the island of Oahu from 1981 to 1983. The subjects and population controls were comprised of five different ethnic groups (i.e., Caucasian, Japanese, Chinese, Philipino, and Hawaiian). All subject interviews were conducted in the home by use of a quantitative dietary history method. The weekly intake of fat, zinc, and vitamins A and C, including supplements was determined for each subject. Among 70 years or older men, but not among younger men, the mean weekly consumption of saturated fat, carotenes, and zinc, adjusted for age and ethnicity, was greater for cases than for controls. In a multiple logistic regression analysis, the odds ratio for the highest quartile of fat intake among the older men was 1.7 (95% confidence interval (CL) 1.0-2.8). The corresponding odds ratios were 1.6 (95% CL1.0-2.5) for carotenes, 1.4 (95% CL 0.9- 2.3) for total vitamin C, and 1.7 (95% CL 1.1-2.7) for total zinc. There were significant linear trends in the odds ratios for saturated fat and zinc, but no synergistic interactions among the nutrients. The results suggest that several different components of the diet may contribute independently to the risk of prostatic cancer in elderly men (Kolonel et al., 1988). In a multicentre hospital based case-control study on prostate cancer, the association between high zinc intake and prostate cancer risk, particularly for advanced cancers was evaluated. The study was conducted between 1991and 2002 and considered 1294 cases and 1451 controls. Zinc intake was computed from a valid and reproducible food frequency questionnaire, with the use of an Italian food composition database. Odds ratios (OR) of dietary intake of zinc and the corresponding 95% confidence intervals (CI) were estimated by unconditional multiple logistic regression models, after allowance for several covariates, including total energy. Compared with the lowest quintile, the OR for the highest quintile was 1.56 (95% CI, 1.07–2.26), with a significant trend in risk (p = 0.04). The trend in risk was significant for advanced cancers only, the OR being 2.02 (95% CI, 1.14–3.59) for prostate cancers with a high Gleason score. In this case-control study, a direct association between high zinc intake and prostate cancer risk, particularly for advanced cancers was observed and thus excluded the favourable effect of zinc on prostate carcinogenesis (Gallus et al., 2007). A population based case-control study was conducted to examine association of dietary supplement use (including zinc) with prostate cancer risk in King County, Washington. 697 incident prostate cancer cases (ages 40–64 yr) identified from the Puget Sound Surveillance, Epidemiology and End Results program registry and 666 controls recruited from the same overall population using random-digit dialling sampling. Participants reported their frequency of use of three types of multivitamins and single supplements of vitamins A, C, and E, calcium, iron, and zinc over the 2 yr before diagnosis. Logistic regression analyses controlled for age, race, education, family history of prostate cancer, body mass index, number of prostate-specific antigen tests in the previous 5 yr, and dietary fat intake. Although zinc use was rare, there was a borderline statistically significant 45 % reduction in risk of prostate cancer among those using zinc daily, with a significant test for trend. Adjusted odds ratios (95% confidence limits) for the contrast of ≥ 7/wk versus no use were 0.55. When cases were stratified by stage of disease at diagnosis, there was no suggestion of different effects among participants with early (stages A and B) and advanced (stages C and D) disease. When stratified by histopathological grade, somewhat stronger protective effect was observed in higher-grade disease, although trends were similar in both groups. The results of this study indicate that use of individual supplement of zinc may be protective against prostate cancer (Kristal et al., 1999). A study was conducted to determine the relationship between supplemental zinc intake and prostate cancer risk among the participants in the Health Professionals Follow-Up Study. The study was approved by the institutional review board on the use of human subjects in research of the Harvard School of Public Health. Follow-Up study was initiated in 51,529 male health professionals aged 40 to 75 years and follow-up questionnaires mailed biennially to cohort members to update information on newly diagnosed illnesses. Dietary intake was assessed with the use of a 131-item semi quantitative food-frequency questionnaire. Supplemental zinc provided 32% of total zinc intake representing the major source of zinc. Compared with nonusers, men who consumed supplemental zinc also consumed more multivitamins, supplemental calcium, supplemental vitamin E, lycopene, copper, iron, folate, and fish, but had lower intakes of red meat, and were slightly less likely to have had a history of prostate specific antigen screening. Non significant associations between supplemental zinc intakes at doses less than or equal to 100 mg/d and the risk of prostate cancer. However, compared with nonusers, men who consumed more than 100 mg/d of supplemental zinc had a relative risk of advanced prostate cancer of 2.29 (95% confidence interval = 1.06 to 4.95; Trend = .003), and men who took supplemental zinc for 10 or more years had a relative risk of 2.37 (95% confidence interval = 1.42 to 3.95; Trend <.001).Residual confounding by supplemental calcium intake or some unmeasured correlate of zinc supplement use cannot be ruled out, so the finding that chronic zinc oversupply may play a role in prostate carcinogenesis, warrant further investigation. Supplemental zinc intake at doses of up to 100 mg/d was not associated with prostate cancer risk.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 112 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

However, excessively high supplemental zinc intake may be associated with an increased risk of advanced prostate cancer (Leitzmann et al., 2003). In an old zinc mining and smelting area in the US, a study was conducted to determine the excess in lung cancer mortality associated with residence. The age- and sex-adjusted mortality rates were compared to state and national rates. Age and sex specific lung cancer mortality rates were calculated for white individuals by county in Missouri (1968-1977) and Kansas (1973-1977) and then age adjusted. Additional lung cancer data were obtained from the Environmental Protection Agency (EPA) for Oklahoma, Kansas, and Missouri. Data were combined for the three counties to form one 'super-county.' The analysis determined that lung cancer mortality was elevated in the region. Quantification of inhabitant’s exposure to zinc was not part of the study. The authors mentioned several possible causes for the increased lung cancer rates such as smoking habits, occupational exposure (e.g. in mining and associated activities) and residence. Ore contaminants were arsenic, cadmium, iron, sulphur, germanium and radioactivity. Tuberculosis and silicosis were commonly seen among the region’s inhabitants. From this study no conclusions on a possible association between exposure to environmental levels of zinc and the increased lung cancer rate could be drawn (Neuberger et al., 1982). A cohort study was conducted on male workers exposed for at least one year in zinc refineries, to determine if the refinery operation is associated with any excess mortality patterns. Employees were incorporated in the study when they had worked in the electrolytic department for at least one year. Age-adjusted standardized mortality ratio’s (SMR) were calculated on the basis of comparison with the mortality rates for the entire population for the year 1970. Of the 1247 workers who were exposed to “zinc” (either alone or in combination with “copper”), 88 died before the end of the follow-up. For 12 of these, the cause of death could not be retrieved. 143 workers were lost to follow-up entirely. Cancer rates were only analysed for the entire cohort of refinery workers (i.e. all 4802 participants). Overall SMRs were calculated to be 92 for the cohort and 83 for the subgroup of zinc refinery workers. Significantly high cause-specific SMRs were as follows: (1) cerebrovascular disease (CBVD) for the cohort; (2) all cancers, cancer of the digestive tract, and CBVD for the copper subgroup; (3) all cancers, cancer of the respiratory tract, and CBVD for one plant that demonstrated a significantly high overall SMR. The significant excess of cancer deaths among the study cohort was largely due to the plant that exhibited the significantly high overall mortality rate, but lack of smoking data qualifies this finding. An association between cancer mortality and employment in zinc and/or copper refinery was not found, under the study conditions. A conclusion about any association between cancer mortality and zinc exposure cannot be drawn, because cancer mortality for “zinc”-workers was not analysed separately from cancer mortality for “copper”-workers (Logue et al., 1982). 5.8.3. Summary and discussion of carcinogenicity No adequate experimental animal studies are available to evaluate the carcinogenicity of zinc compounds in humans. There are a range of epidemiological studies that investigated the association between zinc exposure either through occupational activities or food supplementation and increased cancer risks. While no associations were found between occupational zinc exposure and excess cancer risk, the main association that has been made in this context is related to dietary/supplemental zinc and prostate cancer risk. In contrast to established clinical and experimental evidence that prostate cancer is associated with a decrease in the zinc uptake, numerous epidemiology studies and reports of the effect of dietary and supplemental zinc on the incidence of prostated cancer have provided divergent, inconsistent and inconclusive results which range from adverse effects of zinc, protective effects of zinc and no effect of zinc on the risk of prostate cancer. Clinical and experimental studies have established that zinc levels are decreased in prostate cancer and support a role of zinc as a tumor suppressor agent. Malignant prostate cells in situ are incapable of accumulating high zinc levels from circulation (Franklin et al., 2005; Costello and Franklin, 2006; Franklin and Costello, 2007). In a recent critical assessment of epidemiology studies regarding dietary/supplemental zinc and prostate cancer risk, Costello et al., concluded that epidemiological studies have not provided an established relationship for any effect or lack thereof of dietary/supplemental zinc on the risk of prostate cancer. Proclamations of an association of dietary/supplemental zinc and increased prostate cancer are based on inconclusive and uncorroborated reports (Costello et al., 2007). On the basis of the existing information it can be concluded that there is no conclusive evidence for carcinogenic activity of any of the zinc compounds considered in this chemical safety report.

5.9. Toxicity for reproduction

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 113 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

5.9.1. Effects on fertility

5.9.1.1. Non-human information A range of studies have been conducted to assess the effects of zinc on fertility and reproductive performance, most of them with soluble zinc chloride and zinc sulphate. A complete overview and review of available fertility studies is available in the EU risk assessment of zinc compounds (EU RAR, 2004), the review of the of health effects of zinc compounds by the US Agency for Toxic Substances and Disease Registry (ATSDR, 2005), the toxicological review of zinc and compounds by the US Environmental Protection Agency (US EPA, 2005) or the review by the WHO (WHO, 2001). The results of the key experimental studies addressing potential effects of zinc compounds on fertility are summarised in the following table: Table 41. Overview of experimental studies on fertility Test substance Method Results Remarks Reference Zinc chloride One-generation study As of 3.5 mg Zn/kg bw/day: 2 (reliable with Khan et al., 2001 in rats administered P - Mortality; body weight restrictions) zinc chloride at doses gain; fertility indext; thymus supporting study of 0, 3.6, 7.2, 14.4 mg atrophy Zn/kg bw/d in water F1 - litter size (non significant); over one generation by number of surviving pubs (non gavage. Exposure significant); started 77 days prior to mating As of 7.2 mg Zn/kg bw/day: P – hemosidosis of spleen; lymphocyte deficiency F1 - number of surviving pubs ; BW gain (PND 21) Zinc chloride One-generation study 0.75 resp. 1.5 mg Zn/kg bw/day:2 (reliable with Khan et al., 2001 in mice administered P - Mortality; body weight restrictions) zinc chloride at doses gain; abs./rel. Liver/thymus/ supporting study of 0, 0.75, 1.5 and 3, spleen weight; fertility indext; mg Zn/kg bw/d number pregnancies respectively, 0. 1.5, 3 F1 - litter size (non significant); and 6, mg Zn/kg bw/d number of surviving pubs (non in water with 1.5mL significant); HNO3/l over one generation by gavage. 1.5 resp. 3 mg Zn/kg bw/day: Exposure started 49 P - body weight gain; days prior to mating F1 – 14day survival index;

3 resp. 6 mg Zn/kg bw/day: F1 – only 1 birth; 9 still births. Zinc chloride Two-generation study As of 3.5 mg Zn/kg bw/day: 2 (reliable with Khan et al., 2007 in rats administered P - Mortality; body weight restrictions) zinc chloride at doses gain; abs/rel liver/kidney supporting study of 7.5, 15and 30 mg/kg weight; lesions in GI tract, bw/d in water over two inflammation in prostate successive generations F1 - Mortality; body weight via the oral route. gain; abs/rel Application procedure brain/prostate/spleen weight; not specified but likely F2 – no effects oral gavage. Exposure started 77 days prior to 7.2 mg Zn/kg bw/day: mating. P – abs./rel. brain/seminal vesicle weight; F1 - abs/rel liver/adrenal/seminal vesicle weight F2 – no effects

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 114 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

14.1 mg Zn/kg bw/day: P – abs./rel. Spleen/uterus weight; F1 - body weight gain (PND21); abs/rel kidney weight; litter size and #surviving pubs until PND4; F2 – body weight gain (PND21); abs/rel kidney weight; litter size and number surviving pubs until PND4;

Maternal toxicity at any dose level. The NOAEL for fertility and development toxicity is about 15 mg ZnCl2/kg bw/d, this corresponds to 7.2 mg Zinc/kg bw/day. No NOAEL for systemic toxicity could be derived. Zinc sulphate Charles foster rats fed 200 mg Zn/kg bw/day 2 (reliable with Samanta et al., with a diet containing P – Zn-concentration in testis and restrictions) 1986 4000ppm Zn (in form sperm; sperm mobility; supporting study of zinc sulphate); number of pregnancies exposure equals 200 F1 – number of live births mg Zn/kg bw exposure started 30-32 days prior to mating.

The reproductive toxicity of zinc compounds, represented by soluble zinc chloride and zinc sulphate has been investigated in one and two-generation reproductive studies with zinc chloride and zinc sulphate conducted by Khan et al. (2001, 2003, 2007) and Samanta et al. (1986). Moreover, information on potential effects of zinc compounds on reproductive organs can be derived from subchronic toxicity studies conducted Maita et al. (1981) and Edwards and Buckley (1995). The most recent one and two generation reproductive toxicity studies conducted by Khan et al., (2001, 2003, 2007) in rats and mice with zinc chloride provide the most coherent picture on the effects of zinc. All these studies have in common that while effects on fertility such as reduced litter size in F1 and F2 generation have been determined, these were only noticeable at doses which resulted in toxic effects in the dam. Maita et al., (1981) reported that mice and rats fed with zinc sulphate in dietary concentrations up to 30,000 mg/kg feed did not produce adverse effects on either male or female sex organs after 13 weeks of exposure. This dietary level was equal to ca. 1100 mg or 565 mg Zn/kg bw/day for mice and rats, respectively. Edwards K. and Buckley P (1995) showed that rats exposed to 13 or 60 mg Zn/kg bw/day in the diet over a period of 90 days did not show any detrimental effects on sex organs. In the exposure group of 335 mg Zn/kg bw/day, all males showed hypoplasia in testes and seminiferous tubules in males hypoplastic uterus in females, but these findings are not considered reliable as the animals of this high dose group were generally of poor health conditions and killed for humane reasons prior to study termination. In addition to those key reproductive toxicity studies summarised in Table 41, some additional studies indicating high oral doses of zinc (i.e., exposures greater that 25 mg day/kg bw/day) to impair fertility as indicated by a decreased number of implantations sites and increased number of resorptions are of note: A study was carried out to determine the effect of zinc supplementation on the number of implantation sites and resorptions in pregnant rats. The control group consisting of 12 pregnant females was maintained on 10 % vegetable protein diet (containing 30 ppm zinc) from Day 1 through Day 18 of pregnancy. The experimental group consisting of 13 animals was also maintained on the same diet, but received additionally 150 ppm zinc as a 2% zinc sulphate solution administered daily orally. All the animals were sacrificed on Day 18 of pregnancy, and their uteri examined for implantation sites and resorptions. Of a total number of 101 implantation sites in the 12 control animals there were two resorptions, one in each of two animals. In marked contrast, in the 13 zinc supplemented animals, there were 11 resorptions out of 116 implantations. Eight of the animals had at least one

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 115 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

resorption each. This difference was statistically significant. The result indicates that oral administration of moderately high levels of zinc (150 ppm) may be associated with harmful effects in the course of pregnancy of rat (Kumar et al., 1976). The low protein diet may have affected the physiology of the animals resulting in an increased sensitivity for zinc. As this hypothesis cannot be further and also considering the limited available study information, this study is only of limited validity for the assessment of effects of zinc exposure on fertility (EU RAR, 2004). Another study aimed at determining the effect of post-coitum, and pre- and post-coitum dietary zinc supplementation on the conception in the Charles-Foster rat. In the post-coitum study (test 1), two groups of 15 pregnant rats were fed 0 and 4,000 ppm zinc as zinc sulphate in diet (i.e., approximately 200 mg Zn/kg bw/day) from day 1 through day 18 of pregnancy. In the pre- and post-coitum study (test 2), two groups of 15 female rats were treated with same doses for 21 days pre-mating period, maximum 5 days of mating period and 18 days of post-coitum period. All the females were sacrificed on Day 18 of gestation and uterus content and fetuses were examined. In test 1, significant decrease in the incidences of conception and number of implantation sites per mated female was observed in the treatment group with respect to the control group. However, the difference in implantation sites when considered per pregnant female was not significant. In test 2, no significant difference in incidences of conception and implantation sites was observed in the control and treatment groups. In both the tests, there was no treatment-related change in the fetal and placental weights, stillbirths and malformed fetuses were absent and the number of resorption sites was negligible. Based on these results, dietary zinc supplementation at 4,000 ppm did not affect the fetal growth in pregnant rats. This dose, however, altered the normal conception when started after coitus but showed no effect when initiated sufficient time before coitus (Pal et al., 1987).

5.9.1.2. Human information In reviews by the World Health Organisation in the Environmental Health Criteria for Zinc (WHO, 2001) and by the US Agency for Toxic Substances and Disease Registry in the Toxicity Profile for Zinc (ATSDR, 2005), existing human studies which examined the responses of women to zinc supplementation during pregnancy have been summarised. Studies on large controlled trials that were conducted to investigate the effects of dietary zinc supplementation in healthy pregnant women were peer reviewed. The reviewers concluded that zinc at a rate of 20mg/day and 30 mg/day did not result in any adverse reproductive effects during pregnancy (Hunt et al., 1984; Kynast and Saling et al., 1986).Two exemplar studies are summarised in the following: A double blind trial was conducted in 56 pregnant women at risk of delivering a small for gestational-age baby to determine the effects of dietary zinc supplementation during the last 15-25 weeks of pregnancy following administration of 22.5 mg zinc/day. No adverse reproductive effects were observed (Simmer et al., 1991). Pregnant women who received 0.3 mg zinc/kg/day as zinc sulphate capsules during the last two trimesters did not exhibit any changes in maternal body weight gain, blood pressure, postpartum haemorrhage or infection, indicating no adverse reproductive effects (Mahomed et al., 1989). 5.9.2. Developmental toxicity

5.9.2.1. Non-human information The following table summarizes the key studies addressing the developmental toxicity of zinc compounds in experimental animals: Table 42. Overview of experimental studies on developmental toxicity Test Species Route Method Result Remark Reference substance* Zinc Mouse Oral Females No discernible effects 2 (reliable Food and sulphate CD-1 received daily were seen on or with Drugs doses of 0, 0.3, maternal or foetal restrictions) Research 1.4, 6.5 and 30 survival. No Key study Labs., Inc, mg ZnSO4 difference in number 1973* (unspecified)/k of abnormalities found g bw by oral in foetuses. gavage during NOAEL: days 6-15 of 30 mg/kg bw/day gestation. equalling 12mg Zn/kg bw/d

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 116 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Test Species Route Method Result Remark Reference substance* (anhydrate); 6.8mg Zn/kg bw/d (heptahydrate); Zinc Rat Oral Females No discernible effects 2 (reliable Food and sulphate Wistar received daily were seen on or with Drugs doses of 0, 0.4, maternal or foetal restrictions) Research 2.0, 9.1 and survival. No Key study Labs., Inc, 42.5 mg ZnSO4 difference in number 1973* (unspecified)/k of abnormalities found g bw by oral in foetuses. gavage during NOAEL: days 6-15 of 30 mg/kg bw/day gestation. equalling 17mg Zn/kg bw/d (anhydrate); 9.6 mg Zn/kg bw/d (heptahydrate); Zinc Rat Oral Females No discernible effects 2 (reliable EU RAR, sulphate Charles received daily were seen on or with 2004 Foster doses of 0, and maternal or foetal restrictions) 200 mg Zn/kg survival. A reduced Key study bw (in form of number of ZnSO4) in diet implantations during days 1- observed. No 18 of gestation difference in number of abnormalities found in foetuses. NOAEL: 200 mg/kg bw/day Zinc Hamster Oral Females No discernible effects 2 (reliable Food and sulphate received daily were seen on or with Drugs doses of 0, 0.9, maternal or foetal restrictions) Research 4.1, 19, and 88 survival. No Key study Labs., Inc, mg ZnSO4 difference in number 1973* (unspecified)/k of abnormalities found g bw by oral in foetuses. gavage during NOAEL: days 6-10 of 20 mg/kg bw/day gestation. Zinc Rabbit Oral Females No discernible effects 2 (reliable Food and sulphate Dutch received daily were seen on or with Drugs doses of 0, 0.6, maternal or foetal restrictions) Research 2.8, 13 and 60 survival. No Key study Labs., Inc, mg ZnSO4 difference in number 1974* (unspecified)/k of abnormalities found g bw during in foetuses. days 6-18 of NOAEL: gestation. 13.6 mg/kg bw/day

Zinc Rat Oral Females No discernible effects 2 (reliable Uriu-Hare, carbonate Sprague received daily were seen on or with 1989 Dawley doses of 0, 2.5, maternal or foetal restrictions) and 50 mg survival. No Key study Zn/kg bw (in difference in number form of of abnormalities found ZnCO3) in diet in foetuses. during days 1- NOAEL: 20 of gestation. 50 mg/kg bw/day

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 117 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Test Species Route Method Result Remark Reference substance*

* ZnSO4 form is unspecified. The NOAEL, expressed as Zn cation, has been calculation for both anhydrate- and heptahydrate forms.

Several prenatal toxicity studies are available that examined the developmental toxicity of various zinc compounds in rats, mice, rabbit or hamsters up to dietary exposure levels of 200 mg Zn/kg bw/day or 50 mg Zn/kg bw/day by gavage (for details see Table 42). No developmental toxicity has been observed in these studies and there NOAEL’s have been established at the highest doses tested. Although some developmental effects such as decreases in body weights or decrease in individual organ weights were observed in F1 and/or F2 generations in the one or two generation reproductive toxicity studies conducted by Khan et al. (2007) at high exposure levels, these observations are, however, not suitable for risk assessment or hazard classifications as they were always accompanied with maternal toxicity. Moreover, no developmental toxicity was observed at non-maternally toxic doses in a teratogenicity study in which CF-1 albino mice were administered intraperitoneally 0, 12.5, 20.5 and 25 mg/kg on Day 11 of gestation (test 1) and at 20.5 mg/kg on Days 8 -11 of gestation (test 2) (Chang et al., 1977).

Table 43. Overview of experimental studies on developmental toxicity for ammonium chloride Method Results Remarks Reference rat (Sprague-Dawley) 2 (reliable with Goldman et al. restrictions) (1964) oral: drinking water supporting study NH4Cl: 1/6M, 1 mL/kg b.w. (equivalent to 8.9 mg/kg bw/day) experimental result (nominal in water) Test material (CAS Exposure: From seven to tenth day name): ammonium of gestation (4 days), (once a day) chloride

Ammonium chloride was administered orally dissolved in saline. Number of animals: 10 The fetuses were examined grossly after delivery by cesarean section on day 20. mouse ((C57BL/Ola 3 CBA/Ca)F1 2 (reliable with Sinawat et al. hybrid mice) restrictions) (2003)

see methode description supporting study

0.3 mmol/l ammonium chloride experimental result (nominal conc.) Test material (CAS 0.6 mmol/l ammonium chloride name): ammonium (nominal conc.) chloride

Exposure: see principle of guideline (see principle of guideline)

equivalent or similar to Human IVF

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 118 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

5.9.2.2. Human information In establishing the Environmental Health Criteria for Zinc, the World Health Organisation has reviewed and summarised existing human studies examining the responses of women to zinc supplementation during pregnancy. None of the studies indicated any significant effects on the developing foetus (WHO, 2001). Two exemplar studies are summarised in the following: A study was conducted on pregnant women to determine the effects of nutrients during pregnancy on maternal and fetal outcome. Four hundred fifty women were observed during pregnancy and postpartum. Forty-three variables including 12 laboratory indices of maternal nutrient status were assessed. Maternal plasma zinc levels were inversely correlated with fetal weight. Blood examinations revealed a significant association between the total occurrence of fetomaternal complications or fetal distress, and lowest quartile zinc/albumin and highest quartile folate. Under the study conditions, plasma zinc was determined to be a discriminator for fetomaternal complications only in women in the lowest quartile for plasma zinc (Mukherjee et al., 1984). A double blind trial was conducted on pregnant women to determine the effects zinc supplementation during pregnancy on maternal and fetal outcome. 494 women booking before 20 week of gestation in a hospital were prescribed either 66 mg zinc sulphate (equivalent to 20 mg elemental zinc) capsules or placebo for once daily use, starting from day of booking till delivery. Various adverse outcomes were tested, including maternal bleeding, hypertension, complications of labour and delivery, gestational age, Apgar scores, and neonatal abnormalities. The main outcome measure was birth weight. There were no differences between the mothers and neonates of the zinc supplemented and placebo group. Under the test conditions, zinc supplementation during pregnancy did not affect maternal or fetal outcome (Mahomed et al., 1989). 5.9.3. Summary and discussion of reproductive toxicity Effects on fertility The reproductive toxicity of zinc compounds has been investigated in one and two generation reproductive toxicity studies in which rats or mice were dosed by gavage or via the diet with soluble zinc compounds (i.e., zinc chloride, zinc sulphate) at exposure levels up to 14 mg Zn/kg bw/day (gavage) or 200 mg Zn/kg bw/day (diet) (Khan et al., 2001, 2003, 2007). Further information on potential effects of zinc compounds on male or female reproductive organs could be retrieved from subchronic toxicity studies as conducted by Maita et al. (1981) and Edwards and Buckley (1995). The available information suggests that high oral doses of zinc (i.e., exposure levels greater than 20 mg Zn/kg bw/day) may adversely affect spermatogenesis and result in impaired fertility indicated by decreased number of implantation sites and increased number of resorptions (US EPA, 2005). However, these effects were only observed in the presence of maternal toxicity as seen in the one or two generation studies conducted by Khan et al., (2001, 2003, 2007) or, in case of the study conducted by Kumar et al., (1976), when other study non-zinc relevant study specificities could have impacted the study outcome. In a large number of controlled trials, dietary supplementation with zinc rate of 20 mg/day and 30 mg/day did not result in any adverse reproductive effects in healthy pregnant women as summarised in WHO (2001) and ATSDR (2005). Developmental toxicity The developmental toxicity of zinc compounds can be assessed on the basis of prenatal toxicity studies that have been conducted with soluble zinc sulphate and zinc chloride and slightly soluble zinc carbonate in rats, mice, hamsters or rabbits. Moreover, a total of three one or two generation reproductive toxicity studies conducted by Khan et al,. (2001, 2003, 2007) provide further information on potential teratogenic effects of zinc compounds. No prenatal toxicity was observed with either zinc sulphate, zinc chloride or zinc carbonate at exposure levels up to 50 mg Zn/kg bw/day by oral gavage or 200 mg Zn/kg bw/day if the zinc was dose via the diet. Established NOAELs in these studies were typically at highest dose tested and systemically tolerated by the dams. Developmental effects such as decrease in body or organ weights were, however, observed in F1 and/or F2 generations in the one or two generation reproductive toxicity studies conducted by Khan et al. (2001, 2003, 2007). These studies are not considered suitable for the assessment of teratogenic effects for hazard classification or risk assessment purposes since they were always observed in the presence of maternal toxicity. In studies with women receiving zinc supplementation during pregnancies at levels of approximately ≤ 0.3 mg Zn/kg bw/day, no reproductive or developmental effects were observed (WHO, 2001; SCF, 2003). Evidence of

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 119 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 zinc toxicity during human pregnancy has not been reported, but this may be due to the fact that very high exposures to zinc in human pregnancy are unusual. In contrast, zinc is necessary for normal growth and development (e.g., gene expression, vitamin metabolism) and therefore it is not surprising that zinc deficiency during pregnancy can cause a variety of adverse effects to the foetus or may result in reduced fertility or delayed sexual maturation in animals as well as in humans (EU RAR, 2004; WHO, 2001). In conclusion, there is no experimental evidence that would justify a classification of zinc compounds for hazardous effects for reproductive or developmental toxicity under the Dangerous Substance Directive 67/548/EEC or Regulation (EC) 1272-2008 on the on classification, labelling and packaging of substances and mixtures. The available reproductive and developmental toxicity information has been exclusively generated with soluble zinc compounds zinc chloride or zinc sulphate which ensure maximum bioavailable concentration of zinc and hence, allow the use of the information also for the assessment of the slightly soluble zinc compounds and insoluble zinc metal on a read across basis. No experimental fertility data were identified for these compounds. 5.10. Other effects

5.10.1. Non-human information

5.10.1.1. Neurotoxicity Zinc is an important trace element in the brain. A considerable amount of zinc is accumulated in the brain, particularly in the hippocampus, amygadala, cerebral cortex and olfactory cortex. Although some zinc in the brain firmly binds to metalloproteins or enzymes, a substantial amount of zinc (approximately 10%) forms free zinc ions or is loosely bound and detectable by staining using chelating reagents. Chelatable zinc is stored in the presynaptic vesicles of particular excitatory neurons and is secreted from vesicles to synaptic clefts with excitatory neurotransmitter glutamate during the neuronal excitation (Frederickson, 2000). Synaptically-released zinc is believed to play a crucial role in normal brain function. Therefore, zinc deficiency impairs brain development and capabilities of learning and memory. Notwithstanding, recent studies have indicated excess zinc released in a pathological condition can have adverse effects on the central nervous system and that disruption of zinc homeostasis have been suggested to be implicated in several neurogenerative diseases including Alzheimer’s disease, prion disease, amyotrophic lateral sclerosis (ALS) and Wilson’s disease. However, the mechanisms underlying these diseases are complicated with a range of factors involved and only poorly understood. While the information suggests that metal-metal interactions and the disturbance of zinc homeostasis play a role in these type of diseases, the exact role and contribution of zinc in these processes is still undefined (Konoha et al., 2006).

5.10.1.2. Immunotoxicity Zinc affects multiple aspects of the immune system. It is crucial for normal development and function of cells, mediating innate immunity, neutrophils and NK cells. Macrophages, phagocytosis, intracelluar killing and cytokine production and the growth and function of T and B cells are adversely affected by zinc deficiency. The ability of zinc to function as an antioxidant and stabilize membranes suggests that it has a role in the prevention of free radical-induced injury during inflammatory processes (Prasad, 2008). The mechanistical basis of the role of zinc in the immune system has been reviewed and discussed by Hirano et al., (2008). The results of an exemplar experimental study on the effects of chronic zinc supplementation on circulating levels of peripheral blood leucocytes and lymphocytes in humans is summarised in the following table: Table 44. Overview of experimental studies on immunotoxicity Method Results Remarks Reference The effects of chronic Zn No effect of Zn 2 (reliable with Bonham M, supplementation on circulating supplementation was observed restrictions) O’Connor JM, levels of peripheral blood leucocytes on circulating levels of supporting study Alexander HD, and lymphocyte subsets were studied peripheral blood leucocytes or Coulter J, Walsh in a double-blinded intervention trial on lymphocyte subsets. Cu PM, McAnena in male subjects. status was also unaltered. (2003) Human Independent of supplement, chronic (oral: feed) there appeared to be seasonal 30 mg Zn/d (nominal in diet) variations in selected Vehicle: unchanged (no vehicle) lymphocyte subsets in both

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 120 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Method Results Remarks Reference Exposure: 14 wk placebo and supplemented groups. Alterations in circulating levels of B cells (cluster of differentiation (CD) 19), memory T cells (CD45RO) and expression of the intracellular adhesion molecule- 1 (CD54) on T cells were observed. No adverse effects of Zn supplementation were observed on immune status or Cu status

5.10.1.3. Specific investigations: other studies As discussed in various chapters in this chemical safety report, to maintain a healthy condition it is important to maintain the metal homeostasis. The following reports a study which has been conducted to examine the impact of high zinc diet on the iron balance and associated secondary effects. A study was conducted in rats to elucidate the pathophysiology of zinc-induced iron deficiency anemia. Male Sprague-Dawley rats were fed with a diet containing 0.005% (standard Zn diet group) and 0.2% (high Zn diet group) Zn. After 20 weeks, hematological parameters and histopathological changes in the bone marrow, spleen and liver were examined. The serum Zn concentration in the high Zn diet group was significantly higher than that in the standard diet group. On the other hand, the serum Fe concentration in the high Zn diet group was significantly lower than that in the standard diet group. The high Zn diet group exhibited Hb concentrations, Ht levels and MCV, MCH and MCHC values (microcytic hypochromic anemia) that were significantly lower than those in the standard diet group. On the other hand, the number of circulating reticulocytes was significantly elevated in the high Zn diet group relative to the standard diet group. However, there was no significant difference in the number of RBC between the 2 groups of rats. Serum EPO levels were significantly higher in the high Zn diet group than in the standard diet group. There were no substantial differences in the cellularity and the composition of hematopoietic cells between the bone marrow specimens obtained from the 2 groups of rats. Similarly, there was no obvious proliferation of hematopoietic cells in the liver specimens obtained from the 2 groups of rats, although mild degeneration of hepatocytes was observed in the high Zn diet group as compared with the standard diet group. While atrophy of white pulp and development of matured erythrocytes (extra-medullary hematopoiesis) were observed in the spleens from the high Zn diet group, there were no significant histopathological changes in the spleens from the standard diet group. This extra-medullary hematopoiesis was not observed at least up to 12 weeks after the start of dietary treatment. Under the test conditions, the long-term intake of a high Zn diet caused iron deficiency anemia most likely due to suppression of Fe absorption, accompanied by both reticulocytosis and extra-medullary erythropoiesis (Yanagisawa et al., 2009). 5.10.2. Human information

A study was conducted to determine the effects of chronic Zn supplementation on circulating levels of peripheral blood leucocytes and lymphocyte subsets. Male subjects (n=19) were given 30 mg Zn/d for 14 wk followed by 3 mg Cu/d for 8 wk to counteract adverse effects, if any, of Zn supplementation on immune status resulting from lowered Cu status. Placebo supplements were given to a control group (n=19). The study design was a double-blinded intervention trial. Dietary intakes of Zn approximated 10 mg/d. Blood samples, taken throughout the trial, were assessed for full blood profiles and flow cytometric analyses of lymphocyte subsets. Putative indices of Cu status were also examined. No effect of Zn supplementation was observed on circulating levels of peripheral blood leucocytes or on lymphocyte subsets. Cu status was also unaltered. Independent of supplement, there appeared to be seasonal variations in selected lymphocyte subsets in both placebo and supplemented groups. Alterations in circulating levels of B cells (cluster of differentiation (CD) 19), memory T cells (CD45RO) and expression of the intracellular adhesion molecule- 1 (CD54) on T cells were observed. No adverse effects of Zn supplementation were observed on immune status or Cu status, under the test conditions (Bonham et al., 2003). A study was conducted to evaluate whether a daily high-dose calcium supplement perturbs the zinc status post menopausal women. 23 women (mean age: 63 yr) with low bone mineral density were administered daily oral

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 121 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

calcium (1200 mg) during the first 4 wk. Daily co supplementation with calcium (1200 mg) and zinc (30 mg) was provided daily during subsequent 4 wk. Plasma and erythrocyte zinc concentrations plasma bone-specific alkaline phosphatase (BSAP) and 5′-nucleotidase activities, and urinary zinc and calcium excretion were determined first at the end of first 4 wk period and were measured again at the end of the subsequent second 4 wk exposure period. Mean plasma and erythrocyte zinc concentrations after 4 wk of calcium alone were not significantly different from concentrations after co supplementation of calcium and zinc. Mean plasma BSAP activities before co-supplementation with zinc was significantly higher than that after zinc, whereas plasma 5′- nucleotidase activities were not affected by zinc supplementation. Urinary zinc excretion slightly, but significantly, increased after the supplementation of zinc, whereas calcium excretion remained similar. Daily calcium dose of 1200 mg had no detrimental effect on the zinc status in postmenopausal women with low bone mineral density, under the conditions of the test (Morgan et al., 2005). 5.10.3. Summary and discussion of specific investigations Zinc is essential for growth and development, neurological function, wound healing and immunocompetence (SCF, 2003). The main clinical manifestations of zinc deficiency are growth retardation, delay in sexual maturation or increased susceptibility to infections (SCF, 2003). Important in this context is the maintenance of the physiological zinc homeostasis. Disturbance of this zinc homeostasis through for example excessive zinc exposure have been implicated with neurogenerative diseases like Alzheimer’s or Wilson’s disease undefined (Konoha et al., 2006) or with immunosuppressive effects (Raqib et al., 2007), but the exact mechanisms have not been elucidated. There is at this stage no evidence that zinc has any neurotoxicological or immunotoxicological effects under normal zinc exposure conditions and at recommended zinc intake levels. Zinc deficiency, however, adversely affects neurological function and immune competence.

5.11. Derivation of DNEL(s) / DMEL(s)

NH4Cl The NOEL for NH4Cl was determined to be 684 mg/kg bw day. Zn: The repeated dose toxicity of water soluble zinc sulphate and zinc monoglycerolate has been examined in a total of 3 subchronic oral feeding studies. Due to the different dosing regimens, the lowest NOAEL was determined to be 31.5 mg/kg bw/day of zinc monoglycerolate which equals a total zinc exposure of approximate 13 mg/kg bw/day. The zinc NOAEL derived from the feeding studies with zinc sulphate was determined to be 104 mg Zn/kg bw/day in mice and approximately 53.5 mg/kg bw/day in rats. At higher doses the most important effects in the rats were the development of hypocupremia, and significant changes in the pancreas (i.e., focal acinar degeneration and necrosis) and a decreased number of pigmented macrophages in spleen. So Zn is responsible for toxicity.

For the derivation of the derived no effect levels (DNEL(s)) it is of great importance to consider that occupational exposure limits have been established for soluble (i.e., represented by zinc chloride) as well as slightly soluble/insoluble zinc compounds (i.e., represented by zinc oxide) to manage workers risk in operations where zinc exposure might occur. The following Tables 45 and 46 list the existing OELs for zinc chloride as well as zinc oxide Table 45. OELs for zinc chloride Country/organisation 8 hour-TWA 15 min-STEL References mg/m3 mg/m3 USA 1 2 ACGIH (1991) The Netherlands 1 SZW (1997) UK 1 2 a) HSE (1998) Sweden 1b) National Board of

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 122 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Occupational Safety and Health, Sweden (1993) Denmark 0.5 Arbejdstilsynet, 1992 a) This value is a 10 minutes-STEL b) This TWA is determined for dust Table 46. OELs for zinc oxide Country/organisation 8 hour-TWA 15 min-STEL References mg/m3 mg/m3 USA 5 (fumes) 10 (fumes) ACGIH (1991) (guidance 10 (dust) (ceiling) values) USA 5 (fumes) OSHA (1989) (legal limit 15 (dust; total) values) 5 (dust; respirable) The Netherlands 5 (fumes) SZW (1997) Germany 5 (fumes) DFG (1997) 6 (dust) UK 5 (fumes) HSE (1998) 10 (dust) Sweden 5 (fumes) National Board of Occupational Safety and Health, Sweden (1993) Denmark 4 (fumes) Arbejdstilsynet (1992) 10 (dust)

Moreover, for the establishment of DNEL(s) for consumer exposure it is noteworthy that zinc is essential for human growth and development, neurological functions and immunocompetence. The main clinical manifestations of zinc deficiency are growth retardation, delay in sexual maturation or increased susceptibility to infections (WHO, 2001). Health specialists recommend supplementing the diet with zinc in case human diet is zinc deficient. The maximum allowable daily intake has been established to be 50 mg zinc per day.

5.11.1. Overview of typical dose descriptors for all endpoints The human health endpoints that have been identified to be of concern for the various zinc compounds are  Acute oral and inhalation toxicity  Skin and eye irritation;  Repeat dose toxicity humans (i.e., effect at LOAEL: reduced ESOD activity) and animals (i.e., effect at LOAEL: pancreatic damage) The soluble zinc compounds (i.e., zinc chloride, zinc sulphate, zinc bis(dihydrogen phosphate, diammonium tetrachlorozincate and triammonium pentachlorozincate) demonstrated higher acute toxicity via oral and inhalation routes of exposure compared to the slightly soluble and insoluble zinc compounds for which no classifications for this endpoint is required. zinc chloride and zinc sulphate are classified as Xn; R22 and zinc chloride may be toxic by inhalation. The soluble zinc compounds have also demonstrated severe irritant effects to the skin and eyes and respiratory tract and are classified as corrosive (i.e., zinc chloride) and severe eye irritant (i.e., zinc sulphate). Since systemic effects are dependent on the systemic availability of zinc in form of the zinc cation following oral absorption, the repeat dose toxicity studies conducted in humans and animals serve as the basis to assess any systemic effects of zinc released from soluble, slightly soluble and insoluble zinc compounds. The following Tables 47 and 48 list the relevant and available dose descriptors for soluble, slightly soluble as well as insoluble zinc compounds:

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 123 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Table 47. Available dose-descriptor(s) per endpoint for water soluble zinc compounds (i.e., zinc chloride, zinc sulphate, zinc bis(dihydrogen phosphate), diammonium tetrachlorozincate and triammonium pentachlorozincate). Endpoint Quantitative dose descriptor Associated Remarks (appropriate unit) or qualitative relevant effect on study assessment1 Local Systemic

Acute toxicity Oral N/A LD50 = Mortality; Standard acute LD50 300- 2000 studies on zinc mg/kg bw chloride, zinc sulphate and zinc bis(dihydrogen phosphate)).; classification Xn, R22 required

Dermal N/A LD50 > 2,000 Standard acute dermal mg/kg bw toxicity study on zinc sulphate; no classification required

Inhalation N/A < 2 mg/L Mortality; Acute LC50 study on (animal) zinc chloride however, exposure duration very short – 10 min and particle size tested is not a true reflection of human exposure Irritation/ Skin Non to severely N/A Erythema, In-conclusive corrosion irritating oedema, necrosis information; zinc chloride classified as C, R34; no classification of zinc sulphate; Eye Non to severely N/A Corneal opacity; In-conclusive irritating iritis; effects on information; zinc conjunctivae sulphate classified as Xi, R41; no data, classification of zinc chloride Respiratory tract Insufficient N/A Signs of No information information respiratory suggesting need for distress classification as Xi R37 Sensitization Skin Not a skin N/A No effects Negative LLNA and sensitizer GPMT justifies no classification Respiratory No evidence N/A N/A No information to for respiratory suggest the need for sensitization classification as Xi, properties R42 Repeated dose Oral (human) N/A NOAEL = 0.83 At LOAEL of 2.5 toxicity (sub- mg/kg bw/day mg Zn/kg bw/day acute / sub- decreased ESOD chronic / activity and chronic) effects as a result of copper imbalance Oral (animal) N/A Lowest established At higher NOAEL = 13 mg exposure levels Zn/kg bw/day haematological & biochemical effects; pathological

1 Pooled results from studies conducted on one or several soluble forms of zinc

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 124 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Endpoint Quantitative dose descriptor Associated Remarks (appropriate unit) or qualitative relevant effect on study assessment Local Systemic changes in kidneys, GI tract, thyroid & pancreas Inhalation N/A N/A N/A Dermal N/A N/A N/A Mutagenicity in vitro - - Weight of Overall weight of evidence suggests evidence suggests that absence of zinc compounds do mutagenicity in not have a biologically bacterial and relevant genotoxic mammalian test activity; no systems; classification for clastogenicity mutagenicity required was found at high, often cytotoxic doses in vivo - - Predominantly negative, but some conflicting results in chromosomal aberration assays; Carcinogenicity Oral/dermal/ N/A N/A There is no inhalation evidence for carcinogenic activity of zinc compounds in humans or experimental evidence Reproductive Oral/dermal/ N/A NOAEL > 20 No adverse No evidence exists to toxicity inhalation mg/kg bw/day reproductive justify classification of (fertility effects noted in zinc compounds for impairment) NOAEL (humans) pregnant women reproductive toxicity > 0.83 mg Zn/kg administered zinc bw/day at rates of 20-30 mg/day. Zinc may impair fertility at high exposure levels. In animal experiments these effects were always associated with maternal toxicity Developmental Oral/dermal/ N/A NOAEL >50 No No evidence exists toxicity inhalation mg/kg bw/day developmental that would justify the effects seen in classification of zinc NOAEL (humans) specifically compounds for > 0.83 mg Zn/kg designed developmental toxicity bw/day developmental toxicity studies; some developmental effects seen in two generation reproductive toxicity study but only at maternally

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 125 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Endpoint Quantitative dose descriptor Associated Remarks (appropriate unit) or qualitative relevant effect on study assessment Local Systemic toxic doses

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 126 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Table 48. Available dose-descriptor(s) per endpoint for sparingly or insoluble soluble zinc compounds (i.e., zinc oxide, zinc hydroxide, zinc phosphate, zinc carbonate, zinc metal, zinc sulphide) Endpoint Quantitative dose descriptor Associated Remarks (appropriate unit) or qualitative relevant effect on study assessment2 Local Systemic

Acute toxicity Oral N/A LD50 > 2,000 Mortality; Standard acute LD50 mg/kg bw studies on zinc oxide, zinc phosphate and zinc metal; Dermal N/A LD50 > 2,000 No data identified; mg/kg bw read across from standard acute dermal toxicity study on zinc sulphate; & low acute oral toxicity; no classification required Inhalation N/A LOAEL – 5 mg/m3 Metal fume fever; Experience from (humans) workplace exposures; while a NOEL has not been determined, the effects following exposure at LOAEL disappear within 24hrs

Inhalation N/A > 5.7 mg/L Mortality; Acute LC50 study on (animal) oxide; no classification required Irritation/ Skin Not irritating N/A N/A Data on zinc oxide; no corrosion classification required; Eye Non to N/A Corneal opacity; Data on zinc oxide, minimally iritis; effects on zinc phosphate, and irritating conjunctivae zinc metal; no classification required Respiratory tract Insufficient N/A No signs of No information information respiratory suggesting need for irritation in acute classification as Xi inhalation studies R37 Sensitization Skin Not a skin N/A No effects Negative GPMT of sensitizer zinc oxide; no classification required Respiratory No evidence N/A N/A No information for respiratory suggesting need for sensitization classification as Xi, properties R42 Repeated dose Oral (human) N/A NOAEL = 0.83 At LOAEL of 2.5 Read-across from toxicity (sub- mg/kg bw/day mg Zn/kg bw/day dietary supplement acute / sub- decreased ESOD studies with zinc chronic / activity and sulphate; chronic) effects as a result of copper imbalance Oral (animal) N/A Lowest established At higher Read across from NOAEL = 13 mg exposure levels studies with soluble Zn/kg bw/day haematological & zinc compounds biochemical effects; pathological changes in kidneys, GI tract, thyroid &

2 Pooled results from studies conducted on one or several soluble forms of zinc

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 127 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Endpoint Quantitative dose descriptor Associated Remarks (appropriate unit) or qualitative relevant effect on study assessment Local Systemic pancreas Inhalation N/A NOAEL: 2.7 mg At highest dose Non-standard study, ZnO/m³ total lung 5-day inhalation in capacity guinea pigs not decreased and suitable for wet lung weights were increased. classification. Dermal N/A N/A N/A Mutagenicity in vitro - - Weight of Overall weight of evidence suggests evidence suggests that absence of zinc compounds do mutagenicity in not have a biologically bacterial and relevant genotoxic mammalian test activity; no systems; classification for clastogenicity mutagenicity required was found at high, often cytotoxic doses in vivo - - Predominantly negative, but some conflicting results in chromosomal aberration assays; Carcinogenicity Oral/dermal/ N/A N/A There is no inhalation evidence for carcinogenic activity of zinc compounds in humans or experimental evidence Reproductive Oral/dermal/ N/A NOAEL > 20 No adverse Data suggests no toxicity inhalation mg/kg bw/day reproductive evidence exists to (fertility effects noted in justify the impairment) NOAEL (humans) pregnant women classification of zinc > 0.83 mg Zn/kg administered zinc compounds for bw/day at rates of 20-30 reproductive toxicity mg/day. Zinc may impair fertility at high exposure levels. In animal experiments these effects were always associated with maternal toxicity Developmental Oral/dermal/ N/A NOAEL >50 No No evidence exists toxicity inhalation mg/kg bw/day developmental that would justify the effects seen in classification of zinc NOAEL (humans) specifically compounds for > 0.83 mg Zn/kg designed developmental toxicity bw/day developmental toxicity studies; some developmental effects seen in two generation reproductive

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 128 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Endpoint Quantitative dose descriptor Associated Remarks (appropriate unit) or qualitative relevant effect on study assessment Local Systemic toxicity study but only at maternally toxic doses

5.11.2. Correction of dose descriptors if needed (for example route-to-route extrapolation), application of assessment factors and derivation of the endpoint specific DN(M)EL The most relevant dose descriptors for zinc and zinc compounds are the NOAELs derived from repeated dose toxicity studies in humans and rats. For systemic toxicity the data from all zinc compounds can be used for determining specific systemic toxicity of zinc with the ion release rate of zinc becoming the factor that determines the dose. Since slightly soluble and insoluble zinc compounds (i.e., zinc sulphide, zinc oxide, zinc carbonate, zinc phosphate and zinc metal) have low solubility this will result in a worst-case estimate. The oral NOAEL of 50 mg Zn/day derived from the 10 week oral human volunteer study by Yadrick et al., (1989) will be used as the starting point for deriving DNELs for worker and general population. NOAELs for zinc exposure via the dermal or inhalatory route can be estimated by taking into account the bioavailability of zinc via the different exposure routes (for details see section 5.1). Table below summarises the absorption rates of soluble and slightly soluble/insoluble zinc compounds as derived in section 5.1 ‘Toxicokinetics’. Table 49. Summary of absorption rates through different routes of exposure

Exposure route Zinc compound category Absorption rate Oral Soluble zinc 20% Slightly soluble/insoluble zinc 12% Dermal Soluble zinc 2% Slightly soluble/insoluble zinc 0.2% Inhalation Soluble zinc 40% Slightly soluble/insoluble zinc 20% To derive the endpoint specific NOAELs for workers and consumers on the basis of the established NOAEL of 50 mg zinc/day (0.83 mg/kg bw/day based on a woman’s body weight of 60 kg), the NOAEL has to be corrected by assessment factors to account for the uncertainties of the database that led to the establishment of the NOAEL. As the toxicity of zinc compounds is well understood and the NOAEL has been based on human experience and data following chronic exposure to zinc through food supplementation, the assessment factors to be used for zinc compounds are relatively small. Table below provides an overview of the assessment factors under consideration for zinc compounds

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 129 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Table 50. Assessment factors (AF) for zinc compounds Uncertainties Assessment Factor Justification Interspecies 1 No AF required; NOAEL has been derived from human experience through food supplementation Intraspecies -worker 1 No AF required; NOAEL has been derived from human experience through food supplementation Intraspecies –general population 1 No AF required; NOAEL has been derived from human experience through food supplementation Exposure duration 1 No AF required; NOAEL has been derived from human experience through food supplementation Dose response and endpoint 1 No specific AF required; NOAEL is specific/severity issues considered to be reliable. Quality of database 1 No specific AF required; limitations of all available studies have been identified and accounted for. According to the ECHA guidance on information requirements and chemical safety assessment, correction of the dose descriptor for systemic exposure is necessary if  There is a dose descriptor for a given human exposure route and for the same route in experimental animals but for that particular exposure route there is a difference in bioavailability between experimental animals and humans at the relevant level of exposure;  There is not a dose descriptor for a given human exposure route for the same route (in experimental animals or humans);  There are differences in human and environmental exposure conditions;  There are differences in respiratory volumes between experimental animals and humans.

Derivation of the oral DNEL The most relevant dose descriptor has been derived from oral human volunteer studies and human experience from the use of zinc in food supplementation. Neither correction of the dose descriptor nor the use of an assessment factor is considered necessary. Therefore, the oral DNEL for all zinc compounds (i.e., soluble or slightly soluble/insoluble) for workers and consumers equals the most relevant quantitative external dose descriptor for systemic exposure:

o DNELoral sol Zn = 50 mg Zn/day (i.e., 0.83 mg Zn/kg bw/day)

o DNELoral insol Zn = 50 mg Zn/day (i.e., 0.83 mg Zn/kg bw/day)

This setting of the DNEL is fully in line with the approach and result that was concluded in the EU Risk Assessment- part II Human Health (EU RAR 2004).

Derivation of the dermal DNEL (workers, consumers) The derivation of a dermal DNEL on the basis of an oral NOAEL of 50 mg Zn/day derived from human volunteer studies requires a route to route extrapolation. In this process, the follow steps are required  Derivation of the systemic exposure reflecting the oral NOAEL considering the bioavailability of soluble zinc compounds which have been used in the human volunteer studies (i.e., NOAELsyst = 50 mg Zn/day x 20% = 10 mg Zn/day);  Calculation of a dermal exposure to a soluble or slightly soluble/insoluble zinc compound that results in a systemic exposure of 10 mg Zn/day; assumption: bioavailability of soluble zinc compounds

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 130 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

following dermal exposure – 2%; bioavailability of slightly soluble/insoluble zinc compounds following dermal exposure – 0.2%;

o NOAELdermal sol Zn = 10 mg Zn/day / 2% = 500 mg Zn/day

o NOAELdermal insol Zn = 10 mg Zn/day / 0.2% = 5000 mg Zn/day  No assessment factor is considered to be required as the original dose descriptor has been derived from appropriate human volunteer studies; hence the DNELs are as follows:

o DNELdermal sol Zn = 500 mg Zn/day (i.e., 8.3 mg Zn/kg bw/day)

o DNELdermal insol Zn = 5000 mg Zn/day (i.e., 83 mg Zn/kg bw/day)  No further differentiation between worker and consumer DNELs is considered necessary.

Derivation of the inhalatory DNEL (workers, consumers) The oral NOAEL of 50 mg Zn/day is also the basis for the derivation of the inhalatory DNEL. Hence, the derivation of the inhalatory DNEL requires a route to route extrapolation as described in the following:  Derivation of the systemic exposure reflecting the oral NOAEL considering the bioavailability of soluble zinc compounds which have been used in the human volunteer studies (i.e., NOAELsyst = 50 mg Zn/day x 20% = 10 mg Zn/day);  Calculation of a inhalatory exposure to a soluble or slightly soluble/insoluble zinc compound that results in a systemic exposure of 10 mg Zn/day; the following assumptions are made: bioavailability of soluble zinc compounds following inhalatory exposure – 40%; bioavailability of slightly soluble/insoluble zinc compounds following inhalatory exposure – 20%;

o NOAELinhal sol. Zn = 10 mg Zn/day / 40% = 25 mg Zn/day

o NOAELinhal insol Zn = 10 mg Zn/day / 20% = 50 mg Zn/day . Corrected dose descriptor for workers considering a breathing volume of 10m3 per 8hr shift

 3 3 NOAELinhal sol. Zn = 25 mg Zn/day / 10m /day = 2.5 mg/m

 3 3 NOAELinhal insol. Zn = 50 mg Zn/day / 10m /day = 5 mg/m . Corrected dose descriptor for consumers considering a breathing volume of 20m3 per day

 3 3 NOAELinhal sol. Zn = 25 mg Zn/day / 20m /day = 1.3 mg/m

 3 3 NOAELinhal insol. Zn = 50 mg Zn/day / 20m /day = 2.5 mg/m  No assessment factor is considered to be required as the original dose descriptor has been derived from appropriate human volunteer studies; hence the DNELs are as follows:

3 o DNELinhal sol Zn (worker) = 2.5 mg/m ; 3 o DNELinhal insol Zn (worker) = 5 mg Zn/m ;

3 o DNELinhal sol Zn (consumer) = 1.3 mg/m ; 3 o DNELinhal insol Zn (consumer) = 2.5 mg Zn/m ;

The following Tables 51 and 52 summarize DNELs that have been calculated for worker and consumer exposure to soluble and slightly soluble/insoluble zinc compounds according to the ECHA guidance methodology.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 131 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Table 51. Corrected dose descriptor(s) per endpoint and endpoint-specific DNELs for workers Endpoint Zinc Most Corrected Overall Endpoint- compound relevant external dose Assessme specific category quantitativ descriptor for nt factor DNEL e external systemic (external dose exposure dose) descriptor for systemic exposure (NOEL) Repeated Oral Soluble 50 mg Zn/day Not required 1 50 mg Zn/day dose (0.83 mg/kg (0.83 mg/kg toxicity bw/day) bw/day) Slightly soluble/ 50 mg Zn/day Not required 1 50 mg Zn/day insoluble (0.83 mg/kg (0.83 mg/kg bw/day) bw/day) Dermal Soluble 50 mg Zn/day 500 mg Zn/day 1 500 mg Zn/day (0.83 mg/kg (8.3 mg/kg bw/day) (8.3 mg/kg bw/day) bw/day)

Slightly soluble/ 50 mg Zn/day 5000 mg Zn/day 1 5000 mg Zn/day insoluble (0.83 mg/kg (83 mg/kg bw/day) (83mg/kg bw/day) bw/day) Inhalation Soluble 50 mg Zn/day 2.5 mg Zn/m3 1 2.5 mg Zn/m3 (0.83 mg/kg bw/day) Slightly soluble/ 50 mg Zn/day 5 mg Zn/ m3 1 5 mg Zn/ m3 insoluble (0.83 mg/kg bw/day) Table 52. Corrected dose descriptor(s) per endpoint and endpoint-specific DNELs for consumers Endpoint Zinc Most Corrected Overall Endpoint- compound relevant dose Assessme specific category quantitativ descriptor for nt factor DNEL e dose systemic (external descriptor exposure dose) for systemic exposure Repeated Oral Soluble 50 mg Zn/day Not required 1 50 mg Zn/day dose (0.83 mg/kg (0.83 mg/kg toxicity bw/day) bw/day) Slightly soluble/ 50 mg Zn/day Not required 1 50 mg Zn/day insoluble (0.83 mg/kg (0.83 mg/kg bw/day) bw/day) Dermal Soluble 50 mg Zn/day 500 mg Zn/day 1 500 mg Zn/day (0.83 mg/kg (8.3 mg/kg bw/day) (8.3 mg/kg bw/day) bw/day)

Slightly soluble/ 50 mg Zn/day 5000 mg Zn/day 1 5000 mg Zn/day insoluble (0.83 mg/kg (83 mg/kg bw/day) (83mg/kg bw/day) bw/day) Inhalation Soluble 50 mg Zn/day 1.3 mg Zn/m3 1 1.3 mg Zn/m3

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 132 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Endpoint Zinc Most Corrected Overall Endpoint- compound relevant dose Assessme specific category quantitativ descriptor for nt factor DNEL e dose systemic (external descriptor exposure dose) for systemic exposure (0.83 mg/kg bw/day) Slightly soluble/ 50 mg Zn/day 2.5 mg Zn/ m3 1 2.5 mg Zn/ m3 insoluble (0.83 mg/kg bw/day)

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 133 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

5.11.3. Selection of the critical DNEL(s) for critical health effects In line with the rationale provided in section 5.11.2, the DNEL’s for workers and consumers following oral or dermal exposure to soluble and slightly soluble/insoluble compounds are as follows: o Oral

. DNELoral sol Zn = 50 mg Zn/day (i.e., 0.83 mg Zn/kg bw/day);

. DNELoral insol Zn = 50 mg Zn/day (i.e., 0.83 mg Zn/kg bw/day); o Dermal

. DNELdermal sol Zn = 500 mg Zn/day (i.e., 8.3 mg Zn/kg bw/day);

. DNELdermal insol Zn = 5000 mg Zn/day (i.e., 83 mg Zn/kg bw/day); These DNEL’s appropriately protect workers and consumers for the most sensitive health endpoint, i.e. reduced ESOD activity, observed in humans following repeated exposure to zinc compounds. With regard to establishing the critical DNELs for inhalatory exposure of workers or consumers to zinc compounds, two approaches are considered suitable: a. the derivation of the DNEL on the basis of existing oral human dietary supplement studies requiring route to route extrapolation as illustrated in chapter 5.11.2 and b. the use of existing OELs as the respective DNELs for worker exposure. With regard to the latter, the guidance on information requirements and chemical safety assessment states that the OELs and/or the underlying information used for setting the OELs can be used to derive the DNELs for workers (ECHA, 2008). As presented in chapter 5.11.2, existing data from human supplementary studies results in worker DNELs of 2.5 or 5 mg Zn/m3 for soluble and slightly soluble/insoluble zinc compounds respectively and consumer DNELs of 1.3 or 2.5 mg Zn/m3. Table 45 and 46 provide an overview of existing OELs for soluble zinc compounds represented by zinc chloride (i.e., Table 45) as well as slightly soluble/insoluble zinc compounds represented by zinc oxide (i.e., Table 46). While a detailed scientific justification for the OELs is not available, these values have ensured workers safety for decades which correlates with the DNELs derived from the human volunteer studies. Taking a conservative approach it is proposed that for inhalatory exposure to soluble and slightly soluble/ insoluble zinc compounds, the existing OEL values are used as the respective DNEL against which to judge the adequacy of workplace risk management measures (RMM) to control airborne exposure to zinc compounds: o Inhalation - Worker

3 . DNELinhal soluble Zn (worker) = 1 mg Zn/m ; 3 . DNELinhal insoluble Zn (worker) = 5 mg Zn/m ;

o Inhalation - Consumer

3 . DNELinhal soluble Zn (consumer) = 1.3 mg Zn/m ; 3 . DNELinhal insoluble Zn (consumer) = 2.5 mg Zn/m ;

6. HUMAN HEALTH HAZARD ASSESSMENT OF PHYSICO-CHEMICAL PROPERTIES 6.1. Explosivity

Data waiving: see CSR section 1.3 Physico-chemical properties.

Classification according to GHS

Name: diammonium tetrachlorozincate(2-) Reason for no classification: conclusive but not sufficient for classification

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 134 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Classification according to DSD / DPD

Classification status: 67/548/EEC self classification (diammonium tetrachlorozincate(2-)) Reason for no classification: conclusive but not sufficient for classification

6.2. Flammability

Data waiving: see CSR section 1.3 Physico-chemical properties.

Classification according to GHS

Name: diammonium tetrachlorozincate(2-) Reason for no classification (Flammable gases): conclusive but not sufficient for classification Reason for no classification (Flammable aerosols): conclusive but not sufficient for classification Reason for no classification (Flammable liquids): conclusive but not sufficient for classification Reason for no classification (Flammable solids): conclusive but not sufficient for classification

Classification according to DSD / DPD

Classification status: 67/548/EEC self classification (diammonium tetrachlorozincate(2-)) Reason for no classification: conclusive but not sufficient for classification 6.3. Oxidising potential

The available information on the oxidising potential is summarised in the following table:

Table 53. Overview of information on oxidising potential

Method Results Remarks Reference thermogravimetrical Evaluation of results: no oxidising properties2 (reliable with Liipo J, Karhu J, analysis was used for restrictions) Metsärinta M-L, checking oxidising : No oxidation is observed by Virta K, Wiik E, properties thermogravimetrical analysis weight of evidence Santala M, Pihlasalo (2010) experimental result

Test material (EC name): diammonium tetrachlorozincate(2- )

The following information is taken into account for any hazard / risk assessment: the substance has no oxidising properties.

Classification according to GHS

Name: diammonium tetrachlorozincate(2-)

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 135 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Reason for no classification (Oxidising gases): conclusive but not sufficient for classification Reason for no classification (Oxidising liquids): conclusive but not sufficient for classification Reason for no classification (Oxidising solids): conclusive but not sufficient for classification

Classification according to DSD / DPD

Classification status: 67/548/EEC self classification (diammonium tetrachlorozincate(2-)) Reason for no classification: conclusive but not sufficient for classification

7. ENVIRONMENTAL HAZARD ASSESSMENT

General considerations

Zinc and zinc compounds form a “data rich” substance group: a vast volume of information is available on the effect of zinc on the different ecotoxicity endpoints in the open scientific literature. This vast volume of ecotoxicological information was carefully scrutinised by the Rapporteur (the Netherlands) in the framework of the discussions on the EU risk assessment (RA) made under EU Regulation 793/93/EEC. In that process, the Rapporteur’s analysis of the available chronic ecotoxicity data was extensively discussed by the experts from member states and other stakeholders during the meetings of the “Technical committee on new and existing substances” (TCNES), where the data sets to be used for PNEC derivation were officially approved. The scrutiny and discussion of TCNES were focusing on the chronic data for the PNEC derivations.

For this reason, we will use the data used in the RAR as the main data source for the environmental hazard assessment. We will not come back to the decisions on data quality and relevancy that were approved by TCNES, but use the data as they were used in the RA process. Consequently, we will use for the current analysis the data that were considered useful for PNEC derivation in the RAR as such. At the same time, we consider the data that were found not useful in the RA process also as such and we will not use them for the current analysis, neither. Given the vast amount of data, we have only reported the useful data in the IUCLID V file; the data not considered useful in the RA process have been summarised in the RAR (ECB 2008).

The datasets from the RAR have been complemented with relevant, reliable information that became available after the closure of the RA databases. These data have been checked according to the same principles as set out in the RA. They are also reported under IUCLID V and used for the present hazard assessments. There are two exceptions to this general approach: 1) the analysis of the data for marine ecotoxicity. The marine ecotoxicity data in the RAR were not scrutinised for quality and relevancy since it was not an endpoint considered under the RA process. Therefore the raw dataset from the RAR was carefully checked, and completed with new data of good quality and relevancy. The PNEC for marine waters was subsequently derived.

2) In contrast to the discussion on the chronic freshwater ecotoxicity data, the acute aquatic ecotoxicity data were scrutinised and discussed in less detail during the risk assessment process. Yet, the quality and relevancy of these data is of equal if not even more importance because (in contrast to the PNEC derivation, where all the chronic data are used in a species sensitivity distribution), one single value can define the ecotoxicity reference value for classification. For this reason, the acute aquatic ecotoxicity data mentioned in the RAR were re-checked for quality and relevancy. As such, all data considered useful in the RA and answering the criteria for reliability/relevancy were considered in the present analysis also; data that were considered not useful in the RA were not used, nor reported in IUCLID (they are reported in the RA, (ECB 2008)). Since the acute aquatic toxicity dataset of the RA was closed quite early in the RA process (most recent reference 1996), this dataset was also significantly updated. The change in reference value resulting from this revision is not influencing the classifications for aquatic toxicity effect, that were agreed at EU level. However, it was considered important to re- consider the database and the ecotoxicity reference concentration derived from it, since this may influence the M-factor and, consequently, the classification of preparations.

In accordance to the scientific approach, followed in the EU RA process, two approaches are key to the zinc hazard assessment:

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 136 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

1) A basic assumption made in this hazard assessment and throughout this CSR, (in accordance to the same assumption made in the EU RA process) is that the ecotoxicity of zinc and zinc compounds is due to the Zn++ ion. As a consequence, all aquatic, sediment and terrestrial toxicity data in this report are expressed as “zinc”, not as the test compound as such, because ionic zinc is considered to be the causative factor for toxicity. A further consequence of this is that all ecotoxicity data obtained on different zinc compounds, are mutually relevant for each other. For that reason, the available ecotoxicity databases related to zinc and the different zinc compounds are combined before calculating the PNECs. The only way zinc compounds can differ in this respect is in their capacity to release zinc ions into (environmental) solution. That effect is checked eventually in the transformation/dissolution (T/D) tests and may result in different classifications. But in principle, this section 7 of the CSR is the same for all zinc substances that are considered soluble, eventually confirmed by results of T/D tests.

2) Zinc is a natural component of the earth’s crust and present in natural background concentration in all environmental compartments. Because of the importance of the natural background, the “added risk concept” has been used in the RAR on zinc (ECB 2008). In this approach both the "Predicted Environmental Concentration"(PEC) and the "Predicted No Effect Concentration" (PNEC) are determined on the basis of the added amount of zinc, resulting in an “added Predicted Environmental Concentration” (PECadd) and “added Predicted No Effect Concentration” (PNECadd), respectively. The use of the added risk approach implies that only the anthropogenic amount of a substance, i.e. the amount added to the natural background concentration, is considered to be relevant for the risk assessment. Thus, a possible contribution of the natural background concentration to toxic effects is ignored (RAR; ECB 2008). So, for zinc, all PNECs are expressed as “added” concentration to the background.

The use of the added risk approach implies that for risk characterisation, the natural background needs to be taken into account when evaluating monitored concentrations in the environment. The correct assessment of natural background is thus important.

7.1. Aquatic compartment (including sediment) 7.1.1. Toxicity test results

1. Aquatic toxicity: freshwater, short-term establishing the dataset

In accordance to the approach followed in the RAR, only acute data from standardised test protocols were considered in the analysis for setting the reference value for classification. This is possible because numerous data are available, and it ensures that the tests were performed under well defined and standard conditions.

Still, the quality and some aspects of relevancy should be checked in a critical way when using the extensive datasets from the open literature, available for zinc. It is e. g. important to know the conditions under which the organisms were tested and cultured, because these conditions may result in acclimatisation and deviating toxicity response. The information on these test conditions is often scarce in non-standardised test reports.

The short-term aquatic ecotoxicity data base for zinc was reviewed according to the following principles:

←the data accepted for setting the acute aquatic reference value in the RA (ECB 2008, Annex 1.3.2a, table 1) were as such also accepted and used for the present analysis. Prescriptions from standard protocols were strictly followed, e. g. data from an acute Daphnia test exceeding 48 hrs were not used.

←Data that were rejected for use in the RA (ECB 2008, Annex 1.3.2a, table 2) were also not used for the present analysis. In this respect, data from studies that were accepted for use in the chronic database, but rejected for use in the acute toxicity database were reconsidered; this resulted in the acceptance of a few additional data.

←In accordance to the approach followed in the RA, acute data obtained in natural waters that contained e. g. significant amount of DOC, were not used. Exception to this rule were data obtained on the N. -American Great Lakes waters, which were used, in accordance to the RA.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 137 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

←Fish data mentioned in the RA under “EHC 1996” were not used, since they were from a review, not from original study reports. These data are not influencing the outcome of the analysis, since they are all at the higher concentration level.

←More recent (obtained after 1996 to the present) short-term acute toxicity data on standard organisms were included in the database.

After checking and updating the data base, the data are grouped per species as follows:

-pH: low (6 -<7) - neutral/high (7 -8.5)

-hardness: low/medium (<100mg CaCO3/l) and medium/high (>100 mg CaCO3/l).

If 4 or more data points were available on a same species, the geomean was calculated and used for the analysis.

Acute data – results

The short-term acute aquatic toxicity database covers 10 species (1 algae, 4 invertebrates and 5 fish species). The full set of EC50 values are presented under sections 7.1.1.1.1.(fish), 7.1.1.2.1. (invertebrates) and 7.1.1.3. (algae and plants). In the table below, the data are summarised together with the pH and hardness of the test media. A significant number of data are available at both low and neutral/high pH.

Table 54. Acute aquatic toxicity of zinc by species as a function of pH and hardness. species pH hardness E(L, I)C50 value reference (mg Zn/l) algae Selenastrum 7.4 24 0.136 Van Ginneken, 1994 capricornutum (new name: Pseudokircherniella subcapitata Selenastrum 7.4 24 0.150 Van Woensel, 1994 capricornutum (new name: Pseudokircherniella subcapitata Daphnids Daphnia magna 7.7 45 0.1 Biesinger & Christensen, 7.7 45 0.28 1972 7.7 45 0.28 Cairns et al. 1978 7.2-7.4 45 0.07 Mount & Norberg 1984 7.2 46 0.259 Barata et al 1998 7.2 46 0.131 7.6 50 0.330 Chapman, 1980 7.7 91 1.06 Barata et al, 1998 7.7 91 0.475 7.0 130 0.8 Atar & Maly 1982 8.1 105 0.53 Chapman 1980 7.7 179 0.962 Barata et al, 1998 8.1 179 0601 8.2 196 0.66 Chapman 1980 7.2 242 2.14 De Schamphelaere et al 2005 7.8 250 2.91 Muysen et al 2005 7.8 250 1.83 8.5 180-200 0.86 Magliette et al 1995 Daphnia pulex 7.6 45 0.5 Cairns et al 1978 7.2-7.4 45 0.107 Mount & Norberg 1984 6.3 84 0.425 Clifford & McGeer 2009 7.9 51 0.105

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 138 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

7.9 69 0.190 7.9 86 0.399 7.9 46 0.268 7.9 81 0.399 7.9 84 0.532 7.9 84 0.399 7.9 84 0.706 7.9 84 0.399 7.9 84 0.477 7.9 84 0.392 7.3 84 0.765 8.0 84 0.687 7.9 104 0.321 7.9 131 0.556 7.9 122 0.432 7.9 163 0.353 7.9 241 1.014 7.9 224 0.615 Daphnia carinata 7.5 82 0.340 Cooper et al 2009 Ceriodaphnia dubia 6–6.5 280-300 >0.53 Schubauer-Berrigan et al 1993 7-7.5 280-300 0.36 8-8.5 280-300 0.095 6.5 44 0.413 Hyne et al 2005 7.5 44 0.2 7.5 44 0.155 7.5 82 0.174 Cooper et al 2009 7.3 45 0.076 Mount & Norberg 1984 7.2-7.3 52 0.169 Carlson et al 1986 7.8 280 0.67 Muysen & Janssen 2002 7.8 250 0.416 Muysen et al 2005 Ceriodpahnia 7.8 250 0.937 Muysen et al 2005 reticulata Fish Pimephales promelas 6-6.5 280-300 0.78 Schubauer-Berrigan et al 1993 7-7.5 280-300 0.33 8-8.5 280-300 0.50 Thymallus arcticus 7.1-8.0 41 0.315 Buhl & Hamilton 1990 7.1-8.0 41 0.142 7.1-8.0 41 0.112 7.1-8.0 41 1.58 7.1-8.0 41 0.166 7.1-8.0 41 2.92 7.1-8.0 41 0.168 7.1-8.0 41 0.168 Cottus bairdii 7.5 154 0.439 Brinkman & Woodling 2005 Oncorrhynchus 7.1-8.0 44 0.82 Buhl & Hamilton 1990 kisutch 7.1-8.0 44 1.81 7.1-8.0 44 1.65 7.1-8.0 44 0.727 Oncorrhynchus mykiss 7.1-8.0 44 2.17 Buhl & Hamilton 1990 7.1-8.0 44 0.169

The lowest species values (mg Zn/l) are summarised in table below Table 55. Lowest acute aquatic toxicity data observed for zinc species Low pH/ Low pH, Neutral-high pH/ Neutral-high pH, hardness<100mg hardness>100mg hardness<100mg hardness>100mg CaCO3/l CaCO3/l CaCO3/l CaCO3/l algae

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 139 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Selenastrum / / 0.136 / capricornutum Daphnids Daphnia magna / / 0.244 1.052 Daphnia pulex 0.425 / 0.364 0.507 Daphnia carinata / / 0.34 / Ceriodaphnia dubia 0.413 > 0.530 0.147 0.228 Ceriodaphnia / / / 0.937 reticulata fish Pimephales / 0.780 / 0.33 promelas Thymallus arcticus / / 0.307 / Cottus bairdii / / / 0.439 Oncorrhynchus / / 1.155 / kisutch Oncorrhynchus / / 0.169 / mykiss

Discussion: reference values for short term aquatic ecotoxicity

Tables 54, 55 present an overview of the information available for short-term aquatic toxicity for zinc. It can be seen that significant number of data are available at both low and neutral/high pH.

At low pH, 2 values are available for 2 daphnia species. The values are similar. They were obtained at lower hardness, where the highest sensitivity is expected, which is confirmed by the value >530 µg/l, obtained on Ceriodaphnia dubia at high hardness. Algae are as a rule not tested under standardised conditions at low pH, but from chronic algae data (72 hrs NOECs), it is known that the sensitivity of algae is much lower at lower pH. Simulation with the biotic ligand model gives an aquatic ecotoxicity value for algae at pH 6 which is about 5 times higher than the one observed at neutral/high pH. Fish toxicity at low pH is also not critical in this respect, so the values for the daphnids are representative for the sensitivity of organisms to zinc at low pH. The lowest value observed for Ceriodaphnia dubia is used for the classification at low pH.

At neutral/high pH, the value obtained on the algae Selenastrum capricornutum is the lowest of the dataset. This value is taken forward as reference value for classification at this pH. This value is obtained at low hardness conditions, where sensitivity is highest. The same algae species is also the most sensitive in the chronic aquatic toxicity database (see below) so this sensitivity pattern is consistent. Among the daphnids, Ceriodaphnia dubia is also here the most sensitive, and the lowest value comes close to the one for the alga. From the paired data, it follows that the Daphnids are more sensitive at lower hardness than at the higher hardnesses. The fish are also at this pH less sensitive to zinc, although the lowest value observed on O. Mykiss also comes close to the reference value. All together, the lowest values among the species show also here a consistent pattern, supporting the lowest value identified.

In conclusion, the reference values for the Zn++ion that are used for the acute aquatic toxicity hazard assessment of Zn++are:

←for low pH: 0.413 mg Zn/l (based on single lowest value for Ceriodaphnia dubia)

←for the neutral/high pH: 0.136 mg Zn/l (based on single lowest value for Selenastrum capricornutum (=Pseudokircherniella subcapitata)

2. Aquatic chronic toxicity: freshwater

Chronic data - establishing the dataset

In this analysis, like in the RAR, the results of the chronic aquatic toxicity studies are expressed as either the actual (measured) concentration or as the nominal (added) concentration (Cn). The actual concentrations include

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 140 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 the background concentration (Cb) of zinc. Because of the “added risk approach”, the results based on actual concentrations have been corrected for background, if possible. This correction for background is based on the assumption that only the added concentration of zinc is relevant for toxicity. In case both actual and nominal concentrations were reported, the results are expressed in the RAR (and in this CSR) as nominal concentrations, provided the actual concentrations were within 20% of the nominal concentrations.

Many of the reported aquatic toxicity data (either actual or nominal) represent total-zinc concentrations, i. e. the dissolved plus particulate fraction. However, in accordance to the approach taken in the RA, the results are regarded as being dissolved-zinc concentrations, because under the conditions that were used in the laboratory tests, it is assumed that the greater part of zinc present in the test waters was in the dissolved fraction. This is especially true for the long-term studies, e. g. by using flow-through systems, in which particulate matter (suspended inorganic material and/or organic matter) was removed from the artificial test waters or natural waters. The fact that in ecotoxicity testing the nominal added concentration of zinc is very close to the actually measured zinc concentration, is also demonstrated by the many data reported in the papers of the chronic aquatic ecotoxicity database. Also in static and flow-through acute toxicity studies with several saltwater species, dissolved zinc was greater than 93% of the total zinc. Therefore, the PNECaddvalues derived from the aquatic toxicity studies are considered to be relevant for dissolved zinc.

The chronic aquatic toxicity dataset for zinc was checked according to the general criteria for data quality:

-study design preferably conform to OECD guidelines or equivalent

-Toxicological endpoints, which may affect the species at the population level, are taken into account. In general, these endpoints are survival, growth and reproduction.

- whether or not NOEC values are considered chronic is not determined exclusively by exposure time, but also by the generation time of the test species, e. g. for unicellular algae and other microorganisms (bacteria; protozoa), an exposure time of four days or considerably less already covers one or more generations, especially in water, thus for these kinds of species, chronic NOEC values may be derived from relatively short experiments. For PNEC derivation a full life-cycle test, in which all relevant toxicological endpoints are studied, is normally preferred to a test covering not a full life cycle and/or not all relevant endpoints. However, NOEC values derived from tests with a relatively short exposure time may be used together with NOEC values derived from tests with a longer exposure time if the data indicate that a sensitive life stage was tested in the former tests.

-If for one species several chronic NOEC values (from different tests) based on the same toxicological endpoint are available, these values are averaged by calculating the geometric mean, resulting in the “species mean” NOEC.

-If for one species several chronic NOEC values based on different toxicological endpoints are available, the lowest value is selected. The lowest value is determined on the basis of the geometric mean if more than one value for the same endpoint is available.

-In some cases, NOEC values for different life stages of a specific organism are available. If from these data it appeared that a distinct life stage was more sensitive, the result for the most sensitive life stage is selected. The life stage of the organisms is indicated in the tables as the life stage at start of the test (e. g. fish: yearlings) or as the life stage(s) during the test (e. g. Eggs, larvae, which is a test including the egg and larval stage).

-Only the results of tests in which the organisms were exposed to zinc alone are used, thus excluding tests with metal mixtures.

-Like in the RAR, unbounded NOEC values (i. e. no effect was found at the highest concentration tested) are not used.

-If the NOEC was <100 µg/l, the separation factor between the NOEC and LOEC should not exceed a factor of 3.2.

-If the EC10 was used as NOEC equivalent, the EC10 should not be more than 3.2-times lower than the lowest concentration used in the test.

-Like in the RAR, only the results of tests with soluble zinc salts are used, thus excluding tests with “insoluble” zinc salts (ZnO, ZnCO3), unless dissolved zinc is measured.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 141 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Referring to the EU RA on zinc (ECB 2008), all the data that were accepted for deriving the freshwater PNEC in the RA (ECB 2008, Annex 3.3.2. A. part I) were as such also accepted for the present analysis. On the other hand, the data that were considered not useful for the purpose of PNEC derivation in the RA (ECB 2008, Annex 3 .3.2. A. part II), were also not used for the present analysis.

The relevancy of the long-term aquatic ecotoxicity data base for PNEC derivation was further checked in accordance to the same principles as those applied in the RA (ECB 2008). Relevancy was checked:

1) related to the zinc background: results obtained in unpolluted test media (water, sediment or soil) are used, thus excluding tests that were performed in media containing high to very high background Zn concentrations, i.e. in case the control media contained zinc concentrations that are clearly above Zn concentrations normally encountered in relatively unpolluted environmental compartments. A pragmatic cut-off level of 30µg Zn/l, in accordance to decisions taken in the RA (ECB 2008) was set for this.

In accordance to the RA zinc (ECB 2008), data obtained in tests where the zinc background concentration was much lower than the natural background for EU waters, were also not used for PNEC derivation. In accordance to the RA (ECB 2008), a level of 1µg/l Zn was set as a cut-off for this.

2) related to test medium conditions: Zinc ecotoxicity to aquatic organisms is a function of the physicochemical characteristics of the water. Parameters such as hardness, pH, dissolved organic carbon (DOC) are well-known drivers for zinc ecotoxicity. For this reason, it was considered important in the EU RA to select ecotoxicity data that were obtained under test conditions similar to the conditions observed in EU waters. Based on information related to the parameters mentioned above in EU waters, the following boundaries for EU relevancy for pH, hardness have been used in the RA (ECB 2008) and also in the present analysis for data selection (also considering OECD test guidelines): pH: minimum value: 6, maximum value: 9

Hardness: minimum value: 24 mg/l (as CaCO3), maximum value: 250 mg/l (as CaCO3)

As indicated above, background zinc concentration was also considered in the RA to be a factor influencing the toxicity response of organisms to zinc; to avoid influence of acclimatisation towards very low or very high zinc concentrations (not occurring in the EU waters), a minimum value for soluble zinc was also set in the RAR for data selection: “around 1 µg/l” (ECB 2008).

Data obtained under conditions failing these relevancy criteria were not used for PNEC derivation in the EU RA and in the present analysis. For a detailed description of the relevancy criteria and their application in the RA, see the RAR (ECB 2008).

It is realised that the selected ranges of the three criteria will not cover all European aquatic systems, e. g. specific aquatic systems in the Scandinavian countries. In particularly, e.g. hardness can be much lower in the Scandinavian countries, although also other abiotic parameters may differ from the ‘average’ situation in European freshwaters. Therefore, a “soft water PNECadd, aquatic” has been derived in the RA process, in addition to the generic PNECadd, aquatic. This “softwater PNEC” should be used in situations where it has been documented that the hardness is lower then the low end of the range indicated above (24mg/l). The present analysis however relates to the development of a generic PNEC for EU waters.

In the data selection process of the RA, it was noted that the references used for the current aquatic toxicity dataset usually do not contain data on the background concentration of zinc in the test water and in a number of cases also data on pH and/or hardness were lacking. Thus, a stringent application of the above mentioned (minimum and maximum) limits for all three parameters, especially the zinc concentration, would have very strongly reduced the dataset, which was considered not acceptable from a practical point of view (RAR 2008). Therefore the following approach was followed (EU RA zinc, ECB 2008):  When data were reported on these parameters, the above selection criteria will be used.  When no data were reported on these parameters: - Tests that had been conducted in artificial waters were excluded when data on pH and/or hardness were lacking.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 142 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

- Tests that were conducted in natural waters were maintained, unless there were clear indications that the (above) parameters in the water strongly deviated from real environmental conditions. For example, tests in waters that received special treatment to remove zinc (and other cations such as Ca and Mg) were excluded. On the other hand, tests conducted in untreated natural United States' waters that were reported to contain a background zinc concentrations which may be considerably below 1 µg/l (depending on natural seasonal variations), such as Lake Superior water, were not excluded.

DOC The range of DOC concentrations of natural waters observed in the EU, is 2,1 mg/l (5P) < DOC < 13 mg/l (95P) (EU risk assessment , ECB 2008). When applying DOC as a relevancy criterion, a relevant range for EU waters should thus also be used for DOC, in accordance to the setting of the relevancy criteria on pH, hardness. The relevant DOC range should thus be set within these observed values. Because it is against the logic of setting relevancy criteria corresponding to the ranges observed in natural EU waters, the additional relevancy criterion of DOC <2mg/l that was introduced at a later stage in the RA process is not applied in the present analysis. This DOC range indeed does not reflect the range of DOC observed in EU waters. In fact, the cut-off of <2mg DOC/l corresponds to the lower 5 percentile of DOC concentrations observed in natural EU waters (~=2,1 mg/l; GEMS-A - Heijerick et al 2003, cited in the RAR), and as a result, data obtained under relevant EU conditions would be excluded.

Following the RAR-logic of using realistic ranges for abiotic water parameters in natural EU waters for setting relevancy criteria, a DOC criterion should actually reflect the observed range. Taking into account that artificial test media as a rule do not contain DOC, it is proposed to apply only an upper range for DOC as a relevancy criterion, i.e. the 95P concentration of the EU range. Tests have thus be considered relevant for the present analysis if DOC concentrations in the test media are between 0 mg/l and 13 mg/l.

In practice, however, the DOC criterion was applied in the RAR (2008) only to the ecotoxicity data, generated in natural waters in the conclusion (i) programme on bioavailability. Since these waters were chosen to validate the bioavailability models, their abiotic conditions are rather wide ranging, and some of the parameters, (e.g. pH, hardness) may fall outside of the range, agreed as relevancy criterion. As a result, several of the test data obtained in these natural waters were rightfully excluded from the PNEC analysis in the RAR, and are excluded also from the present analysis. However, a few results in the RAR database were not considered for the PNEC analysis because of the DOC criterion. Some of these have been included in the analysis for the reason mentioned above. In practice, this only applies to a few data on:  Daphnia magna: a few results obtained under conditions of DOC < 13mg/l are included (Heijerick et al.,2005; De Schamphelaere et al., 2005a). Since they are pooled with the numerous data entries already considered for this species, the effect of this inclusion on the geomean NOEC value for this species is limited (see below). Since moreover Daphnia is only one of 24 species in the SSD, the effect on the HC5 is insignificant.

 For Oncorhynchus mykiss, 1 data point of the RAR database that was removed for DOC considerations was added. For this species, two entries that were in the RAR were removed because not answering the relevancy criteria of the RAR. Also for this species the effect these changes is very limited because of the numerous other data.

The extensive dataset on chronic aquatic toxicity in the RA (ECB 2008) was also updated with new information screened for the same criteria as those described above, for the species already figuring in the RAR (2008): - a few new data were added for D. Magna, and for O. mykiss (De Schamphelaere and Janssen, 2004; De Schamphelaere et al., 2005a). As a result, the geomean for D. Magna and O. mykiss are now 98 and 146µg/l instead of 88 and 189µg/l (RAR 2008), respectively. -for Pseudokircherniella subcapitata, 2 additional NOECs were obtained from a recent study (Muyssen et al 2003); as a result, the geomean changes slightly (19 µg Zn/l instead of 17 µg Zn/l in the RAR). -The results on Brachydanio rerio (Dave et al 1987) were not used for PNEC derivation, since the test suffered from major quality problems (see explanation below table on long-term fish data).

In addition to the dataset of the RA, high quality chronic toxicity data were added to the dataset for six new species: Chlorella sp. (unicellular alga, Wilde et al., 2006); Daphnia longispina (cladoceran, Muyssen et al., 2003); Anuraeopsis fissa and Brachionus rubens (two rotifer species, Azuara-Garcia et al., 2006); Cottus bairdii (fish, Brinkman et al., 2005); Salmo trutta (fish, Kjallqvist et al., 2003). These data were also screened for the same criteria as those described above.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 143 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Chronic aquatic toxicity data freshwater – results

The dataset on chronic aquatic toxicity is presented in detail under sections 7.1.1.1.2. (long-term fish), 7.1.1.2.1. (invertebrates), and 7.1.1.3. (algae and plants).

The 23 distinct chronic species ecotoxicity values that were used for the SSD in the present analysis are summarised in the table below. The “species mean” NOEC values used for PNEC derivation (freshwater PNECadd, aquatic), range from 19 to 530 µg/l.

Table 56. Summary of chronic “species mean” NOEC values that are used as input values for the SSD for deriving the 5th percentile values as a basis for the freshwater PNECadd¸ aquatic. Species geomean values from the RAR (ECB 2008) that were revised are indicated in italics. New values added after the closure of the RAR database are indicated in bold. Taxonomic groups species “Species mean” NOECadd values (µg/l) Algae (unicellular) Pseudokirchneriela subcapitata 19 Chlorella sp. 48 Algae (multicellular) Cladophora glomerata 60 Poriferans Ephydatia fluviatilis 43 Ephydatia muelleri 43 Spongilus lacustris 65 Eunapius fragilis 43 Molluscs Potamopyrgus jenkinsi 75 Dreissena polymorpha 400 Crustaceans Ceriodaphnia dubia 37 Daphnia magna 98 Daphnia longispina 128 Hyalella azteca 42 Rotifera Anuraeopsis fissa 50 Brachionus rubens 50 Insects Chironomus tentans 137 Fish Jordanella floridae 44 Phoxinus phoxinus 50 Pimephales promelas 78 Oncorhynchus mykiss 146 Salvelinus fontinalis 530 Cottus bairdii 169 Salmo trutta 112

Discussion on the SSD freshwater chronic aquatic toxicity. A comparison of the database of freshwater “species mean” NOEC values with the criteria for using statistical extrapolation shows the following:  The number of species chronic NOEC entries (n = 23) meets largely the general requirement for the number of input data (minimum requirement: 10 species NOEC values, preferably more than 15 NOEC values).  The requirement for the number of taxonomic groups is met: Chronic species NOEC values are available for 1 unicellular algal species, 1 multicellular algal species (macro alga), 4 sponge species, 2 mollusc species, 4 crustacean species, 2 rotiferan species, 1 insect species and 7 fish species.  Regarding the coverage of taxonomic groups, the 8 taxonomic groups represented in the database correspond to the requirements set out in chapter R.10:  A fish: the database has 7 fish species  A second family in the phylum chordate (fish, amphibian,..): the database has 7 fish species  A crustacean: the database has 4 species  An insect: there is 1 species in the database

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 144 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

 A family of a phylum other than arthropoda or chordate, e.g; rotifer, annelid, mollusca,...: the database has 2 rotifers, 2 molluscs  A family in any order of insect or any other phylum not already represented: the database has 4 porifera  Algae: the database has 2 algae species  Higher plants: in the database of accepted NOEC values, data for algae are included, but data for higher plants are lacking. However, data for freshwater higher plants are included in the database of rejected NOEC values of the RAR (ECB 2008). The rejected NOEC values for higher plants are from the following studies :  A long-term study with four different species of freshwater higher plants (Elodea nuttallii, Callitrische platycarpa, Spirodela polyrhiza and Lemna gibba) resulted in unbounded NOEC values of > 650 µg/l for all four plant species (endpoints: survival and growth). The plants used in this study were obtained from unpolluted ditches or ponds in the Netherlands and grown in filtered ditch water with a pH of 8.0 (Van der Werff & Pruyt, 1982). The study was not used in the RAR because it had only an unbounded NOECs.

 A study with duckweed Lemna minor resulted in a NOEC of 160 µg/l (endpoint: growth) at pH 5 and hardness of 310 mg/l in artificial medium (Jenner & Janssen-Mommen, 1993). The study was not used in the RAR because the test conditions were outside the relevancy ranges for pH and hardness.  Tests with duckweed Lemna pauciscostata resulted in a NOEC of 5000 µg/l (endpoint: growth) at pH 4 or 5 and hardness of 700 mg/l in an artificial medium and tests in another artificial medium resulted in about 60-80% growth inhibition at 1000 µg/l at pH 6 or 7 and hardness of 120 mg/l (Nasu & Kugimoto, 1981). The study was not used in the RAR because the test conditions were outside the relevancy ranges for pH and hardness. From the data for these six plant species it was however concluded in the RAR that aquatic higher plants do not appear to be very sensitive to zinc toxicity in comparison with algae or animals and thus the lack of useful NOEC values for higher plants was acceptable. Furthermore, the RAR stressed that “the database of accepted NOEC values includes a relatively high NOEC (60 µg/l) for the macro alga Cladophora glomerata and macro algae ressemble higher plants” (RAR 2008).

It was concluded in the RAR that the taxonomic coverage requirements for applying an SSD were largely met by the RAR dataset. The present analysis has added 6 species to the one of the RAR, including an additional taxonomic group. So, a wealth of information on different species is available, and statistical extrapolation will also be used in the present analysis for PNEC freshwater derivation.

Statistics on the species sensitivity distribution (SSD) Given the multitude of relevant high quality data, statistical extrapolation was used for PNEC determination. Following the RIP R.10. guidance, “different distributions may be used” for the SSD. We tested the lognormal distribution (default option), as calculated with the “ETX” software, and subsequently several other distributions with the “@Risk” software. The statistics of the curve–fitting on the chronic NOEC data are summarised in table below.

Table 57. Summary statistics for SSD on chronic NOEC values for zinc in freshwater (N= 23). distribution HC5 Lower Median A-D A-D K-S K-S Acceptance estimate HC5/ statistic significance statistic significance for PNEC on HC5 lower level level setting 95% C.I. Lognormal 20.6 12.3 1.68 0.89 0.01 0.89 p = 0.05 accepted (ETX) (accepted) (accepted) Lognormal 20.9 Not Not 0.98 0.01<= p 0.16 p < 0.15 accepted (@risk) available available <0.025 (accepted) (accepted) Extreme 27.2 Not Not 0.63 0.05 0.1 accepted values available available (accepted) (accepted) (@risk)

Using the Anderson-Darling (A-D)-test for normality, the default distribution (lognormal) does fit significantly at a level of 1 %. Using the Kolmogorov-Smirnov test, the lognormal is accepted at 5% level. This analysis indicates that the fit of the lognormal distribution is not very good at the lower tail of the distribution (low A-D acceptance). This outcome of the SSD-statistics is the same as the one observed for the lognormal distribution in

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 145 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 the RAR, where it was stipulated that “the Anderson-Darling test indicated that there was only goodness-of-fit for the log-normal distribution at a low significance level (1%). The Kolmogorov-Smirnov (K-S)-test accepted both the log-normal and log-logistic distribution at a higher significance level (5%)” (RAR 2008). Since the statistical significance levels on the lognormal distribution are the same as those under which the lognormal was accepted for PNEC setting in the RAR (2008), the lognormal distribution is accepted in the present analysis also.

The acceptance of the lognormal distribution at the same significance level as in the RAR is expected, since the species data that were added to the distribution are quite close to the average value of the distribution (79µg/l); they are lower in 4 cases (50, 50, 50,68), and higher in 2 cases (119,138). The relatively low goodness-of-fit is related to the lower tail of the SSD (where the input data were not changed as compared to the RAR) and where the fit with the lognormal distribution is not very good (figure 2). For the same reason, the K-S test gives acceptance at 5% level, for the revised database, too (K-S is more related to the values near the middle of the distribution).

Figure 2. Lognormal distribution curve fitting to the freshwater chronic toxicity data for zinc (ETX graphics).

Other distributions fit better to the data. Of the other distributions tested with @risk, the “Extreme values” (EV)- distribution provided the best fit, as demonstrated by the lowest A-D statistic. The fit was highly significant both with the A-D and the K-S test (p values > 0.05).

To be conform with the approach taken in the RAR (2008), the lognormal distribution is used to provide a basis for setting the PNEC freshwater, in spite of a better fit to the data provided by the extreme values distribution. It is noted that the HC5 resulting from the extreme values distribution is significantly higher than the HC5 calculated with the lognormal distribution (20.6 µg/l versus 27.3 µg/l).

It is noted also that the PNEC is a PNECadd., i.e. to be added to the background when using monitored water concentrations.

The 5th percentile value of the SSD (the HC5) is set at the 50% confidence level, using the log-normal

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 146 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

distribution function, which results in an HC5 value of 20.6 µg Znadded/l.

The reference values for chronic aquatic toxicity were determined: -at pH 8: from the extensive chronic ecotoxicity data available for algae, invertebrates and fish (section 7.1.1., 2.). The standard species NOEC values for each taxonomic group for which a bioavailability model is available were taken at pH 8, and the lowest of the 3 was selected as a reference value at pH 8. -at pH 6: the corresponding aquatic toxicity at pH 6 was calculated from the same database for the standard species for which bioavailability models were available, and the lowest of the 3 was selected as a reference value at pH 6. . The results are summarised below: -for algae, the NOEC of the BLM-species Pseudokircherniella subcapitata is the lowest of the SSD at pH 8 (19 µg/l). This value corresponds to a water of pH 8,0, hardness 24 mg CaCO3 and DOC 2.0 mg/l. With the BLM, a corresponding species NOEC of 142 µg/l is calculated for this species at pH 6 (other water conditions same). -for invertebrates, the BLM-species Daphnia magna gives a species mean at pH 8 of 98 µg/l, corresponding to a water of pH 8, hardness 24 mg CaCO3/l and DOC 1,2 mg/l. The Dapnia magna-BLM predicts at pH 6 (other water conditions same) a species NOEC of 82 µg/l. -for O. Mykiss, the species mean at pH 8 is 146 µg/l (hardness 45 mg/l, DOC 2 mg/l). Using the corresponding species BLM gives a species NOEC of 146 µg/l at pH 6 (other conditions same).

From these data, the following reference values for chronic zinc aquatic toxicity are derived: -at pH 8.0: 19 µg Zn/l (Pseudokircherniella subcapitata) -at pH 6.0: 82µg Zn/l (Daphnia magna)

3. Aquatic chronic toxicity: marine waters For zinc in marine waters, a specific effects assessment was made and a specific PNEC was derived, since there is a vast dataset available on marine ecotoxicity. This specific approach is also more relevant for the toxicity of zinc in the marine environment given the different speciation and bioavailability of zinc in salt – and freshwater, and differences in physiology of saltwater organisms. Given the vast amount of available toxicity data, statistical extrapolation was used to derive the marine PNEC. This marine effects assessment is following an added risk approach, as applied for the freshwater.

Establishing the dataset

Sources of data This report analyses the available chronic zinc toxicity data for marine organisms. The ecotoxicological data were derived from original papers, published in peer-reviewed international journals. Literature and environmental databases, including AQUIRE (US EPA), MARITOX, ECETOC, and BIOSIS, as well as review articles covering zinc in marine waters were searched and reviewed for sources of relevant and reliable chronic toxicity data on zinc. Only original literature was used.

Data reliability and relevancy Selection of ecotoxicity data for quality was done according to a systematic approach as presented by Klimisch et al. 1997. Standardized tests, as prescribed by organizations such as ASTM, OECD and US EPA, are used as a reference when test methodology, performance and data treatment/reporting are considered. A detailed description of methods and conditions employed in the study should be provided. The thorough description of key requirements guarantees the high reliability (Q1) of the reported ecotoxicity data. Non-standardized test data, may also have a high reliability, but required a more thorough check on their compliance with reliability criteria before being used for deriving a PNEC. As for data relevancy, tests should be performed in media that reflect natural environmental conditions (e.g. salinity, zinc background concentrations and other abiotic conditions). A set of criteria for checking reliability and relevancy has been defined in this work and is presented below. Those criteria were used to discriminate between data accepted with restrictions (Q2) and unreliable data (Q3).

Test medium Both natural or artificial sea water were accepted as test medium. In case EDTA is present in the test medium, the study is considered not reliable. Chelators other than EDTA (e.g. NTA, citrate,…) can however be added.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 147 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Only the results of tests in which the organisms were exposed to zinc alone were used, thus tests with metal mixtures were not considered for this evaluation. Only the results of tests with soluble zinc salts were used.

Salinity The TGD does not define the salinity range of sea water. The Water Framework Directive allows for using the saltwater EQS at salinities ≥ 5‰, which sets the limit between freshwater and brackish waters. Therefore, tests performed at salinity levels down to a value of 5 g/kg were accepted for this marine dataset.

Background concentrations Adaptation to high zinc concentrations may influence the sensitivity to zinc. To ensure that test organisms are adapted to the test conditions, the culture and test conditions should be similar. In addition, organisms acclimatized to elevated background in culture media or collected at contaminated sites were not used in the analysis. Only results from unpolluted test media were used, thus excluding tests that were performed in media containing high background Zn concentrations (> 10 µg Zn/l). Low background values were not discarded. In some references, no data have been reported on the culture conditions. In those cases it was assumed that culture and test conditions were similar, as is common practice.

Control data Tests were rated as not reliable if control data was not provided (Q3). Effect levels derived from toxicity tests using only one test concentration always result in unbounded and therefore not assignable data. Therefore, only the results from toxicity tests using one control and at least two zinc concentrations were retained for this evaluation. Data reporting mortality rates higher than 20% in the control were not used.

Test statistics Because effect concentrations are statistically derived values, information concerning the statistics was used as a criterion for data selection. In that respect L(E)C10 values are considered as equivalent to NOEC. Studies that do not report test statistics but present trust-worthy data are rated as reliable with restrictions (Q2). Studies that do not report test statistics and do not present a dose-response analysis are rated as unreliable (Q3).

Test design Data without treatment replication or with pseudo-replication were not used (Q3).

Type of test (duration, endpoints covered) Only reliable endpoints from properly conducted chronic tests are being considered. Toxicological endpoints which affect the species at population level were taken into account (i.e. survival, development, reproduction and growth). Historically, chronic exposure has been defined as > 4 days for all invertebrates and fish. With respect to this assessment, the arbitrary selection of this exposure period has been reviewed in light of the sensitivity of the endpoint and the duration of the life stage under assessment. For example, early life stages tests (embryos, larvae) of 24-48 hours have been included in this assessment. Sperm cells and fertilized eggs tests of a few hours were also considered as chronic data. Indeed, abnormal development can be observed within this time frame (e.g. molluscs, echinoderms), and the continuation of these tests would derive no additional information which could provide protection for the environment. Tests performed on adults over 96-h were considered as acute toxicity tests for invertebrates and fish. For algae, and according to ASTM guidelines, data reporting growth rates < 1 in the control were not used.

Test species: endemic versus non-endemic species The culture and test conditions are considered more relevant than the geographical origin of the species and the OECD guidelines recommend the use of a number of “standard” species which do not have a world-wide distribution. Moreover, using the origin of species as criterion would considerably reduce the dataset and limit the data to only a few species / taxa, which may obscure variation in sensitivity. Therefore, the geographical origin of the test species has not been used as selection criterion.

Measured concentrations Zinc is a natural element with typical background levels in seawater ranging from 0.5-1 ug/L (e.g. Laane et al. 1992). Because of the importance of understanding the true exposure concentrations (including the background concentration in the culture media), any study not supported by analytical data would automatically be excluded from the high quality studies (Q1 data). Therefore, data not supported by measured concentrations are excluded from the Q1 dataset. These studies have been rated as reliable with restrictions (Q2) or not reliable (Q3), depending on the data quality as well as the availability of zinc background levels in the test media.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 148 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

The results of the marine aquatic toxicity studies are expressed either as measured concentration, or usually as nominal concentration. The measured concentrations include the background zinc concentration. Because of the added risk approach, measured concentrations have been corrected for background. In case specific background concentration was not mentioned in the paper, a default background concentration of 0.5 µg zinc dissolved/L was subtracted to the total measured concentration. The nominal concentrations were used as such for the PNECadd derivation. If it is not mentioned whether the NOEC/L(E)C10 values are based on measured or nominal concentrations, they were considered as nominal concentrations.

In the reported data, zinc has been used as the test material with several salts being used as the precursor. As with other risk assessments on metals, it is generally recognized that under laboratory conditions almost all the zinc is present in the dissolved fraction, therefore these results can be regarded as being dissolved zinc concentrations.

Derivation of NOEC/LOEC values (methods)

The toxicological variables are estimated based on measured or nominal NOECs or EC 10 values. There has to be a concentration-effect relationship. In the past, the NOEC was determined directly from the concentration-effect curve by consideration of the deviation of the control (e.g. 10%) or it was derived on the basis of ANOVA (analysis of variance) and a subordinate test (e.g. Dunett's). This method to derive the NOEC with the ANOVA is criticized. Pack et al., 1993 recommends the calculation of the ECX point as a preferable alternative. In older investigations, it may be difficult to find out how the NOEC was generated unless test reports or raw data are available. NOEC values without any information about the concentration-dependent response were excluded from PNEC analysis.

Unbounded NOEC values (i.e. no effect was found at the highest concentration tested) were not used in this analysis in accordance to the Zn RAR, 2008.

In a few cases, no NOEC or LOEC value was provided, but raw data were reported. When possible, the raw data were used to derive an EC10 using linear interpolation (LI). The EC10(LI) values were used with restrictions (Q2).

In case only a LOEC is given in the report, it was used to derive a NOEC with the following procedures according to the Zn RAR (2008): - LOEC ≥ 10 and < 20% effect: NOEC can be calculated as LOEC/2. - LOEC ≥ 20 and < 30% effect: NOEC can be calculated as LOEC/3. - If the effect percentage of the LOEC is higher than 30% or if it is unknown, no NOEC can be derived.

In aquatic toxicity the MATC (maximal acceptable toxicant concentration) is often calculated. This is the geometric mean of the NOEC and the LOEC. If in the test report only the MATC is presented, the MATC can be divided by √2 to derive a NOEC.

If for one species, several chronic NOEC values (from different tests) based on the same endpoint are available, these values are averaged by calculating the geometric mean, resulting in a “species mean NOEC”. If several toxicity endpoints are reported for the same species, the most sensitive one, so the lowest NOEC value is selected for PNEC derivation. The lowest value is determined on the basis of the geometric mean if more than one value for the same endpoint exists.

Chronic ecotoxicity in the marine environment – data

The marine zinc database largely fulfils the species and taxonomic requirements for input chronic toxicity data as explained in the RIP R. 10 guidance (at least 10 species NOECs and 8 taxonomic groups). Indeed, 39 species mean NOECs based on 48 NOEC values, from 9 taxonomic groups covering three trophic levels were found to fulfil the relevancy and reliability requirements as explained above. The marine zinc database includes 4 micro- and 5 macro-algae species, 4 annelid species, 6 crustacean species, 5 echinoderm species, 9 mollusc species, 1

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 149 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

nematod species, 1 cnidarian species and 1 fish species. The detailed results per study record are given under sections 7.1.1.1.2. (fish), 7.1.1.2.2. (invertebrates) and 7.1.1.3. (algae and plants).

The geometric mean values of the species NOECs together with their reliability scoring are presented in Table below. Most of the effects data are ranked as reliability 2 (Q2). Most of the data were reported as nominal concentrations.

Table 58. Geomean species NOECs of the marine zinc effects database presented by taxonomic group, species name and family name. Taxonomic Species name Family Geomean Reliability group NOECadd value (according to (µg Zndiss/L) Klimisch et al. 1997) Micro-Algae  Asterionella japonica Fragilariaceae 13.3 2 (4)  Chaetoceros compressum Chaetocerotaceae 11.2 2  Nitzschia closterium Bacillariaceae 65.9 1 and 2  Skeletonema costatum Skeletonemaceae 26.4 2 Macro-Algae  Ascophyllum nodosum Fucaceae 564.8 2 (8)  Ceramium tenuicore Ceramiaceae 7.8 2  Fucus serratus Fucaceae 488 2  Fucus spiralis Fucaceae 613.2 2  Fucus vesiculosus Fucaceae 100 2  Macrocystis pyrifera Lessoniaceae 189.7 1  Pelvetia canaliculata Fucaceae 670.8 2 Ulvaceae 313 2  Ulva pertusa Annelids  Capitella capitata Capitellidae 100 2 (4)  Ctenodrilus serratus Ctenodrilidae 100 2  Neanthes arenaceadontata Nereididae 33.3 2  Ophryotrocha diadema Dorvilleidae 70.7 2 Cnidarians  Eirene viridula Eirenidae 300 2 (1) Crustaceans  Allorchestes compressa Dogielinotidae 62.5 2 (6)  Holmesimysis costata Mysidae 5.6 1  Mysidopsis bahia Mysidae 101 2  Mysidopsis intii Mysidae 101 2  Paragraspus quadridentatus Grapsidae 294.5 2  Tigriopus brevicornis Harpacticidae 297 2 Echinoderms  Arbacia lixula Arbaciidae 10 2 (5)  Asterias amurensis Asteriidae 50 2  Paracentrotus lividus Echinidae 15.98 2  Sphaerechinus granularis Toxopneustidae 10 2  Sterechinus neumayeri Echinidae 160 2 Molluscs  Crassostrea cucullata Ostreidae 18.4 2 (9)  Crassostrea gigas Ostreidae 39.6 2  Crassostrea margaritacea Ostreidae 11.8 2  Haliotis rubra Haliotidae 20.4 2  Haliotis rufescens Haliotidae 13.8 1  Ilyanassa obsoleta Nassariidae 20.7 2  Mya arenaria Myidae 900 2 Mytilidae 84.9 2  Mytilus galloprovincialis Veneridae 55 2  Ruditapes decussatus Nematods  Monhystera disjuncta Monhysteridae 250 2 (1) Fish  Clupea harengus Clupeidae 25 2 (1) TOTAL 39 species 27 families 39 species

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 150 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

mean NOECs

Statistics on the species sensitivity distribution (SSD) Given the multitude of relevant high quality toxicity data, statistical extrapolation was used for PNEC determination. As the approach taken is based on added risks, the results of the toxicity tests based on measured concentrations were corrected for background zinc concentration. Given the wealth of experimental data, no alternative method i.e. assessment factor approach was applied for the PNEC determination.

Following the RIP R. 10 guidance, different distributions may be used for the SSD. Fitting of the chronic zinc toxicity data was assessed towards the log-normal frequency distribution (default distribution), obtained using the RIVM program ETX2.0 (Van Vlaardingen et al. 2004). Several distributions were subsequently calculated with the “@risk” (Palisade Inc. USA) software. The statistics of the curve-fitting on the chronic NOEC data are summarized in Table below.

Table 59. Summary statistics for the SSD on chronic NOEC values for zinc in saltwater (n=39). Distribution Media Lower Median A-D A-D K-S K-S Acceptance n HC5 estimate HC5/lower statistic significance statistic significance for PNEC on HC5 95% C. I. level level setting Lognormal 6.09 3.09 2 0.49 P > 0.1 0.61 P > 0.1 Accepted (ETX) Lognormal 6.2 Not Not 0.48 P > 0.1 0.1 P > 0.1 Accepted (@risk) available available Best fit = 8.5 Not Not 0.44 N/A 0.08 N/A Accepted Weibull available available (@risk)

The goodness of fit tests reports a measure of the deviation of the fitted distribution from the input data. Preference is given to Anderson-Darling (A-D) test because it highlights differences between the tails of the fitted distribution and input data. The Kolmogorov-Smirnov (K-S) can also be used for goodness of fit purposes but it does not detect tail discrepancies very well.

Using the A-D test for normality, the lognormal distribution does fit significantly at all levels. The lognormal distribution is also accepted at all levels using the K-S test. The lognormal distribution results in an HC5 at 50% confidence value (HC5 5% - 95% confidence values) of 6.09 (3.09 - 10.26) µg Zn/L . The observed Q1-2 data are presented on log-scale together with their fitted normal distribution curve in figure below.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 151 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Figure 3. Cumulative distribution of the 39 species mean NOEC values from Zn toxicity tests in the marine organisms database. Observed data and normal distribution curve fitted on the data.

Other distributions fit better to the data. Using @risk, The Weibull distribution turned out to be the best fit with an HC5-50 value of 8.5 µg Zn/L , as indicated by the lowest A-D statistic (table 59). The difference between both distributions is however minimal.

To be conform with the approach taken in the Zn RAR 2008, and because the lognormal distribution provides a significant fit to the data, it is the lognormal distribution which was used to provide a basis for setting the PNEC saltwater, in spite of a better fit with the Weibull statistical distribution.

It is noted also that the PNEC is a PNECadd., i.e. the background concentration needs to be considered in the compliance assessment exercise. The 5th percentile value of the SSD (the HC5), set at 50% confidence value, using the lognormal distribution (ETX 2.0) function, results in a value of 6.09 µg zincadded/L.

Mesocosm studies Davies and Sleep (1979) investigated the influence of zinc upon carbon fixation rates of natural phytoplankton communities present in the English Channel (near Plymouth). A series of three samples of varying biological composition (100% diatoms, 60% dinoflagellates-40% diatoms, and 60% diatoms-40%dinoflagellates) were taken in July at the same location and the tests with zinc were done in natural sea water. The zinc background concentration in seawater varied from 0.4 to 7.6 µg/L. A pre-incubation period of phytoplankton assemblages to zinc levels was designed in order to equilibrate the populations with the experimentally added zinc before measuring their carbon fixation rates. Fixation rates less than 90% of the mean control value were attributed to inhibition caused by the presence of zinc. The lowest concentrations of zinc which caused detectable inhibition of carbon fixation, i.e. rates lower than 90% of the mean control values, were in the range of 10 to 15 µg/L.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 152 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

From the dose-response curves, the EC10 levels were in the range of 7 to 13 µg/L. This means that the HC5 derived from the normal distribution of the log-transformed data is protective for those phytoplankton assemblages. When looking at single species toxicity tests, a number of the diatoms and dinoflagellates are among the more sensitive species in the chronic NOEC database (e.g. Asterionella japonica, Chaetoceros compressum, ...). Also in the freshwater database, unicellular algae and algal communities were among the more sensitive organisms. Davies and Sleep (1979) were already mentioned in the RAR (2008) and used among other references to derive a provisional PNEC saltwater.

Another study on phytoplankton communities from three different areas was carried out by Wolter et al. (1984). Influence of metal to carbon fixation rate of phytoplankton and to glucose incorporation by bacteria was determined. Water samples coming from Kiel Fjord, North Sea and a coastal area of the North Atlantic Ocean were investigated after addition of varying concentrations of metals including zinc. Surface samples were taken in Kiel Fjord during the spring and autumn plankton bloom. Zinc was added to subsamples to give concentrations in the range of 4.3 to 304.3 µg/L. The North Sea and Atlantic samples were collected during a cruise in 1981. The added zinc concentrations were 0.29 to 1.45 µg/L. The added metal concentrations were lower than in the Kiel Fjord experiments due to lower concentrations in the water compared to Baltic Sea water. In all cases the samples contained mixed phytoplankton populations which were dominated by diatoms. From the graph and the text, zinc reduced plankton activity in the Kiel Fjord samples only at concentrations above 100 µg/L. Moreover, carbon fixation measurements carried out four and 24 hours after metal addition to the North Sea and Atlantic samples were not reduced at any test concentration, which then gives unbounded values (see table below for summary of results). The same was true for bacterial glucose incorporation.

Table 60. Results of field experiments made on phytoplankton communities coming from various natural sea waters Location Endpoint Kiel Fjord North Sea Atlantic NOEC (µg Zn/L) >100 >1.45 (unbounded) >1.45 (unbounded) C fixation rate after 4 or 24 hours exposure NOEC (µg Zn/L) / >1.45 (unbounded) >1.45 (unbounded) Bacterial glucose after 4 or 24 hours incorporation exposure

Although this study has nothing inherently wrong in the design, it has its limitations since important details are lacking (no accurate information about test organisms and test conditions, no information on statistics and on control data, …). The unbound NOEC results in North Sea and Atlantic are not useful. The results should be interpreted with care, but notably, the results obtained in Kiel fjord suggest that the HC5 level described above would be protective.

After the registration of the substance in 2010, an outdoor state-of-the-art marine mesocosm study was perfomed on dissolved zinc, and results were reported recently (Foekema et al 2012). In this study, an ecosystem assembly containing a.o. phytoplankton, periphyton, macro algae, zooplankton, molluscs, and annelids, was exposed to dissolved zinc concentrations between 2.7µg/l and 91µg/l, maintained at constant level by regular monitoring and spiking. The dataset showed a consistent picture of the impact of continuous exposure of dissolved zinc on the mesocosm community during the 83 days of the test (figure below). The No observed ecological adverse effect concentration (NOEAEC) resulting from the study was 12 µg Zn/l. At treatment levels 5.6, 7.5, and 12 µg Zn/l, no negative effects were observed, but indications were found of a slight stimulation of the productivity, that might be related to zinc essentiality. Since this did not result in substantial changes in the colmposition or functioning of the mesocosm ecosystem these effects were not considered as being ecologically adverse (Foekema et al 2012).

Figure X / overview of endpoints with statistical significant differences between treated and untreated (2.7µg Zn/l) mesocosms. Effect class “A”: no effects, “B”: slight effects, “C” pronounced temporal effects, “D”: pronounced durable effects. A positive or negative difference from the control is indicated with “+” or “-“, respectively (taken from Foekema et al 2012).

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 153 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

7.1.1.1. Fish

7.1.1.1.1. Short-term toxicity to fish

The results are summarised in the following table:

Table 61. Overview of short-term effects on fish

Method Results Remarks Reference Thymallus arcticus LC50 (96 h): 315 µg/L 2 (reliable with Buhl K. and dissolved (nominal) restrictions) Hamilton S. (1990) freshwater LC50 (96 h): 142 µg/L key study static dissolved (nominal) read-across based on American Society for testing matrials LC50 (96 h): 112 µg/L grouping of 1988: Standard practice for conducting dissolved (nominal) substances (category acute toxicity tests with fishes, approach) macroinvertebrates and amphibians, LC50 (96 h): 1580 µg/L ASTM, E-729-88, Philadelphia dissolved (nominal) Test material (IUPAC name): zinc LC50 (96 h): 166 µg/L chloride (See dissolved (nominal) endpoint summary for justification of LC50 (96 h): 2920 µg/L read-across) dissolved (nominal)

LC50 (96 h): 168 µg/L dissolved (nominal)

LC50 (96 h): 168 µg/L dissolved (nominal) Oncorhynchus kisutch LC50 (96 h): 820 µg/L 2 (reliable with Buhl K. and dissolved (nominal) restrictions) Hamilton S. (1990)

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 154 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 freshwater LC50 (96 h): 1810 µg/L key study dissolved (nominal) static read-across based on LC50 (96 h): 1650 µg/L grouping of American Society for testing matrials dissolved (nominal) substances (category 1988: Standard practice for conducting approach) acute toxicity tests with fishes, LC50 (96 h): 727 µg/L macroinvertebrates and amphibians, dissolved (nominal) Test material ASTM, E-729-88, Philadelphia (IUPAC name): zinc chloride (See endpoint summary for justification of read-across) Oncorhynchus mykiss LC50 (96 h): 2170 µg/L 2 (reliable with Buhl K. and dissolved (nominal) restrictions) Hamilton S. (1990) freshwater LC50 (96 h): 169 µg/L key study static dissolved (nominal) read-across based on American Society for testing materials grouping of 1988: Standard practice for conducting substances (category acute toxicity tests with fishes, approach) macroinvertebrates and amphibians, ASTM, E-729-88, Philadelphia Test material (IUPAC name): zinc chloride (See endpoint summary for justification of read-across) Cottus bairdii LC50 (96 h): 439 µg/L 2 (reliable with Brinkman S and dissolved (meas. (not restrictions) Woodling J (2005) freshwater specified)) key study flow-through read-across based on lab-designed dose-response test grouping of substances (category approach)

Test material (IUPAC name): zinc (See endpoint summary for justification of read- across) Pimephales promelas LC50 (96 h): 780 µg/L 2 (reliable with Schubauer-Berrigan dissolved (meas. (not restrictions) M.k., Dierkes J.R., freshwater specified)) Monson P.D. and key study Ankley, G.T. static LC50 (95 h): 330 µg/L (1993a) dissolved (meas. (not read-across based on lab-designed dose-response test at specified)) grouping of varying pH and at relatively high substances (category hardness conditions. LC50 (96 h): 500 µg/L approach) dissolved (meas. (not specified)) Test material (IUPAC name): zinc sulphate (See endpoint summary

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 155 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

for justification of read-across) Oncorhynchus mykiss (reported as LC50 : ca. 0.481 mg/L NH3 2 (reliable with Johnson et al. Salmo gairdneri) diss. (meas. (geom. mean)) restrictions) (1984) freshwater supporting study flow-through experimental result methode in Ambient water criteria for Test material (CAS ammonia not reported, see primary name): Ammonium literature chloride Pimephales promelas LC50 : ca. 1.344 mg/L 2 (reliable with Johnson et al. dissolved (NH3) (meas. restrictions) (1984) freshwater (geom. mean)) supporting study flow-through experimental result methode in Ambient water criteria for ammonia not reported, see primary Test material (CAS literature name): ammonium chloride mornone americana LC50 : ca. 0.279 mg/L 2 (reliable with Johnson et al. dissolved (NH3) (meas. restrictions) (1984) freshwater (geom. mean)) supporting study static experimental result methode in Ambient water criteria for ammonia not reported, see primary Test material (CAS literature name): ammonium chloride

Discussion

Good quality and relevant data for 5 species. Tests were done according to standard protocol or equivalent.

Data are grouped per species according to

-pH: low (6 -<7) - neutral/high (7 -8.5)

-and hardness: low/medium (<100mg CaCO3/l) and medium/high (>100 mg CaCO3/l).

Fish are generally less sensitive than invertebrates and algae.

The following information is taken into account for acute fish toxicity for the derivation of PNEC:

Key data (lowest LC50 values) are: -for Oncorrhynchus Mykiss: 0.169 mg Zn/l (single value) at neutral/high pH and low hardness -for Pimephales promelas (single values): 0.780 mg Zn/l at low pH (high hardness) and 0.330 mg Zn/l at neutral/high pH, high hardness -for Pimephales promelas: LC50 0.780 mg Zn/l (at low pH); 0.33mg Zn/l at neutral/high pH

7.1.1.1.2. Long-term toxicity to fish

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 156 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

The results are summarised in the following table:

Table 62. Overview of long-term effects on fish

Method Results Remarks Reference Salmo gairdneri (new name: NOEC (72 d): 440 µg/L 2 (reliable with Cairns M.A. Oncorhynchus mykiss) dissolved (meas. (not restrictions) ,Garton R.R. and specified)) based on: Tubb R.A. (1982) freshwater mortality key study survival of early life stages (eggs to read-across based on early juveniles grouping of substances (category flow-through approach) lab -designed dose response test with Test material smolts (2-year old salmon) (IUPAC name): zinc chloride (See endpoint summary for justification of read-across) Oncorhynchus mykiss NOEC (30 d): 39 µg/L 1 (reliable without De Schamphelaere dissolved (estimated) based restriction) K.A.C., Heijerick freshwater on: mortality D.G. and Janssen, key study CR. (2003a) juvenile fish: growth NOEC (30 d): 95 µg/L dissolved (meas. (arithm. read-across based on flow-through mean)) based on: mortality grouping of substances (category OECD Guideline 215 (Fish, Juvenile NOEC (30 d): 45 µg/L approach) Growth Test) dissolved (meas. (arithm. mean)) based on: mortality Test material (IUPAC name): zinc NOEC (30 d): 151 µg/L chloride (See dissolved (meas. (arithm. endpoint summary mean)) based on: mortality for justification of read-across) NOEC (30 d): 159 µg/L dissolved (meas. (arithm. mean)) based on: mortality

NOEC (30 d): 256 µg/L dissolved (meas. (arithm. mean)) based on: mortality

NOEC (30 d): 157 µg/L dissolved (meas. (arithm. mean)) based on: mortality

NOEC (30 d): 974 µg/L dissolved (meas. (arithm. mean)) based on: mortality

NOEC (30 d): 696 µg/L dissolved (meas. (arithm. mean)) based on: mortality

NOEC (30 d): 324 µg/L dissolved (meas. (arithm. mean)) based on: mortality

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 157 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

NOEC (30 d): 79 µg/L dissolved (meas. (arithm. mean)) based on: mortality

NOEC (30 d): 169 µg/L dissolved (meas. (arithm. mean)) based on: mortality

NOEC (30 d): 48 µg/L dissolved (meas. (arithm. mean)) based on: mortality Oncorhynchus mykiss NOEC (30 d): 199 µg/L 1 (reliable without De Schamphelaere dissolved (meas. (arithm. restriction) K.A.C., Lofts, S. freshwater mean)) based on: mortality and Janssen C.R. key study (2005a) juvenile fish: growth and mortality read-across based on flow-through grouping of substances (category OECD Guideline 215 (Fish, Juvenile approach) Growth Test) Test material (IUPAC name): zinc chloride (See endpoint summary for justification of read-across) Cottus bairdi NOEC (30 d): 172 µg/L 2 (reliable with Brinkman S. and dissolved (meas. (not restrictions) Woodling, J. (2005) freshwater specified)) based on: mortality key study recently emerged fish survival NOEC (30 d): 169 µg/L read-across based on flow-through dissolved (meas. (not grouping of specified)) based on: substances (category lab-designed dose-response test mortality approach) Test material (IUPAC name): zinc (See endpoint summary for justification of read- across) Jordanella floridae NOEC (30 d): 26 µg/L 2 (reliable with Spehar R.L. (1976) dissolved (estimated) based restrictions) freshwater on: length key study P (larvae) to F1 (larvae) ; 1day NOEC (30 d): 75 µg/L dissolved (estimated) based read-across based on flow-through on: length grouping of substances (category lab-designed dose-response test over LOEC (30 d): 51 µg/L approach) long time period dissolved (meas. (not specified)) based on: length Test material (IUPAC name): zinc LOEC (30 d): 139 µg/L sulphate (See dissolved (meas. (not endpoint summary specified)) based on: length for justification of read-across)

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 158 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Salmo gairdneri (new name: NOEC (22 mo): 130 µg/L 2 (reliable with Sinley J.R., Goettl Oncorhynchus mykiss) dissolved (meas. (not restrictions) J.P.Jr. and Davies specified)) based on: P.H. (1974) freshwater mortality key study two test periods considered, see details NOEC (25 d): 25 µg/L read-across based on in results dissolved (estimated) based grouping of on: mortality substances (category flow-through approach) lab-designed dose-response test over a Test material long exposure period , spanning life (IUPAC name): zinc stages from eyed eggs to fish of sexual sulphate (See maturity, and adult fish, respectively endpoint summary (see results) for justification of read-across) Salvelinus fontinalis NOEC (36 mo): 530 µg/L 2 (reliable with Holcombe G.W., dissolved (meas. (not restrictions) Benoit, D.A. and freshwater specified)) based on: number Leonard E.N. hatched key study (1979) different early life stages and adult fish tested over prolonged periods. NOEC (12 wk): 720 µg/L read-across based on dissolved (meas. (not grouping of flow-through specified)) based on: substances (category mortality approach) lab-designed dose-response test on different early life stages and adult fish NOEC (8 wk): 720 µg/L Test material over prolonged periods. dissolved (meas. (not (IUPAC name): zinc specified)) based on: sulphate (See mortality endpoint summary for justification of read-across) Salmo trutta NOEC (116 d): 250 µg/L 1 (reliable without Källqvist T., dissolved (meas. (arithm. restriction) Rosseland B., freshwater mean)) based on: time to Hytterod S. and hatch key study Kristiansen T. early-life stage: reproduction, (2003) (sub)lethal effects NOEC (116 d): 57 µg/L read-across based on dissolved (meas. (arithm. grouping of flow-through mean)) based on: time to substances (category hatch approach) OECD Guideline 210 (Fish, Early-Life Stage Toxicity Test) NOEC (116 d): 56 µg/L Test material dissolved (meas. (arithm. (IUPAC name): zinc mean)) based on: time to sulphate (See hatch endpoint summary for justification of NOEC (116 d): 61 µg/L read-across) dissolved (meas. (arithm. mean)) based on: time to hatch Pimephales promelas NOEC (8 mo): 78 µg/L 2 (reliable with Benoit G.A. and dissolved (meas. (not restrictions) Holcombe G.W. freshwater specified)) based on: (1978) reproduction key study early-life stage: reproduction, (sub)lethal effects NOEC (8 mo): 145 µg/L read-across based on dissolved (meas. (not grouping of flow-through specified)) based on: substances (category survival, hatchability of approach)

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 159 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Dose response study on the toxicity of offspring, developmental Test material zinc to Pimephales promelas effects (IUPAC name): zinc sulphate (See NOEC (8 mo): 295 µg/L endpoint summary dissolved (meas. (not for justification of specified)) based on: read-across) mortality

NOEC (8 mo): > 575 µg/L dissolved (meas. (not specified)) based on: growth rate Pimephales promelas NOEC (7 d): 85 µg/L 2 (reliable with Norberg, T. J. and dissolved (meas. (not restrictions) Mount, D.I. (1985) freshwater specified)) based on: mortality supporting study newly hatched larvae (<1d); survival and growth test NOEC (7 d): 184 µg/L read-across based on dissolved (meas. (not grouping of semi-static specified)) based on: length substances (category approach) equivalent or similar to fish early life stage toxicity test EPA 560/6-82-002 Test material (IUPAC name): zinc (See endpoint summary for justification of read- across) Pimephales promelas NOEC (32 d): 129 µg/L 2 (reliable with Norberg-King T. dissolved (meas. (not restrictions) (1989) freshwater specified)) based on: mortality supporting study Eight tests with zinc (as ZnSO4.7H2O) as test compound: one full Early Life NOEC (7 d): 128 µg/L read-across based on Stage Toxicity Test (according to dissolved (meas. (not grouping of OECD 210), and seven 5-7 days specified)) based on: length substances (category “subchronic” larval tests, to validate approach) the latter short-term alternative for the NOEC (7 d): 117 µg/L full ELS-test. dissolved (meas. (not Test material specified)) based on: (IUPAC name): zinc flow-through survival and growth sulphate (See endpoint summary OECD Guideline 210 (Fish, Early-Life NOEC (7 d): 129 µg/L for justification of Stage Toxicity Test) (folowed for one dissolved (meas. (not read-across) of the test results) specified)) based on: length

NOEC (7 d): 277 µg/L dissolved (meas. (not specified)) based on: survival and growth

NOEC (7 d): 291 µg/L dissolved (meas. (not specified)) based on: survival and growth

NOEC (7 d): 291 µg/L dissolved (meas. (not specified)) based on:

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 160 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

survival and growth

NOEC (5 d): 128 µg/L dissolved (meas. (not specified)) based on: length

NOEC (5 d): 117 µg/L dissolved (meas. (not specified)) based on: mortality Pimephales promelas LOEC (6 d): 240 µg/L 2 (reliable with Dawson, D.A., dissolved (meas. (not restrictions) Stebler, E.F., Burks freshwater specified)) based on: S.L. and Bantle J.A. morphology supporting study (1988) early-life stage: reproduction, (sub)lethal effects Test material (IUPAC name): zinc static sulphate lab-designed dose-response test Clupea harengus NOEC (27 d): 25 µg/L 2 (reliable with Somasundaram B, dissolved (zinc) (estimated) restrictions) King PE & saltwater based on: development Shackley SE (1984) key study embryo and sac-fry stage: (sub)lethal LOEC (27 d): 50 µg/L effects dissolved (nominal) based read-across based on on: larval development grouping of semi-static substances (category approach) 27-d early life stage of fish development test Test material (EC name): zinc sulfate (See endpoint summary for justification of read- across) Clupea harengus NOEC (17 d): 500 µg/L 2 (reliable with Somasundaram B, dissolved (zinc) (nominal) restrictions) King PE & saltwater based on: Development Shackley S (1985) (ultrastructure of posterior key study embryo and sac-fry stage: (sub)lethal gut and pronephric duct effects read-across based on grouping of semi-static substances (category approach) 17-d postfertilization development test Test material (EC name): zinc sulfate (See endpoint summary for justification of read- across) see details on test organism IC25 (7 d): ca. 1.3 mg/L 1 (reliable without Dwyer et al. (2005) dissolved (NH4) (meas. restriction) freshwater (geom. mean)) based on: growth rate supporting study see details on test organism IC25 (7 d): ca. 7.2 mg/L experimental result semi-static dissolved (NH4) (meas.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 161 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

U.S. Environmental Protection Agency (geom. mean)) based on: Test material (CAS (USEPA) standard effluent test growth rate name): Ammonium procedures (USEPA 1994) phosphate IC25 (7 d): ca. 13.4 mg/L dissolved (NH4) (meas. (geom. mean)) based on: growth rate

IC25 (7 d): ca. 11 mg/L dissolved (NH4) (meas. (geom. mean)) based on: growth rate

IC 25 (7 d): ca. 8.8 mg/L dissolved (NH4) (meas. (geom. mean)) based on: growth rate

IC25 (7 d): ca. 8.9 mg/L dissolved (NH4) (meas. (geom. mean)) based on: growth rate

IC25 (7 d): ca. 15.8 mg/L dissolved (NH4) (meas. (geom. mean)) based on: growth rate

IC25 (7 d): ca. 24.1 mg/L dissolved (NH4) (meas. (geom. mean)) based on: growth rate Pimephales promelas NOEC (28 d): 11.8 mg/L 2 (reliable with Mayes M.A. et al (meas. (not specified)) restrictions) (1986) freshwater LOEC (28 d): 18.7 mg/L supporting study embryo and sac-fry stage: (sub)lethal (meas. (not specified)) effects experimental result flow-through Test material (EC name): Ammonium - American Society for Testing and chloride Materials 1980 Standard practice for conducting acute toxicity tests with fishes, macro-invertebrates and amphibians. Standard E729-80. Philadelphia, PA pp. 1-25 and - Benoit et al., 1983 A fathead minnow Pimephales prom Menidia beryllina NOEC (28 d): 8 mg/L (meas.4 (not assignable) Miller,D.C. et al. (not specified)) (1990) saltwater supporting study LOEC (28 d): 16 mg/L embryo and sac-fry stage: (sub)lethal (meas. (not specified)) experimental result effects Test material (EC not mentioned name): Ammonium chloride Lepomis cyanellus NOEC (44 d): 23.9 mg/L 4 (not assignable) McCormick, J.H.,

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 162 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

(meas. (not specified)) (1984) embryo and sac-fry stage: (sub)lethal supporting study effects LOEC (44 d): 53.2 mg/L (meas. (not specified)) experimental result not mentioned Test material (EC name): Ammonium chloride Salmo clarki LC50 (36 d): 123 mg/L test 4 (not assignable) Thurston, R.V. and mat. (meas. (not specified)) Russo, R. C.: embryo and sac-fry stage: (sub)lethal based on: mortality supporting study (1978) effects experimental result no data Test material (EC name): ammonium chloride

Discussion

Extensive high quality chronic data were available on 7 (freshwater) and 1 (marine) fish species. These data were all screened for relevancy to the environment under study.

The freshwater species are part of 4 families: Cyprinodontidae (Jordanella), Cyprinidae (Phoxinus, Pimephales), Salmonidae (Oncorrhynchus, Salvelinus, Salmo trutta) andCottidae (Cottus). The sensitivity of these species is equally distributed over the species sensitivity distribution. The fish species NOECs are combined with the other freshwater chronic data in the SSD to give the HC5 from which the PNEC is derived.

The saltwater species comes from the family Clupeidae (Clupea harengus). This species is not among the most sensitive ones in thespecies sensitivity distribution. The fish species NOEC is combined with the other marine chronic data in the SSD to give the HC5 from which the PNEC saltwater is derived.

The following information is taken into account for long-term fish toxicity for the derivation of PNEC: freshwater: Data on 7 species available. Species NOECs range between 0.044 and 0.530 mg Zn/l (dissolved concentrations) Marine: Data on 1 species available. NOEC = 0.025 mg Zn/l (dissolved concentrations) Species NOECs have been put into the respective species sensitivity distributions.

7.1.1.2. Aquatic invertebrates

7.1.1.2.1. Short-term toxicity to aquatic invertebrates

The results are summarised in the following table:

Table 63. Overview of short-term effects on aquatic invertebrates

Method Results Remarks Reference Daphnia magna EC50 (48 h): 860 µg/L 1 (reliable without Magliette R.J. dissolved (meas. (arithm. restriction) (1995) freshwater mean)) based on: effect parameter not explicitly key study static

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 163 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

mentioned US EPA/600/4-85/013: methods for read-across based on measuring the acute toxicity of LC50 (48 h): 1220 µg/L grouping of effluents to freshwaterand marine dissolved (meas. (arithm. substances (category organisms mean)) based on: mortality approach)

Test material (IUPAC name): zinc bromide (See endpoint summary for justification of read-across) Ceriodaphnia dubia EC50 (48 h): 360 µg/L 1 (reliable without Magliette R.J. dissolved (meas. (arithm. restriction) (1995) freshwater mean)) based on: effect parameter not explicitly key study static mentioned read-across based on US EPA/600/4-85/013: methods for LC50 (48 h): 500 µg/L grouping of measuring the acute toxicity of dissolved (meas. (arithm. substances (category effluents to freshwaterand marine mean)) based on: mortality approach) organisms Test material (IUPAC name): zinc bromide (See endpoint summary for justification of read-across) Daphnia magna EC50 (48 h): 1833 µg/L 1 (reliable without Muysen B.T.A., dissolved (meas. (arithm. restriction) Bossuyt B.T.A. and freshwater mean)) based on: mobility Jansen, C.R. (2005) key study static EC50 (48 h): 2909 µg/L dissolved (meas. (arithm. read-across based on OECD Guideline 202 (Daphnia sp. mean)) based on: mobility grouping of Acute Immobilisation Test) substances (category approach)

Test material (IUPAC name): zinc (See endpoint summary for justification of read- across) Ceriodaphnia dubia EC50 (48 h): 416 µg/L 1 (reliable without Muysen B.T.A., dissolved (meas. (arithm. restriction) Bossuyt B.T.A. and freshwater mean)) based on: mobility Jansen, C.R. (2005) key study static read-across based on OECD Guideline 202 (Daphnia sp. grouping of Acute Immobilisation Test) substances (category approach)

Test material (IUPAC name): zinc (See endpoint summary for justification of read- across)

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 164 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 other aquatic crustacea: Ceriodaphnia EC50 (48 h): 937 µg/L 1 (reliable without Muysen B.T.A., reticulata dissolved (meas. (arithm. restriction) Bossuyt B.T.A. and mean)) based on: mobility Jansen, C.R. (2005) freshwater key study static read-across based on grouping of OECD Guideline 202 (Daphnia sp. substances (category Acute Immobilisation Test) approach)

Test material (IUPAC name): zinc (See endpoint summary for justification of read- across) Ceriodaphnia dubia EC50 (48 h): 413 µg/L 1 (reliable without Hyne R.V., Pablo F, dissolved (meas. (arithm. restriction) Moreno J; , freshwater mean)) based on: mobility Markisch S.J. key study (2005) static EC50 (48 h): 200 µg/L dissolved (meas. (arithm. read-across based on US EPA EPA 821-R-02-012. Methods mean)) based on: mobility grouping of for measuring the acute toxicity of substances (category effluents and receiving waters to EC50 (48 h): 155 µg/L approach) freshwater and marine organisms, 5th dissolved (meas. (arithm. Ed. (2002) mean)) based on: mobility Test material (IUPAC name): zinc (See endpoint summary for justification of read- across) Daphnia magna EC50 (48 h): 2140 µg/L 1 (reliable without De Schamphelaere dissolved (meas. (arithm. restriction) K.A.C., Lofts, S. freshwater mean)) based on: mobility and Janssen (2005) key study static read-across based on OECD Guideline 202 (Daphnia sp. grouping of Acute Immobilisation Test) substances (category approach)

Test material (IUPAC name): zinc (See endpoint summary for justification of read- across) Daphnia magna LC50 (48 h): 800 µg/L 2 (reliable with Attar E.N., and dissolved (meas. (arithm. restrictions) Maly E.J. (1982) freshwater mean)) based on: mortality key study flow-through read-across based on lab designed flow-through test system grouping of for testing acute toxicity to daphnia substances (category magna. approach)

Test material (IUPAC name): zinc

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 165 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

chloride (See endpoint summary for justification of read-across) Daphnia magna LC50 (48 h): 100 µg/L 2 (reliable with Biesinger K.E., & dissolved (meas. (not restrictions) Christensen G.M. freshwater specified)) based on: (1972) mortality key study static LC50 (48 h): 280 µg/L read-across based on dose response test with Daphnia magna dissolved (meas. (not grouping of in beakers, similar to the later specified)) based on: substances (category developed standard D. magna chronic mortality approach) test. linear series of concentrations were used for delineating reproductive Test material impairment levels. (IUPAC name): zinc chloride (See endpoint summary for justification of read-across) Ceriodaphnia dubia EC50 (48 h): 670 µg/L 1 (reliable without Muyssen B.T.A. dissolved (meas. (initial)) restriction) and Jansseen C.R. freshwater based on: mobility (2002) key study static read-across based on equivalent or similar to OECD grouping of Guideline 202 (Daphnia sp. Acute substances (category Immobilisation Test) approach)

Test material (IUPAC name): zinc chloride (See endpoint summary for justification of read-across) Daphnia magna LC50 (48 h): 330 µg/L 2 (reliable with Chapman G, Ota S dissolved (meas. (arithm. restrictions) and Recht F. (1980) freshwater mean)) based on: mobility key study static LC50 (48 h): 530 µg/L dissolved (meas. (arithm. read-across based on equivalent or similar to OECD mean)) based on: mobility grouping of Guideline 202 (Daphnia sp. Acute substances (category Immobilisation Test) LC50 (48 h): 660 µg/L approach) dissolved (meas. (arithm. mean)) based on: mobility Test material (IUPAC name): zinc chloride (See endpoint summary for justification of read-across) Ceriodaphnia dubia LC50 (48 h): 169 µg/L 2 (reliable with Carlson A.R., dissolved (meas. (not restrictions) Nelson, H. and freshwater specified)) based on: Hammermeister D. mortality key study (1986) static read-across based on lab-designed dose response test along grouping of

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 166 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 the lines of later standard cladoceran substances (category test protocols approach)

Test material (IUPAC name): zinc chloride (See endpoint summary for justification of read-across) Daphnia magna LC50 (48 h): 259 µg/L 1 (reliable without Barata C, Baird D, dissolved (meas. (not restriction) Markisch S. (1998) freshwater specified)) based on: mobility key study semi-static LC50 (48 h): 1060 µg/L read-across based on equivalent or similar to OECD dissolved (meas. (not grouping of Guideline 202 (Daphnia sp. Acute specified)) based on: substances (category Immobilisation Test) mobility approach)

LC50 (48 h): 962 µg/L Test material dissolved (meas. (not (IUPAC name): zinc specified)) based on: sulphate (See mobility endpoint summary for justification of LC50 (48 h): 131 µg/L read-across) dissolved (meas. (not specified)) based on: mobility

LC50 (48 h): 457 µg/L dissolved (meas. (not specified)) based on: mobility

LC50 (48 h): 601 µg/L dissolved (meas. (not specified)) based on: mobility Ceriodaphnia dubia LC50 (48 h): > 530 µg/L 2 (reliable with Schubauer-Berrigan dissolved (meas. (not restrictions) M.k., Dierkes J.R., freshwater specified)) based on: Monson P.D. and mortality key study Ankley, G.T. lab-designed dose-response test at (1993b) varying pH and at relatively high LC50 (48 h): 360 µg/L read-across based on hardness conditions. dissolved (meas. (not grouping of specified)) based on: substances (category mortality approach)

LC50 (48 h): 95 µg/L Test material dissolved (meas. (not (IUPAC name): zinc specified)) based on: sulphate (See mortality endpoint summary for justification of read-across) Daphnia magna LC50 (48 h): 280 µg/L 2 (reliable with Cairns J., Buikema, dissolved (nominal) based restrictions) A.L., Heath A.G. freshwater on: mortality &nd Parker, B.C. key study (1978) static

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 167 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 lab-designed dose -response test along read-across based on the lines of later developed standard grouping of test protocols substances (category approach)

Test material (IUPAC name): zinc sulphate (See endpoint summary for justification of read-across) Daphnia pulex LC50 (48 h): 500 µg/L 2 (reliable with Cairns J., Buikema, dissolved (nominal) based restrictions) A.L., Heath A.G. freshwater on: mortality &nd Parker, B.C. key study (1978) static read-across based on lab-designed dose -response test along grouping of the lines of later developed standard substances (category test protocols approach)

Test material (IUPAC name): zinc sulphate (See endpoint summary for justification of read-across) Daphnia magna EC50 (48 h): 1.4 mg/L 1 (reliable without Blinova I, Ivask A dissolved (meas. (not restriction) Heinlaan M, freshwater specified)) based on: Mortimer M and mobility key study Kahru A. (2010) static EC50 (48 h): 1.8 mg/L read-across based on OECD Guideline 202 (Daphnia sp. dissolved (meas. (not grouping of Acute Immobilisation Test) specified)) based on: substances (category mobility approach)

EC50 (48 h): 2 mg/L Test material (CAS dissolved (meas. (not name): zinc sulphate specified)) based on: (See endpoint mobility summary for justification of read- EC50 (48 h): 1.6 mg/L across) dissolved (meas. (not specified)) based on: mobility

EC50 (48 h): 2.5 mg/L dissolved (meas. (not specified)) based on: mobility

EC50 (48 h): 1.4 mg/L dissolved (meas. (not specified)) based on: mobility

EC50 (48 h): 1.7 mg/L dissolved (meas. (not

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 168 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

specified)) based on: mobility other aquatic crustacea: EC50 (24 h): 0.22 mg/L 1 (reliable without Blinova I, Ivask A Thamnocephalus platyurus dissolved (meas. (not restriction) Heinlaan M, specified)) based on: Mortimer M and freshwater mortality key study Kahru A. (2010) static EC50 (24 h): 0.92 mg/L read-across based on dissolved (meas. (not grouping of test done with "Thamnotoxkit F" specified)) based on: substances (category mortality approach)

EC50 (24 h): 1.6 mg/L Test material (CAS dissolved (meas. (not name): zinc sulphate specified)) based on: (See endpoint mortality summary for justification of read- EC50 (24 h): 0.61 mg/L across) dissolved (meas. (not specified)) based on: mortality

EC50 (24 h): 0.75 mg/L dissolved (meas. (not specified)) based on: mortality

EC50 (24 h): 1.1 mg/L dissolved (meas. (not specified)) based on: mortality

EC50 (24 h): 1.7 mg/L dissolved (meas. (not specified)) based on: mortality other aquatic crustacea: Tetrahymena EC50 (24 h): 7.1 mg/L 1 (reliable without Blinova I, Ivask A thermophila dissolved (meas. (not restriction) Heinlaan M, specified)) based on: growth Mortimer M and freshwater inhibition key study Kahru A. (2010) static EC50 (24 h): 21.1 mg/L read-across based on dissolved (meas. (not grouping of test according to standardised specified)) based on: growth substances (category "Protoxkit F" inhibition approach)

EC50 (24 h): 18.6 mg/L Test material (CAS dissolved (meas. (not name): zinc sulphate specified)) based on: growth (See endpoint inhibition summary for justification of read- EC50 (24 h): > 22 mg/L across) dissolved (meas. (not specified)) based on: growth inhibition Daphnia magna LC50 (48 h): 68 µg/L 2 (reliable with Mount DI and dissolved (nominal) restrictions) Norberg TJ (1984) freshwater supporting study

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 169 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 static read-across based on grouping of lab-designed dose-response test substances (category approach)

Test material (IUPAC name): zinc (See endpoint summary for justification of read- across) Daphnia pulex LC50 (48 h): 107 µg/L 2 (reliable with Mount DI and dissolved (nominal) restrictions) Norberg TJ (1984) freshwater supporting study static read-across based on lab-designed dose-response test grouping of substances (category approach)

Test material (IUPAC name): zinc (See endpoint summary for justification of read- across) Ceriodaphnia dubia LC50 (48 h): 76 µg/L 2 (reliable with Mount DI and dissolved (nominal) restrictions) Norberg TJ (1984) freshwater supporting study static read-across based on lab-designed dose-response test grouping of substances (category approach)

Test material (IUPAC name): zinc (See endpoint summary for justification of read- across) Daphnia pulex EC50 (48 h): 105 µg/L 2 (reliable with Clifford M and dissolved (meas. (arithm. restrictions) McGeer J.C. (2009) freshwater mean)) based on: mobility supporting study static EC50 (48 h): 190 µg/L dissolved (meas. (arithm. read-across based on Environment Canada EPS/RM/11 test mean)) based on: mobility grouping of guideline for "Biological test method: substances (category acute lethality test using Daphnia spp. EC50 (48 h): 399 µg/L approach) dissolved (meas. (arithm. mean)) based on: mobility Test material (IUPAC name): zinc EC50 (48 h): 321 µg/L Sulphate (See dissolved (meas. (arithm. endpoint summary mean)) based on: mobility for justification of read-across)

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 170 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

EC50 (48 h): 556 µg/L dissolved (meas. (arithm. mean)) based on: mobility

EC50 (48 h): 1014 µg/L dissolved (meas. (arithm. mean)) based on: mobility

EC50 (48 h): 268 µg/L dissolved (meas. (arithm. mean)) based on: mobility

EC50 (48 h): 399 µg/L dissolved (meas. (arithm. mean)) based on: mobility

EC50 (48 h): 432 µg/L dissolved (meas. (arithm. mean)) based on: mobility

EC50 (48 h): 353 µg/L dissolved (meas. (arithm. mean)) based on: mobility

EC50 (48 h): 615 µg/L dissolved (meas. (arithm. mean)) based on: mobility

EC50 (48 h): 425 µg/L dissolved (meas. (arithm. mean)) based on: mobility

EC50 (48 h): 765 µg/L dissolved (meas. (arithm. mean)) based on: mobility

EC50 (48 h): 687 µg/L dissolved (meas. (arithm. mean)) based on: mobility

EC50 (48 h): 523 µg/L dissolved (meas. (arithm. mean)) based on: mobility

EC50 (48 h): 399 µg/L dissolved (meas. (arithm. mean)) based on: mobility

EC50 (48 h): 706 µg/L dissolved (meas. (arithm. mean)) based on: mobility

EC50 (48 h): 399 µg/L dissolved (meas. (arithm. mean)) based on: mobility

EC50 (48 h): 477 µg/L dissolved (meas. (arithm. mean)) based on: mobility

EC50 (48 h): 392 µg/L

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 171 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

dissolved (meas. (arithm. mean)) based on: mobility Ceriodaphnia dubia LC50 (48 h): 174 µg/L 2 (reliable with Cooper N.L., dissolved (meas. (initial)) restrictions) Bidwell J.R., freshwater based on: mobility Kumar, A. (2009) supporting study static read-across based on US EPA -821-R-02-012: Methods for grouping of measuring the acute toxicity of substances (category effluents and receiving waters to approach) freshwater and marine organisms (2002) Test material (IUPAC name): zinc sulphate (See endpoint summary for justification of read-across) other aquatic crustacea: Daphnia LC50 (48 h): 340 µg/L 2 (reliable with Cooper N.L., carinata dissolved (meas. (initial)) restrictions) Bidwell J.R., based on: mobility Kumar, A. (2009) freshwater supporting study static read-across based on grouping of US EPA -821-R-02-012: Methods for substances (category measuring the acute toxicity of approach) effluents and receiving waters to freshwater and marine organisms Test material (2002) (IUPAC name): zinc sulphate (See endpoint summary for justification of read-across) physa gyrina LC50 : ca. 1.94 mg/L 2 (reliable with Johnson et al. dissolved (NH3) (meas. restrictions) (1984) freshwater (geom. mean)) based on: mortality supporting study flow-through experimental result methode in Ambient water criteria for ammonia not reported, see primary Test material (CAS literature name): ammonium chloride Daphnia magna LC50 : ca. 1.5 mg/L 2 (reliable with Johnson et al. dissolved (NH3) (meas. restrictions) (1984) freshwater (geom. mean)) based on: mortality supporting study static experimental result methode in Ambient water criteria for ammonia not reported, see primary Test material (CAS literature name): ammonium chloride Daphnia pulicaria LC50 : ca. 1.16 mg/L 2 (reliable with Johnson et al. dissolved (NH3) (nominal) restrictions) (1984) freshwater based on: mortality supporting study flow-through

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 172 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 methode in Ambient water criteria for experimental result ammonia not reported, see primary literature Test material (CAS name): ammonium chloride Daphnia magna EC50 (48 h): 101 mg/L test 2 (reliable with Gersich, F.M. and mat. (nominal) based on: restrictions) Hopkins, D. L., freshwater mortality (1986) supporting study static experimental result ASTM E729-80 Test material (EC name): Ammonium chloride other aquatic crustacea: Homarus EC50 (8 d): 152 mg/L test 4 (not assignable) Delistraty,D.A. et americanus mat. based on: mortality al. (1977) supporting study saltwater experimental result static Test material (EC no data name): Ammonium chloride other aquatic mollusc: Helicoma EC50 (96 h): 221 mg/L test 4 (not assignable) Arthur ,J.W. et al., trivolvus mat. based on: mortality (1987) supporting study semi-static experimental result E07-04:APHA(1980) "Standard methods for the examination of water Test material (EC and waste water" name): Ammonium chloride Daphnia magna EC50 (96 h): 139 mg/L test 4 (not assignable) Dowden,B.F., mat. based on: mortality Bennett,H.J. (1965) static supporting study not specified experimental result

Test material (EC name): Ammonium chloride Mulinia laterails (Bivalve) EC50 (10 d): 42 mg/L 4 (not assignable) Huber, M.C. et al (1997) semi-static EC50 (10 d): 82.9 mg/L supporting study not indicated experimental result

Test material (EC name): Ammonium chloride

Discussion

Many good quality and relevant data for 5 standard species. Tests were done according to standard protocol or

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 173 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 equivalent.

Data are grouped per species according to

-pH: low (6 -<7) - neutral/high (7 -8.5)

-and hardness: low/medium (<100mg CaCO3/l) and medium/high (>100 mg CaCO3/l).

Ceriodaphnia dubia is for all test conditions the most sensitive species.

The following information is taken into account for short-term toxicity to aquatic invertebrates for the derivation of PNEC:

Key data (lowest EC50 values) are: -for Ceriodapnia dubia: 0.413 mg Zn/l (single value) at low pH and low hardness -for Ceriodapnia dubia: >0.53 mg Zn/l (single value) at low pH and high hardness -for Ceriodapnia dubia: 0.147 mg Zn/l (geomean value) at neutral/high pH and low hardness -for Ceriodapnia dubia: 0.228 mg Zn/l (geomean value) at neutral/high pH and high hardness

7.1.1.2.2. Long-term toxicity to aquatic invertebrates

The results are summarised in the following table:

Table 64. Overview of long-term effects on aquatic invertebrates

Method Results Remarks Reference Daphnia magna NOEC (50 d): 31 µg/L 2 (reliable with Palauskis J.D. and dissolved (nominal) based restrictions) Winner R.W. freshwater on: reproduction (1988) key study semi-static NOEC (50 d): 33 µg/L dissolved (nominal) based read-across based on chronic tests were performed for an on: reproduction grouping of extended time period (50 days). test substances (category solutions contained varying levels of NOEC (50 d): 84 µg/L approach) hardness and DOC. organisms were dissolved (nominal) based adapted to different hardnesses. on: reproduction Test material reproduction was evaluated as in the (IUPAC name): zinc standard Daphnia magna test. NOEC (50 d): 83 µg/L sulphate dissolved (nominal) based heptahydrate (See on: reproduction endpoint summary for justification of NOEC (50 d): 159 µg/L read-across) dissolved (nominal) based on: reproduction

NOEC (50 d): 208 µg/L dissolved (nominal) based on: reproduction Ceriodaphnia dubia NOEC (1 wk): 25 µg/L 2 (reliable with Belanger SE and dissolved (nominal) based restrictions) Cherry DS (1990) freshwater on: reproduction key study semi-static NOEC (1 wk): 25 µg/L dissolved (nominal) based read-across based on USEPA 1002.0 on: reproduction grouping of substances (category

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 174 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

NOEC (1 wk): 25 µg/L approach) dissolved (nominal) based on: reproduction Test material (IUPAC name): zinc NOEC (1 wk): 40 µg/L (See endpoint dissolved (nominal) based summary for on: reproduction justification of read- across) NOEC (1 wk): 50 µg/L dissolved (nominal) based on: reproduction

NOEC (1 wk): 45 µg/L dissolved (nominal) based on: reproduction

NOEC (1 wk): 29 µg/L dissolved (nominal) based on: reproduction

NOEC (1 wk): 50 µg/L dissolved (nominal) based on: reproduction

NOEC (1 wk): 33 µg/L dissolved (nominal) based on: reproduction Daphnia magna NOEC (3 wk): 100 µg/L 2 (reliable with Münzinger A. and dissolved (nominal) based restrictions) Monicelli F. (1991) freshwater on: reproduction and survival key study semi-static NOEC (3 wk): 100 µg/L read-across based on Dose-response test on D. magna over dissolved (nominal) based grouping of same exposure period as the standard on: reproduction and substances (category D. magna test (21d). survival approach)

NOEC (3 wk): 100 µg/L Test material dissolved (nominal) based (IUPAC name): zinc on: reproduction and (See endpoint survival summary for justification of read- across) Dreissena polymorpha NOEC (10 wk): 400 µg/L 2 (reliable with Kraak M.H.S. et al dissolved (nominal) based restrictions) (1994) freshwater on: survival key study semi-static read-across based on dose response experiments over 10 grouping of week exposure period. Measured zinc substances (category concentrations (3-38-101-382-1,266- approach) 2,739 µg/l) within 10% of nominal zinc concentrations (0-40-100-400- Test material 1,400-3,000 µg/l) in exposure groups. (IUPAC name): zinc chloride (See endpoint summary for justification of read-across)

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 175 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 other aquatic mollusc: Potamopyrgus NOEC (16 wk): 75 µg/L 2 (reliable with Dorgelo J. , jenkisi dissolved (nominal) based restrictions) Meester, H. and on: growth Van Velzen, C. freshwater key study (1995) semi-static read-across based on grouping of dose-response experiments with 5 substances (category nominal zinc concentrations added approach) (including control). Shell length increment of juveniles evaluated every Test material week. (IUPAC name): zinc chloride (See endpoint summary for justification of read-across) Ceriodaphnia dubia NOEC (4 d): 50 µg/L 1 (reliable without Masters J.A. , dissolved (nominal) based restriction) Lewis, M.A., freshwater on: reproduction Davidson, D.H. and key study Bruce, R.D. (1991) semi-static NOEC (4 d): 14 µg/L dissolved (nominal) based read-across based on USEPA chronic survival and on: reproduction grouping of reproduction of Ceriodaphnia substances (category NOEC (7 d): 50 µg/L approach) dissolved (nominal) based on: reproduction Test material (IUPAC name): zinc NOEC (7 d): 100 µg/L chloride (See dissolved (nominal) based endpoint summary on: reproduction for justification of read-across) Daphnia magna NOEC (21 d): 97 µg/L 2 (reliable with Chapman G., Ota, S dissolved (meas. (not restrictions) and Recht, F. freshwater specified)) based on: (1980) reproduction key study semi-static NOEC (21 d): 43 µg/L read-across based on equivalent or similar to EPA OPPTS dissolved (meas. (not grouping of 850.1300 (Daphnid Chronic Toxicity specified)) based on: substances (category Test) reproduction approach)

NOEC (21 d): 42 µg/L Test material dissolved (meas. (not (IUPAC name): zinc specified)) based on: chloride (See reproduction endpoint summary for justification of read-across) Daphnia magna NOEC (3 wk): 35 µg/L 2 (reliable with Biesinger K.E. and dissolved (nominal) based restrictions) Christensen G.M. freshwater on: reproduction (1972) key study semi-static read-across based on dose response test with Daphnia magna grouping of in beakers, similar to the later substances (category developed standard D. magna chronic approach) test. linear series of concentrations were used for delineating reproductive Test material impairment levels. (IUPAC name): zinc

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 176 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

chloride (See endpoint summary for justification of read-across) Daphnia magna NOEC (3 wk): 74 µg/L 2 (reliable with Biesinger et al dissolved (meas. (not restrictions) (1986) freshwater specified)) based on: reproduction key study semi-static read-across based on Study was done according to a grouping of renewed static testprocedure. For each substances (category test 4 replicates beakers were used approach) each containing 5 daphnids. The test was a dose-response study. Test material (IUPAC name): zinc chloride (See endpoint summary for justification of read-across) Daphnia magna NOEC (3 wk): 37 µg/L 2 (reliable with Enserink E.L., dissolved (estimated) based restrictions) Maas-Diepeveen freshwater on: growth J.L. and Van key study Leeuwen C.J. semi-static and intermittent flow- NOEC (3 wk): 310 µg/L (1991) through experimental setup dissolved (estimated) based read-across based on on: reproduction grouping of Dose-response test on D. magna over substances (category same exposure period as the standard NOEC (3 wk): 310 µg/L approach) D. magna test (21d). dissolved (estimated) based on: survival Test material (IUPAC name): zinc LOEC (3 wk): 120 µg/L chloride (See dissolved (nominal) based endpoint summary on: growth for justification of read-across) LOEC (3 wk): 1000 µg/L dissolved (nominal) based on: reproduction

LOEC (3 wk): 1000 µg/L dissolved (nominal) based on: survival

EC10 (3 wk): 420 µg/L dissolved (nominal) based on: reproduction

EC10 (3 wk): 420 µg/L dissolved (nominal) based on: survival and emergence Daphnia magna NOEC (3 wk): 82 µg/L 1 (reliable without De Schamphelaere dissolved (meas. (arithm. restriction) K.A.C., Heijerick freshwater mean)) based on: D.G. and Janssen, reproduction and survival key study CR. (2003b) semi-static NOEC (3 wk): 50 µg/L read-across based on OECD Guideline 211 (Daphnia magna dissolved (meas. (arithm. grouping of Reproduction Test) mean)) based on: substances (category

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 177 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

reproduction and survival approach)

NOEC (3 wk): 54 µg/L Test material dissolved (meas. (arithm. (IUPAC name): zinc mean)) based on: chloride (See reproduction and survival endpoint summary for justification of NOEC (3 wk): 92 µg/L read-across) dissolved (meas. (arithm. mean)) based on: reproduction and survival

NOEC (3 wk): 48 µg/L dissolved (meas. (arithm. mean)) based on: reproduction and survival

NOEC (3 wk): 152 µg/L dissolved (meas. (arithm. mean)) based on: reproduction and survival

NOEC (3 wk): 155 µg/L dissolved (meas. (arithm. mean)) based on: reproduction and survival

NOEC (3 wk): 156 µg/L dissolved (meas. (arithm. mean)) based on: reproduction and survival

NOEC (3 wk): 143 µg/L dissolved (meas. (arithm. mean)) based on: reproduction and survival

NOEC (3 wk): 136 µg/L dissolved (meas. (arithm. mean)) based on: reproduction and survival

NOEC (3 wk): 143 µg/L dissolved (meas. (arithm. mean)) based on: reproduction and survival Daphnia magna NOEC (3 wk): 39 µg/L 1 (reliable without Heijerick D.G, De dissolved (meas. (arithm. restriction) Schamphelaere freshwater mean)) based on: K.A.C. Van Sprang reproduction key study P.A. and Janssen semi-static C.R. (2005) NOEC (3 wk): 48 µg/L read-across based on OECD Guideline 211 (Daphnia magna dissolved (meas. (arithm. grouping of Reproduction Test) mean)) based on: substances (category reproduction approach)

NOEC (3 wk): 79 µg/L Test material dissolved (meas. (arithm. (IUPAC name): zinc mean)) based on: chloride (See reproduction endpoint summary

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 178 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

NOEC (3 wk): 168 µg/L for justification of dissolved (meas. (arithm. read-across) mean)) based on: reproduction

NOEC (3 wk): 161 µg/L dissolved (meas. (arithm. mean)) based on: reproduction

NOEC (3 wk): 154 µg/L dissolved (meas. (arithm. mean)) based on: reproduction

NOEC (3 wk): 133 µg/L dissolved (meas. (arithm. mean)) based on: reproduction

NOEC (3 wk): 117 µg/L dissolved (meas. (arithm. mean)) based on: reproduction Daphnia magna NOEC (3 wk): 155 µg/L 1 (reliable without De Schamphelaere dissolved (meas. (arithm. restriction) K.A.C., Lofts, S. freshwater mean)) based on: and Janssen C.R. reproduction key study (2005b) semi-static NOEC (3 wk): 95 µg/L read-across based on OECD Guideline 211 (Daphnia magna dissolved (meas. (arithm. grouping of Reproduction Test) mean)) based on: substances (category reproduction approach)

NOEC (3 wk): 244 µg/L Test material dissolved (meas. (arithm. (IUPAC name): zinc mean)) based on: chloride (See reproduction endpoint summary for justification of NOEC (3 wk): 251 µg/L read-across) dissolved (meas. (arithm. mean)) based on: reproduction

NOEC (3 wk): 143 µg/L dissolved (meas. (arithm. mean)) based on: reproduction

NOEC (3 wk): 73 µg/L dissolved (meas. (arithm. mean)) based on: reproduction other aquatic crustacea: Daphnia NOEC (21 d): 91 µg/L 1 (reliable without Muyssen B., longispina dissolved (meas. (not restriction) Bossuyt B. and specified)) based on: Janssen C.R. freshwater reproduction key study (2003a) semi-static NOEC (21 d): 209 µg/L read-across based on

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 179 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

OECD Guideline 211 (Daphnia magna dissolved (meas. (not grouping of Reproduction Test) specified)) based on: substances (category reproduction approach)

NOEC (21 d): 42 µg/L Test material dissolved (meas. (not (IUPAC name): zinc specified)) based on: chloride (See reproduction endpoint summary for justification of NOEC (21 d): 48 µg/L read-across) dissolved (meas. (not specified)) based on: reproduction other aquatic arthropod: Chironomus NOEC (8 wk): 166 µg/L 2 (reliable with Sibley P. K., tentans dissolved (meas. (arithm. restrictions) Ankley, G.T., mean)) based on: survival, Cotter A.M. and freshwater growth, emergence and key study Leonard E.N. reproduction (1996) static read-across based on grouping of lab-designed dose response test . substances (category approach)

Test material (IUPAC name): zinc chloride (See endpoint summary for justification of read-across) poriferans: Ephydatia fluviatilis NOEC (7 d): 43 µg/L 2 (reliable with Richelle-Maurer E. dissolved (nominal) based restrictions) and Van de Vyver freshwater on: developments effects in G. (2001) sponges (measured by cell key study static aggregation, settlement and formation of functional read-across based on lab-designed dose-response test sponges) grouping of substances (category approach)

Test material (IUPAC name): zinc chloride (See endpoint summary for justification of read-across) poriferans: Ephydatia muelleri NOEC (7 d): 43 µg/L 2 (reliable with Richelle-Maurer E. dissolved (nominal) based restrictions) and Van de Vyver freshwater on: developments effects in G. (2001) sponges (measured by cell key study static aggregation, settlement and formation of functional read-across based on lab-designed dose-response test sponges) grouping of substances (category approach)

Test material (IUPAC name): zinc chloride (See endpoint summary

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 180 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

for justification of read-across) poriferans: Songila lacustris NOEC (7 d): 65 µg/L 2 (reliable with Richelle-Maurer E. dissolved (nominal) based restrictions) and Van de Vyver freshwater on: developments effects in G. (2001) sponges (measured by cell key study static aggregation, settlement and formation of functional read-across based on lab-designed dose-response test sponges) grouping of substances (category approach)

Test material (IUPAC name): zinc chloride (See endpoint summary for justification of read-across) poriferans: Eunapius fragilis NOEC (7 d): 43 µg/L 2 (reliable with Richelle-Maurer E. dissolved (nominal) based restrictions) and Van de Vyver freshwater on: developments effects in G. (2001) sponges (measured by cell key study static aggregation, settlement and formation of functional read-across based on lab-designed dose-response test sponges) grouping of substances (category approach)

Test material (IUPAC name): zinc chloride (See endpoint summary for justification of read-across) rotifera: Anuraeopsis fissa NOEC (25 d): 50 µg/L 2 (reliable with Azuara-Gracia R, dissolved (nominal) based restrictions) Sarma S.S., and freshwater on: population growth rate Nandini S. (2006) key study static read-across based on lab-designed dose response test grouping of substances (category approach)

Test material (IUPAC name): zinc chloride (See endpoint summary for justification of read-across) rotifera: Brachionus rubens NOEC (25 d): 50 µg/L 2 (reliable with Azuara-Gracia R, dissolved (nominal) based restrictions) Sarma S.S., and freshwater on: population growth rate Nandini S. (2006) key study static read-across based on lab-designed dose response test grouping of substances (category approach)

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 181 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Test material (IUPAC name): zinc chloride (See endpoint summary for justification of read-across) other aquatic mollusc: Haliotis NOEC (10 d): 10 µg/L 1 (reliable without Conroy PT, Hunt rufescens, Red Abalone, Haliotidae dissolved (zinc) (nominal) restriction) JW & Anderson BS based on: development (1996) saltwater key study flow-through read-across based on grouping of Anderson BS, Hunt JW, Turpen SL, substances (category Coulon AR, Martin M, McKeown DL approach) & Palmer FH (1990) Procedures manual for conducting toxicity tests Test material developed by the Marine Bioassay (IUPAC name): zinc Project. 90-10WQ. State Water sulfate (See endpoint Resources Control Board, Sacramento, summary for CA, USA. justification of read- across) other aquatic mollusc: Haliotis NOEC (9 d): 19 µg/L 2 (reliable with Hunt JW & rufescens, Red Abalone, Haliotidae dissolved (zinc) (meas. (not restrictions) Anderson BS specified)) based on: (1989) saltwater development key study flow-through read-across based on grouping of Anderson BS, Hunt JW, Martin M, substances (category Turpen SL, Palmer FH, 1988. Marine approach) Bioassay Project. 3rd Report. Protocol Development: Reference Toxicant and Test material (EC Initial Complex Effluent Testing. name): zinc sulfate Div.of Water Qual.Rep.No.88-7WQ, (See endpoint State Water Resources Control Board, summary for State of California, Sacramento, CA : justification of read- 154 p. across) other aquatic mollusc: Mytilus EC10 (2 d): 90 µg/L 2 (reliable with Pavicic J, Skreblin galloprovincialis, Blue mussel, dissolved (zinc) (nominal) restrictions) M, Kregar I, Mytilidae based on: development Tusekznidaric M & key study Stegnar P (1994) saltwater read-across based on static grouping of substances (category 2-d larval development of marine approach) mollusks, lad designed test for dose- response Test material (EC name): zinc sulfate (See endpoint summary for justification of read- across) Arbacia lixula, Sea Urchin, Arbaciidae NOEC (38 h): 10 µg/L 2 (reliable with Cesar A, Marín- dissolved (zinc) (estimated) restrictions) Guirao L, Vita R & saltwater based on: development Marín A (2002) key study static

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 182 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 equivalent or similar to EPA/600/ R- read-across from 95-136, supporting substance (structural analogue equivalent or similar to Environment or surrogate) Canada EPS 1/RM/27 Test material (EC equivalent or similar to CETESB name): zinc sulfate L5.250 (See endpoint summary for justification of read- across) Paracentrotus lividus, Mediterranean NOEC (28 h): 10 µg/L 2 (reliable with Cesar A, Marín- Sea Urchin, Echinidae dissolved (zinc) (estimated) restrictions) Guirao L, Vita R & based on: development Marín A (2002) saltwater key study static read-across based on grouping of equivalent or similar to EPA/600/ R- substances (category 95-136, approach) equivalent or similar to Environment Test material (EC Canada EPS 1/RM/27 name): zinc sulfate (See endpoint equivalent or similar to CETESB summary for L5.250 justification of read- across) Sphaerechinus granularis, Sea Urchin, NOEC (38 h): 10 µg/L 2 (reliable with Cesar A, Marín- Toxopneustidae dissolved (zinc) (estimated) restrictions) Guirao L, Vita R & based on: development Marín A (2002) saltwater key study static read-across based on grouping of equivalent or similar to EPA/600/ R- substances (category 95-136, approach) equivalent or similar to Environment Test material (EC Canada EPS 1/RM/27 name): zinc sulfate (See endpoint equivalent or similar to CETESB summary for L5.250 justification of read- across) Sterechinus neumayeri, Antarctic Sea EC10 (23 d): 80 µg/L 2 (reliable with King CK & Riddle Urchin, Echinidae dissolved (zinc) (estimated) restrictions) MJ (2001) based on: development saltwater key study static read-across based on grouping of 20-23d development of sea urchin substances (category larvae approach)

Test material (EC name): zinc sulfate (See endpoint summary for justification of read- across)

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 183 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 other aquatic crustacea: Allorchestes LC10 (28 d): 63 µg/L 2 (reliable with Ahsanullah M & compressa Dana, Amphipod, dissolved (zinc) (meas. (not restrictions) Williams AR Dogielinotidae specified)) based on: (1991) mortality key study saltwater read-across based on flow-through grouping of substances (category 28-d amphipod chronic toxicity test approach)

Test material (EC name): zinc sulfate (See endpoint summary for justification of read- across) other aquatic crustacea: Holmesimysis NOEC (24 d): 5.6 µg/L 1 (reliable without Hunt JW, Anderson costata, Mysid shrimp, Mysidae dissolved (zinc) (nominal) restriction) BS, Turpen SL, based on: mortality Englund MA & saltwater key study Piekarski W (1997) NOEC (7 d): 31.8 µg/L semi-static dissolved (estimated) based read-across based on on: mortality grouping of EPA/600/R-95-136 substances (category approach)

Test material (EC name): zinc sulfate (See endpoint summary for justification of read- across) Mysidopsis bahia (new name: NOEC (7 d): 101 µg/L 2 (reliable with Harmon VL & Americamysis bahia) dissolved (zinc) (nominal) restrictions) Langdon CJ (1996) based on: mortality saltwater key study semi-static read-across based on grouping of EPA/600/4-87/028 substances (category approach)

Test material (EC name): zinc sulfate (See endpoint summary for justification of read- across) other aquatic crustacea: Mysidopsis NOEC (7 d): 101 µg/L 2 (reliable with Harmon VL & intii, Pacific Mysid, Mysidae dissolved (zinc) (nominal) restrictions) Langdon CJ (1996) based on: growth (, saltwater mortality) key study semi-static read-across based on grouping of EPA/600/4-87/028 substances (category approach)

Test material (EC name): zinc sulfate

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 184 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

(See endpoint summary for justification of read- across) other aquatic crustacea: Tigriopus NOEC (10 d): 297 µg/L 2 (reliable with Le Dean L & brevicornis, Copepod, Harpacticidae dissolved (zinc) (nominal) restrictions) Devineau J (1987) based on: reproduction (; saltwater larval development) key study static read-across based on grouping of 10-d reproductive toxicity and larval substances (category development of copepods test approach)

Test material (EC name): zinc sulfate (See endpoint summary for justification of read- across) Eirene viridula, Cnidaria, Medusa, NOEC (3 mo): 300 µg/L 2 (reliable with Karbe L (1972) Eirenidae dissolved (zinc) (nominal) restrictions) based on: development saltwater key study semi-static read-across based on grouping of 3-mo hydroid development test substances (category approach)

Test material (IUPAC name): zinc sulfate (See endpoint summary for justification of read- across) other aquatic crustacea: Holmesimysis NOEC (96 h): 100 µg/L 2 (reliable with Anderson BS, Hunt costata (opposum shrimp) dissolved (meas. (not restrictions) JW, Martin M, specified)) based on: Turpen SL & saltwater mortality key study Palmer FH (1988) static NOEC (96 h): 89 µg/L read-across based on dissolved (meas. (not grouping of 96-h mortality test on Holmesimysis specified)) based on: substances (category costata mortality approach)

NOEC (96 h): 66 µg/L Test material (EC dissolved (meas. (not name): zinc sulfate specified)) based on: (See endpoint mortality summary for justification of read- NOEC (96 h): 84 µg/L across) dissolved (estimated) based on: mortality other aquatic crustacea: Holmemysis NOEC (4 d): 68.4 µg/L 2 (reliable with Martin M, Hunt costata dissolved (estimated) based restrictions) JW, Anderson BS, on: survival, growth Turpen SL & saltwater key study Palmer FH (1989)

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 185 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 renewal (48h) read-across based on grouping of 4 days survival and growth tests on substances (category Holmemysis costata approach)

Test material (EC name): zinc sulfate (See endpoint summary for justification of read- across) Paracentrotus lividus, Mediterranean EC10 (3 d): 23 µg/L 2 (reliable with Novelli AA, Losso Sea Urchin, Echinidae dissolved (zinc) (nominal) restrictions) C, Ghetti PF & based on: development Volpi Ghirardini A saltwater key study (2003) static read-across based on grouping of equivalent or similar to EPA 600/ R- substances (category 95/136 approach)

Test material (EC name): zinc nitrate (See endpoint summary for justification of read- across) Paracentrotus lividus, Lmk (sea EC10 (2 d): 17.7 µg/L 2 (reliable with Radenac G, Fichet urchin) dissolved (zinc) (estimated) restrictions) D & Miramand P based on: development (2001) saltwater key study static read-across based on grouping of 2-d larval development of sea urchin substances (category approach)

Test material (EC name): zinc nitrate (See endpoint summary for justification of read- across) Crassostrea gigas NOEC (48 h): 100 µg/L 1 (reliable without Chapman PM & dissolved (meas. (not restriction) McPherson C saltwater specified)) based on: larval (1993) development key study static read-across based on ASTM (1989) grouping of substances (category approach)

Test material (EC name): zinc nitrate (See endpoint summary for justification of read- across)

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 186 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 other aquatic mollusc: Crassostrea EC10 (4 d): 39.6 µg/L 2 (reliable with Watling HR (1982) gigas, Oyster, Ostreidae dissolved (zinc) (estimated) restrictions) based on: growth (valve saltwater width) key study semi-static read-across based on grouping of 4-d growth inhibition test of oyster substances (category larvae, test designed for dose-response approach)

Test material (EC name): zinc chloride (See endpoint summary for justification of read- across) other aquatic mollusc: Crassostrea EC10 (4 d): 18.4 µg/L 2 (reliable with Watling HR (1982) cucullata, Oyster, Ostreidae dissolved (zinc) (estimated) restrictions) based on: growth (valve saltwater width) key study semi-static read-across based on grouping of 4-d growth inhibition test of oyster substances (category larvae, tests designed for dose- approach) response Test material (EC name): zinc chloride (See endpoint summary for justification of read- across) other aquatic mollusc: Crassostrea EC10 (4 d): 11.8 µg/L 2 (reliable with Watling HR (1982) margaritacea , Oyster, Ostreidae dissolved (zinc) (estimated) restrictions) based on: growth (valve saltwater width) key study semi-static read-across based on grouping of 4-d growth inhibition test of oyster substances (category larvae, test designed for dose-response approach)

Test material (EC name): zinc chloride (See endpoint summary for justification of read- across) other aquatic mollusc: Mytilus NOEC (2 d): 80 µg/L 2 (reliable with Beiras R & galloprovincialis, Blue mussel, dissolved (zinc) (nominal) restrictions) Albentosa M (2004) Mytilidae based on: development key study saltwater read-across based on static grouping of substances (category 2-d larval development of marine approach) mollusks, dose-response designed test Test material (EC name): zinc chloride

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 187 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

(See endpoint summary for justification of read- across) other aquatic mollusc: Ruditapes EC10 (2 d): 55 µg/L 2 (reliable with Beiras R & decussatus, Grooved Carpet shell dissolved (zinc) (nominal) restrictions) Albentosa M (2004) clam, Veneridae based on: development key study saltwater read-across based on static grouping of substances (category 2-d larval development of marine approach) mollusks, dose-response designed test Test material (EC name): zinc chloride (See endpoint summary for justification of read- across) other aquatic mollusc: Ilyanassa NOEC (142 min): 20.7 µg/L 2 (reliable with Conrad GW (1988) obsoleta (Say), Eastern Mudsnail, dissolved (zinc) (estimated) restrictions) Nassariidae based on: development key study saltwater NOEC (142 min): > 0.1 — < 1 µmol/L dissolved read-across based on static (nominal) based on: grouping of development substances (category 142-min larval development test, lab approach) designed test for dose-response Test material (EC name): zinc chloride (See endpoint summary for justification of read- across) Asterias amurensis, Northern Pacific NOEC (80 min): 50 µg/L 2 (reliable with Lee CH, Ryu TK, Seastar, Asteriidae dissolved (zinc) (nominal) restrictions) Chang M & Choi based on: fertilization: JW (2004) saltwater presence of fertilization key study membrane static read-across based on grouping of equivalent or similar to EPA/600/ R- substances (category 95-136 approach)

Test material (EC name): zinc chloride (See endpoint summary for justification of read- across) other aquatic crustacea: Paragrapsus NOEC (4 d): 295 µg/L 2 (reliable with Ahsanullah M & quadridentatus, Toothed Shore Crab, dissolved (zinc) (estimated) restrictions) Arnott GH (1978) Grapsidae based on: mortality key study saltwater read-across based on semi-static grouping of

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 188 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

4-d survival test on crab larvae substances (category approach)

Test material (EC name): zinc chloride (See endpoint summary for justification of read- across) other aquatic worm: Capitella NOEC (28 d): 320 µg/L 2 (reliable with Reish DJ, Gerlinger capitata, Annelida, Polychaete worm, dissolved (nominal) based restrictions) TV, Phillips CA & Capitellidae on: reproduction Schmidtbauer PD key study (1977) saltwater NOEC (28 d): 560 µg/L dissolved (nominal) based read-across based on semi-static on: reproduction grouping of substances (category 28-d annelid reproduction test EC10 (28 d): 100 µg/L approach) dissolved (zinc) (estimated) based on: reproduction Test material (EC name): zinc chloride (See endpoint summary for justification of read- across) other aquatic worm: Ctenodrilus NOEC (21 d): 100 µg/L 2 (reliable with Reish DJ & Carr RS serratus, Annelid, Polychaete, dissolved (zinc) (nominal) restrictions) (1978) Ctenodrilidae based on: reproduction key study saltwater read-across based on static grouping of substances (category 21-d annelid reproduction test approach)

Test material (EC name): zinc chloride (See endpoint summary for justification of read- across) other aquatic worm: Ophryotrocha NOEC (28 d): 50 µg/L 2 (reliable with Reish DJ & Carr RS diadema, Annelid, Polychaete, dissolved (zinc) (nominal) restrictions) (1978) Dorvilleidae based on: reproduction key study saltwater read-across based on static grouping of substances (category 28-d annelid reproduction test approach)

Test material (EC name): zinc chloride (See endpoint summary for justification of read- across) other aquatic worm: Neanthes NOEC (7 mo): 100 µg/L 2 (reliable with Reish DJ, Gerlinger

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 189 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 arenaceaodentata, Annelida, dissolved (nominal) based restrictions) TV, Phillips CA & Polychaeta, Nereididae on: reproduction Schmidtbauer PD key study (1977) saltwater NOEC (7 mo): 100 µg/L dissolved (nominal) based read-across based on semi-static on: reproduction grouping of substances (category 7-mo annelid reproduction test EC10 (7 mo): 33.3 µg/L approach) dissolved (zinc) (estimated) based on: reproduction Test material (EC name): zinc chloride (See endpoint summary for justification of read- across) other aquatic worm: Ophryotrocha NOEC (35 d): 33.3 µg/L 2 (reliable with Reish DJ, Gerlinger diadema, Annelida, Polychaeta, dissolved (zinc) (estimated) restrictions) TV, Phillips CA & Dorvilleidae based on: reproduction (, Schmidtbauer PD early development) key study (1977) saltwater NOEC (35 d): 100 µg/L read-across based on semi-static dissolved (nominal) based grouping of on: reproduction substances (category 35-d annelid reproduction and early approach) development test NOEC (35 d): 100 µg/L dissolved (nominal) based Test material (EC on: reproduction name): zinc chloride (See endpoint NOEC (35 d): 100 µg/L summary for dissolved (nominal) based justification of read- on: reproduction across)

NOEC (35 d): 320 µg/L dissolved (nominal) based on: reproduction other aquatic worm: Neanthes NOEC (9 mo): 33.3 µg/L 2 (reliable with Reish DJ & arenaceaodentata, Annelid, dissolved (zinc) (estimated) restrictions) Gerlinger TV Polychaete, Nereididae based on: reproduction (1984) key study saltwater NOEC (9 mo): 100 µg/L dissolved (nominal) based read-across based on semi-static on: reproduction grouping of substances (category 9-months annelid reproduction test NOEC (6 mo): 100 µg/L approach) dissolved (nominal) based on: reproduction Test material (EC name): zinc chloride LOEC (9 mo): 320 µg/L (See endpoint dissolved (nominal) based summary for on: reproduction justification of read- across) LOEC (6 mo): 320 µg/L dissolved (nominal) based on: reproduction Monhystera disjuncta, Nematoda, NOEC (4 d): 250 µg/L 2 (reliable with Vranken G, Monhysteridae dissolved (zinc) (estimated) restrictions) Vanderhaeghen R based on: reproduction & Heip C (1991) saltwater key study static read-across based on

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 190 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

4-d nematode reproduction and early grouping of development test substances (category approach)

Test material (EC name): zinc chloride (See endpoint summary for justification of read- across) Mya arenaria NOEC (7 d): 900 µg/L 2 (reliable with Eisler R (1977) dissolved (nominal) based restrictions) saltwater on: mortality key study static NOEC (7 d): 10000 µg/L dissolved (nominal) based read-across based on lab designed test for dose-response on: mortality grouping of substances (category NOEC (7 d): 25000 µg/L approach) dissolved (nominal) based on: mortality Test material (EC name): zinc chloride NOEC (21 d): 1750 µg/L (See endpoint dissolved (nominal) based summary for on: mortality justification of read- across) NOEC (21 d): 25000 µg/L dissolved (nominal) based on: mortality other aquatic mollusc: Haliotis rubra, EC10 (2 d): 20.4 µg/L 2 (reliable with Gorski J & Blacklip Abalone, Haliotidae dissolved (zinc) (nominal) restrictions) Nugegoda D (2006) based on: development saltwater key study static read-across based on grouping of Similar to: Hunt JW, Anderson BS. substances (category 1990. Abalone larval development: approach) Short-term toxicity test protocol. In: Anderson BW, Hunt JW, Turpen SL, Test material Coulon AR, Martin M, McKeown DL, (IUPAC name): zinc Palmer FH, eds, Procedures Manual chloride (See for Conducting Toxicity Tests endpoint summary Developed by the Marine Bioassay for justification of Project. 90-10WQ. California State read-across) Water Resources Control Board, Sacramento, CA, USA, pp 17–48. Ceriodaphnia dubia EC20 (7 d): ca. 12.82 mg/L 2 (reliable with Huff et al. (2009) dissolved (N @ pH8, 25°C) restrictions) freshwater (meas. (geom. mean)) based on: reproduction supporting study static experimental result equivalent or similar to OECD Guideline 211 (Daphnia magna Test material (CAS Reproduction Test) name): Ammonium chloride Daphnia magna EC20 (21 d): ca. 12.83 mg/L 2 (reliable with Huff et al. (2009) dissolved (N @ pH8, 25C) restrictions)

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 191 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

(meas. (geom. mean)) based freshwater on: reproduction supporting study static experimental result equivalent or similar to OECD Test material (CAS Guideline 211 (Daphnia magna name): ammonium Reproduction Test) chloride mussel (div. see details) EC50 (4 d): 6.7 mg/L 1 (reliable without Wang et al (2007) dissolved (NH3) (meas. restriction) freshwater (geom. mean)) based on: mortality and growth supporting study flow-through EC50 (4 d): 6.5 mg/L experimental result see publication dissolved (NH3) (meas. (http://www.fishwild.vt.edu/mussel/PD (geom. mean)) based on: Test material (CAS Ffiles/chronic_toxicity.pdf) mortality and growth name): ammonium chloride EC50 (4 d): 7.4 mg/L dissolved (NH3) (meas. (geom. mean)) based on: mortality and growth

IC20 (28 d): < 0.4 mg/L dissolved (NH3) (meas. (initial)) based on: mortality and growth

IC20 (28 d): < 0.49 mg/L dissolved (NH3) (meas. (initial)) based on: mortality and growth

IC20 (28 d): < 1.02 mg/L dissolved (NH3) (nominal) based on: mortality and growth Daphnia magna NOEC (21 d): 14.6 mg/L test2 (reliable with Gersich, F.M. and mat. (meas. (not specified)) restrictions) Hopkins, D. L., freshwater based on: Mortality , (1986) reproduction supporting study semi-static LOEC (21 d): 30.2 mg/L testexperimental result not mentioned mat. (meas. (not specified)) Test material (EC name): Ammonium chloride other aquatic crustacea: Penaeus EC50 (56 d): 38.8 mg/L test 4 (not assignable) Chen, J.C., penicillatus mat. (meas. (not specified)) Lin,C.Y., (1992) based on: mortality supporting study equivalent or similar to APHA (1985) experimental result

Test material (EC name): ammonium chloride

Discussion

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 192 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Extensive high quality chronic data were available on 13 different freshwater invertebrate species and 26 marine invertebrate species. These data were all screened for relevancy to the environment under study.

The freshwater species are part of several different taxonomic groups: poriferans (4 species), molluscs (2 species), crustaceans (4 species), rotifers (2 species) and insects (1 species). The sensitivity of these species is equally distributed over the species sensitivity distribution.

The marine species are part of several different taxonomic groups: annelids (4 species), cniderians (1 species), crustaceans (6 species), echinoderms (5 species), molluscs (9 species) and nematods (1 species). The sensitivity of these species is equally distributed over the species sensitivity distribution.

The invertebrate species NOECs are combined with the other freshwater and marine chronic data in the SSD to give the HC5 from which the respective PNECs are derived.

The following information is taken into account for long-term toxicity to aquatic invertebrates for the derivation of PNEC: freshwater: Data on 13 species available. Species NOECs range between 0.037 and 0.400 mg Zn/l (dissolved concentrations) Marine: Data on 26 species available. Species NOECs range between 0.0056 and 0.9 mg Zn/l (dissolved concentrations) Species NOECs have been put into their respective species sensitivity distributions.

7.1.1.3. Algae and aquatic plants

The results are summarised in the following table:

Table 65. Overview of effects on algae and aquatic plants

Method Results Remarks Reference Chlorella sp. (algae) EC10 (48 h): 350 µg/L 2 (reliable with Wilde K. L. , dissolved (meas. (not restrictions) Stauber J.L., freshwater specified)) based on: growth Markich, S.J., rate key study Franklin N. M. and static Brown, P.L. (2006) EC10 (48 h): 105 µg/L read-across based on lab-designed dose-response test at dissolved (meas. (not grouping of different pH. specified)) based on: growth substances (category rate approach)

EC10 (48 h): 93 µg/L Test material dissolved (meas. (not (IUPAC name): zinc specified)) based on: growth sulphate (See rate endpoint summary for justification of EC10 (48 h): 16 µg/L read-across) dissolved (meas. (not specified)) based on: growth rate

EC10 (48 h): 5.9 µg/L dissolved (meas. (not specified)) based on: growth rate Pseudokirchnerella subcapitata (algae) NOEC (72 h): 5.4 µg/L 1 (reliable without De Schamphelaere dissolved (meas. (initial)) restriction) K.A.C., Heijerick

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 193 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

based on: growth rate D.G., and Janssen, freshwater key study C.R. (2003a) NOEC (72 h): 5.2 µg/L static dissolved (meas. (initial)) read-across based on based on: growth rate grouping of OECD Guideline 201 (Alga, Growth substances (category Inhibition Test) NOEC (72 h): 5.5 µg/L approach) dissolved (meas. (initial)) based on: growth rate Test material (IUPAC name): zinc NOEC (72 h): 5.5 µg/L chloride (See dissolved (meas. (initial)) endpoint summary based on: growth rate for justification of read-across) NOEC (72 h): 5.2 µg/L dissolved (meas. (initial)) based on: growth rate

NOEC (72 h): 8.6 µg/L dissolved (meas. (initial)) based on: growth rate

NOEC (72 h): 7.7 µg/L dissolved (meas. (initial)) based on: growth rate

NOEC (72 h): 8.5 µg/L dissolved (meas. (initial)) based on: growth rate

NOEC (72 h): 6.8 µg/L dissolved (meas. (initial)) based on: growth rate

NOEC (72 h): 7.9 µg/L dissolved (meas. (initial)) based on: growth rate

NOEC (72 h): 7.4 µg/L dissolved (meas. (initial)) based on: growth rate

NOEC (72 h): 4.9 µg/L dissolved (meas. (initial)) based on: growth rate

NOEC (72 h): 124 µg/L dissolved (meas. (initial)) based on: growth rate

NOEC (72 h): 74 µg/L dissolved (meas. (initial)) based on: growth rate

NOEC (72 h): 41 µg/L dissolved (meas. (initial)) based on: growth rate

NOEC (72 h): 15 µg/L dissolved (meas. (initial)) based on: growth rate

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 194 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

NOEC (72 h): 10 µg/L dissolved (meas. (initial)) based on: growth rate

NOEC (72 h): 9.4 µg/L dissolved (meas. (initial)) based on: growth rate Pseudokirchnerella subcapitata (algae) NOEC (72 h): 58 µg/L 1 (reliable without De Schamphelaere dissolved (meas. (initial)) restriction) K.A.C., Heijerick freshwater based on: growth rate D.G., and Janssen, key study C.R. (2003b) static NOEC (72 h): 91 µg/L dissolved (meas. (initial)) read-across based on OECD Guideline 201 (Alga, Growth based on: growth rate grouping of Inhibition Test) substances (category NOEC (72 h): 73 µg/L approach) dissolved (meas. (initial)) based on: growth rate Test material (IUPAC name): zinc NOEC (72 h): 27 µg/L chloride (See dissolved (meas. (initial)) endpoint summary based on: growth rate for justification of read-across) NOEC (72 h): 105 µg/L dissolved (meas. (initial)) based on: growth rate Pseudokirchnerella subcapitata (algae) NOEC (72 h): 81 µg/L 1 (reliable without Muyssen B., dissolved (meas. (arithm. restriction) Bossuyt B. and freshwater mean)) based on: growth rate Janssen C.R. key study (2003b) static NOEC (72 h): 93 µg/L dissolved (meas. (arithm. read-across based on OECD Guideline 201 (Alga, Growth mean)) based on: growth rategrouping of Inhibition Test) substances (category NOEC (72 h): 50 µg/L approach) dissolved (meas. (arithm. mean)) based on: growth rateTest material (IUPAC name): zinc NOEC (72 h): 86 µg/L chloride (See dissolved (meas. (arithm. endpoint summary mean)) based on: growth ratefor justification of read-across) Selenastrum capricornutum (new IC50 (72 h): 136 µg/L 1 (reliable without Van Ginneken I. name: Pseudokirchnerella dissolved (meas. (not restriction) (1994) subcapitata) (algae) specified)) based on: growth rate key study freshwater NOEC (3 d): 24 µg/L read-across based on static dissolved (meas. (not grouping of specified)) based on: growth substances (category OECD Guideline 201 (Alga, Growth rate approach) Inhibition Test) Test material (IUPAC name): zinc oxide (See endpoint summary for justification of read- across) Selenastrum capricornutum (new IC50 (3 d): 150 µg/L 1 (reliable without Van Woensel M.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 195 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 name: Pseudokirchnerella dissolved (meas. (not restriction) (1994) subcapitata) (algae) specified)) based on: growth rate key study freshwater NOEC (3 d): 50 µg/L read-across based on static dissolved (meas. (not grouping of specified)) based on: growth substances (category OECD Guideline 201 (Alga, Growth rate approach) Inhibition Test) Test material (IUPAC name): zinc (See endpoint summary for justification of read- across)

Ceramium tenuicore (algae) EC10 (7 d): 7.8 µg/L 2 (reliable with Eklund B (2005) dissolved (nominal) based restrictions) brackish water on: biomass key study static EC10 (7 d): 8 µg/L dissolved (nominal) based on: biomass read-across based on Lab designed test for dose-response. grouping of According to Eklund, the test will EC10 (7 d): 4.2 µg/L substances (category become an international standard dissolved (nominal) based approach) within ISO (Growth inhibition test on: biomass with the marine and brackish water Test material (EC macroalga Ceramium tenuicore. ITM- EC10 (7 d): 2.5 µg/L name): zinc metal rapport 131). dissolved (nominal) based (See endpoint on: biomass summary for justification of read- EC10 (7 d): 5.1 µg/L across) dissolved (estimated) based on: biomass Macrocystis pyrifera (Giant Kelp), NOEC (16 d): 1071 µg/L 1 (reliable without Anderson BS & Macroalga, Lessoniaceae (algae) dissolved (zinc) (meas. (not restriction) Hunt JW (1988) specified)) based on: saltwater Reproduction (sporophyte key study Anderson BS, Hunt production) JW, Martin M, semi-static read-across based on Turpen SL & NOEC (2 d): 190.2 µg/L grouping of Palmer FH (1988) 16-d and 2-d toxicity test to early life dissolved (estimated) based substances (category stages of macroalgae designed for on: length of germination approach) dose-response tubes Test material (EC LOEC (2 d): 589 µg/L name): zinc sulfate dissolved (meas. (not (See endpoint specified)) based on: length summary for of germination tube justification of read- across) LOEC (2 d): 553 µg/L dissolved (meas. (not specified)) based on: length of germination tubes

LOEC (2 d): 1090 µg/L dissolved (meas. (not specified)) based on: length of germination tubes

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 196 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Asterionella japonica, Diatom, EC10 (3 d): 8.6 µg/L 2 (reliable with Fisher NS & Frood Fragilariaceae (algae) dissolved (zinc) (estimated) restrictions) D (1980) based on: growth rate (cell saltwater division) key study static read-across based on grouping of 3-d growth inhibition test with marine substances (category diatom, designed for dose-response approach)

Test material (EC name): zinc sulfate (See endpoint summary for justification of read- across) Nitzschia closterium, Diatom, NOEC (3 d): 40 µg/L 2 (reliable with Fisher NS & Frood Bacillariaceae (algae) dissolved (nominal) based restrictions) D (1980) on: growth rate saltwater key study EC10 (3 d): 52 µg/L static dissolved (zinc) (estimated) read-across based on based on: growth rate (cell grouping of 3-d growth inhibition test with marine division) substances (category diatom, designed for dose-response approach)

Test material (IUPAC name): zinc sulfate (See endpoint summary for justification of read- across) Chaetoceros compressum, Diatom, LOEC (3 d): 20 µg/L 2 (reliable with Fisher NS & Frood Chaetocerotaceae (algae) dissolved (nominal) based restrictions) D (1980) on: growth rate saltwater key study EC10 (3 d): 11.2 µg/L static dissolved (zinc) (estimated) read-across based on based on: growth rate (cell grouping of 3-d growth inhibition test with marine division) substances (category diatom, test designed for dose-response approach)

Test material (IUPAC name): zinc sulfate (See endpoint summary for justification of read- across) Skeletonema costatum, Diatom, NOEC (3 d): 20 µg/L 2 (reliable with Fisher NS & Frood Skeletonemaceae (algae) dissolved (nominal) based restrictions) D (1980) on: growth rate saltwater key study LOEC (3 d): 20 µg/L static dissolved (nominal) based read-across based on on: growth rate (growth grouping of 3-d growth inhibition test with marine response in UV irradiated substances (category diatom, test designed for dose-response seawater) approach)

EC10 (3 d): 26.4 µg/L Test material (EC dissolved (zinc) (estimated) name): zinc sulfate

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 197 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

based on: growth rate (cell (See endpoint division) summary for justification of read- across) Asterionella japonica, Diatom, NOEC (4 d): 20 µg/L 2 (reliable with Fisher NS, Jones GJ Fragilariaceae (algae) dissolved (nominal) based restrictions) & Nelson DM on: growth rate (1981) saltwater key study EC10 (4 d): 20.6 µg/L static dissolved (zinc) (estimated) read-across based on based on: growth rate (cell grouping of 4-d growth inhibition of marine algae, division) substances (category test designed for dose-response approach)

Test material (EC name): zinc sulfate (See endpoint summary for justification of read- across) Ulva pertusa, Green macroalga, NOEC (5 d): 313 µg/L 2 (reliable with Han T & Choi G-W Ulvaceae (algae) dissolved (zinc) (nominal) restrictions) 2005 (2005) based on: sporulation saltwater (reproduction) key study static read-across based on grouping of 5-d sporulation-inhibition test with substances (category marine macroalgae, designed for dose- approach) response Test material (EC name): zinc nitrate (See endpoint summary for justification of read- across) Ascophyllum nodosum, Brown EC10 (10 d): 564.8 µg/L 2 (reliable with Stromgren T 1979 Macroalga, Fucaceae (algae) dissolved (zinc) (estimated) restrictions) (1979) based on: growth rate saltwater key study flow-through read-across based on grouping of 10-d growth inhibition of macroalgae substances (category designed for dose-response approach)

Test material (EC name): zinc chloride (See endpoint summary for justification of read- across) Fucus serratus, Toothed wrack, Brown EC10 (10 d): 488 µg/L 2 (reliable with Stromgren T 1979 Macroalga, Fucaceae (algae) dissolved (zinc) (estimated) restrictions) (1979) based on: growth rate saltwater key study flow-through read-across based on grouping of

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 198 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

10-d growth inhibition of macroalgae substances (category designed for dose-response approach)

Test material (EC name): zinc chloride (See endpoint summary for justification of read- across) Fucus vesiculosus, Bladder wrack, NOEC (10 d): 100 µg/L 2 (reliable with Stromgren T 1979 Brown Macroalga, Fucaceae (algae) dissolved (zinc) (nominal) restrictions) (1979) based on: growth rate saltwater key study flow-through read-across based on grouping of 10-d growth inhibition of macroalgae substances (category designed for dose-response approach)

Test material (EC name): zinc chloride (See endpoint summary for justification of read- across) Fucus spiralis, Brown Macroalga, EC10 (10 d): 613.2 µg/L 2 (reliable with Stromgren T 1979 Fucaceae (algae) dissolved (zinc) (estimated) restrictions) (1979) based on: growth rate saltwater key study flow-through read-across based on grouping of 10-d growth inhibition of macroalgae substances (category designed for dose-response approach)

Test material (EC name): zinc chloride (See endpoint summary for justification of read- across) Pelvetia canaliculata, Brown EC10 (10 d): 670.8 µg/L 2 (reliable with Stromgren T 1979 Macroalga, Fucaceae (algae) dissolved (zinc) (estimated) restrictions) (1979) based on: growth rate saltwater key study flow-through read-across based on grouping of 10-d growth inhibition of macroalgae substances (category designed for dose-response approach)

Test material (EC name): zinc chloride (See endpoint summary for justification of read- across) Nitzschia closterium, Diatom, EC10 (3 d): 84 µg/L 1 (reliable without Johnson HL,

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 199 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Bacillariaceae (algae) dissolved (zinc) (meas. (not restriction) Stauber JL, Adams specified)) based on: growth MS & Jolley DF saltwater rate (cell division rate) key study (2007) static read-across based on grouping of 72 h growth inhibition of algae, test substances (category designed for dose-response approach)

Test material (EC name): zinc chloride (See endpoint summary for justification of read- across) Ceramium tenuicore (algae) EC10 (7 d): 16 µg/L 2 (reliable with Eklund B (2005) dissolved (nominal) based restrictions) saltwater on: length key study static EC10 (7 d): 7.7 µg/L dissolved (nominal) based read-across based on Lab designed test for dose-response. on: length grouping of According to Eklund, the test will substances (category become an international standard EC10 (7 d): 18 µg/L approach) within ISO (Growth inhibition test dissolved (nominal) based with the marine and brackish water on: length Test material (EC macroalga Ceramium tenuicore. ITM- name): zinc metal rapport 131). EC10 (7 d): 15 µg/L (See endpoint dissolved (nominal) based summary for on: length justification of read- across) EC10 (7 d): 7.2 µg/L dissolved (nominal) based on: length

EC10 (7 d): 11.9 µg/L dissolved (estimated) based on: length

Cladophora glomerata (aquatic plants) NOEC (3 d): 60 µg/L 2 (reliable with Whitton B.A. dissolved (nominal) based restrictions) (1967) freshwater on: growth rate key study semi-static read-across based on lab-designed dose-response test grouping of substances (category approach)

Test material (IUPAC name): zinc sulphate (See endpoint summary for justification of read-across) Lemna gibba (aquatic plants) NOEC (70 d): > 650 µg/L 2 (reliable with Van der Werff M., dissolved (nominal) based restrictions) and Pruyt M.J. freshwater on: groth rate and survival (1982) supporting study

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 200 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 semi-static read-across based on grouping of lab-designed dose-response test substances (category approach)

Test material (IUPAC name): zinc sulphate (See endpoint summary for justification of read-across) Elodea nuttallii (aquatic plants) NOEC (70 d): > 650 µg/L 2 (reliable with Van der Werff M., dissolved (nominal) based restrictions) and Pruyt M.J. freshwater on: groth rate and survival (1982) supporting study semi-static read-across based on lab-designed dose-response test grouping of substances (category approach)

Test material (IUPAC name): zinc sulphate (See endpoint summary for justification of read-across) Callitriche platycarpa (aquatic plants) NOEC (70 d): > 650 µg/L 2 (reliable with Van der Werff M., dissolved (nominal) based restrictions) and Pruyt M.J. freshwater on: groth rate and survival (1982) supporting study semi-static read-across based on lab-designed dose-response test grouping of substances (category approach)

Test material (IUPAC name): zinc sulphate (See endpoint summary for justification of read-across) Spirodela polyrhiza (aquatic plants) NOEC (70 d): > 650 µg/L 2 (reliable with Van der Werff M., dissolved (nominal) based restrictions) and Pruyt M.J. freshwater on: groth rate and survival (1982) supporting study semi-static read-across based on lab-designed dose-response test grouping of substances (category approach)

Test material (IUPAC name): zinc sulphate (See endpoint summary for justification of read-across)

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 201 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Navicula sp. (algae) EC50 (10 d): 90.4 mg/L test 2 (reliable with Admiraal,W., mat. (meas. (not specified)) restrictions) (1977) saltwater based on: growth rate supporting study static NOEC (10 d): 26.8 mg/L test mat. (meas. (not specified)) experimental result no data based on: growth rate Test material (EC LOEC (10 d): 53.5 mg/L testname): ammonium mat. (meas. (not specified)) chloride based on: growth rate Chlorella vulgaris (algae) EC50 (5 d): 1300 mg/L test 2 (reliable with Przytocka-Jusiak, mat. based on: growth rate restrictions) M. et al, (1977) freshwater supporting study static read-across from no data supporting substance (structural analogue or surrogate)

Test material (EC name): Ammonium carbonate (See endpoint summary for justification of read-across)

Data waiving

Information requirement: Growth inhibition study with algae / cyanobacteria Reason: other justification Justification: Unsuitable test system/does not meet important criteria of today standard methods (no control data) Wikfors, G. H. and R. Ukeles. 1982. Growth and adaptation of estuarine unicellular algae in media with excess copper, cadmium or zinc, and effects of metal-contaminated algal food on Crassostrea virginica larvae. Mar. Ecol. Prog. Ser. 7:191-206.

Discussion

Effects on algae / cyanobacteria

Acute freshwater toxicity tests of high quality and relevancy performed according to standard protocol. Information is on 1 species which is in both the acute and chronic aquatic database on zinc the most sensitive. The lowest IC50 value is taken as reference value for classification for acute effect at neutral/high pH.

Chronic freshwater toxicity tests of high quality and relevancy according to standard protocol or equivalent. Data on 2 species, one of which the most sensitive of all freshwater organisms, second species is less sensitive.

Chronic seawater tests of high quality and relevancy according to standard protocol or equivalent. Data on 12 species, for which 3 species are in the low part of the species sensitivity distribution. One species of macroalgae is the second most sensitive of all seawater organisms.

The following information is taken into account for effects on algae / cyanobacteria for the derivation of PNEC:

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 202 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 acute toxicity to freshwater algae: lowest IC50 0.136 mg Zn/l (Selenastrum capricornutum; single value) (neutral/high pH) chronic toxictiy to freshwater algae: lowest NOEC 0.019 mg Zn/l (Pseudokircherniella subcapitata =Selenastrum capricornutum; geomean of 27 data) chronic toxicity to marine algae: 12 species available which NOECs range between 0.0078 and 0.67 mg/l (dissolved concentrations)

Effects on aquatic plants other than algae

The EU risk assessment on zinc (ECB 2008) used one chronic NOEC on Cladophora glomerata in the PNEC database. Additional information on long term experiments on 4 higher aquatic plant species was used as supportive evidence, demonstrating the low sensitivity of this taxonomic group.

The following information is taken into account for effects on aquatic plants other than algae for the derivation of PNEC: chronic NOEC for one multicellular algae species available (0.06mg Zn/l). Additional supportive information on 4 higher aquatic plant species available; chronic NOEC >650µg Zn/l on all 4 species tested.

7.1.1.4. Sediment organisms

Freshwater sediments chronic PNEC - establishing the dataset In this CSR, the results of the chronic benthic toxicity studies are all expressed as the actual (measured) concentration. The actual concentrations include the background concentration (Cb) of zinc. Because of the “added risk approach”, the results based on actual concentrations have been corrected for background. This correction for background is based on the assumption that only the added concentration of zinc is relevant for toxicity. Consistent with approved methodology, the reported benthic toxicity data presented here represent total (bulk)-zinc concentrations, i.e. the dissolved plus particulate fraction. The chronic benthic toxicity literature for zinc was checked according to the general criteria for data quality:  Study design preferably conform to OECD guidelines or equivalent  Toxicological endpoints, which may affect the species at the population level, are taken into account. In general, these endpoints are survival, growth and reproduction.  For PNEC derivation a full life-cycle test, in which all relevant toxicological endpoints are studied, is normally preferred to a test covering not a full life cycle and/or not all relevant endpoints.  If for one species several chronic NOEC values (from different tests) based on the same toxicological endpoint are available, these values are averaged by calculating the geometric mean, resulting in the “species mean” NOEC.  If for one species several chronic NOEC values based on different toxicological endpoints are available, the lowest value is selected. The lowest value is determined on the basis of the geometric mean if more than one value for the same endpoint is available.  In some cases, NOEC values for different life stages of a specific organism are available. If from these data it appeared that a distinct life stage was more sensitive, the result for the most sensitive life stage is selected.  Only the results of tests in which the organisms were exposed to zinc alone are used, thus excluding tests with metal mixtures.  Like in the RAR, unbounded NOEC values (i.e. no effect was found at the highest concentration tested) are not used.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 203 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Ecotoxicity data for freshwater sediment The available database of chronic freshwater sediment toxicity tests for zinc provides information on seven different species (single-species exposures) and two long-term field colonization studies. They are detailed in table 67

Single-species tests The available studies were for tests with crustaceans (Gammarus pulex and Hyallela azteca ), insects (Ephoron virgo, Hexagenia sp. and Chironomus tentans) and worms (Lumbriculus variegates and Tubifex tubifex).  The available data for benthic invertebrates (sediment-dwelling organisms) includes only long-term tests (3 to 8 weeks exposure), which sufficiently provide information on survival, growth and reproductive effects. These species represent the most frequently utilized freshwater benthic species for assessing the toxicity of substances in spiked freshwater sediments or in field-collected freshwater sediments and for which species-specific standard sediment test protocols have been developed by e.g. ASTM, U.S. EPA and OECD.

 The seven species with chronic endpoints considered to be useful for PNEC derivation (PNEC add, sediment) include four species that were not used in the PNEC calculation in the RAR (2008). In addition, a recent study with H. azteca (Norwood et al., 2009) demonstrated that the growth endpoint was more sensitive than the survival endpoint used for evaluation during the Zn RAR (2008). As such, the species mean value for H. azteca is also considered new data.  The seven species NOEC or EC10 (effective concentration to reduce performance by 10% relative to controls) values were obtained in unpolluted sediments with a background zinc concentration (Cb) of 23 to 55 mg/kg d.w. In addition to survival at least one other endpoint (growth, emergence and/or reproduction) was studied in each test. The “species mean” chronic values used for PNECadd, sediment derivation range from 146 to 1101 mg/kg d.w. Note that the following NOEC values from all six studies are based on the added concentration, in this case being the actual concentration measured minus Cb, the background concentration. A summary of the seven species and endpoints are provided: o A 5-w test with G. pulex (Nguyen et al., 2005) resulted in a NOEC value based on growth of 146 mg/kg d.w. o Two 4-w tests with H. azteca (Norwood et al., 2009) were conducted with sediment from two sites. The EC10 values based on growth were 224 and 1250 mg/kg d.w., resulting in a geometric species mean value of 529 mg/kg d.w., respectively. o A 3-w test with E. virgo (Nguyen et al., 2005) resulted in a NOEC value based on growth of 164 mg/kg d.w. o A 3-w test with Hexagenia sp. (Norwood et al., 2008) resulted in a NOEC value based on growth of 570 mg/kg d.w. o Two C. tentans (Sibley et al., 1996; Farrar & Bridges, 2002) resulted in NOEC values based on growth, emergence and/or reproduction of 795 and 609 mg/kg d.w., respectively. This results in a geometric species mean value of 696 mg/kg d.w. o The 4-w test with L. variegates (Nguyen et al., 2005) resulted in a NOEC value based on survival, growth and reproduction of 878 mg/kg d.w. o A 4-w test with T. tubifex (Farrar & Bridges, 2003) resulted in a NOEC value based on reproduction of 1101 mg/kg d.w.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 204 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Hyalella azteca 529 1 Insects Ephoron virgo 164 1 Hexagenia sp. 570 1 Chironomus tentans 696 1 Worms Lumbriculus variegates 878 1 Tubifex tubifex 1101 1

 Because of the inclusion of four additional species, the species sensitivity distribution (SSD) was preferred to calculate an HC5 from the log-normal distribution of the data, and derive the PNEC add, sediment. This is different from the approach taken in the RAR (2008), where an Assessment Factor of 10 was applied to the lowest NOEC value of the dataset, containing three species. With the inclusion of four more species in the dataset, the lognormal distribution now provides a significant fit to the data. Other conditions to apply statistical extrapolation were also met (see below, discussion on the safety factor to be applied to the HC5).

Field colonization tests In the last decade, two long-term field colonization studies have been performed in sediments spiked with zinc (Liber et al., 1996; Burton et al., 2003), using in total five different freshwater sediments:  Liber et al. (1996): In the 1-year study by Liber et al. (1996), performed at one site (with five consecutive sampling dates), “minor” effects were observed at added (measured-Cb) SEMZn concentrations of 310 mg/kg d.w. (after 2.5 months exposure) and at 725 mg/kg d.w. (between 3 and 11 months exposure), the highest two concentrations tested. Overall, the highest concentration tested (725 mg/kg d.w.) was considered to be the overall NOECecosystem.

 Burton et al. (2005): In the study by Burton et al. (2005), performed at four European sites (with one to three consecutive sampling dates per site; exposure time ranged from 11 to 37 weeks), only two zinc concentrations, nominal 400 and 1200 mg/kg d.w., were tested along with the control, thus reliable NOECecosystem values could not be derived from this study. The range of the actual zinc concentrations in Burton et al. (2005) were 119-255 mg/kg d.w. (added) at the low-zinc treatment and 214-782 mg/kg d.w. (added) at the high-zinc treatment. The site at Schmallenberg, Germany (sampled over the entire 37-week exposure period) exhibited no significant effects on ecosystem parameters at added sediment-zinc concentrations of 205 mg/kg d.w. (23 weeks) or 226 mg/kg d.w. (37 weeks), and the ratio of Simultaneous Extractable Metal (SEM) to Acid Volatile Sulfide (AVS) concentrations (see Accounting for bioavailability section below) were less than 1 (0.47 and 0.58, respectively). Minor effects were observed in the low-zinc treatment at two of the sites; Biesbosch, The Netherlands (at 37 weeks following field deployment, 178 mg/kg d.w. for added zinc) and Pallanza, Italy (at 23 weeks following field deployment, 119 mg/kg d.w. for added zinc). The SEM:AVS ratio at these sites were 2.3 and 43, respectively, suggesting enhanced bioavailability in these sediments. This finding supports assumptions regarding SEM:AVS, where ratios greater than 2 can be associated with observed biological effects. Despite the presence of minor effects at the low zinc treatment for Pallanza, the sulfide (1.9 mg/kg d.w.) and total organic carbon (0.34 mg/kg d.w.) characteristics of this site represent less than 5% (5P) of sediments throughout Europe (FOREGS database; http://www.gsf.fi/publ/foregsatlas/index.php). As such, unique conditions such as these should be considered within the context of site-specific bioavailability (as described in the “Accounting for bioavailability” section below), and not subjected to the generic PNECsediment presented here.

Marine chronic sediment data - establishing the dataset In this CSR, the results of the chronic marine toxicity studies were all expressed as the actual (measured) or

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 205 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 nominal concentration (Cn). Using the “added risk approach”, results based on actual or nominal concentrations were corrected for background concentrations (Cb). This correction for background is based on the assumption that only the added concentration of zinc is relevant for toxicity. Consistent with approved methodology, the reported benthic toxicity data presented here represent total (bulk)-zinc concentrations, i.e. the dissolved plus particulate fraction. The chronic marine benthic toxicity literature for zinc was checked according to the general criteria for data quality:  Study design preferably conform to OECD guidelines or equivalent  Toxicological endpoints, which may affect the species at the population level, are taken into account. In general, these endpoints are survival, growth and reproduction.  For PNEC derivation a full life-cycle test, in which all relevant toxicological endpoints are studied, is normally preferred to a test covering not a full life cycle and/or not all relevant endpoints.  If for one species several chronic NOEC values (from different tests) based on the same toxicological endpoint are available, these values are averaged by calculating the geometric mean, resulting in the “species mean” NOEC.  If for one species several chronic NOEC values based on different toxicological endpoints are available, the lowest value is selected. The lowest value is determined on the basis of the geometric mean if more than one value for the same endpoint is available.  In some cases, NOEC values for different life stages of a specific organism are available. If from these data it appeared that a distinct life stage was more sensitive, the result for the most sensitive life stage is selected.  Only the results of tests in which the organisms were exposed to zinc alone are used, thus excluding tests with metal mixtures.  Like in the RAR, unbounded NOEC values (i.e. no effect was found at the highest concentration tested) are not used.  Like in the RAR, only the results of tests with soluble zinc salts are used, thus excluding tests with more sparingly soluble zinc salts (ZnO, ZnCO3).

Ecotoxicity data for marine sediment Two single-species tests in a marine water-sediment system with zinc were found. The available studies were for a benthic crustacean (Melita plumulosa) and a higher plant (grey mangrove; Avicennia marina). The available studies represent long-term tests (7 weeks for M. plumulosa and 6 months for A. marina), which sufficiently provides information on survival, growth and reproductive effects. The tests were performed in unpolluted sediment with a background Zn concentration (Cb) of 240 and 42.9 mg/kg d.w. for M. plumulosa and A. marina, respectively. The lowest NOEC values obtained from these studies were 490 mg/kg d.w. (reproduction; added) and 207.1 mg/kg d.w. (emergence; added) for M. plumulosa and A. marina, respectively.

The available database of chronic marine sediment toxicity tests are detailed in table below.:

Table 67. Overview of long-term effects on sediment organisms

Method Results Remarks Reference Chironomus tentans NOEC (56 d): 13.07 µmol/g 1 (reliable without Sibley P. K., G. T. dw (meas. (arithm. mean)) restriction) Ankely, A. M. freshwater based on: survival, growth, Cotter and E. N. emergence, reproduction key study Leonard (1996) long-term toxicity (laboratory study) NOEC (56 d): 850 mg/kg read-across based on semi-static sediment dw (estimated) grouping of based on: survival, growth, substances (category 56 day test on insect Chironomus

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 206 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 tentans, designed for dose-response emergence and reproduction approach)

Test material (IUPAC name): zinc dichloride (See endpoint summary for justification of read-across) Chironomus tentans NOEC (20 d): 639 mg/kg 1 (reliable without Farrar J. D. and sediment dw (meas. (not restriction) Bridges T. S. freshwater specified)) based on: growth (2003) key study long-term toxicity (laboratory study) NOEC (20 d): 2420 mg/kg sediment dw (meas. (not read-across based on flow-through specified)) based on: grouping of mortality substances (category Toxicity and bioaccumulation of approach) sediment-associated contaminants with freshwater invertebrates (USEPA, Test material 2000) (IUPAC name): zinc dichloride (See endpoint summary for justification of read-across) Hyalella azteca NOEC (28 d): 936 mg/kg 1 (reliable without Farrar J. D. and sediment dw (meas. (not restriction) Bridges T. S. freshwater specified)) based on: (2003) mortality key study long-term toxicity (laboratory study) NOEC (28 d): 32 mg/kg read-across based on flow-through sediment dw (meas. (not grouping of specified)) based on: growth substances (category Toxicity and bioaccumulation of approach) sediment-associated contaminants with freshwater invertebrates (USEPA, Test material 2000) (IUPAC name): zinc dichloride (See endpoint summary for justification of read-across) Tubifex tubifex NOEC (28 d): 1135 mg/kg 1 (reliable without Farrar J. D. and sediment dw (meas. (not restriction) Bridges T. S. freshwater specified)) based on: (2003) reproduction key study long-term toxicity (laboratory study) NOEC (28 d): 2610 mg/kg read-across based on flow-through sediment dw (meas. (not grouping of specified)) based on: substances (category following test guidelines described in mortality approach) Test Methods for measuring the toxicity of sediment-associated Test material contaminants with freshwater (IUPAC name): zinc invertebrates (ASTM, 2000) dichloride (See endpoint summary for justification of read-across) Lumbriculus variegatus NOEC (28 d): 933 mg/kg 1 (reliable without Nguyen LTH, YE sediment dw (meas. (not restriction) Roman, M freshwater

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 207 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

specified)) based on: Vandegehuchte and long-term toxicity (laboratory study) Survival, growth, key study CR Janssen (2005a) reproduction semi-static read-across based on grouping of equivalent or similar to culturing substances (category conditions as described in US EPA approach) (2000) and OECD (2005) Test material (IUPAC name): zinc dichloride (See endpoint summary for justification of read-across) Gammarus pulex NOEC (35 d): 369 mg/kg 1 (reliable without Nguyen LTH, YE sediment dw (meas. (not restriction) Roman, M freshwater specified)) based on: Vandegehuchte and Survival key study CR Janssen (2005a) long-term toxicity (laboratory study) NOEC (35 d): 201 mg/kg read-across based on semi-static sediment dw (meas. (not grouping of specified)) based on: growth substances (category equivalent or similar to culturing approach) conditions as described in US EPA (2000) and OECD (2005) Test material (IUPAC name): zinc dichloride (See endpoint summary for justification of read-across) Hyalella azteca NOEC (42 d): 455 mg/kg 1 (reliable without Nguyen LTH, YE sediment dw (meas. (not restriction) Roman, M freshwater specified)) based on: Vandegehuchte and Survival key study CR Janssen (2005b) long-term toxicity (laboratory study) NOEC (42 d): 928 mg/kg read-across based on semi-static sediment dw (meas. (not grouping of specified)) based on: growth substances (category equivalent or similar to The tests were and reproduction approach) conducted according to the ASTM (1994) and US EPA (2000) standard Test material guidelines. (IUPAC name): zinc dichloride (See endpoint summary for justification of read-across) Ephoron virgo NOEC (21 d): 306 mg/kg 1 (reliable without Nguyen LTH, YE sediment dw (meas. (not restriction) Roman, M freshwater specified)) based on: Vandegehuchte and Survival key study CR Janssen (2005c) long-term toxicity (laboratory study) NOEC (21 d): 219 mg/kg read-across based on semi-static sediment dw (meas. (not grouping of specified)) based on: growth substances (category equivalent or similar to The tests were approach) conducted according to the ASTM (1994), US EPA (2000) and OECD Test material (2005) standard guidelines. (IUPAC name): zinc dichloride (See endpoint summary

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 208 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

for justification of read-across) Avicennia marina NOEC (30 wk): 250 mg/kg 1 (reliable without MacFarlane G. R. sediment dw (nominal) restriction) and M. D. Burchett saltwater based on: growth and (2002) survival key study long-term toxicity (laboratory study) read-across based on semi-static grouping of substances (category equivalent or similar to OECD approach) guideline 208 Test material (IUPAC name): zinc dichloride (See endpoint summary for justification of read-across) multi-species test NOEC (55 wk): 776 mg/kg 1 (reliable without Liber K., D. Call, T. sediment dw (meas. (not restriction) P. Markee, K. L. freshwater specified)) based on: Schmude, M. D. abundance key study Balcer, F. (1996) long-term toxicity (field study) read-across based on static grouping of substances (category 13 months field study on approach) macroinvertebrates dominated by chironomids, oligochaetes and bivalves Test material (IUPAC name): zinc dichloride (See endpoint summary for justification of read-across) Melita plumulosa NOEC (42 d): > 1770 mg/kg 2 (reliable with Gale SA, CK King sediment dw total particulate restrictions) and RV Hyne saltwater metal concentration (meas. (2006) (not specified)) based on: weight of evidence long-term toxicity (laboratory study) survival read-across based on semi-static NOEC (42 d): 1280 mg/kg grouping of sediment dw total particulate substances (category 42 d chronic sublethal sediment metal concentrations (meas. approach) toxicity test using the estuarine (not specified)) amphipod Melita plumulosa, designed Test material for dose-response NOEC (42 d): 730 mg/kg (IUPAC name): zinc sediment dw total particulate (See endpoint metal concentration (meas. summary for (not specified)) based on: justification of read- reproduction across) macroinvertebrate assemblages "effects" (12 wk): 119 mg/kg3 (not reliable) Burton A. G., sediment dw (meas. (not Nguyen LTH, freshwater specified)) based on: benthic weight of evidence Janssen C., Baudo macroinvertebrate effects R., McWilliam R., long-term toxicity (field study) (species richness and read-across based on Bossuyt (2005) macroinvertebrate density) grouping of Long term field study on benthic substances (category macroinvertebrate communities approach)

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 209 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Test material (IUPAC name): zinc (See endpoint summary for justification of read- across) Chironomus riparius EC10 (10 d): 80 mg/kg 1 (reliable without Norwood W. P., T. sediment dw (meas. (not restriction) watson-Leung and freshwater specified)) based on: growth D. Milani (2009) (wet weight) supporting study long-term toxicity (laboratory study) EC10 (10 d): 232 mg/kg read-across based on Zn spiked sediment chronic toxicity sediment dw (meas. (not grouping of tests on 4 species of freshwater aquatic specified)) based on: growth substances (category invertebrate species: Hyalella azteca approach) (28d survival and growth tests), LC10 (10 d): 269 mg/kg Chironomus riparius (10d survival and sediment dw (meas. (not Test material (EC growth tests), Hexagenia spp. (21d specified)) based on: growth name): zinc metal survival and growth tests) and Tubifex (See endpoint tubifex (28d reproduction and adult LC10 (10 d): 850 mg/kg summary for survival tests) sediment dw (meas. (not justification of read- specified)) based on: growth across) Hexagenia sp. EC10 (21 d): 608 mg/kg 1 (reliable without Norwood W. P., T. sediment dw (meas. (not restriction) watson-Leung and freshwater specified)) based on: growth D. Milani (2009) (wet weight) supporting study long-term toxicity (laboratory study) read-across based on Zn spiked sediment chronic toxicity grouping of tests on 4 species of freshwater aquatic substances (category invertebrate species: Hyalella azteca approach) (28d survival and growth tests), Chironomus riparius (10d survival and Test material (EC growth tests), Hexagenia spp. (21d name): zinc metal survival and growth tests) and Tubifex (See endpoint tubifex (28d reproduction and adult summary for survival tests) justification of read- across) Hyalella azteca EC10 (28 d): 250 mg/kg 1 (reliable without Norwood W. P., T. sediment dw (meas. (not restriction) watson-Leung and freshwater specified)) based on: growth D. Milani (2009) (dry weight) supporting study long-term toxicity (laboratory study) EC10 (28 d): 1288 mg/kg read-across based on Zn spiked sediment chronic toxicity sediment dw (meas. (not grouping of tests on 4 species of freshwater aquatic specified)) based on: growth substances (category invertebrate species: Hyalella azteca approach) (28d survival and growth tests), LC10 (28 d): 526 mg/kg Chironomus riparius (10d survival and sediment dw (meas. (not Test material (EC growth tests), Hexagenia spp. (21d specified)) based on: name): zinc metal survival and growth tests) and Tubifex mortality (See endpoint tubifex (28d reproduction and adult summary for survival tests) LC10 (28 d): 2111 mg/kg justification of read- sediment dw (meas. (not across) specified)) based on: mortality R. abronius, E. estuarius, A. abdita, G. LC50 (96 h): ca. 78.7 mg/L 2 (reliable with Kohn et al. (1994) japonica total ammonia (meas. (geom.restrictions)

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 210 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

mean)) based on: mortality saltwater supporting study LC50 (96 h): ca. 125.5 mg/L short-term toxicity (laboratory study) total ammonia (meas. (geom.experimental result mean)) based on: mortality equivalent or similar to EPA OPPTS Test material: 850.1740 (Whole Sediment Acute LC50 (96 h): ca. 49.8 mg/L aqueous ammonia Toxicity of Invertebrates, marine) total ammonia (meas. (geom. mean)) based on: mortality

LC50 (96 h): ca. 148.3 mg/L total ammonia (meas. (geom. mean)) based on: mortality Ampelisca abdita NOEC (10 d): ca. 30 mg/L 3 (not reliable) Mueller (1995) dissolved (NH4) (meas. saltwater (geom. mean)) based on: supporting study mortality long-term toxicity (laboratory study) experimental result static Test material (CAS name): unknown inorganic ammonium compound Sphaerium novaezelandiae NOEC (60 d): ca. 0.8 mg/L 2 (reliable with Hickey, (1999) total nitrogen (meas. (geom. restrictions) freshwater mean)) based on: mortality supporting study long-term toxicity (laboratory study) LOEC (60 d): ca. 5.4 mg/L total nitrogen (meas. (geom. experimental result static mean)) based on: mortality Test material (CAS - 2-L polyethylene pots LC50 (60 d): ca. 3.8 mg/L name): ammonium - 300 g of sediment total nitrogen (meas. (geom. chloride - 1.5 L of water mean)) based on: mortality - thermostated water bath (20°C) - a 16:8 light:dark cycle - hardness 26.9 mg/L (as CaCO3), calcium 1.2 mg/L, magnesium 5.7 mg/L, and ammonia , 1.0 μ g (N)/L - The ammonia toxicant (BDH Analar ammonium chloride) was dosed at four nominal concentrations (1.2, 3.6, 11, and 33 mg/L total ammonia) using a peristaltic pump - All tubing was replaced and calibration tested at weekly intervals. Clams were fed daily with a digested trout pellet-yeast diet (US EPA 1985b), modified by omission of cerophyll. The volume of food (6.2 mL/pot) provided a concentration of about 10 mg/L of suspended solids. Every second day 10 mg of a dry food (48% Tetra Mint, 24% dried ground alfalfa, 24% ground dried wheat grass, and 4% Neonovumt)

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 211 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 was added to each container - time frame: 60 d Lumbriculus variegatus, Chironomus LC50 (10 d): ca. 471.7 mg/L 1 (reliable without WHITEMAN tentans, Hyalella azteca total nitrogen (meas. (geom. restriction) (1995) mean)) based on: survival freshwater supporting study LC50 (10 d): ca. 704 mg/L long-term toxicity (laboratory study) total nitrogen (meas. (geom. experimental result mean)) based on: survival static Test material (CAS LC50 (4 d): ca. 82 mg/L name): Aqueous 300-ml high form beakers with 150 ml total nitrogen (meas. (geom. ammonia of sand/sediment and 10 organisms per mean)) based on: survival beaker. Eight replicates were tested at each exposure concentration with five LC50 (10 d): ca. 316.6 mg/L beakers in each treatment used for total nitrogen (meas. (geom. biological endpoints, and three for mean)) based on: survival pore-water chemistry. Lumbriculus variegatus, Chironomus LC50 (10 d): ca. 532.1 mg/L tentans: 10 d total nitrogen (meas. (geom. mean)) based on: survival Hyalella azteca: 4 d LC50 (4 d): ca. 9.2 mg/L total nitrogen (meas. (geom. mean)) based on: survival Amphipod (Hyalella azteca), LC50 (96 h): ca. 126 mg/L 2 (reliable with Besser et al. (1998) Oligochaete (Lumbriculus variegatus), total nitrogen (meas. (geom. restrictions) Midge (Chironomus tentans) mean)) based on: mortality supporting study freshwater LC50 (96 h): ca. 286 mg/L total nitrogen (meas. (geom. experimental result short-term toxicity (laboratory study) mean)) based on: mortality Test material (CAS static LC50 (96 h): ca. 564 mg/L name): ammonium total nitrogen (meas. (geom. chloride mean)) based on: mortality

LC50 (96 h): ca. 117 mg/L total nitrogen (meas. (geom. mean)) based on: mortality

LC50 (96 h): ca. 302 mg/L total nitrogen (meas. (geom. mean)) based on: mortality

LC50 (96 h): ca. 430 mg/L total nitrogen (meas. (geom. mean)) based on: mortality Hyalella azteca LC50 (4 wk): ca. 1.1 mg/kg 2 (reliable with Borgmann et al. sediment dw total NH3 restrictions) (1993) freshwater (meas. (geom. mean)) based on: mortality supporting study long-term toxicity (laboratory study) LC50 (4 wk): ca. 13 mg/L experimental result static total NH3 (meas. (geom. mean)) based on: mortality Test material (CAS Borgmann, U., Norwood, W. P. & name): ammonium Babirad, I. M. (1991). Relationship LC50 (4 wk): ca. 13 mg/L chloride between chronic toxicity and total NH3 (meas. (geom. bioaccumulation of cadmium in

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 212 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Hyalella azteca. Can J. Fish. Aquat. mean)) based on: mortality ScL, 48, 1055-60 LC50 (4 wk): ca. 12 mg/L total NH3 (meas. (geom. mean)) based on: mortality

Discussion

Extensive high quality chronic data were available on 7 different freshwater sediment species and 2 marine sediment species. In addition, two freshwater field studies are available for freshwater sediments. These data were all screened for relevancy to the environment under study.

The following information is taken into account for sediment toxicity for the derivation of PNEC:

Freshwater: chronic toxicity data are available for 7 species. Species mean NOECs used for PNEC derivation range from 201 to 1135 mg/kg dw. The species mean NOEC added range between 146 to 1101 mg/kg dw (after correction for background).

In addition, two field studies are available for freshwater sediments. Liber et al. 1996 have reported an overall NOECecosystem of 725 mg/kg d. w. added zinc. Burton et al. 2005 have observed minor effects on species richness and macroinvertebrate density at concentration of 119 mg/kg d. w. added zinc. Marine water: chronic toxicity data are available for 2 marine species: the amphipod Melita plumulosa with a NOEC reproduction of 730 mg/kg dw and the mangrove Avicennia marina with a NOEC emergence of 250 mg/kg dw. After background correction the NOEC values then become 490 mg/kg dw and 207.1 mg/kg dw for M. plumulosa and A. marina, respectively. No field studies were found for marine sediments.

7.1.1.5. Other aquatic organisms

The results are summarised in the following table:

Table 68. Overview of effects on other aquatic organisms: communities

Method Results Remarks Reference macroinvertebrate communities and NOEC (wk): > 20 — < 27 1 (reliable without Crane M, Kwok families of Ephemeroptera, Plecoptera µg/L dissolved (meas. restriction) KWH, Wells C, and Trichoptera were assessed. (arithm. mean)) based on: Whitehouse P and benthic macroinvertebrate key study Lui GCS. (2007) freshwater structure and insect family richness read-across based on field study with benthic grouping of macroinvertebrates and insects substances (category approach) Spatially matched measurements of benthic macroinvertebrate family Test material richness and measured dissolved metal (IUPAC name): zinc concentrations were compared over (See endpoint two sampling periods spanning all summary for regions from England and Wales. justification of read- across) microcosm/mesocosm NOEC (4 wk): 22.8 µg/L 1 (reliable without Rand GM, Hoang dissolved (meas. (TWA)) restriction) TC, Brausch JM. freshwater based on: Phytoplakton: (2010) community abundance, key study flow-through richness and diversity

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 213 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

The study design and performance was NOEC (4 wk): > 60.4 µg/L read-across based on based on SETAC workshop dissolved (meas. (TWA)) grouping of discussions in Potsdam (EWOFFT: based on: zooplankton: substances (category Hill et al. 1994), Lacanau (HARAP: community abundance, approach) Campbell et al. 1998), and richness and diversity Schmallenberg (CLASSIC: Giddings Test material (CAS et al. 2002), which are summarized name): zinc guidance documents. dichloride (See endpoint summary for justification of read-across) microcosm/mesocosm NOEC (14 wk): 14 µg/L 2 (reliable with Rand GM, Hoang dissolved (meas. (TWA)) restrictions) TC, Brausch JM. freshwater based on: chlorophyta (2012) eveness key study flow-through NOEC (14 wk): 14 µg/L read-across based on The study design and performance was dissolved (meas. (TWA)) grouping of based on SETAC workshop based on: Zooplankton substances (category discussions in Potsdam (EWOFFT: eveness approach) Hill et al. 1994), Lacanau (HARAP: Campbell et al. 1998), and NOEC (14 wk): 21 µg/L Test material (CAS Schmallenberg (CLASSIC: Giddings dissolved (meas. (TWA)) name): zinc et al. 2002), which are summarized based on: chlorophyll a in dichloride (See guidance documents. periphyton endpoint summary for justification of read-across) multispecies test NOEC (18 h): >= 7 — <= 132 (reliable with Davies AG & Sleep µg/L dissolved (estimated) restrictions) JA (1979) saltwater based on: C fixation rate key study static LOEC (18 h): >= 10 — <= 15 µg/L dissolved (nominal) read-across based on Photosynthetic inhibition (C fixation) based on: C fixation rate grouping of test on phytoplancton communities in substances (category the field, designed for dose-response approach)

Test material (IUPAC name): zinc (See endpoint summary for justification of read- across) Species of macro-algae, crustaceae, No Observed Ecological 1 (reliable without Foekema EM, sponges, mollusca and annelids were Adverse Effect restriction) Kramer KJM, Kaag introduced. Zoo- and phytoplankton Concentration (83 d): 12 NHBM, Sneekes and other macro invertebrates were µg/L dissolved (meas. key study AC, Bierman S, introduced with the water and (TWA)) based on: primary Hoornsman (2012) sediment. production, grwoth of read-across based on Littorina littorea grouping of saltwater substances (category approach) static Test material (CAS Custom-designed study. Each name): zinc mesocosm study is designed to answer dichloride (See specific questions; nonetheless various endpoint summary guidance documents that describe the for justification of basic principles of this kind of studies read-across)

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 214 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 when performed for risk assessment Form: powder were used as general guidance. Although these guiding documents concern fresh water mesocosm studies, the addressed issues were taken into account as much as possible during the marine mesocosm study that is described in this report. natural phytoplancton communities NOEC (24 h): 1.45 µg/L 2 (reliable with Wolter K, U. from field dissolved (meas. (not restrictions) Rabsch, P. specified)) based on: C Krischker and A. G. saltwater fixation rate (growth) supporting study Davies (1984) static LOEC (4 h): > 100 µg/L read-across based on dissolved (meas. (not grouping of Field experiment on natural specified)) based on: C substances (category phytoplancton communities looking at fixation rate approach) C fixation rates as the endpoint, designed for dose-response Test material (IUPAC name): zinc dichloride (See endpoint summary for justification of read-across)

7.1.2. Calculation of Predicted No Effect Concentration (PNEC)

7.1.2.1. PNECwater: freshwater

All adequate chronic data on fish, invertebrates, algae and plants were considered together in a species sensitivity distribution (SSD), and the PNEC was calculated by means of statistical extrapolation, using all available chronic NOEC values as input. The database is indeed sufficiently large and answers the basic requirements to use an SSD, since it covers the required 8 different taxonomic groups and > 10 test organisms.

Since the log-normal distribution significantly fits the data, this distribution was used for the SSD (like in the RAR). Other conditions to apply statistical extrapolation were also met (see discussion below on the safety factor to be applied to the HC5.

Because of the inclusion of 6 additional species, the species sensitivity distribution (SSD) that was calculated for the present analysis is slightly different from the one of the RAR (2008).

Discussion on the assessment factor to be applied on the HC5 for PNEC derivation. Based on uncertainty considerations an assessment factor between 1 and 5 should be applied to the 50% th th confidence value of the 5 percentile value (thus PNEC = 5 percentile value/AF). The AF is to be judged on a case by case basis. The following considerations are made on the uncertainty around the HC5 and for determining the size of the assessment factor:  The chronic NOEC database of 23 species entries covers more than the requirements for taxonomic groups (8) and species (at least 10, preferably more than 15) set out in the guidance. Based on this, there is no need for assessment factor higher than 1.  The large number of species in the SSD results in a low uncertainty on the HC5 value, as is shown by the small difference between the 50% confidence level and the 95% confidence limits found for the lognormal distribution which is less than a factor of 2.5. Based on this, there is no need for an assessment factor higher than 1.  The lognormal distribution that was used for PNEC derivation (in spite of other distributions giving a better fit to the data) resulted in an HC5 of 20.6 µg/l, which is markedly lower than the HC5 value

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 215 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

calculated from the extreme values distribution (27.2mg/l), which provided the best fit. So the distribution that is used for the PNEC derivation gives a conservative HC5 value. Based on this observation, there is no need for an assessment factor higher than 1.  The chronic data are from tests in a variety of natural freshwaters, covering a considerable part of the wide range of freshwater types and freshwater characteristics (pH value, hardness and background zinc concentration) that are found in European freshwaters. Characteristic ranges for given abiotic conditions (pH, hardness, zinc background of culture and test media) were defined in the EU RA, and applied also in the present analysis. Data obtained outside these boundaries, were not used, in accordance to the RA (ECB 2008). Therefore, the data properly reflect the range of abiotic conditions found in the European aquatic compartments. Based on these observations, there is no need for an assessment factor higher than 1.  There are general indications that the bioavailability of metals under real life conditions is lower than the bioavailability in laboratory toxicity tests. On the one hand this is taken into account by comparing the dissolved concentrations at both the PEC and PNEC side. On the other hand, the dissolved fractions under real-life environmental conditions may contain DOC and other complexing agents that are absent in laboratory tests. This complexxation further reduces the bioavailability of zinc. In the chronic NOEC dataset, tests performed in laboratory waters without DOC are the majority. As a consequence, the PNEC derived from these data is conservative, as compared to the natural environment, where DOC is always present. In the risk characterisation, the bioavailability related to DOC and other abiotic factors is being taken into account in the (conservative) bioavailability factors that are applied on the PEC. Based on this observation there is no need for an assessment factor higher than 1.  Some specific NOEC values of the database are below the HC5: 15 of the 25 values for the alga Pseudokirchneriella subcapitata and one of the 13 values for the crustacean Ceriodaphnia dubia. The “species mean” NOEC value for P. subcapitata (19 µg/l) is slightly lower than the HC5 (20.6µg/l); the one for C. Dubia (37 µg/l) is above the HC5. On statistical considerations, the chance of having a value below the HC5 is significant when the SSD includes > 20 data points. So, having one or more values below the HC5 is inherent to bigger datasets and is not an issue as such. o When the data on P. Subcapitata are considered in more detail, it becomes obvious that all 15 tests with P. subcapitata that resulted in a NOEC below the HC5 value (all from the study by De Schamphelaere et al., 2003) were performed in artificial test water with a very low DOC concentration, while DOC was is an important mitigating factor for the toxicity/bioavailability of aquatic organisms and this algal species in particular. So, these distinct low values can be explained by the artificial conditions of the laboratory test media and have limited significance for the situation in the real environment. For algae, there is information on another species (Chlorella sp.) which demonstrates a rather average sensitivity to zinc (NOEC: 50 µg/l). So, there is no indication that algae as a taxonomic group are particularly sensitive to zinc. o There is one NOEC value (14µg/l) for Ceriodaphnia dubia that is below the HC5 value. The organisms in this test were cultured in "Little Miami river water". The characteristics of this water are not specified in the paper, but the river’s zinc content is documented in Shiller & Boyle (1985) to be very low, i.e. < 1 µg/l (0.85 µg/l dissolved zinc). The observed low NOEC can thus be explained by the fact that the organisms were cultured and conditioned under low zinc conditions, lower than those accepted in the RAR to be relevant for the EU. Such pre-test culturing conditions may indeed induce a level of zinc sensitivity, which is not relevant for EU waters. o Thus, since the few NOEC values observed below the HC5 can be explained by the artificial test conditions (deviating from the natural environment), by very low zinc background in the test and culture medium, and further by the intrinsic characteristics of the 5% value when the SSD has a higher number of data, this consideration does not justify application of an additional assessment factor.  The RAR did reject a few results obtained under very low background conditions, but only when the tests were performed in artificial test media; so, the chronic NOEC database included still NOECs obtained in e.g. North American natural waters with very low zinc background. For reasons of consistency, these data are also used for the present analysis. There are 2 groups of data to be mentioned in this respect: o data obtained in Great Lakes waters. Documented total zinc background levels for Great Lakes waters are indicated to be “around” 2 µg Zn/l or lower. The corresponding soluble zinc background is probably in the order of 1 µg Zn/l or only slightly lower, considering that in these waters the dissolved zinc background has been estimated as being the major fraction of the total (Parametrix, pers. communication), and/or applying the Kd and default suspended

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 216 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

matter content of EU waters (15 mg/kg DW), which would yield a soluble zinc background of 0,75 µg/l. o Stronger reservations remain however on the relevancy of the 2 NOEC entries obtained in “New River” water. This river has been identified as being not representative for the EU waters, because of documented very low background zinc concentration of 0,35 µg/l (Shiller & Boyle 1985 - see detailed discussion on the relevancy of the “New river” below under section “mesocosm studies”). For reasons of consistency with the RAR database, the data are included, but with strong reservation. It has been demonstrated that ecotoxicity results obtained under very low zinc background show consistently a higher sensitivity of organisms to zinc than is observed for organisms cultivated under conditions relevant for the natural background in EU waters (Muyssen and Janssen 2002, Muyssen and Janssen 2005). The inclusion of these data into the NOEC database thus introduces a significant level of conservatism into the SSD and the PNEC derivation for EU waters. This additional level of conservatism added to the PNEC analysis should be considered when evaluating the protective power of the HC5 arising from the generic SSD. It provides no justification for application of an assessment factor higher than 1, rather on the contrary.

 For zinc, a chronic Biotic Ligand Model (BLM) is available that allows to predict the aquatic zinc toxicity occurring under realistic worst case conditions for EU waters. These realistic worst case conditions have been defined in the RA (ECB 2008). With this model, it can be demonstrated that the NOECs predicted for the realistic worst case conditions of EU waters, correspond well with the generic species mean NOEC values observed for the BLM organisms and used in the SSD (see table below). This demonstrates that the generic NOECs and HC5 derived from the SSD correspond to the realistic worst case bioavailability conditions of EU waters, and are thus conservative.

Table 69. Comparison of NOECs normalised to realistic worst case conditions and the measured NOECs from the generic SSD 3 4 Species NOECgeneric NOECrwc NOECrwc/NOECgeneric O. mykiss 146 184 1,26 D. magna 98 86 0.88 R. subcapitata 19 21 1.11 Average =1.08

There are several explanations for this observation:  The generic SSD contains a number of data that were obtained in artificial waters, which are, in terms of bioavailability, very worst case waters, e.g. they contain no DOC or any other complexing agent, which will always be present in natural waters and will reduce bioavailability. This is e.g. the case for the algae database (most of the distinct entries obtained in artificial water).  The artificial laboratory waters are also often characterised by worst case conditions related to other

parameters such as hardness (e.g. the OECD algae test is performed at 24 mg CaCO3/l, which is the lower boundary of relevance for the ecotoxicity values), and general low nutrient status. Among the invertebrates, the low NOECs observed on Ceriodaphnia dubia are N-American studies performed in river water characterised by very low Zn background (< 1µg/l); conditioning of Daphnids to Zn background, resulting in higher sensitivity after culturing in low zinc background medium, has been documented (Muyssen and Janssen, 2002).  Most of the fish data of the generic NOEC database obtained in natural waters are N-American studies, performed in water from the N-American Great Lakes. These waters are characterised by low

3 Species geometric means including all NOECs from the conclusion (i) programme on zinc bioavailability with pH between 6-9 and hardness between 24 and 250 mg CaCO3/l, also including data obtained in natural waters.

4 Realistic worst case (rwc) following from GEMS-B database (Heijerinck et al 2003). All organisms: 10th percentile of DOC (2,0 mg/l); D. magna and O. mykiss: 10th percentile of inorganic parameters (including pH (7.36) and hardness (42 mg CaCO3/l); P. subcapitata: 90th percentile of inorganic parameters (including pH (8.24) and hardness (308 mg CaCO3/l)

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 217 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

background zinc level (around 1µg/l and lower), low hardness (around 40-50 mg CaCO3/l)) and also low DOC.  From these observations, it follows that the NOECs of the generic SSD are for the greater part obtained under conditions that maximise zinc bioavailability or the sensitivity of organisms to zinc, which explains why the generic SSD corresponds to the SSD calculated for realistic worst case EU conditions. Based on this observation there is no need for an assessment factor higher than 1.

 Mesocosm data. The RAR (2008) mentions a number of field studies and laboratory mesocosm studies, published in the open literature and discusses them in relation to the protective characteristics of the HC5. Before considering these studies in the light of the derived HC5, the test conditions under which these tests were performed were checked for their quality and relevancy for the EU environment and the generic HC5. Indeed, the single species test results were strictly evaluated in the RA (ECB 2008) for quality and relevancy for the EU environment by checking the conditions under which the tests were performed against well-defined criteria of water conditions. As such, a number of test results were rejected in the RAR, even when they were obtained under only slightly deviating conditions and even when performed in natural water (ECB 2008). Consequently, before using results from mesocosm studies for checking the protective capacity of the HC5, these data should also be checked against the same criteria for relevancy and with the same rigour. At the same time, a number of specific quality criteria have been developed for mesocosm studies (Van Leeuwen et al 1994) which should also be applied to these studies before their use. Apart from criteria that are similar to those applied to assess the quality of single species tests (e.g.: clear dose-response, exposure concentration checked during test, description of statistics), it is notably important for these tests that real replicates are used i.e. different parallel set-ups of a given treatment. If this criterion is not met, the test is considered insufficient quality (Q3). It is noted that such rigorous check for relevancy and quality was not made in the RAR. As a result, results of mesocosm studies were considered in the RAR which were not relevant for the generic PNEC: they were indeed obtained under conditions that deviate from the relevancy criteria, set for the single species tests. In this respect, it is noted that the RAR derived a specific PNEC for waters with very low hardness (<24 mg CaCO3/l). This PNEC”softwater” is factor 2.5 lower than the generic PNEC for Europe (see below). Hardness of the test medium is thus a key aspect to consider when assessing the relevancy of test data, both from single species tests and mesocosms. Several of the studies mentioned in the RAR were also performed on N-American waters with a documented zinc background that was much lower than the one, considered representative of EU waters (< 1µg dissolved zinc/l). In agreement with the approach followed in the RAR (and the present analysis) for the single species test results obtained under such conditions, these mesocosm studies were also considered not relevant for the generic PNEC in the present analysis.

The outcome of the relevancy/quality check of the mesocosm studies discussed in relation to the protection level of the HC5 are summarised in table below. Based on this check, it is indicated if they are of sufficient quality and if they are relevant for the generic or softwater PNEC discussion.

Table 70. Summary of Mesocosm studies reported in the risk assessment study Conditions of test/remarks Relevancy for PNEC Reported discussion NOEC/LOEC (µg Zn/l) Marshall et al Natural Zn background 0,6 µg/l Not relevant for LOEC : 16 (dissolved: 1983 -no data on water physico- generic EU-PNEC according to the RA chemistry reported; according to (ECB 2008), “a reliable the RA (ECB 2008), “a reliable LOEC/NOEC could not LOEC/NOEC could not be be derived”) derived” Belanger et al Performed in natural zinc Not relevant for NOECgrowth: 25 1986 background of 0.35µg/l generic EU-PNEC NOECsurvival : 500 (nominal) Genter et al 1987 Performed in natural zinc Not relevant for LOEC: 50 (nominal) background of 0.35µg/l generic EU-PNEC Farris et al 1989 Performed in natural zinc Not relevant for LOEC: 34 (actual background of 0.35µg/l generic EU-PNEC concentration)

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 218 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Farris et al 1994 Performed in natural zinc Not relevant for NOEC: 25 (nominal) background of 0.35µg/l generic EU-PNEC Niederlehner & Organisms were grown in water Sensitivity of 73 (actual, dissolved; Cairns 1983 with (too) low hardness (13 mg organisms related to multi-species-LOEC) CaCO3/l). softwater  data relevant for PNEC softwater Pratt et al 1987 Organisms were grown in water Sensitivity of 4.2 (actual, dissolved with (too) low hardness (13 mg organisms related to LOEC for total biomass CaCO3/l). softwater  data (DW); -No replicates  Q3 study would be relevant for 4.2 (actual NOEC for PNEC softwater, but algal biomass); -Q3 study 11 (actual multi-species NOEC) Paulson et al 2000 Test in low water hardness (23 mg Not relevant for NOECs*: 18,3 for dry CaCO3/l), and under P-deficiency. generic PNEC due to weight, chlorophyll a, Authors attributed effects to zinc too low hardness and photosynthetic activity, induced P-deficiency in this low P- P-deficiency of bacterial activity. water testwaters - 40.5 for species richness - 98 fo species composition * NOECs recalculated by Van Sprang et al (2009) Gächter 1976 Test in 2 pre-alpin lakes. Q3 study 14 (total nominal multi- Test of insufficient quality (Q3): species phytoplankton -actual concentrations not NOEC) measured, -results graphically 11 (Total nominal multi- presented over the year (no dose- species phytoplankton response reported); NOEC) -no replicates, -no statistics. Clements 2004 Macro-invertebrate communities Relevant for PNEC 502 (total, measured) generic NOEC for abundance, structural endpoints

These studies are extensively discussed in the RAR (ECB 2008). In addition to the evaluation in the RAR, the following points are made:  New river water studies. A number of the model ecosystem studies on periphyton, zooplankton and invertebrates were performed in New River water (Belanger et al 1986, Genter et al 1987, Farris et al 1989, Farris et al 1994). The New River is part of the Ohio river basin, which was studied in detail by Shiller & Boyle (1985) for dissolved zinc concentrations. The New river is among the tributaries located south of the Ohio, which are characterised by very low (<0,4µg/l) zinc concentrations. Actually, the New River is also called the Kanawha river after its junction with the Gauley river (http://www.wvexp.com/index.php/New_River#Maps: “New River is formed by the union of its north and south forks in Watauga County, North Carolina (NC). From this junction, the New River flows northeastward across the state line into Virginia, and at Radford, VA, turns abruptly northwestward across western Virginia and West Virginia to its junction with Gauley River, in Fayette County, just above Kanawha Falls, in Kanawha County. Below the junction with the Gauley, the river is known as Kanawha River”). The Kanahwa river has a documented zinc concentration of 0,35 µg/l (Shiller & Boyle 1985). During the discussions on the RAR (2008), it was not known that New River gave into Kanawha river; therefore the range observed for all tributaries of the Ohio (0,3-3µg/l) was applied to the New River, and the argument on zinc background was not considered for relevancy of the studies done in New River water. However, the higher values (up to 3µg/l) are reported for the tributaries located north of the Ohio, which are assumed to be influenced by antropogenic zinc input (Shiller & Boyle 1985). The New River is part of the tributaries located south, which are categorised as being more pristine (Shiller & Boyle, 1985)) and all show a very low zinc content. This observation has now been confirmed by identifying the relationship between Kanawha (documented zinc) and the New River. In conclusion, the studies performed on New river water are, according to the relevancy criterion

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 219 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

of background Zn > 1 µg/l agreed in the RAR (2008), not relevant for the EU freshwater environment, and thus not for the generic PNEC. A similar lack of relevancy can be attributed to the Marshall et al (1983) data, which were also obtained under conditions of very low background.

From the overview presented here, it is concluded that very limited reliable and relevant information is available from mesocosm studies in the open literature, for consideration of the generic HC5.

Van Sprang et al (2009) also discussed in detail the available evidence on mesocosms in the zinc risk assessment, and investigated the protective capacity of the single species HC5 value towards effects in aquatic mesocosms after normalisation of the single-species NOEC values to the abiotic conditions occurring in de mesocosm experiments. These authors mention also several issues related to the quality and relevancy of the studies, and conclude that “it is difficult to draw definitive conclusions as to whether or not the HC5 is conservative enough to protect ecosystems”. They recommended that a) additional exposures of multi-species systems (or model ecosystems) to zinc should be carried out to get to more reliable conclusions, and b) that special care should be taken in the design of the studies to avoid the problems related to those available in the literature (Van Sprang et al 2009).

A large scale chronic mesocosm/microcosm study has been performed with a study design and performance that strictly followed established protocols for microcosm systems. Control treatments were included for all statistical comparisons and characteristics of sediments and water composition (background and treatments) were analytically verified. Dissolved zinc concentrations were monitored regularly and were maintained by spiking treatments every four days (nominal concentrations of 8, 20, 40, 80, and 160 μg/L.

The study was comprising:  a spectrum of species of different taxonomic groups and trophic levels,

 all life stages of the included organisms,

 realistic exposures, with three replicates for each treatment.

 a food web including indirect effects due to competition or predation, and

 ecosystem function endpoints, and the study had an exposure time of 14 weeks. At the time of registration (October 2010), the evaluation had been reported for up to 4 weeks exposure (Rand GM, et al, 2010). Based on these first (28 days) results, it could be provisionally concluded that effects on community abundance, richness and diversity showed no relevant direct effects (NOEC) up to 22.8 μg/L for phytoplankton and >60.4 μg/L for zooplankton, following four weeks of exposure. These provisional results thus suggested that the HC5 as derived in the 2010 analysis was protective for microcosm/mesocosm scale communities.

After the registration, the full study results were reported (Rand et al 2012). The microcosm NOEC was set at 14 µg Zn/l, based on effects on the most sensitive taxonomic group, phytoplankton..

Related to this NOEC, the following considerations can be made: In spite of efforts to keep original conditions, a significant deviation towards high pH (>pH 9) was observed during the test. This effect has been observed in other microcosm systems, and is important for the results of the zinc microcosm study. Indeed, the microcosm NOEC was set at 14 µg Zn/l, based on effects on the most sensitive taxonomic group, phytoplankton. It is noted in this respect that unicellular algae are also the most sensitive species in the SSD (Pseudokircherniella subcapitata), and that the sensitivity of these algae to zinc is highly pH-dependent (figure below), in a sense that toxicity is highest at higher pH.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 220 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Figure 5. NOEC of algae (µg Zn/L) modelled with the Biotic ligand Model (BLM;Heijerick et al. (2005) for different pH and Ca combinations with DOC of 1 mg C/L.

The BLM-model that was developed for the EU waters does not cover the high pH values of the microcosm study; however, high sensitivity of algae can be assumed at this pH range. Due to this factor, the microcosm study can be considered as very conservative. In addition, the temperature during the microcosm study was around 30°C. Kwok et al (2007) observed a higher sensitivity of tropical species (measured at a median T of 25.8°C) for zinc, than temperate species (measured at median T of 20°C). The HC10 ratio temperate / tropical species in this study was 2 (Kwok et al 2007). This observation also suggests that the microcosm study was performed under sensitive conditions.

The PNEC derived for the REACH scenarios is a generic PNEC, i.e. based on the HC5 of the species sensitivity distribution (SSD) as following from the reported ecotoxicity data, not corrected for bioavailability. This corresponds with the generic nature of the exposure scenarios that are described under section 9. Normalising this SSD to very sensitive conditions (e.g. very low (1mg/l) DOC) gives HC5 values as low as 10- 12µg/l; DOC values of 3mg/l combined with low pH (<6.5) and Ca (10mg/l) give a HC5 of 13µg/l. It is noted that the generic character of the PNEC for water used in the present analysis does not affect the assessment of the risk related to the emissions to the aquatic environment, since the driver for this assessment is the sediment PNEC, not the water PNEC (see exposure scenarios section 9).

The above analysis on the SSD-related HC5 of the present report indicates that the microcosm NOEC, which was observed under very sensitive conditions, concurs with the predicted HC5 of the SSD under sensitive conditions. In conclusion, the microcosm study supports the protective character of the SSD-derived HC5/PNEC derived in the present analysis.

Evidence from freshwater field survey data As a last, important element in this discussion, an extensive field study has come available after the closure of the risk assessment. This study enables to check the protective capacity of the HC5 for the real aquatic environment at an even higher level.

Recently, (after the closure of the EU RAR), an extensive large scale field study was performed on UK waters to evaluate the protective capacity of water quality standards for metals (Crane et al 2007). The survey checked the capacity of water quality standards (the PNEC from the EU RA for zinc), for protecting biodiversity of benthic invertebrate communities, which are known as being sensitive to metals (Crane et al 2007). Matched metal concentrations and benthic invertebrate field monitoring data were compared for a significant number of sites (291 for zinc). The conclusion of the study was that the PNEC for zinc as derived from the risk assessment, was overly protective with about a factor 2. The survey indeed showed that the PNEC for zinc should be in the range 20-27µg Zn dissolved/l (Crane et al 2007).

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 221 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

This extensive piece of field evidence is an important additional element in the discussion on the safety factor to be applied on the HC5, that was not available at the time of the closure of the RAR. Its outcome does not support the application of the additional assessment factor of 2 as applied in the RAR.

PNECadd freshwater: conclusion The assessment of the freshwater PNECadd for zinc is largely based on the chronic aquatic toxicity data that were presented in the EU RA on zinc (ECB 2008). From the knowledge available at the time of closure, the RAR derived a PNEC using an application factor of 2. This factor was however subject of significant debate, as the European Scientific Committee for Health and Environmental risks (SCHER) concluded in its opinion, following the approval of the RA in 2007: “SCHER recognises that the selection of this factor is a matter of judgement and is not based on definitive scientific evidence” (SCHER 2007). The present analysis contains, as compared to the knowledge available at the closure of the RA, significant additional information, notably: -additional high quality NOEC values for 7 additional species in the SSD -results from a highly relevant extensive mesocosm/microcosm study and -results from a large scale field study checking ecological biodiversity against ecotoxicity information in the real environment

The introduction of these major new elements in the uncertainty analysis that were not present at the time the RA was concluded, form the basis for the present update of the freshwater PNECadd. Taking into account the elements discussed above, it is considered that application of an additional assessment factor on the HC5 following from the SSD is not justified by the evidence. Consequently, the PNEC is set at the level of the HC5 which is considered as protective for EU freshwater ecosystems: PNECadd freshwater: 20.6 µg (dissolved) zinc/l.

PNECadd for freshwaters with very low hardness. In the RAR, a specific “softwater” PNEC was derived for zinc in waters of very low hardness (<24 mg CaCO3/l). This PNEC was derived using a water effect ratio (WER) approach, based on experimental data (RAR 2008). The WER for generic waters/softwaters was found out to be 2.5 (RAR 2008). Accordingly, the “softwater” PNEC in the present analysis can be set at 20.6 µg Zn/l/2.5) = 8.2 µg Zn/l. Comparison with the results of reliable mesocosm studies performed in waters of very low hardness (table 70) confirms that this PNEC softwater is protective.

In the RAR, a number of studies were rejected for use in PNEC derivation (RAR 2008). These studies were rejected after detailed consideration by the Rapporteur and TCNES (Table 3.3.2.a, Part II from the RAR) (ECB 2008). These studies have the annotation Q(uality) or R(elevancy), indicating the reason for not using them. With respect to the annotation R, the specific relevance criterion or criteria used to reject the study can be derived directly from the reported items in Table 3.3.2.a, Part II from the RAR, for example the lack of data on pH and/or hardness in an artificial test medium, or a hardness value that is below the minimum value used as selection criterion. In some cases the reason(s) for not using a study cannot be derived directly from Table 3.3.2.a, especially in case of the annotation Q. In all cases, specific information on the reason(s) to reject a study can be found in the footnotes of Table 3.3.2.a in the RAR (ECB 2008). When a study is used for PNEC derivation despite it does not meet all Q and/or R criteria, specific information can be found in the footnote as well. The footnote also provides additional information, on e.g. the culture and test conditions (including background zinc concentrations), if such information was reported by the original study

Bioavailability considerations The EU risk assessment discussed in detail the evidence on bioavailability of zinc in freshwater and concluded that the consideration of bioavailability was an integral and essential part of the procedure to assess risks for zinc in water: ”The PNECs for zinc metal have been derived solely for the purposes of this risk assessment. They must not be used for other purposes, such as setting environmental quality standards or sanitation levels, without further in-depth consideration as to whether they are fit for that purpose. In every case the bioavailability correction, which has been used in the present RAR, should be incorporated as an essential part of the process.”(EU RA zinc, overall conclusions, ECB 2008).

There are 2 ways of quantifying zinc bioavailability in water, and integrating it in the risk assessment:  In the RA, a procedure for quantifying and integrating bioavailability of zinc in water was described, called the “BioF” approach. In this system, biotic ligand models that were developed for 3 species (for

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 222 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

algae (Pseudokircherniella subcapitata), invertebrates ( Daphnia magna) and fish (Oncorrhynchus mykiss, respectively) are used to predict the toxicity of zinc to these organisms, under the given physicochemical conditions of the water. The resulting toxicity levels are compared to the toxicity, observed for these organisms under maximum (reference) bioavailability conditions. The ratio between both toxicity values gives the bioavailability factor, i.e. the percentage of zinc available under the given water conditions. The BioF is subsequently used to correct the PEC for getting the bioavailable PEC. For reasons of conservatism, the BioF is set at the level of the model giving the highest bioavailability of the 3 (for details see the zinc RA, ECB 2008). An excel spreadsheet calculator has been developed, applying the principles agreed in the risk assessment, that allows for a calculation of the bioavailability related to a given local combination of abiotic conditions in the water (go to: http://www.reach- zinc.eu/). In absence of specific local data, regional data e.g. whole river data, can be used for making the calculations. However, preference is given to specific data for this analysis.  Since the closure of the RA, the science supporting the BLM models has been expanded to a level, where it is now possible to normalize the whole of the species sensitivity distribution (SSD) towards the conditions of a given water. This system now allows to calculate a BioF based on the ratio between the HC5 of a reference SSD (obtained under conditions maximizing zinc bioavailability) and the HC5 of the SSD, obtained after normalising all species data towards the conditions of a given water. Also for this more advanced approach, an excel calculator has been developed (which is combines BLM models for Zn, Cu and Ni, and gives BioFs for these 3 metals) that can also be found on : http://www.reach-zinc.eu/.

Both approaches for calculating zinc bioavailability in freshwater are using the same 3 physico-chemical parameters that drive the bioavailability of zinc in water, i.e.: DOC, pH and hardness. It is emphasised that the BLM models mentioned above work are only valid for freshwater and are only applicable for calculating bioavailability work within given boundaries of these parameters (covering the vast majority of physchem conditions of EU waters). These boundaries are indicated in the excel calculators.

7.1.2.2. PNECwater: marine

Discussion on the assessment factor to apply on the HC5 for PNEC derivation

Based on uncertainty considerations, an assessment factor (AF) between 1 and 5 should be applied to 5th percentile value of the species sensitivity distribution curve at 50% confidence value (thus PNEC = 5th percentile value / AF). The AF is to be judged on a case by case basis.

As a minimum, the following points have to be considered when determining the size of the assessment factor: 1. the overall quality of the database and the endpoints covered, e.g., if all the data are generated from “true” chronic studies (e.g., covering all sensitive life stages); 2. the diversity and representativity of the taxonomic groups covered by the database, and the extent to which differences in the life forms, feeding strategies and trophic levels of the organisms are represented; 3. knowledge on presumed mode of action of the chemical (covering also long-term exposure). Details on justification could be referenced from structurally similar substances with established mode of action; 4. statistical uncertainties around the 5th percentile estimate, e.g., reflected in the goodness of fit or the size of confidence interval around the 5th percentile, and consideration of different levels of confidence (e.g. by a comparison between the 5% of the SSD (50%) with the lower confidence interval on the 5% of the SSD); 5. comparison between field and mesocosm studies, where available, and the 5th percentile and mesocosm/field studies to evaluate the laboratory to field extrapolation.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 223 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Based on the available chronic NOEC data for the marine environment, the following points were considered when determining the size of the assessment factor:

1. The overall quality of the database and the end-points covered, e.g., if all the data are generated from “true” chronic studies (e.g. covering all sensitive life stages); The marine Zn-database covered only ecological relevant endpoints. The selected endpoints were all relevant for potential effects at population level: mortality, reproduction, development (including e.g. fertility of sperm cells, metamorphosis), and growth. All data were carefully screened for quality and representativity, and only adequate data were used for the PNEC derivation. ‘Chronic’ exposure times or relevant exposure periods for sensitive life-stages are also achieved for the nine taxonomic groups covered in the Zn database. The exposure times for algae were between 2 days to 10 days on zoospores. For the invertebrates, exposure times went from a few hours on very sensitive life-stages (e.g. sperm cell toxicity tests on echinoderms) up to 9 months (reproduction tests on annelids), and for fish, an exposure time of up to 27 days on fertilized egg-early life stage test was reported. Sensitive life stages were covered in the database. Fish experiments were performed with fertilized eggs. Chronic tests with crustaceans/molluscs/nematods were initiated with fertilized eggs, larvae or juveniles, while tests with echinoderms were performed with fertilized eggs only. From the chronic tests using polychaetes, young adults or adults were used mainly over a very long exposure time (usually several months). As for the algae, diatom tests were reported to be sometimes more sensitive than tests performed on young life stages, i.e. zoosopores.

2. The diversity and representativeness of the taxonomic groups covered by the database; The figure below illustrates the marine biodiversity in terms of percentage of species across the various phyla (ECETOC, 2001). The Molluscs group is the largest taxonomic group in marine waters, followed by the Crustaceans group (ECETOC, 2001). These groups are the ones for which a large number of toxicity data are available for zinc and so both groups were found to be broadly represented in the marine zinc effects database. The database also includes typical marine groups such as echinoderms and cnidarians. In addition to the inter- taxonomic diversity, the zinc marine database covers in each group a number of families which reflects a high level of intra-taxonomic diversity (see e.g. the group of annelids and echinoderms).

Figure 6. Species diversity in the marine environment (from ECETOC 2001). The stars highlight taxonomic groups represented in the zinc marine database.

The algae database contains macro- and micro-algae data. Data exist for the three representative groups in macro-algae, which are the green, brown and red macro-algae. Micro-algae are represented by diatoms.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 224 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Dinoflagellates data were also found but are reported in the Q3 database. Reported values for dinoflagellates indicate that this group is rather insensitive towards zinc. This database contains chronic toxicity for one fish species. However, the Q3 database contains additional information on fish data, including acute toxicity which indicates that fish are less sensitive to zinc than invertebrates or algae. From the extracted data, the Zn-database does fulfil largely the requirement of 10-15 different NOEC values (48 individual NOEC values resulting in 39 different species NOEC values).

3. Statistical uncertainties around the 5th percentile estimate, e.g., reflected in the goodness-of-fit or the size of confidence interval around the 5th percentile; A log-normal distribution was calculated and accepted at all significance levels. A factor of 1.7 was observed between the lower confidence limit of the HC5 and the 50th percentile of the HC5 (HC5-50). The Weibull distribution presented a slightly lower A-D test value meaning that there is a slightly better fit to the input toxicity data.

4. Comparisons between field and mesocosm studies and the 5th percentile to evaluate the laboratory to field extrapolation. The available mesocosm data demonstrate that the no effect concentration level observed for phytoplankton communities is higher than the HC5 value calculated from the lognormal distribution.

5. NOEC values lower than the HC5-50 The only NOEC value that is slightly lower than the HC5-50 is observed for the mysid shrimp, Holmesimysis costata (NOEC mortality = 5.5 µg zinc/L; Hunt et al. 1997). However, the other NOEC values within the crustacean group where in the range of one to two orders of magnitude higher indicating that this group is not particularly sensitive to zinc. Moreover, if there is a high number of data in the SSD the chance of having a NOEC which is similar or below the HC5 is significant. This surely applies to the marine SSD counting 39 entries.

Conclusion on the Assessment Factor (AF)

The following considerations are made on the uncertainty around the HC5:  The chronic NOEC database is extensive and contains 39 species entries that cover much more than the requirements for taxonomic groups (8) and species (at least 10, preferably more than 15) set out in the guidance;  no justification to put an AF  Sensitive life stages or long chronic exposure periods (a few months) are represented in each taxonomic group as set out in the guidance;  no justification to put an AF  The large number of species in the SSD results in a low uncertainty on the HC5 value, as is shown by the small difference between the 50% confidence level and the 95% confidence limits found for the lognormal distribution: less than a factor of 2.5;  no justification to put an AF  The lognormal distribution that was used for PNEC derivation resulted in an HC5 of 6.09 µg/l, which is markedly lower than the HC5 value calculated from the Weibull distribution (8.5 µg/l), which provided the best fit. So the HC5 that is used for the PNEC derivation is a conservative value;  no justification to put an AF  The HC5 value from the log-normal SSD is protective for mesocosm data;  no justification to put an AF There is no indication for a particular sensitive group in the SSD. In addition, whenever an SSD includes > 20 data points, the chance of having a value below the HC5 is significant. So, having one or more values below the HC5 is inherent to bigger datasets and is not an issue as such;  no justification to put an AF>1.

Based on these observations, there is no need for an assessment factor higher than 1. The PNEC saltwater for zinc is thus set at 6.1 µg/L.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 225 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Table 71. PNEC aquatic Value Assessment Remarks/Justification factor PNEC aqua – freshwater (µg/l) 20.6 1 Evidence does not support assessment factor >1, see text

PNEC aqua - marine water (µg/l) 6.1 1 Evidence does not support assessment factor >1, see text

PNEC aqua – intermittent releases Not relevant (µg/l)

7.1.2.3. PNEC sediment

In the RA on zinc, PNEC add was derived for sediment using a single Hyallela NOEC divided by 10. Insufficient data were available to use SSD. The EU SCHER committee in its subsequent opinion (SCHER 2007) noted “that there are some inconsistencies and lack of transparency in the RARs that lead us to question the PNEC (sediment) that was used. Bounded NOECs are available for three species. There are several values for survival, growth and reproduction of H. azteca: a high value from a study by Borgman and Norwood (1997) – ultimately rejected by the RAR because of very high background and a study by Farrar and Bridges (2003) at 900 mg/kg. The choice of the application factor of 10 was based on lab studies for 3 species, of which Hyalella has the highest sensitivity. But several field studies which included multi-species and multiple endpoints were also available. One by Liber et al (1996) reports a NOEC of 725mg/kg but is not used in the RAR because “minor” effects were observed at all test concentrations. Another study by Burton et al (2003) reports effects at all test concentrations but is not considered appropriate for use in the risk characterisation since the studies were not designed to give a NOEC and PNEC. SCHER has not been able to look at all the original reports used in the RARs – because despite several requests they were not forthcoming - but we are persuaded that there should be a serious reconsideration of the endpoints and the application factor used in the analysis and hence the PNEC. We have a sense from the weight of evidence that it is currently too low.” The above statement from SCHER underlines the uncertainty, related to the setting of the PNECadd sediment in the EU RA. Significant effort has been done therefore to generate more data to the issue. That has resulted in additional sediment toxicity data have become available after the closure of the RA, and that make e.g. the use of a statistical approach possible. Deriving the PNEC

The toxicity of zinc to freshwater benthic organisms was evaluated to develop the PNECadd, sediment. The ecotoxicity database for freshwater sediment of the EU risk assessment was updated and used to estimate an HC5 (concentration estimated for the 5th percentile of the distribution) by statistical extrapolation using accepted regression methods (log-normal distribution). In addition, ‘best-fit’ regressions were also evaluated for comparison. The estimated PNECadd, sediment value was considered alongside published studies regarding 1) consensus-based sediment quality guidelines for threshold effect concentrations, 2) a world-wide compilation of sediment quality values and background levels for metals, 3) field/colonization studies regarding zinc toxicity in benthos, and 4) background concentrations reported in the Zinc RAR. Moreover, alternative approaches for estimating PNECadd, sediment values (Equilibrium Partitioning [EqP] and Assessment Factor [AF]) were evaluated for comparison.

Deriving the PNEC with statistical extrapolation. Since there were only three species NOECs available in the RAR, it was concluded in the RAR that the taxonomic coverage requirements for applying an SSD were not met by the RAR dataset. The present analysis has added four species to the database to provide a better representation of sensitive taxonomy in benthic systems. Moreover, the RAR dataset only

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 226 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

represented a 2.5-fold difference in sensitivity among species (432 to 1101 mg/kg d.w.), whereas the current dataset represents a 7.5-fold difference in sensitivity among species (146 to 1101 mg/kg d.w.). The range of toxicity values obtained with inclusion of four new species now represents better taxonomic diversity, and thus, it is possible to apply statistical extrapolation. Given the number of relevant high quality data, statistical extrapolation was used for PNEC determination. Following the RIP R.10. guidance, “different distributions may be used” for the SSD. We tested the lognormal distribution (default option), as calculated with the “ETX” software, and subsequently several other distributions with the “@RISK” software. The statistics of the curve–fitting on the chronic NOEC data are summarised in table below.

Table 72. Summary statistics for the SSD on chronic NOEC values for zinc in freshwater sediment. Distribution Lognormal Logistic (@RISK) Extreme Value (ETX) (@RISK) HC5 117.8 mg/kg 141.4 mg/kg d.w. 140.0 mg/kg d.w. d.w. A-D Statistic 0.57 0.51 0.68

A-D Significance 0.01 (accepted) 0.1≤ p ≤ 0.25 (accepted) 0.05≤ p ≤ 0.1 (accepted) Level K-S Statistic 0.82 0.23 0.32

K-S Significance Level 0.01 (accepted) >0.1 (accepted) 0.025≤ p ≤ 0.05 (accepted) Statistical acceptance Accepted Accepted Accepted

Using both the Anderson-Darling (A-D) and Kolmogorov-Smirnov (K-S) tests for normality, the default distribution (lognormal) fits significantly at a level of 1%. The A-D and K-S tests also accepted the logistic and extreme value distributions at higher significance levels (5- 25%). For purposes of comparing estimated HC5 values, the logistic and extreme value distributions do fit the data well in the region of interest. The log normal SSD is presented in figure below.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 227 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Figure 7. Lognormal distribution curve fitting to the freshwater sediment chronic toxicity data for zinc (ETX graphics).

To conform with the approach prescribed in the TGD, the lognormal distribution is used to provide a basis for setting the PNECadd, sediment. It is noted that the HC5 resulting from the logistic and extreme value distributions, although not as significant using both A-D and K-S tests, are higher than the HC5 calculated with the lognormal distribution (141.4, 140.0 and 117.8 mg/kg d.w., respectively). It is also noted that the PNEC is expressed as the “added” concentration, i.e. to be added to the background when using monitored water concentrations. The 5th percentile value of the SSD (the HC5) is set at the 50% confidence level, using the lognormal distribution function, which results in an HC5 value of 117.8 mg/kg d.w. (equivalent to 25.9 mg/kg w.w.).

Discussion on the uncertainty of the PNEC derivation

The following considerations are made on the uncertainty around the PNECadd, sediment and for determining the size of the assessment factor:

 Although the guidance on PNECadd, sediment derivation does not specifically detail the number or diversity of species required to calculate a sediment HC5 from an SSD, the chronic NOEC database of seven species entries covers species that are among the benthic organisms most frequently used for assessing sediment toxicity. They represent also different taxonomic groups, and these taxonomic groups are among the most important for the sediment ecosystem. Moreover, the RAR dataset only represented a 2.5-fold difference in sensitivity among species (432 to 1101 mg/kg d.w.), whereas the current dataset represents a 7.5-fold difference in sensitivity among species (146 to 1101 mg/kg d.w.). Based on this, there is no need for assessment factor higher than 1.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 228 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

 The lognormal distribution that was used for freshwater PNECadd, sediment derivation provides a significant fit to the data and resulted in an HC5 of 117.8 mg/kg d.w., which is lower than the HC5 value calculated from the logistic or extreme value distributions (141.4 and 140.0 mg/kg d.w., respectively). So the distribution that is used for the PNECadd, sediment derivation gives a conservative HC5 value. Based on this observation, there is no need for an assessment factor higher than 1.  In an effort to focus on the agreement among various published sediment quality guidelines (SQGs), consensus based SQGs were developed for 28 chemicals of concern in freshwater sediments (MacDonald et al. 2000). For each contaminant of concern, including zinc, a threshold effect concentration (TEC) and a probable effect concentration (PEC) SQG was developed. From five published studies reporting SQGs, the geometric mean was used as a consensus based value. In addition, the resultant SQGs for each chemical was evaluated for reliability using matching sediment chemistry and toxicity data from various field studies conducted throughout the United States. For zinc, the consensus based TEC-SQG and PEC-SQG, with ranges from the five studies used, was 121 (98-150) and 459 (270-820) mg/kg d.w., respectively. Validation results taken from 347 different toxicity studies demonstrated that 82% (133 of 163) of tests conducted at concentrations below the TEC resulted in no effect. Similarly, 90% (108 of 120) tests conducted at concentrations above the PEC resulted in significant effects. As such, the PNECadd, sediment derived here (117.8 mg/kg d.w.) is nearly identical to the consensus based TEC (121 mg/kg d.w.). So the value that is used for the PNECadd, sediment derivation gives a conservative HC5 value. Based on this observation, there is no need for an assessment factor higher than 1.  In a publication by Chapman et al. (1999), more than 50 sediment quality values (SQV) for 22 metals and metalloids were summarized from different jurisdictions in the U.S.A., Canada, The Netherlands, Norway, Australia, New Zealand, and China. For almost every metal or metalloid, SQVs from different jurisdictions varied over several orders of magnitude. For zinc, species-specific threshold effects levels (TEL; analogous to a NOEC) ranged from 98 mg/kg d.w. (28-d amphipod) to 10,100 mg/kg d.w. (chironomid). It is emphasized that the proposed HC5 of 117.8 mg/kg d.w., estimated by statistical extrapolation, is within the lower range of the world-wide distribution of TEL values. So the distribution that is used for the PNECadd, sediment derivation gives a conservative HC5 value. Based on this observation, there is no need for an assessment factor higher than 1.

 From the field colonization studies discussed previously, the proposed PNECadd, sediment (117.8 mg/kg d.w.) is lower than the observed NOEC values reported for five different study sites. These field studies demonstrate that the PNECadd, sediment derivation gives a conservative HC5 value. Based on this observation, there is no need for an assessment factor higher than 1.

Alternative approaches for estimating PNECadd, sediment values were investigated for comparison to the statistical extrapolation technique. Here, the Equilibrium Partitioning (EqP) and Assessment Factor (AF) approaches were evaluated for comparison.

 The EqP method in which the PNECadd. sediment has been estimated from the PNECadd,

aquatic (20.6 g/L; revised value as compared to the EU RA, resulting from the update of the chronic aquatic database), results in a PNECadd, sediment of 2,237 mg/kg d.w. This value is nearly 15-times higher than the lowest NOEC for benthic species (146 mg/kg d.w.). It is emphasised, however, that the EqP method has limitations for the derivation of a reliable PNECadd, sediment, especially for metals, because of the

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 229 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

uncertainties (assumptions) that exist. This approach is therefore not recommended further in the PNECsediment derivation.  Using the AF approach, the size of the AF is considered on a case-by-case assessment of the number, type and representativeness (long-term tests with sub-lethal endpoints) of available studies. Given the number of long-term tests for freshwater benthic organisms (n=7, see above), the TGD (2003; Table 21) prescribes that an AF = 10 be applied for datasets comprising at least “Three long-term tests (NOEC or EC10) with species representing different living and feeding conditions”. As such, the lowest NOEC (added) of the chronic dataset (G. pulex; 146 mg/kg d.w.) is divided by an AF of 10. This results in a PNECadd, sediment = 146 mg/kg d.w. / 10 = 14.6 mg/kg d.w. (equivalent to 3.22 mg/kg w.w.). Given the established range (70 to 175 mg/kg d.w.) and median value (140 mg/kg d.w.) for natural background zinc concentrations for freshwater sediment, as presented in the Zn RA, a PNECadd, sediment of 14.6 mg/kg d.w. could not reliably be distinguished from the natural variation in background zinc concentrations. This approach is therefore not recommended further in the PNECsediment derivation. The concentrations of zinc in freshwater sediment are dependent on natural conditions; thus, it is difficult to determine experimentally a natural background concentration in Europe. Due to geochemical differences, the natural background concentrations will differ. In addition, since the concentrations that are measured in the environment are the sum of an anthropogenic and a ‘natural’ source, one cannot simply distinguish the ‘natural’ part from the anthropogenic part. Below a number of different estimates for background zinc values in freshwater sediment are summarised. All currently available natural background data for freshwater sediment are in the same order of magnitude as identified in the RA (range 70 to 175 mg/kg d.w.), with a median value of 140 mg/kg d.w. These background estimates demonstrate that the estimated PNECadd, sediment (117.8 mg/kg d.w.) is a conservative value that when added to sediments, could be distinguished from the natural variation in background zinc concentrations. If available monitoring data can unequivocally be linked with a particular natural background value in an area, preference should be given to that specific background value.  A large dataset of sediment is available from the Belgium Flanders monitoring network (VMM, 2003). A total of 1083 sampling locations distributed over various water types have been monitored in Flanders during the period 1994-2001. Results show that 61% (n=662) of the sampling stations have a zinc sediment level lower than 200 mg/kg d.w., about 25% (n=277) are between 200 and 500 mg/kg d.w., about 10% (n=108) are between 500 and 1000 mg/kg d.w., and 3% (n=36) exceed 1000 mg/kg d.w. According to ‘Desire for Levels’ (Van de Meent, 1990) the provisionally natural background level for zinc in Dutch sediment can be set on a value of 68 mg/kg. In the Netherlands the applied natural background concentration for the Dutch ‘standard’ sediment is set at 140 mg/kg d.w. (value also applied in the RA)  Sediment data from Sweden have reported median values of 150 and 240 mg/kg d.w. for Northern Sweden and Southern Sweden, respectively (Landner and Lindeström, 1998). In an earlier report by the Swedish Environmental Protection Agency (1993) a ‘preliminary background’ concentration (based on upper quartile of available data from pre-industrial sediment layers) of 175 mg/kg d.w. was given. In Finland natural background concentrations of total zinc for stream sediments were reported between 20 and 140 mg/kg d.w. (90P), with a median value of 46 mg/kg d.w. (Lahermo et al. 1996). A survey in Norway of metal concentrations in 231 lake sediments was carried out in 1996-97 (Rognerud et al., 1999). Samples were taken from 231 lakes distributed over the whole country. The range of Zn concentrations was 22-919 mg/kg d.w. in the upper sediment and 13-884 mg/kg d.w. in the deep sediment. The deep- sediment samples are considered to reflect pre-industrial background levels. The median values were 136 and 106 mg/kg d.w., respectively. The Norwegian authorities

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 230 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

propose that for lake sediments, the natural background concentrations may be derived from the 75 percentile for near surface and deep sediments respectively (similar to water). This leads to a natural background value of 150 mg/kg d.w. in sediment.  In a publication by Chapman et al. (1999), reported natural background concentrations for 22 metals and metalloids were summarized from different jurisdictions in the U.S.A., Canada, The Netherlands, Norway, Australia, New Zealand, and China. For zinc, site-specific background zinc concentrations in freshwater sediments ranged from 50 to 143 mg/kg d.w. with a median value of 93 mg/kg d.w.

Freshwater PNECadd, sediment: Conclusion

The assessment of the freshwater PNECadd, sediment for zinc is based on the chronic benthic toxicity data that were presented in the EU RAR for zinc (2008), complemented with four new high quality studies (species). From the knowledge available at the time of closure, the RAR derived a PNEC using an application factor of 10 on the lowest available NOEC value (488 mg/kg d.w.). Because of the significant increase of information on chronic benthic toxicity, the PNEC from the RAR has been revised. Taking into account the weight of evidence provided by the elements discussed above, it is considered that use of the HC5 from the SSD, using statistical extrapolation techniques, is justified for PNEC derivation, and that no additional assessment factor needs to be applied. Consequently, the PNEC is set at the level of the HC5 which is considered as protective for EU freshwater ecosystems: freshwater PNECadd, sediment = 117.8 mg/kg d.w. (equivalent to 25.9 mg/kg w.w.). It is emphasized that this is an added PNEC, i.e. natural background needs to be taken into account when characterising the risk from monitored data.

Accounting for bioavailability Different approaches for characterizing the bioavailable fraction of metals in sediment have been studied for nearly 20 years. Examples of these alternative approaches include consideration of organic matter content as well as acid volatile sulfide (AVS) and simultaneous extractable metals (SEM). It is well-known that the AVS in sediment reacts with the SEM (i.e., the metal that is measured in the acid extract used to measure AVS) to form an insoluble metal sulphide. This metal sulphide form is essentially non-bioavailable to benthic organisms. The amount of AVS in sediments therefore serves as a critical parameter in determining metal bioavailability and toxicity in sediments. Metals, in essence, will exist in the form of their respective metal sulphide if the AVS is present in excess of the reactive forms of the sediment metals (SEM). On the other hand, if the total concentration of the metals is greater than the concentration of the AVS, then potentially, some fraction of the metals may exist as bioavailable metal and cause toxicity. The AVS/SEM approach was investigated in detail in the EU risk assessment on zinc (ECB 2008). It allows for making a correction on the exposure assessment for bioavailability. In the RA, a general, conservative bioavailability correction of 50% was applied. This is also used in the present analysis. In addition, a more specific 2correction for AVS-bound zinc can be made at the local scale, if the necessary data are available. R2 = 0.38 )

Co-varianceW 1 AVS SEM Zn D

g /

In thel RA (ECB 2008), a number of industrial sites were reporting local AVS/SEM levels. These data show that o

at thesem sites, elevated zinc concentration in sediment was related to higher AVS levels in these local sediments, µ

( 0 too (RA zinc, table 3.4.66.). This phenomenon of co-variance betweenFlanders SEMZn and AVS was furthere n documentedZ by an alalysis of coupled sediment data from the Netherlands, falan,drs (Belgium) and other places M The Netherlands (VangheluweE et al 2003). Figure 8 below illustrates the observed trend between and SEMZn for those data points S Rest of Europe

whereg -1AVSZn > SEMZn. Sediments where SEMZn > AVSZn, were excluded from the analysis since in that case

o Δ Δ Lineair (Flanders) l

2013-05-27-2 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 231 -1 0 1 2 3 4

log excess AVSZn (µmol/g DW) EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

part of the measured SEMZn was not bound to AVS and can therefore affect the identification of a possible relationship between both parameters.

Figure 8. Covariance between AVSZn and SEMZn in European sediments (taken from Vangheluwe et al 2003).

This figure clearly shows a trend indicating that SEMZn increases with increasing Δ AVSZn. Covariance between SEMZn and AVS has been suggested in literature and has been explained by the fact that Zn-sulfides are more stable than Fe-sulfides (Liber et al., 1996). So, in general, it seems like elevated zinc levels in ediment would be related to higher AVS levels. This observation is important when considering limited exceedances of PNEC on a local scale, as following from default conservative calculations. Considering the observed co-variance between AVS and SEMZn it is recommended to use measured coupled data to maintain the ecological relevance of the analysis i.e. the coupled SEM and AVS data generated for Flemish sediments (Vangheluwe et al, 2003).

Marine PNEC sediment derivation Deriving the PNEC

The toxicity of zinc to marine benthic organisms was evaluated to develop the PNECadd, sediment. The available ecotoxicity database for marine sediment was limited to two species so a statistical extrapolation approach was not justified for estimating an HC5 (concentration estimated for the 5th percentile of the distribution). Instead, the Equilibrium Partitioning (EqP) approach was used to translate the HC5 estimated for the robust aquatic marine water dataset (39 species) into a PNECadd, sediment. The estimated PNECadd, sediment value was considered with published studies regarding world-wide compilation of sediment quality values and background levels for metals. In addition, an SSD was constructed using ecotoxicity data for both freshwater and marine sediments for estimation of an HC5 using statistical extrapolation. The Assessment Factor [AF] approach was also evaluated for comparison.

Deriving the PNECadd, sediment using the EqP approach Aquatic Ecotoxicity Data The marine aquatic database zinc, used for the EqP approach largely fulfils the species and taxonomic requirements for chronic toxicity data as explained in the RIP R. 10 guidance (at least 10 species NOECs and 8 taxonomic groups). Indeed, 39 species mean NOECs based on 48 NOEC values, from 9 taxonomic groups covering three trophic levels were found to fulfil the relevancy and reliability requirements as explained by Klimisch et al. 1997. The marine zinc database includes 4 micro- and 5 macro-algae species, 4 annelid species, 6 crustacean species, 5 echinoderm species, 9 mollusk species, 1 nematode species, 1 cnidarian species and 1

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 232 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 fish species. A lognormal distribution of the aquatic dataset resulted in an HC5 (5% - 95% confidence values) of 6.09 (3.09 - 10.26) µg Zn/L.

EqP Calculation

In conformity with the calculation of the PECadd for sediment (RAR 2008), the properties of suspended matter are used to calculate the PNECadd for sediment, i.e., PNECadd, sediment = PNECadd, suspended matter. Studies characterizing the equilibrium partitioning of zinc to suspended matter in estuarine and marine systems were compiled to determine a median partitioning constant (Kpsusp) for zinc (Turner 1993, 1996 and 2002). Briefly, according to the TGD, the Ksusp-water and PNECadd, sed are calculated using the following equations:

1. Ksusp-water : Fwatersusp + (Fsolidsusp x Kpsusp x RHOsolid)

2. PNECadd, sed = PNECadd, susp : (Ksusp-water / RHOsusp) x PNECadd, aquatic Where:

3 3 Ksusp-water = volumetric suspended matter / water partition coefficient (m /m )

3 3 Fwatersusp = volume fraction water in suspended matter (m /m )

3 3 Fsolidsusp = volume fraction solids in suspended matter (m /m )

3 Kpsusp = suspended matter / water partition coefficient (m /kg)

3 RHOsolid = density of the solid fraction (kg/m )

PNECadd, sed = Predicted No Effect Concentration in sediment (mg/kg wet sediment)

PNECadd, susp = Predicted No Effect Concentration in suspended matter (mg/kg wet suspended matter)

3 RHOsusp = bulk density of wet suspended matter (kg/m )

3 PNECadd, aquatic = Predicted No Effect Concentration in water (mg/m )

The range in Kpsusp values from 18 separate natural marine sediment studies was 300-37,000 L/kg (Turner 1993, 1996 and 2002). A median Kpsusp value was calculated to be 9,399 L/kg. Using this median value, the EqP approach resulted in a PNECadd, sediment of 12.5 mg/kg w.w., as follows (according to the TGD):

1. Ksusp-water : Fwatersusp + (Fsolidsusp x Kpsusp x RHOsolid) =

3 3 3 3 3 3 0.9 m /m + (0.1 m /m x 9.399 m /kg x 2,500 kg/m ) = 0.9 m3/m3 + 2,350 m3/m3 = 2,351 m3/m3

2. PNECadd, sed = PNECadd, susp : (Ksusp-water / RHOsusp) x PNECadd, aquatic = (2,351 m3/m3 / 1,150 kg/m3) x 6.09 mg/m3 = 12.5 mg/kg wet sediment

The above marine PNECadd, sediment of 12.5 mg/kg w.w. (22% solids by weight) is equivalent to a marine PNECadd, sediment of 56.5 mg/kg d.w.

Discussion on the uncertainty of the PNEC derivation

The following considerations are made on the uncertainty around the PNECadd, sediment and for determining the size of the assessment factor:

 The marine dataset (PNECaquatic) used for the calculation of the PNECadd, sediment in the EqP approach is extensive and of high quality and relevancy for the marine environment. Similarly, the Kp dataset used for deriving a median Kp value for use in the EqP approach is extensive and robust, including data from both EU and N. American waters. As such it is comparable in size and representativity with the dataset on freshwater Kp,

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 233 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

used in the EU RA. Therefore the marine Kp is considered reliable for the derivation of the PNEC sediment in the present exercise.

 Two long-term chronic toxicity tests with zinc are available, the lowest NOECadded value (A. Marina; emergence; 207.1 mg/kg d.w.) is nearly fourfold higher than the PNEC calculated using the EqP approach (56.5 mg/kg d.w.). This difference suggests that the calculated PNECadd, sediment is sufficiently conservative to provide adequate protection for benthic organisms in marine systems and there is no need for an assessment factor At the end of the RAR (2008), it was concluded that the PNECadd, sediment, which was based on sediment toxicity data for freshwater benthic organisms, would be applied for both the freshwater and saltwater environment. This was concluded since no PNECadd, sediment could be derived for the saltwater environment due to the lack of toxicity data for benthic marine organisms. As such, it was satisfactorily argued that the PNECadd, sediment reported in the RAR (49 mg/kg d.w.) was acceptable for marine sediments. The consistency between the EqP-calculated PNECadd, sediment (56.5 mg/kg d.w.) and the RAR result (49 mg/kg d.w.), taken with consideration of the NOEC values from the literature, suggests that the calculated PNEC is sufficiently conservative to provide adequate protection for benthic organisms in marine systems and there is no need for an assessment factor.  In a publication by Chapman et al. (1999), more than 50 sediment quality values (SQV) for 22 metals and metalloids were summarized from different jurisdictions in the U.S.A., Canada, The Netherlands, Norway, Australia, New Zealand, and China. For almost every metal or metalloid, SQVs from different jurisdictions varied over several orders of magnitude. For zinc, apparent/threshold effects levels (analogous to a NOEC) ranged from 120 to 1,600 mg/kg d.w. It is emphasized that the proposed HC5 of 56.5 mg/kg d.w., estimated by EqP, is considerably lower than the entire world-wide distribution of apparent/threshold effects values. So the approach that is used for the PNECadd, sediment derivation gives a conservative value and there is no need for an assessment factor.

An alternative approach for estimating a PNECadd, sediment value was investigated for comparison to the EqP technique. Here, an SSD was constructed using ecotoxicity data for both freshwater and marine sediments for estimation of an HC5 using statistical extrapolation (log-normal distribution). Along with the two studies reported above, seven chronic single- species freshwater sediment toxicity tests with zinc are available for crustaceans (Gammarus pulex and Hyallela azteca ), insects (Ephoron virgo, Hexagenia sp. and Chironomus tentans) and worms (Lumbriculus variegates and Tubifex tubifex). These include only long-term tests, which sufficiently provide information on survival, growth and reproductive effects. The analysis of the combined dataset presents a 7.5-fold difference in sensitivity among species (146 to 1101 mg/kg d.w.) compared to a 2.4-fold difference in sensitivity among marine species alone. Although the outcome of this analysis is in agreement with the EqP approach (see above), preference is given to using data from the marine compartment only, rather than combining them with data from the freshwater compartment:  The lognormal distribution (default option), as calculated with the “ETX” software was used to estimate the HC5 and curve–fitting statistics for the combined dataset. The 5th percentile value of the SSD (the HC5) is set at the 50% confidence level, using the lognormal distribution function, which results in an HC5 (5 and 95 P) value of 120.9 (45-206) mg/kg d.w. Using both the Anderson-Darling (A-D) and Kolmogorov-Smirnov (K-S) tests for normality, the default distribution (lognormal) fits significantly at a level of 1%.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 234 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

 When estimating an HC5 using an SSD, an Assessment Factor (AF) may be applied based on consideration of general factors such as the diversity and representativity of the taxonomic groups covered by the database, the overall quality of the database and the endpoints covered, statistical uncertainties around the 5th percentile estimate, and comparisons between field and mesocosm studies. Since the reliability of the combined dataset is high, an AF = 2 can be applied because of the absence of field/microcosm data for marine sediments. The PNECadd, sediment that would be derived as such would be 120.9 mg/kg d.w. / 2 = 60.5 mg/kg d.w. (equivalent to 13.3 mg/kg w.w.). This PNEC is similar to the PNECadd, sediment estimated using the EqP approach (56.5 mg/kg d.w.). So the EqP approach provides a defensible estimation of the PNECadd, sediment and there is no need for an assessment factor. For comparison to the EqP technique, the Assessment Factor (AF) approach for estimating PNECadd, sediment values was investigated. This AF approach results in an overly conservative estimate of the marine PNECadd, sediment which cannot reliably be distinguished from the natural variation in background zinc concentrations:  Using the AF approach, the size of the AF is considered on a case-by-case assessment of the number, type and representativeness (long-term tests with sub-lethal endpoints) of available studies. Given the number of long-term tests for freshwater (n=6) and marine benthic organisms (n=2, see above), the RIP guidance (Table R.10-9), prescribes that an AF = 10 be applied for datasets comprising at least “Three long-term tests with species representing different living and feeding conditions including a minimum of two tests with marine species”. As such, the lowest NOECadded for zinc is from a freshwater sediment test with the amphipod G. pulex (146 mg/kg d.w.). This results in a PNEC add, sediment = 146 mg/kg d.w. / 10 = 14.6 mg/kg d.w. (equivalent to 3.22 mg/kg w.w.). Given the established range (15 to 195 mg/kg d.w.) and median value (60 mg/kg d.w.) for natural background zinc concentrations in marine sediment, a PNECadd, sediment of 14.6 mg/kg d.w. could not reliably be distinguished from the natural variation in background zinc concentrations (discussed below). In fact, a PNECadd, sediment of 14.6 mg/kg d.w. would be close to the 10 P concentration established in the Belgian Marine Data Center database for open sea sediments. The concentrations of zinc in marine sediment are dependent on natural conditions; thus, it is difficult to determine experimentally a natural background concentration in Europe. Due to geochemical differences, the natural background concentrations will differ. In addition, since the concentrations that are measured in the environment are the sum of anthropogenic and ‘natural’ sources, one cannot simply distinguish the ‘natural’ part from the anthropogenic part. A number of different estimates for background zinc values in marine sediment are summarised below. Although many researchers have reported background concentrations of zinc in coastal and estuarine sediments, measurements from sediments collected in the open sea likely provide the best approximation of natural background. As such, natural background for marine sediment is likely within the range of 15 to 195 mg/kg d.w.), with a median value of 60 mg/kg d.w. These background estimates demonstrate that the estimated PNECadd, sediment (56.5 mg/kg d.w.) is a conservative value that when added to sediments, could be distinguished from the natural variation in background zinc concentrations. If available monitoring data can unequivocally be linked with a particular natural background value in an area, preference should be given to that specific background value:  An extensive monitoring program in Belgium (Belgian Marine Data Center) is available online with measured data for coastal, estuarine and open sea sediment measurements covering years 2000-2008. Results show that the median zinc concentrations (10 and 90 P) for coastal, estuarine and open sea sediments are 117 (17.4-189), 359 (137-640) and 64.9 (9.2-200) mg/kg d.w., respectively. Similarly, an older study by van Eck et al., (1991) reported measured zinc concentrations in

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 235 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

estuarine and coastal sediments throughout Europe that ranged between 7 and 1400 mg/kg d.w.  In a publication by Chapman et al. (1999), reported natural background concentrations for 22 metals and metalloids were summarized from different jurisdictions in the U.S.A., Canada, The Netherlands, Norway, Australia, New Zealand, and China. For zinc, site-specific background zinc concentrations in marine sediments ranged from 40 to 191 mg/kg d.w. with a median value of 110 mg/kg d.w.

Marine PNECadd, sediment: Conclusion

The assessment of the marine PNECadd, sediment for zinc identified two long-term chronic toxicity studies. However, an EqP (using partitioning coefficients and a robust aquatic toxicity database) approach provided a reliable derivation for the marine benthic compartment. The resulting value is considered protective for EU marine ecosystems: marine PNECadd, sediment of 56.5 mg/kg d.w. (equivalent to 12.5 mg/kg w.w.). It is emphasized that this is an added PNEC, i.e. natural background needs to be taken into account when characterising the risk from monitored data.

Accounting for bioavailability Different approaches for characterizing the bioavailable fraction of metals in sediment have been studied for nearly 20 years. Examples of these alternative approaches include consideration of organic matter content as well as acid volatile sulphide (AVS) and simultaneous extractable metals (SEM). It is well-known that the AVS in sediment reacts with the SEM (i.e., the metal that is measured in the acid extract used to measure AVS) to form an insoluble metal sulphide. This metal sulphide form is essentially non-bioavailable to benthic organisms. The amount of AVS in sediments therefore serves as a critical parameter in determining metal bioavailability and toxicity in sediments. Metals, in essence, will exist in the form of their respective metal sulphide if the AVS is present in excess of the reactive forms of the sediment metals (SEM). On the other hand, if the total concentration of the metals is greater than the concentration of the AVS, then potentially, some fraction of the metals may exist as bioavailable metal and cause toxicity. The AVS/SEM approach was investigated in detail in the EU risk assessment on zinc (ECB 2008). It allows for making a correction on the exposure assessment for bioavailability. In the RA, a general, conservative bioavailability correction of 50% was applied. This is also used in the present analysis. In addition, a more specific correction for AVS-bound zinc can be made at the local scale, if the necessary data are available.

Co-variance AVS SEM Zn In the RA (ECB 2008), a number of industrial sites were reporting local AVS/SEM levels. These data show that at these sites, elevated zinc concentration in sediment was closely related to higher AVS levels in these local sediments, too (RA zinc, table 3.4.66.). This phenomenon of co-variance between SEMZn and AVS was described above (Figure 7).

Figure 7 clearly shows a trend indicating that SEMZn increases with increasing Δ AVSZn. Covariance between SEMZn and AVS has been suggested in literature and has been explained by the fact that Zn-sulfides are more stable than Fe-sulfides (Liber et al., 1996). So, in general, it seems like elevated zinc levels in ediment would be related to higher AVS levels. This observation is important when considering limited exceedances of PNEC on a local scale, as following from default conservative calculations. Considering the observed co-variance between AVS and SEMZn it is recommended to use measured coupled data to maintain the ecological relevance of the analysis i.e. the coupled SEM and AVS data generated for Flemish sediments (Vangheluwe et al, 2003).

Bioavailability considerations Approaches for characterizing the bioavailable fraction of metals in sediment have been studied for nearly 20 years. Examples of these alternative approaches include consideration of organic matter content as well as acid

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 236 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 volatile sulfide (AVS) and simultaneous extractable metals (SEM). It is well-known that the AVS in sediment reacts with the SEM (i.e., the metal that is measured in the acid extract used to measure AVS) to form an insoluble metal sulfide. This metal sulfide form is essentially not bioavailable to benthic organisms. The amount of AVS in sediments therefore serves as a critical parameter in determining metal bioavailability and toxicity in sediments. Metals, in essence, will exist in the form of their respective metal sulfide if the AVS is present in excess of the reactive forms of the sediment metals (SEM). On the other hand, if the total concentration of the metals is greater than the concentration of the AVS, then potentially, some fraction of the metals may exist as bioavailable metal and cause toxicity. Although the AVS/SEM approach was investigated in detail in the EU risk assessment on zinc (ECB 2008), the appropriateness of this approach for assessing bioavailability in marine sediments was not addressed. In a study by Casas (1994), a clean marine sediment was spiked with zinc to determine if AVS was an adequate measure of the metal-binding capacity of the sediment. A 10-d, flow-through, acute bioassay using the marine polychaete Cupitellu cupitutu was conducted to examine the prediction of toxicity from zinc and sediment chemistry data. In all treatments where AVS exceeded SEM Zn concentrations, significant mortality was not observed. In addition, two studies conducted by the National Oceanic and Atmospheric Administration in the U.S. (Long, 1995; Wolfe, 1994) on sediment toxicity in marine and estuarine environments concluded that consideration of AVS and SEM improved assessment of biological effects from metals, including zinc, in these systems. It was suggested that the same threshold of effects proposed for freshwater sediments (i.e., no effects observed when AVS concentrations exceeded SEM concentrations) applied for marine and estuarine sediments. Similar to analyses conducted for freshwater sediments in the EU risk assessment on zinc (ECB 2008), three robust datasets in marine environments were evaluated to determine the likelihood and extent of bioavailable zinc (SEM Zn) exceeding AVS concentrations in natural sediment samples. Results demonstrated that SEM Zn in most sites did not exceed AVS concentrations, suggesting bioavailable zinc was not present. Of the 23 sites (of a total of 192 sites sampled) where SEM Zn exceeded AVS concentrations, the BioF (fraction of bioavailable zinc relative to total zinc) averaged 0.25 and only exceeded 0.50 at one site. Furthermore, based on 192 data from the U.S., the values of the 10th-, 50th- and 90th-percentiles of the BioF ratios are 0, 0 and 0.054, which results in an average value of 0.018. It is concluded that the SEM-Zn/AVS concept applies in marine sediments as well. Observed Zn-bioavailability in marine sediments is generally very low. However, similar to the approach taken in the RA for freshwater sediments, a general, conservative bioavailability correction of 50% is applied by default in the present analysis, when no information on SEM-Zn/AVS is available. A more specific correction for AVS-bound zinc can be made at the local scale, if the necessary data are available. The co-variance between SEM Zn and AVS, as observed in freshwater sediment, is also expected to occur also in marine sediments.

Table 73. SEM-Zn and AVS in marine sediments Sampling location SEM Zn – ∆ AVS ≤ 0 SEM Zn – ∆ AVS > 0 BioF Reference (# of samples with (# of samples with (range) no bioavailability) bioavailability) Long Island Sound, 66 3 0.009-0.29 Wolfe, 1994 NY, USA Hudson-Raritan 40 3 0.37-0.46 Long, 1995 Estuary, NY, USA Virginia Coastline, 63 17 0.004-0.60 U.S. EPA, 1996 USA SEM = simultaneous extractable metal. AVS = acid volatile sulfide. ∆ AVS = AVS fraction available for zinc binding once higher affinity metals (SEM Cd, SEM Cu, SEM Pb) have been subtracted. BioF = bioavailability factor.

Table 74. PNEC sediment PNEC Assessment Remarks/Justification factor PNEC sediment 1 Extrapolation method: statistical extrapolation (freshwater): 117.8 mg/kg sediment dw It is emphasized that the reported PNEC for freshwater sediments is an added PNEC, i.e. natural background needs to be taken into account when

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 237 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

characterizing the risks from monitored data. PNEC sediment 1 Extrapolation method: partition coefficient (marine water): 56.5 mg/kg sediment dw It is emphasized that the reported PNEC for saltwater sediments is an added PNEC, i.e. natural background needs to be taken into account when characterizing the risks from monitored data.

7.2. Terrestrial compartment

In accordance to the EU risk assessment, and in a same approach as for aquatic ecotoxicity, the toxicity of the zinc ion is considered in this section on soil. As a consequence, the analysis below is relevant for all zinc compounds. It is noted that the scope of the terrestrial effect assessment under REACH is restricted to soil organisms in a narrow sense, i.e. non-vertebrate organisms living the majority of their lifetime within the soil and being exposed to substances via the soil pathway.

7.2.1. Toxicity test results

1. Sources of ecotoxicological data

In the EU risk assessment on zinc, an extensive analysis was made of the available terrestrial toxicity data, available at that time (ECB 2008). The data were carefully scrutinised for quality and relevancy by the Rapporteur and member states. In the present exercise, all data that were considered useful for deriving the PNEC soil in the EU risk assessment, are used. In addition, an update of the terrestrial toxicity data that became available after the closure of the EU RA, has been made. Based on this update, the PNEC derivation for Zn in soils has been revised. This PNEC derivation is now based on the data and bioavailability models presented in the Risk Assessment Report (RAR) for Zn under the existing substances regulation, the comments of the SCHER on this RAR and new reliable data not yet included in the RAR.

All toxicity data judged reliable and relevant in the RAR for Zn are included (171 NOEC or EC10 values). On March 2 2010, an additional literature search was performed covering the scientific literature since 2000 for new reliable toxicity data for Zn on terrestrial organisms (plants, invertebrates and micro-organisms). This new data search resulted in 43 new NOEC or EC10 values.

2. Selection of ecotoxicological data

The toxicity data on invertebrates and plants are from single-species tests that study common ecotoxicological parameters such as survival, growth and/or reproduction. The toxicity data on micro-organisms are from tests in which microbe-mediated soil processes, such as C- and N- mineralisation were studied. These microbial toxicity tests are multiple species tests because these microbe-mediated processes reflect the action of many species in soil microbial communities.

Relevance

Biological relevancy

The toxicity data on terrestrial organisms are from ecotoxicity tests that study relevant ecotoxicological parameters such as survival, growth, reproduction, litter breakdown, abundance. Relevant endpoints for soil micro-organisms focused on functional parameters (such as respiration, nitrification, mineralisation) and microbial growth, but also enzymatic processes are considered relevant.

Relevancy of the test media

Only data from observations in natural and artificial (OECD) soil media have been used in this report, tests performed in substrates that were judged as not representative for soils (e. g. nutrient solution, agar, pure quartz

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 238 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 sand and farmyard manure) were not included in this effects assessment.

The data used in the effect assessment should be based on organisms and exposure conditions relevant for Europe. Excluding all data derived in non-EU soils would, however, considerably reduce the amount of data to be used. Therefore, also data based on soils collected outside Europe have been used when the soil properties were within the representative range for Europe.

Test duration

What comprises “chronic exposure” is a function of the life cycle of the test organisms. A priori fixed exposure durations are therefore not relevant. The duration should be related to the typical life cycle and should ideally encompass the entire life cycle or, for longer-lived species the most sensitive life stage. Retained exposure durations should also be related to recommendations from standard ecotoxicity (e. g. ISO, OECD, ASTM) protocols.

Typically chronic test duration for the higher plants are within the range of 4 (e. g. the barley root elongation test based on ISO 11269-1 (1995)) and 21 days (e. g. the tomato shoot yield test based on ISO 11269-2 (1995)). OECD n° 208 (plant seedling emergence and growth test, 1984) recommended a test duration of at least 14 days after emergence of the seedlings. For soil invertebrates, assessing the chronic effects of substances on sub-lethal endpoints such as reproduction on oligochaetes has a typical exposure duration of 3 to 6 weeks for the standard organism Enchytraeus albidus (OECD, 2000; ISO 16387). For another standard species Folsomia candida survival and reproduction is typically assessed after 28 days of exposure (ISO 11267, 1999). Reported test duration using soil micro-organisms vary largely but standard tests last 28 days for carbon transformation (OECD n° 216) and for nitrogen transformation (OECD n° 217).

Reliability

Type of test

Both standard test organisms and non-standard species can be used in the framework of a risk assessment. In general, toxicity data generated from standardized tests, as prescribed by organizations such as OECD and USEPA will need less scrutiny than non-standardized test data, which will require a more thorough check on their compliance with reliability criteria before being used. GLP and non-GLP tests can be used provided that the latter fulfil the stipulated requirements.

Concentration-effect relationships

Because effect concentrations are statistically derived values, information concerning the statistics should be used as a criterion for data selection. If no methodology is reported or if values are ‘visually’ derived, the data were considered unreliable. Effect levels derived from toxicity tests using only 1 test concentration always results in unbounded and therefore unreliable data. Therefore, only the results from toxicity tests using 1 control and at least 2 Zn concentrations were retained.

Tests that do not comply with the above-mentioned stipulations are rated as not reliable and are not recommended for use in the risk assessment exercise.

3. Derivation of EC10/NOEC values

According to the REACH Guidance on information requirements and chemical safety assessment (Chapter R.10.2.2.1), there is a preference to use EC10 values as calculated from the concentration-effect relationship, for derivation of the Predicted No Effect Concentration (PNEC). In some cases no reliable EC10 can be derived because e. g. no significant dose-response curve can be fitted or the EC10 is outside the concentration range tested. When in these cases a bounded NOEC value can be derived, this NOEC value will be used instead of the EC10 for PNEC derivation. No unbounded NOEC (i. e. no effect at highest dose tested) or LOEC (i. e. significant effect at lowest dose tested) values or EC10 values extrapolated outside the concentration range tested are used for derivation of the PNEC.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 239 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

4. Toxicity data

In accordance to the EU RA, all toxicity data are expressed as added Zn concentration in soil, based on either the nominal dose added or on measured, background corrected soil Zn concentrations.

For plants, in total 45 individual high quality NOEC or EC10 values are selected for the PNEC derivation, representing 18 different species. NOEC or EC10 values vary from 32 mg Zn/kg dw for Trifolium pratense and Vicia sativa (Van der Hoeven and Henzen, 1994) to 5855 mg Zn/kg dw for Triticum aestivum (Warne et al., 2008a).

Information on soil properties allowing bioavailability correction for plants (eCEC and pH) is only available for 31 NOEC or EC10 values, representing 9 different plant species and including the same minimum and maximum values as the total dataset for plants.

In total 61 individual high quality NOEC or EC10 values for reproduction of terrestrial invertebrates are selected for the PNEC derivation. Twenty-four NOEC/EC10 values are available for toxicity of Zn to reproduction of terrestrial arthropods, representing 2 different species and ranging between 14.6 and 1000 mg Zn/kg dw (both for Folsomia candida; Lock and Janssen, 2001c and Lock et al., 2003). The other 37 NOEC or EC10 values cover 6 different worm species and vary from 35.7 mg Zn/kg for Enchytraeus albidus (Lock and Janssen, 2001c) to 1634 mg Zn/kg dw for Lumbricus terrestris (Spurgeon et al., 2000).

For all 61 reliable toxicity thresholds, the information on soil properties allowing bioavailability correction for plants (eCEC) is available.

For microbial assays, in total 108 individual high quality NOEC/EC10’s are selected for the PNEC derivation. These values represent 4 nitrogen transformation processes, 5 carbon transformation processes and 8 enzymatic processes and range from 17 mg Zn/kg dw for respiration (Chang and Broadbent, 1981 and Lighthart et al., 1983) to 2623 mg Zn/kg dw for phosphatase (Doelman and Haanstra, 1989).

Information on the background Zn concentration, allowing correction for differences in bioavailability among soils, is only available for 76 NOEC or EC10 values, representing 13 microbial processes (4 for N cycle, 5 for C cycle and 4 enzymatic processes). The total range in NOEC/EC10 values for the dataset with results for background Zn concentration is the same as for the total dataset for micro-organisms.

All study records used in the analysis are detailed in tables 79 and 80.

5. Calculation of the HC5-50

The available ecotoxicity database for the effect of Zn to soil organisms is large. Therefore, the use of the statistical extrapolation method is –as specified by the Guidance document on information requirements and chemical safety assessment Chapter R.10.3.1.3– preferred for PNEC derivation rather than the use of an assessment factor on the lowest NOEC. The PNEC will be based on the 50% confidence value of the 5th percentile value (HC5-50) and an additional assessment factor taking into account the uncertainty on the HC5- 50 (thus PNEC = HC5-50/AF).

5.1. Generic, non-normalised HC5-50

The non-normalised terrestrial HC5-50 was derived based on either all individual reliable NOEC/EC10 values or the species mean NOEC/EC10 values for the most sensitive endpoint. Two different approaches were used: 1) taking into account all data and; 2) only taking into account the data with information on soil properties allowing correction for differences in bioavailability among soils.

It must be stressed that, considering the important influence of soil properties on bioavailability and toxicity of zinc in soils, these approaches are less ecologically relevant for HC5-50 derivation compared to the HC5-50 values taking into account correction for bioavailability (both ageing and effect of variation in soil properties). The cumulative frequency distribution (SSD) of the non-normalised species mean NOEC values for Zn is presented in Figures 9 to 12. Using statistical extrapolation and the log-normal distribution results in a HC5-50 of 35.6 mg Zn/kg based on all individual NOEC/EC10 data and a HC5-50 of 33.7 mg Zn/kg when using all individual NOEC/EC10 data with

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 240 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

information on soil properties allowing correction for bioavailability among soils (Table 75). Based on the Anderson-Darling goodness-of-fit statistics, the log-normal distribution is accepted for all distributions based on both all individual reliable observations and the species/process mean values.

Table 75. Generic HC5 and HC5-50 (with 5% and 95% confidence interval) values for toxicity of Zn to the terrestrial environment based on a log-normal distribution of non- normalised NOEC/EC10added values Scenario All individual NOEC/EC10 values Species/process mean values N HC5 HC5-50 N HC5 HC5-50 (mg Zn/kg) (mg Zn/kg) (mg/kg) (mg Zn/kg) All data 214 35.6 35.6 43 51.9 51.3 (29.1-42.5) (34.7-69.4) Data allowing 168 33.8 33.7 30 36.5 35.8 bioavailability (26.8-41.2) (20.4-54.0) correction

Using a species/process mean approach for non-normalised data yields higher HC5-50 values compared to SSD based on all individual values: 51.3 and 35.8 mg Zn/kg for the total dataset or the data with information on soil properties allowing correction for bioavailability among soils, respectively. Averaging (geomean) all the results available for one species/process avoids over-representation of commonly tested species or processes, e.g. Eisenia fetida (29 data) or nitrification (20 data). This species/process mean approach is preferred for data corrected for the differences in soil properties when the intra-species variation can be considered as the main source of variation among data for a given species/process. However, this is not the case for the generic approach (non-normalised data) where variation between toxicity data for a certain species or process is also caused by differences in bioavailability among soils.

Table 76. Generic species/process mean values. All data Data allowing bioavailability correction Species/microbial process Mean Species/microbial process Mean NOEC/EC10 NOEC/EC10 mg Zn/kg mg Zn/kg 1 Vicia sativa 32 1 Vicia sativa 32 2 Trifolium pratense 45 2 Hordeum vulgare 33 3 Denitrification 62 3 Denitrification 39 4 Glutamic acid 4 mineralization 64 Trifolium pratense 45 5 5 Glutamic acid Nitrate reductase 67 mineralization 55 6 Urease 72 6 Urease 73 7 Respiration 89 7 Zea mais 83 8 Hordeum vulgare 89 8 Respiration 83 9 Enchytraeus albidus 94 9 Enchytraeus albidus 94 10 Vigna mungo L. 100 10 Nitrification 120 11 Nitrification 120 11 Sinella curviseta 180 12 Dehydrogenase 128 12 Dehydrogenase 195 13 Sorghum bicolor 141 13 Allium cepa 200 14 Zea mais 171 14 Avena sativa 200 15 15 Trigonella poenum Sinella curviseta 180 graceum 200 16 N-mineralization 185 16 Glucose mineralization 204 17 Triticum vulgare 200 17 N-mineralization 211 18 18 Maize residue Spinacea oleracea 200 mineralization 241 19 Avena sativa 200 19 Folsomia candida 246 20 Allium cepa 200 20 Eisenia fetida 284

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 241 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

21 Trigonella poenum 21 graceum 200 Brassica rapa 300 22 Amidase 200 22 Acetate mineralization 303 23 Glucose mineralization 204 23 Eisenia andrei 320 24 Maize residue 24 mineralization 241 Aporrectodea caliginosa 342 25 Folsomia candida 246 25 Arylsulphatase 406 26 Eisenia fetida 284 26 Lumbricus terrestris 520 27 Medicago sativa 300 27 Triticum aestivum 584 28 Beta vulgaris 300 28 Phosphatase 826 29 Brassica rapa 300 29 Ammonification 1000 30 Acetate mineralization 303 30 Lumbricus rubellus 1634 31 Eisenia andrei 320 32 Aporrectodea caliginosa 342 33 Arylsulphatase 378 34 Lactuca sativa 400 35 Pisum sativum 400 36 Lycopersicon esculentum 400 37 Phosphatase 444 38 Lumbricus terrestris 520 39 Triticum aestivum 584 40 Phytase 590 41 Ammonification 1000 42 Lumbricus rubellus 1634 43 Pyrophosphatase 1640

There is no clear distinction among the three trophic levels (plants, invertebrates and micro-organisms) in their sensitivity to Zn in soil (Table 76). Both individual and species/process mean toxicity data for the various plant and invertebrates species and microbial processes strongly overlap (Figures 9 to 12). Therefore, all data are pooled together into one species sensitivity distribution for the derivation of the PNEC value.

For 46 NOEC or EC10 values, no information is available on soil properties allowing correction for differences in bioavailability among soils (CEC and pH for plants and background Zn concentration for microbial processes). The toxicity data with information on these soil properties available cover 30 species/processes compared to a total of 43 for the total dataset. Toxicity data for 9 plant species and 4 microbial processes (all enzymatic processes) do not have information on the soil properties required. The reduced dataset however still covers the same extreme values (both minimum and maximum) as the total dataset (Table 76). The HC5-50 values as calculated from a log-normal distribution are consistently lower for the reduced dataset compared to the total dataset (both based on all individual values as based on species/process mean values, Table 75). It can therefore be concluded that the dataset with information on soil properties is both representative for the total dataset and conservative for the assessment of toxicity of Zn to terrestrial organisms.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 242 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Figure 9. The species sensitivity distribution based on all individual EC10 and NOEC values selected for PNEC derivation.

Figure 10. The species sensitivity distribution based on all individual EC10 and NOEC values with information on soil properties allowing correction for bioavailability among soils.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 243 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Figure 11. The species sensitivity distribution based on all species/process mean values.

Figure 12. The species sensitivity distribution based on species/process mean values for data with information on soil properties allowing correction for bioavailability among soils.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 244 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

4.2. Normalised HC5-50 for soils: implementation of bioavailability

In the zinc risk assessment (ECB 2008), a detailed analysis of bioavailability of zinc to soil organisms was made. As a result, an approach for making a bioavailability correction on the ecotoxicity values and PNEC was developed and applied. The same approach is applied in the present analysis.

The general frame work for implementation of bioavailability into derivation of PNEC values is presented in Figure 13 and uses the following steps: 1. Select the reliable NOEC/EC10 data with information on soil properties allowing correction for bioavailability among soils. 2. Derive the NOEC/EC10added values by subtracting the Zn background concentration of the tested control soils from the total NOEC/EC10 values (measured NOEC/EC10) or use the NOEC/EC10added values from nominal NOEC/EC10 values. 5 3. Apply an “ageing” factor of 3 to all NOEC/EC10added values. This is a conservative value, applied to all soil types (cfr EU risk assessment , ECB 2008). In the added risk approach, application of a generic lab-to-field correction factor (i.e. a constant factor independent of soil properties) can also be done after steps 4, 5 or 6 instead of after step 2. This will not affect the final result of the HC5-50. It is here proposed after step 2, in accordance to the approach followed in the EU Risk assessment. This approach is also consistent with the total risk approach as used for other metals (e.g. Cu and Ni). 4. Normalise each (lab-to-field corrected) NOEC/EC10added value towards a reference soil using the appropriate slope.

s l o p e a b i o t i c f a c t o r r e f  N O E C / E C 1 0 r e f  N O E C / E C 1 0    a b i o t i c f a c t o r 

The available knowledge on the soil-type dependent bioavailability resulted in the following equations that are proposed for correcting bioavailability in soil (EU risk assessment, ECB 2008):   LogEC50=1.4+1.14*logCEC (F. candida)  LogEC50=1.9+0.79*logCEC (E. fetida)  LogEC50=1.1+0.87*logCEC + 0.12*pH (wheat)  LogEC50=1.2+0.76*logZnBG (nitrification)  LogEC50=1.7+0.76*logZnBG (respiration)

Where  EC50 is the 50% effect concentration (mg Zn/kg)

 CEC is the effective cation exchange capacity (cmolc/kg), i.e. the CEC measured at the pH of the soil  ZnBG is the zinc background concentration in soil (mg Zn/kg). Note that background concentrations refer to ambient concentrations, not natural background. The soils used in the experiments were sampled from agricultural areas (not all), i.e. the tests and the assessments are made on soils that may already contain zinc from diffuse sources.

The slope derived for the springtail Folsomia candida will be used for all terrestrial arthropods, while the slope derived for Eisenia fetida will be applied for all toxicity data for soft-bodied terrestrial invertebrates. All plant data will be normalised with the slopes derived for wheat and all NOEC or EC10 data for microbial processes will be normalised with the same slope derived for both nitrification and respiration. The CEC at soil pH is usually not reported but can be predicted from %clay, %OM and soil pH based on an existing multivariate model that is calibrated on natural soils (Helling et al., 1964): CEC=(30.4+4.4*pH)*%clay/100 + (-35+30*pH)*%OM/100

For OECD soils, it is assumed that the clay has no CEC contribution since, unlike prevailing clay types in Europe, kaolin clay has no permanent charge and its variable charge is inferior to that of the organic

5 The factor 3 is the generic default and is applied for the generic soil in the usual case that ageing has occurred for one year or longer. A ratio of 2 should only be used in cases where a rapid increase in zinc soil concentration could occur, e.g. due to the melting of snow, when ageing has occurred for less than one year (EU RA, ECB 2008).

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 245 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

matter which is at least 5% in these studies. This assumption yields CEC=14.4 cmolc/kg at pH 6. 5. Where multiple data are available for the same species/process and endpoint, calculate the species/process mean value for the most sensitive endpoint for each species or process. 6. Build the species sensitivity distribution (SSD) from the species/process geomean NOEC/EC10 values and derive the HC5/ HC5-50

Soils with larger CEC (large clay content and OM content) also tend to contain larger zinc concentrations. In the 15 soils collected for the research program on bioavailability of Zn in soils, there was a positive correlation 2 between these two parameters (LogZnBG=-0.1+0.72*log CEC; p<0.001; R =0.67). Taking into account the effect of soil properties, allows to normalise all individual EC10 or NOEC values and to calculate soil-specific HC5-50 values. As an example, this approach is applied for 8 different soil types (table 77). These soils are the same as those referred to in the EU risk assessment. For all soil types, the log-normal distribution results in a good fit according to the Anderson-Darling goodness-of-fit test. The HC5-50 values for these soil scenarios clearly stress the importance of soil properties on the predicted hazard of zinc in soils (Table 77, Figure 14). The HC5-50 varies almost 10-fold between soils (between 30 mg Zn/kg for the sandy forest soil and 282 mg Zn/kg for the river clay soil). Normalised species/process mean values for the 8 soil scenarios are presented in Table 78. The bioavailability factors (BioF, i.e. ratio of normalised and generic species/process mean HC5 values) based on the SSD including plants, invertebrates and micro-organisms agree well with the minimum BioF factors reported in the RAR for zinc for either plants and invertebrates or microbial processes as based on the mean slopes.

Figure 13. Flow chart for the implementation of bioavailability factors into PNEC derivation for Zn.

Table 77. HC5 and HC5-50 (with 5% and 95% confidence interval) for the terrestrial environment based on lab-field corrected and normalised NOEC/EC10 values and a lognormal distribution.

Scenario pH CEC at soil Soil total HC5 HC5 HC5-50 BioFsoil (water pH Zn All Species Species $ ) (cmolc/kg) (mg/kg) values mean mean Generic HC5* for 101 109 107 selected data for which (61-162) abiotic factors exist Cattle farms, sandy soil 5.83 10.97 28 94 93 91 0.8

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 246 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

(extensive) (1993) (54-134) Cattle farms, sandy soil 5.94 6.98 32 85 72 71 0.7 (intensive) (1993) (41-106) Cattle farms, sandy soil 5.9 7.7 31 88 77 76 0.7 (1994) (44-112) Forest, sandy soil 3.7 5.03 10.1 40 31 30 0.3 (1994) (17-47) Arable farm, sandy soil 5.96 16.53 31 110 120 118 1.1 (1995) (71-173) Cattle farm, peaty soil 5.93 33.04 124 279 264 260 2.4 (1995) (155-381) Arable farms – marine 8.09 14.42 68 167 188 185 1.7 clay soil (1996) (113-265) Cattle farms – river 6.59 28.88 172 316 287 282 2.6 clay soils (1996) (168-415) $ : BioF = HC5/HC5 generic for species mean values *: including lab-field factor of 3

Figure 14. The species sensitivity distributions for the 8 soil scenarios as fitted by the lognormal distribution.

The soils listed in table 77 and figure 14 are examples. Calculations can be done when soil characteristics are documented using the soil bioavailability calculator.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 247 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Table 78. Normalised and lab-field corrected species/process mean NOEC/EC10 values for the 8 soil scenarios. Cattle farm, sandy Cattle farm, sandy Cattle farm, sandy Forest, sandy soil Arable farm, sandy Cattle farm, peaty Arable farm, Cattle farm, river soil (extensive) soil (intensive) soil soil soil marine clay soil clay soil Species/ Mean Species/ Mean Species/ Mean Species/ Mean Species/ Mean Species/ Mean Species/ Mean Species/ Mean process NOEC process NOEC process NOEC process NOEC process NOEC process NOEC process NOEC process NOEC /EC10 /EC10 /EC10 /EC10 /EC10 /EC10 /EC10 /EC10 Hordeum 69 Hordeum 48 Hordeum 52 Hordeum 20 Denitrificati 88 Hordeum 186 Denitrificati 160 Hordeum 199 vulgare vulgare vulgare vulgare on vulgare on vulgare Denitrificati 82 Vicia sativa 75 Vicia sativa 81 Vicia sativa 30 Hordeum 103 Denitrificati 253 Hordeum 165 Vicia sativa 310 on vulgare on vulgare Vicia sativa 108 Denitrificati 90 Denitrificati 88 Denitrificati 38 Urease 157 Vicia sativa 290 Vicia sativa 256 Denitrificati 324 on on on on Urease 145 Allium cepa 144 Allium cepa 155 Allium cepa 58 Vicia sativa 160 Urease 449 Urease 285 Enchytraeus 514 albidus Allium cepa 207 Trigonella 144 Trigonella 155 Trigonella 58 Nitrification 234 Allium cepa 556 Enchytraeus 297 Urease 576 poenum poenum poenum albidus Trigonella 207 Trifolium 153 Urease 157 Trifolium 62 Glutamic 273 Trigonella 556 Sinella 370 Allium cepa 594 poenum pratense pratense acid poenum curviseta mineralizati on Nitrification 217 Urease 160 Trifolium 165 Urease 67 Respiration 291 Enchytraeus 572 Nitrification 425 Trigonella 594 pratense albidus poenum Trifolium 220 Sinella 162 Enchytraeus 181 Brassica 71 Allium cepa 307 Trifolium 590 Allium cepa 491 Trifolium 630 pratense curviseta albidus rapa pratense pratense Enchytraeu 239 Enchytraeus 167 Sinella 181 Zea mais 87 Trigonella 307 Nitrification 672 Trigonella 491 Brassica 726 s albidus albidus curviseta poenum poenum rapa Glutamic 253 Brassica 176 Brassica 190 Nitrification 100 Trifolium 326 Brassica 680 Glutamic 496 Sinella 816 acid rapa rapa pratense rapa acid curviseta mineralizati mineralizati on on Brassica 253 Zea mais 214 Zea mais 230 Sinella 111 Enchytraeus 331 Glutamic 782 Trifolium 521 Nitrification 861 rapa curviseta albidus acid pratense mineralizati on Respiration 269 Nitrification 240 Nitrification 234 Glutamic 116 Brassica 375 Zea mais 824 Respiration 528 Zea mais 880 acid rapa mineralizati on Sinella 271 Glutamic 279 Glutamic 273 Respiration 124 Sinella 432 Respiration 834 Aporrectode 574 Aporrectode 994 curviseta acid acid curviseta a caliginosa a caliginosa mineralizati mineralizati on on Zea mais 307 Respiration 298 Respiration 291 Enchytraeus 129 Zea mais 455 Sinella 952 Brassica 600 Glutamic 1003 albidus curviseta rapa acid

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 248 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

mineralizati on Maize 441 Aporrectode 324 Aporrectode 350 Avena 194 Maize 477 Aporrectode 1106 Zea mais 728 Respiration 1069 residue a caliginosa a caliginosa sativa residue a caliginosa mineralizati mineralizati on on Aporrectod 463 Folsomia 418 Folsomia 467 Maize 203 Glucose 502 Maize 1368 Maize 867 Eisenia 1667 ea candida candida residue mineralizati residue residue andrei caliginosa mineralizati on mineralizati mineralizati on on on Glucose 464 Avena 478 Maize 477 Glucose 214 Aporrectode 640 Glucose 1439 Glucose 911 Eisenia 1679 mineralizati sativa residue mineralizati a caliginosa mineralizati mineralizati fetida on mineralizati on on on on Acetate 656 Maize 489 Glucose 502 Aporrectode 250 Acetate 709 Avena 1845 Folsomia 956 Maize 1754 mineralizati residue mineralizati a caliginosa mineralizati sativa candida residue on mineralizati on on mineralizati on on Avena 688 Glucose 514 Avena 515 Folsomia 288 Arylsulphata 866 Eisenia 1854 Eisenia 963 Lumbricus 1822 sativa mineralizati sativa candida se andrei andrei terrestris on Folsomia 700 Eisenia 543 Eisenia 587 Acetate 302 N- 894 Eisenia 1867 Eisenia 970 Glucose 1845 candida andrei andrei mineralizati mineralizati fetida fetida mineralizati on on on Eisenia 776 Eisenia 547 Eisenia 591 Arylsulphata 370 Avena 1018 Lumbricus 2027 Lumbricus 1053 Avena 1970 andrei fetida fetida se sativa terrestris terrestris sativa Eisenia 782 Lumbricus 593 Lumbricus 641 N- 381 Eisenia 1073 Acetate 2033 Acetate 1288 Folsomia 2109 fetida terrestris terrestris mineralizati andrei mineralizati mineralizati candida on on on Arylsulphat 802 Acetate 726 Acetate 709 Eisenia 419 Eisenia 1081 Folsomia 2459 Arylsulphata 1574 Acetate 2607 ase mineralizati mineralizati andrei fetida candida se mineralizati on on on N- 827 Arylsulphata 888 Arylsulphata 866 Eisenia 422 Folsomia 1117 Arylsulphata 2485 N- 1624 Arylsulphata 3187 mineralizati se se fetida candida se mineralizati se on on Lumbricus 848 N- 916 N- 894 Lumbricus 458 Lumbricus 1173 N- 2563 Avena 1629 N- 3287 terrestris mineralizati mineralizati terrestris terrestris mineralizati sativa mineralizati on on on on Phosphatas 1191 Phosphatas 1318 Phosphatas 1286 Phosphatas 549 Phosphatas 1286 Phosphatas 3689 Phosphatas 2337 Phosphatas 4730 e e e e e e e e Dehydrogen 1193 Dehydrogen 1321 Dehydrogen 1289 Dehydrogen 550 Dehydrogen 1289 Dehydrogen 3698 Dehydrogen 2342 Dehydrogen 4742 ase ase ase ase ase ase ase ase Ammonifica 1748 Triticum 1394 Triticum 1502 Triticum 565 Ammonificat 1888 Triticum 5377 Lumbricus 3308 Lumbricus 5726 tion aestivum aestivum aestivum ion aestivum rubellus rubellus Triticum 2004 Lumbricus 1865 Ammonificat 1888 Ammonificat 805 Triticum 2968 Ammonificat 5416 Ammonificat 3431 Triticum 5740 aestivum rubellus ion ion aestivum ion ion aestivum Lumbricus 2665 Ammonificat 1935 Lumbricus 2015 Lumbricus 1440 Lumbricus 3685 Lumbricus 6368 Triticum 4748 Ammonificat 6945

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 249 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 rubellus ion rubellus rubellus rubellus rubellus aestivum ion

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 250 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

7.2.1.1. Toxicity to soil macro-organisms

The results are summarised in the following table:

Table 79. Overview of effects on soil macro-organisms

Method Results Remarks Reference Eisenia fetida (annelids) NOEC (21 d): 350 mg/kg 2 (reliable with Spurgeon D.J., soil dw element (nominal) restrictions) Tomlin M.A. & long-term toxicity (laboratory study) based on: reproduction Hopkin S.P. (1997) (cocoons) key study Substrate: artificial soil NOEC (21 d): 350 mg/kg read-across based on equivalent or similar to OECD soil dw element (nominal) grouping of Guideline 207 (Earthworm, Acute based on: reproduction substances (category Toxicity Tests) (cocoons) approach) Test material (IUPAC name): zinc sulphate (See endpoint summary for justification of read-across) Eisenia fetida (annelids) NOEC (56 d): 199 mg/kg 2 (reliable with Spurgeon D J, soil dw element (nominal) restrictions) Hopkin S P and long-term toxicity (laboratory study) based on: reproduction Jones D T (1994) (cocoons) key study Substrate: artificial soil read-across based on equivalent or similar to OECD grouping of Guideline 207 (Earthworm, Acute substances (category Toxicity Tests) approach)

Test material (IUPAC name): zinc dinitrate (See endpoint summary for justification of read-across) Eisenia fetida (annelids) NOEC (21 d): 97 mg/kg soil 2 (reliable with Spurgeon D.J. & dw element (nominal) based restrictions) Hopkin S.P. (1996) long-term toxicity (laboratory study) on: reproduction (cocoon numbers) key study Substrate: artificial soil NOEC (21 d): 553 mg/kg read-across based on Nine toxicity tests were conducted soil dw element (nominal) grouping of using a procedure based on the OECD based on: reproduction substances (category (1984) artificial foil earthworm toxicity (cocoon numbers) approach) test. For each test, Eisenia fetida were exposed to zinc in a range of artificial NOEC (21 d): 484 mg/kg Test material soils with differing pH and/or organic soil dw element (nominal) (IUPAC name): zinc matter content. based on: reproduction dinitrate (See (cocoon numbers) endpoint summary for justification of NOEC (21 d): 85 mg/kg soil read-across) dw element (nominal) based on: reproduction (cocoon numbers)

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 251 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

NOEC (21 d): 183 mg/kg soil dw element (nominal) based on: reproduction (cocoon numbers)

NOEC (21 d): 414 mg/kg soil dw element (nominal) based on: reproduction (cocoon numbers)

NOEC (21 d): 115 mg/kg soil dw element (nominal) based on: reproduction (cocoon numbers)

NOEC (21 d): 161 mg/kg soil dw element (nominal) based on: reproduction (cocoon numbers)

NOEC (21 d): 223 mg/kg soil dw element (nominal) based on: reproduction (cocoon numbers) Eisenia fetida (annelids) NOEC (42 d): 714 mg/kg 2 (reliable with Spurgeon D.J. et al soil dw element (meas. (not restrictions) (2000) long-term toxicity (laboratory study) specified)) based on: reproduction (cocoons) key study Substrate: mixture of a commercially available sandy loam soil and read-across based on commercially available Sphagnum peat grouping of substances (category Life-cycle (sur vival, weight change, approach) and cocoon production rate) and biomarker(neutral- Test material (EC redretentionbycoelomocyteslysosomes) name): zinc nitrate responses to zinc in four earthworm (See endpoint species were measured in laborator y summary for tests. justification of read- across) Lumbricus terrestris (annelids) NOEC (42 d): 1634 mg/kg 2 (reliable with Spurgeon D.J. et al soil dw element (meas. (not restrictions) (2000) long-term toxicity (laboratory study) specified)) based on: reproduction (cocoons) key study Substrate: mixture of a commercially available sandy loam soil and read-across based on commercially available Sphagnum peat grouping of substances (category Life-cycle (sur vival, weight change, approach) and cocoon production rate) and biomarker(neutral- Test material (EC redretentionbycoelomocyteslysosomes) name): zinc nitrate responses to zinc in four earthworm (See endpoint species were measured in laborator y summary for tests. justification of read- across) Lumbricus rubellus (annelids) NOEC (42 d): 520 mg/kg 2 (reliable with Spurgeon D.J. et al soil dw element (meas. (not restrictions) (2000) long-term toxicity (laboratory study) specified)) based on:

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 252 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

reproduction (cocoons) Substrate: mixture of a commercially key study available sandy loam soil and commercially available Sphagnum peat read-across based on grouping of Life-cycle (sur vival, weight change, substances (category and cocoon production rate) and approach) biomarker(neutral- redretentionbycoelomocyteslysosomes) Test material (EC responses to zinc in four earthworm name): zinc nitrate species were measured in laborator y (See endpoint tests. summary for justification of read- across) Aporrectodea caliginosa (annelids) NOEC (42 d): 195 mg/kg 2 (reliable with Spurgeon D.J. et al soil dw element (meas. (not restrictions) (2000) long-term toxicity (laboratory study) specified)) based on: reproduction (cocoons) key study Substrate: mixture of a commercially available sandy loam soil and read-across based on commercially available Sphagnum peat grouping of substances (category Life-cycle (sur vival, weight change, approach) and cocoon production rate) and biomarker(neutral- Test material (EC redretentionbycoelomocyteslysosomes) name): zinc nitrate responses to zinc in four earthworm (See endpoint species were measured in laborator y summary for tests. justification of read- across) Aporrectodea caliginosa (annelids) NOEC (56 d): 600 mg/kg 2 (reliable with Khalil M.A. et al. soil dw element (nominal) restrictions) (1996) long-term toxicity (laboratory study) based on: reproduction key study Substrate: natural soil read-across based on Motality and cocoon production were grouping of measured over 56 days to determine substances (category LC50, EC50 and EC10 values for zinc. approach)

Test material (IUPAC name): zinc sulfate (See endpoint summary for justification of read- across) Eisenia fetida (annelids) NOEC (21 d): 237 mg/kg 2 (reliable with Spurgeon D.J. and soil dw element (nominal) restrictions) Hopkin S.P. (1995) long-term toxicity (laboratory study) based on: reproduction (cocoons) key study Substrate: artificial soil read-across based on equivalent or similar to OECD grouping of Guideline 207 (Earthworm, Acute substances (category Toxicity Tests) approach)

Test material (IUPAC name): zinc dichloride (See endpoint summary

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 253 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

for justification of read-across) Eisenia sp. (annelids) NOEC (21 d): 320 mg/kg 2 (reliable with Van Gestel C.A.M. soil dw element (nominal) restrictions) et al (1993) long-term toxicity (laboratory study) based on: reproduction key study Substrate: artificial soil read-across based on Accumulation and elimination of zinc grouping of and effects on the growth and substances (category reproduction of the earthworm Eisenia approach) andrei were determined in an artificial soil. Test material (EC name): zinc chloride (See endpoint summary for justification of read- across) Eisenia fetida (annelids) NOEC (28 d): 180 mg/kg 1 (reliable without Lock et al. (2003) soil dw element (nominal) restriction) long-term toxicity (extended laboratory based on: reproduction study) (based on the number of key study cocoons) Substrate: natural soil read-across based on NOEC (28 d): 100 mg/kg grouping of ISO 11268-2 (Effects of Pollutants on soil dw element (nominal) substances (category Earthworms. 2. Determination of based on: reproduction approach) Effects on Reproduction) (1996) (based on the number of cocoons) Test material (IUPAC name): zinc NOEC (28 d): 1000 mg/kg dichloride (See soil dw element (nominal) endpoint summary based on: reproduction for justification of (based on the number of read-across) cocoons)

NOEC (28 d): 320 mg/kg soil dw element (nominal) based on: reproduction (based on the number of cocoons)

NOEC (28 d): 560 mg/kg soil dw element (nominal) based on: reproduction (based on the number of cocoons)

NOEC (28 d): 320 mg/kg soil dw element (nominal) based on: reproduction (based on the number of cocoons)

NOEC (28 d): 560 mg/kg soil dw element (nominal) based on: reproduction (based on the number of cocoons)

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 254 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

NOEC (28 d): 1000 mg/kg soil dw element (nominal) based on: reproduction (based on the number of cocoons)

NOEC (28 d): 560 mg/kg soil dw element (nominal) based on: reproduction (based on the number of cocoons)

NOEC (28 d): 180 mg/kg soil dw element (nominal) based on: reproduction (based on the number of cocoons)

EC10 (28 d): 350 mg/kg soil dw element (nominal) based on: reproduction (based on the number of cocoons) Eisenia fetida (annelids) EC10 (21 d): 438 mg/kg soil 2 (reliable with Lock K. & Janssen dw element (nominal) based restrictions) C.R. (2001) long-term toxicity (extended laboratory on: reproduction (cocoon study) production) key study

Substrate: natural soil EC10 (21 d): 127 mg/kg soil read-across based on dw element (nominal) based grouping of OECD Guideline 207 (Earthworm, on: reproduction (cocoon substances (category Acute Toxicity Tests) production) approach) Test material (EC name): zinc chloride (See endpoint summary for justification of read- across) Enchytraeus albidus (annelids) EC10 (42 d): 132 mg/kg soil 2 (reliable with Lock K. & Janssen dw element (nominal) based restrictions) C.R. (2001) long-term toxicity (extended laboratory on: reproduction (juveniles) study) key study EC10 (42 d): 35.7 mg/kg soil Substrate: natural soil dw element (nominal) based read-across based on on: reproduction (juveniles) grouping of OECD Guideline 220 (Enchytraeidae substances (category Reproduction Test) (draft) approach)

Test material (EC name): zinc chloride (See endpoint summary for justification of read-

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 255 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

across) Enchytraeus albidus (annelids) EC10 (20 wk): 178 mg/kg 2 (reliable with Lock K. & Janssen soil dw element (nominal) restrictions) C.R. (2002) long-term toxicity (laboratory study) based on: reproduction (juveniles) key study Substrate: artificial soil read-across based on OECD Guideline 220 (Enchytraeidae grouping of Reproduction Test) substances (category approach)

Test material (EC name): zinc chloride (See endpoint summary for justification of read- across) Eisenia fetida (annelids) NOEC (28 d): 250 mg/kg 2 (reliable with Owojori O.J., soil dw element (nominal) restrictions) Reinecke A.J. and long-term toxicity (laboratory study) based on: reproduction Rozanov A.B. (cocoons) key study (2008) Substrate: artificial soil read-across based on Zn, added as ZnCl2 in a range of sub- grouping of lethal concentrations (0, 250, 500, 750 substances (category mg/kg Zn) was combined with 0, 2000 approach) or 4000 mg/kg NaCl. The endpoints: mortality, weight change, and the Test material (EC internal zinc concentration were name): zinc chloride assessed at day 1, 7, 14 and 28 while (See endpoint cocoon production was assessed only summary for at day 28. justification of read- across) Folsomia candida (Collembola (soil- NOEC (4 wk): NOEC 2 (reliable with Sandifer R.D. & dwelling springtail)) element (nominal) based on: restrictions) Hopkin S.P. (1996) reproduction (juvenile long-term toxicity (laboratory study) production) key study

EC50's for zinc were determined for NOEC (4 wk): NOEC read-across based on juvenile production of Folsomia element (nominal) based on: grouping of candida at pH 6.0, 5.0 and 4.5 in a reproduction (juvenile substances (category standard laboratory test system. production) approach)

NOEC (4 wk): NOEC Test material (EC element (nominal) based on: name): zinc nitrate reproduction (juvenile (See endpoint production) summary for justification of read- across) Folsomia candida (Collembola (soil- NOEC (6 wk): NOEC 2 (reliable with Sandifer R.D. & dwelling springtail)) element (nominal) based on: restrictions) Hopkin S.P. (1997) reproduction (juvenile long-term toxicity (laboratory study) production) key study

EC50's for zinc were determined for read-across based on juvenile production of Folsomia grouping of candida at 15, 20 and 25 °C in a substances (category standard laboratory test system. approach)

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 256 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Test material (EC name): zinc nitrate (See endpoint summary for justification of read- across) Folsomia candida (Collembola (soil- NOEC (4 wk): NOEC 2 (reliable with Smit E.C. & Van dwelling springtail)) element (nominal) based on: restrictions) Gestel C.A.M. reproduction (juvenile (1998) long-term toxicity (laboratory study) numbers) key study

The influence of soil characteristics NOEC (4 wk): NOEC read-across based on and way of contamination on the element (nominal) based on: grouping of bioaccumulation and toxicity of zinc reproduction (juvenile substances (category was investigated for the springtail numbers) approach) Folsomia candida. NOEC (4 wk): NOEC Test material element (nominal) based on: (IUPAC name): zinc reproduction (juvenile chloride (See numbers) endpoint summary for justification of read-across) Folsomia candida (Collembola (soil- EC10 (4 wk): EC10 element 2 (reliable with Van Gestel C.A.M dwelling springtail)) (nominal) based on: restrictions) & Hensbergen P.J. reproduction (: juveniles) (1997) long-term toxicity (laboratory study) key study

The effects of Cd and Zn, alone or in read-across based on combination, on the survival, growth grouping of and reproduction of the collembolan substances (category Folsomia candida were determined approach) after 2,4 and 6 weeks of exposure in an artificial soil. Test material (IUPAC name): zinc dichloride (See endpoint summary for justification of read-across) Folsomia candida (Collembola (soil- NOEC (28 d): NOEC 1 (reliable without Lock et al. (2003) dwelling springtail)) element (nominal) based on: restriction) reproduction (based on the long-term toxicity (extended laboratory number of juveniles) key study study) NOEC (28 d): NOEC read-across based on ISO 11267 (Inhibition of Reproduction element (nominal) based on: grouping of of Collembola by Soil Pollutants) reproduction (based on the substances (category number of juveniles) approach)

NOEC (28 d): NOEC Test material element (nominal) based on: (IUPAC name): zinc reproduction (based on the dichloride (See number of juveniles) endpoint summary for justification of NOEC (28 d): NOEC read-across) element (nominal) based on: reproduction (based on the number of juveniles)

NOEC (28 d): NOEC

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 257 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

element (nominal) based on: reproduction (based on the number of juveniles)

NOEC (28 d): NOEC element (nominal) based on: reproduction (based on the number of juveniles)

NOEC (28 d): NOEC element (nominal) based on: reproduction (based on the number of juveniles) Folsomia candida (Collembola (soil- EC10 (28 d): EC10 element 2 (reliable with Smit C.E. & Van dwelling springtail)) (nominal) based on: restrictions) Gestel C.A.M reproduction (1997) long-term toxicity (extended laboratory key study study) EC10 (42 d): EC10 element (nominal) based on: read-across based on Experiment I: effect of constant (18°C, reproduction grouping of 75% relative humidity) or variable substances (category (24h cycle of 12 to 24°C and 95 to approach) 50% RH) climate conditions (both treatments 12/12 light/dark cycle) on Test material (EC reproduction of Folsomia candida. name): zinc chloride (See endpoint Experiment II: effect of exposure summary for temperature (13, 16, 19 and 22°C (75% justification of read- RH and 12/12 light/dark cycle) on across) reproduction of Folsomia candida. Folsomia candida (Collembola (soil- EC10 (28 d): EC10 element 2 (reliable with Lock K. & Janssen dwelling springtail)) (nominal) based on: restrictions) C.R. (2001) reproduction (juveniles) long-term toxicity (extended laboratory key study study) EC10 (28 d): EC10 element (nominal) based on: read-across based on ISO 11267 (Inhibition of Reproduction reproduction (juveniles) grouping of of Collembola by Soil Pollutants) substances (category EC10 (28 d): EC10 element approach) (nominal) based on: reproduction (juveniles) Test material (EC name): Zinc chloride (See endpoint summary for justification of read-

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 258 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

across) Sinella curviseta (Collembola (soil- EC10 (28 d): EC10 element 2 (reliable with Xu J. et al (2009) dwelling springtail)) (nominal) based on: restrictions) reproduction (juveniles) long-term toxicity (extended laboratory key study study) read-across based on Laboratory studies evaluated the grouping of sensitivity of Sinella curviseta Brook substances (category (Collembola: Entomobryidae) to approach) selected heavy metals (Cu, Pb and Zn). Survival, reproduction and growth of Test material (EC S. curviseta were determined in a 4- name): zinc chloride week exposure test in an agricultural (See endpoint soil amended with metals to summary for concentrations of 100, 200, 400, 800, 1 justification of read- 600 and 3 200 mg/kg. across) Eisenia fetida (annelids) LC50 (14 d): 163 mg/kg soil 4 (not assignable) Yeardley,R.B. et al, dw test mat. based on: (1995) short-term toxicity (laboratory study) mortality supporting study Substrate: artificial soil experimental result

EPA/600/3-88/029 (1988) Test material (EC name): Ammonium chloride

Discussion of effects on soil macro-organisms except arthropods

37 NOEC or EC10 values cover 6 different worm species and vary from 35.7 mg Zn/kg for Enchytraeus albidus (Lock and Janssen, 2001c) to 1634 mg Zn/kg dw for Lumbricus terrestris (Spurgeon et al., 2000).

All toxicity data are expressed as added Zn concentration in soil, based on either the nominal dose added or on measured, background corrected soil Zn concentrations.

The following information is taken into account for effects on soil macro-organisms except arthropods for the derivation of PNEC:

37 NOEC or EC10 values on soil macroorganisms are available covering 6 different worm species and vary from 35.7 mg Zn/kg for Enchytraeus albidus to 1634 mg Zn/kg dw for Lumbricus terrestris

Discussion of effects on soil arthropods

All toxicity data are expressed as added Zn concentration in soil, based on either the nominal dose added or on measured, background corrected soil Zn concentrations.

The following information is taken into account for effects on soil arthropods for the derivation of PNEC:

Twenty-four NOEC/EC10 values are available for toxicity of Zn to reproduction of terrestrial arthropods, representing 2 different species and ranging between 14.6 and 1000 mg Zn/kg dw (both for Folsomia candida).

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 259 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

7.2.1.2. Toxicity to terrestrial plants

The results are summarised in the following table:

Table 80. Overview of effects on terrestrial plants

Method Results Remarks Reference Zea mays (Monocotyledonae Zea mays: NOEC (6 wk): 83 2 (reliable with MacLean A.J. (monocots)) mg/kg soil dw element restrictions) (1974) (nominal) based on: growth long-term toxicity (extended laboratory (based on weight of shoots) key study study) read-across based on vegetative vigour test grouping of substances (category Substrate: natural soil approach)

Tests were conducted in the soil Test material (EC samples both with and without name): zinc sulphate pretreatment of the soil with 500 ppm (See endpoint P summary for justification of read- across) Hordeum vulgare (Monocotyledonae Hordeum vulgare: EC10 (45 2 (reliable with Aery N.C. & (monocots)) d): 215 mg/kg soil dw restrictions) Jagetiya B.L. element (nominal) based on: (1997) long-term toxicity (laboratory study) growth (based on root key study weight) vegetative vigour test read-across based on grouping of Substrate: natural soil substances (category approach) 45 days shoot and root yield test with wheat plants (Triticum aestivum) Test material (IUPAC name): zinc sulfate (See endpoint summary for justification of read- across) Trigonella foenum-graecum Trigonella foenum-graecum: 2 (reliable with Dang et al. (1990) NOEC (8 wk): 200 mg/kg restrictions) Allium cepa (Monocotyledonae soil dw element (nominal) (monocots)) based on: growth (based on key study whole plant) long-term toxicity (extended laboratory read-across based on study) Allium cepa: NOEC (8 wk): grouping of 200 mg/kg soil dw element substances (category vegetative vigour test (nominal) based on: growth approach) (based on whole plant) Substrate: natural soil Test material (IUPAC name): zinc Onion and fenugreek growth was sulfate (See endpoint measured in glasshouse on a slightly summary for alkaline clay-loam soil from Northern justification of read- India. Zn was applied at the rate of 0, across) 50, 100, 200 and 400 mg/kg soil. Vigna mungo (Dicotyledonae (dicots)) Vigna mungo, L.: NOEC (45 2 (reliable with Kalyanaraman S.B. d): 100 mg/kg soil dw restrictions) & Sivagurunathan

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 260 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

element (nominal) based on: P. (1993) long-term toxicity (extended laboratory growth (based on roots and key study study) stems) read-across based on vegetative vigour test grouping of substances (category Substrate: natural soil approach)

The effect of increasing Zn Test material conentration on Blackgram growth was (IUPAC name): zinc measured in a 45-day glasshouse pot sulfate (See endpoint experiment. summary for justification of read- across) Triticum aestivum (Monocotyledonae Triticum aestivum: EC10 (212 (reliable with Warne M. et al (monocots)) d): 755 mg/kg soil dw restrictions) (2008) element (meas. (not short-term toxicity (laboratory study) specified)) based on: growth key study seedling emergence toxicity / Triticum aestivum: EC10 (21read-across based on vegetative vigour test d): 275 mg/kg soil dw grouping of element (meas. (not substances (category Substrate: natural soil specified)) based on: growth approach)

OECD Guideline 208 (Terrestrial Triticum aestivum: EC10 (21Test material (EC Plants Test: Seedling Emergence and d): 235 mg/kg soil dw name): zinc sulphate Seedling Growth Test) element (meas. (not (See endpoint specified)) based on: growth summary for justification of read- Triticum aestivum: EC10 (21across) d): 5855 mg/kg soil dw element (meas. (not specified)) based on: growth

Triticum aestivum: EC10 (21 d): 655 mg/kg soil dw element (meas. (not specified)) based on: growth

Triticum aestivum: EC10 (21 d): 965 mg/kg soil dw element (meas. (not specified)) based on: growth

Triticum aestivum: EC10 (21 d): 875 mg/kg soil dw element (meas. (not specified)) based on: growth

Triticum aestivum: EC10 (21 d): 565 mg/kg soil dw element (meas. (not specified)) based on: growth

Triticum aestivum: EC10 (21 d): 250 mg/kg soil dw element (meas. (not specified)) based on: growth

Triticum aestivum: EC10 (21 d): 505 mg/kg soil dw

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 261 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

element (meas. (not specified)) based on: growth

Triticum aestivum: EC10 (21 d): 530 mg/kg soil dw element (meas. (not specified)) based on: growth

Triticum aestivum: EC10 (21 d): 620 mg/kg soil dw element (meas. (not specified)) based on: growth

Triticum aestivum: EC10 (21 d): 430 mg/kg soil dw element (meas. (not specified)) based on: growth

Triticum aestivum: EC10 (21 d): 335 mg/kg soil dw element (meas. (not specified)) based on: growth Brassica rapa (Dicotyledonae (dicots)) Brassica rapa: NOEC : 300 2 (reliable with Sheppard SC, mg/kg soil dw element restrictions) Evenden WG, long-term toxicity (extended laboratory (nominal) based on: first Abboud SA and study) bloom key study Stephenson M (1993) vegetative vigour test Brassica rapa: NOEC : 300 read-across based on mg/kg soil dw element grouping of Substrate: natural soil (nominal) based on: first substances (category bloom approach) Plant growth test with Brassica rapa Test material (EC name): zinc sulphate (See endpoint summary for justification of read- across) Medicago sativa (Dicotyledonae Medicago sativa: NOEC (67 2 (reliable with Boawn L.C. & (dicots)) d): 300 mg/kg soil dw restrictions) Rasmussen P.E. element (nominal) based on: (1971) Zea mays (Monocotyledonae growth (based on the weight key study (monocots)) of the whole plant) read-across based on Lactuca sativa (Dicotyledonae Zea mays: NOEC (28 d): grouping of (dicots)) 300 mg/kg soil dw element substances (category (nominal) based on: growth approach) Hordeum vulgare (Monocotyledonae (based on the weight of the (monocots)) whole plant) Test material (IUPAC name): zinc Sorghum bicolor (Monocotyledonae Zea mays: NOEC (28 d): dinitrate (See (monocots)) 200 mg/kg soil dw element endpoint summary (nominal) based on: growth for justification of Triticum vulgare (Monocotyledonae (based on the weight of the read-across) (monocots)) whole plant)

Pisum sativum (Dicotyledonae Lactuca sativa: NOEC (40 (dicots)) d): 400 mg/kg soil dw element (nominal) based on: Spinacea olerancea (Dicotyledonae

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 262 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

(dicots)) growth (based on the weight of the whole plant) Beta vulgaris (Dicotyledonae (dicots)) Hordeum vulgare: NOEC Lycopersicon esculentum (33 d): 100 mg/kg soil dw (Dicotyledonae (dicots)) element (nominal) based on: growth (based on the weight long-term toxicity (extended laboratory of the whole plant) study) Sorghum bicolor: NOEC (35 vegetative vigour test d): 100 mg/kg soil dw element (nominal) based on: Substrate: natural soil growth (based on the weight of the whole plant) Eighteen plant species were grown under uniform conditions in a growth Sorghum bicolor: NOEC (35 chamber in alkaline soil treated with d): 200 mg/kg soil dw 10, 100, 200, 300, 400 and 500 ppm element (nominal) based on: zinc. growth (based on the weight of the whole plant)

Triticum vulgare: NOEC (33 d): 200 mg/kg soil dw element (nominal) based on: growth (based on the weight of the whole plant)

Pisum sativum: NOEC : 400 mg/kg soil dw element (nominal) based on: growth (based on the weight of the whole plant)

Spinacea olerancea: NOEC : 200 mg/kg soil dw element (nominal) based on: growth (based on the weight of the whole plant)

Beta vulgaris: NOEC (42 d): 300 mg/kg soil dw element (nominal) based on: growth (based on the weight of the whole plant)

Lycopersicon esculentum: NOEC : 400 mg/kg soil dw element (nominal) based on: growth (based on the weight of the whole plant) Hordeum vulgare (Monocotyledonae Hordeum vulgare: NOEC 2 (reliable with Luo Y. & Rimmer (monocots)) (48 d): 33 mg/kg soil dw restrictions) D.L. (1995) element (nominal) based on: long-term toxicity (extended laboratory growth (based on weight of key study study) shoots) read-across based on weight of shoots grouping of substances (category Substrate: natural soil approach)

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 263 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

In order to asses the effects of metal Test material interactions on plant growth, a (IUPAC name): zinc greenhouse experiment was conduced, dichloride (See in which spring barley was grown for endpoint summary 48 days in a soil to which zinc was for justification of added. read-across) Trifolium pratense (Dicotyledonae Trifolium pratense: NOEC 2 (reliable with Hooftman, R.N. and (dicots)) (24 d): 100 mg/kg soil dw restrictions) R. Henzen (1996) element (nominal) based on: Vicia sativa (Dicotyledonae (dicots)) growth (root and shoot yield)key study Van der Hoeven, N. and L. Henzen long-term toxicity (laboratory study) Trifolium pratense: NOEC read-across based on (1994a) (24 d): 84 mg/kg soil dw grouping of Substrate: OECD soil and natural soil element (nominal) based on: substances (category Van der Hoeven, N. growth (root yield) approach) and L. Henzen OECD Guideline 208 (Terrestrial (1994b) Plants Test: Seedling Emergence and Trifolium pratense: NOEC Test material Seedling Growth Test) (25 d): 32 mg/kg soil dw (IUPAC name): zinc Van der Hoeven, N. element (nominal) based on: dichloride (See and L. Henzen growth (root and shoot yield)endpoint summary (1994c) for justification of Trifolium pratense: NOEC read-across) (25 d): 32 mg/kg soil dw element (nominal) based on: growth (root and shoot yield)

Trifolium pratense: NOEC (25 d): 32 mg/kg soil dw element (nominal) based on: growth (root and shoot yield)

Vicia sativa: NOEC (24 d): 32 mg/kg soil dw element (nominal) based on: growth (root yield) Avena sativa (Monocotyledonae Avena sativa: NOEC (5 mo):2 (reliable with De Haan et al. (monocots)) 100 mg/kg soil dw element restrictions) (1985) (nominal) based on: growth long-term toxicity (extended laboratory (based on grain yield) key study study) Avena sativa: NOEC (5 mo):read-across based on vegetative vigour test 200 mg/kg soil dw element grouping of (nominal) based on: growth substances (category Substrate: natural soil (based on grain yield) approach)

5 months plant growth with oat (Avena Avena sativa: NOEC (5 mo):Test material sativa) 200 mg/kg soil dw element (IUPAC name): zinc (nominal) based on: growth diacetate (See (based on grain yield) endpoint summary for justification of Avena sativa: NOEC (5 mo):read-across) 400 mg/kg soil dw element (nominal) based on: growth (based on grain yield)

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 264 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Discussion

For plants, 45 individual high quality NOECs/EC10 values are selected for PNEC derivation, representing 18 different species. NOEC and EC10 values vary between 32 mg Zn/kgdw for Trifolium pratenseand Vicia sativa (Van der Hoeven and Henzen, 1994) to 5855 mg Zn/kg dw for Triticum aestivum (Warne et al., 2008a).

All toxicity data are expressed as added Zn concentration in soil, based on either the nominal dose added or on measured, background corrected soil Zn concentrations.

The following information is taken into account for toxicity on terrestrial plants for the derivation of PNEC:

NOEC and EC10 values are available for 18 different species. TheNOEC/EC10 values vary between 32 mg Zn/kgdw for Trifolium pratense andVicia sativa to 5855 mg Zn/kg dw for Triticum aestivum.

7.2.1.3. Toxicity to soil micro-organisms

The results are summarised in the following table:

Table 81. Overview of effects on soil micro-organisms

Method Results Remarks Reference Species/Inoculum: soil NOEC (12 wk): 17 mg/kg 2 (reliable with Chang F-H and soil dw element (nominal) restrictions) Broadbent F E carbon dioxide production was based on: respiration rate (1981) measured after the addition of metal key study solutions to silt loam soils read-across from supporting substance (structural analogue or surrogate)

Test material (EC name): zinc sulphate (See endpoint summary for justification of read- across) Species/Inoculum: soil NOEC (45 d): 110 mg/kg 2 (reliable with Lighthart et al. soil dw element (nominal) restrictions) (1983) Soil microbial respiration was based on: respiration rate measured on five soils after treatment key study with metal salt additions to evaluate NOEC (45 d): 327 mg/kg the effect of metal toxicity soil dw element (nominal) read-across based on based on: respiration rate grouping of substances (category NOEC (45 d): 165 mg/kg approach) soil dw element (nominal) based on: respiration rate Test material (IUPAC name): zinc NOEC (45 d): 110 mg/kg sulfate (See endpoint soil dw element (nominal) summary for based on: respiration rate justification of read- across) NOEC (45 d): 17 mg/kg soil dw element (nominal) based on: respiration rate Species/Inoculum: soil NOEC (12 wk): 100 mg/kg 2 (reliable with Chang F-H and soil dw element (nominal) restrictions) Broadbent F E

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 265 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

based on: N-mineralization (1982) Influence of trace metals on N- key study immobilisation, N mineralisation and nitrification is investigated read-across based on grouping of substances (category approach)

Test material (EC name): zinc sulphate (See endpoint summary for justification of read- across) Species/Inoculum: soil NOEC (3 wk): 164 mg/kg 2 (reliable with Liang & Tabatabai soil dw element (nominal) restrictions) (1977) They studied the effets of trace based on: N-mineralization elements commonly found in sludge key study samples on nitrogen mineralisation in NOEC (3 wk): 164 mg/kg soils. In this study, the amounts of soil dw element (nominal) read-across based on ammonium-N, nitrate-N, and nitrite-N based on: N-mineralization grouping of produced were determined in four soils substances (category incubated with either a trace element NOEC (3 wk): 164 mg/kg approach) solution or water. soil dw element (nominal) based on: N-mineralization Test material (IUPAC name): zinc NOEC (3 wk): 164 mg/kg sulfate (See endpoint soil dw element (nominal) summary for based on: N-mineralization justification of read- across) Species/Inoculum: soil NOEC (3 wk): 1000 mg/kg 2 (reliable with Premi and Cornfield soil dw element (nominal) restrictions) (1969) This paper reports on the effects of based on: Ammonification adding zinc as sulphate and carbonate key study on ammonification and nitrification on subsequent incubation of soil. read-across based on grouping of substances (category approach)

Test material (IUPAC name): zinc sulfate (See endpoint summary for justification of read- across) Species/Inoculum: soil NOEC (10 d): 109 mg/kg 2 (reliable with Liang & Tabatabai soil dw element (nominal) restrictions) (1978) The effets of trace elements on based on: Nitrification nitrification were studied by comparing key study the amounts of NH4+-N oxidized by trace element-amended soils and read-across based on unamended soils. grouping of substances (category approach)

Test material (IUPAC name): zinc sulfate (See endpoint summary for

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 266 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

justification of read- across) Species/Inoculum: soil NOEC (3 wk): 100 mg/kg 2 (reliable with Premi and Cornfield soil dw element (nominal) restrictions) (1969) This paper reports on the effects of based on: Nitrification adding zinc as sulphate and carbonate key study on ammonification and nitrification on subsequent incubation of soil. read-across based on grouping of substances (category approach)

Test material (IUPAC name): zinc sulfate (See endpoint summary for justification of read- across) Species/Inoculum: soil NOEC (7 wk): 100 mg/kg 2 (reliable with Wilson (1977) soil dw element (nominal) restrictions) Zinc was added to the soils at rates of based on: Nitrification 0, 10 ,100 and 1000 µg Zn/g soil. The key study soils were uniformly treated with 100 NOEC (7 wk): 100 mg/kg µg N/g as NH4Cl and (NO3- + NO2-)- soil dw element (nominal) read-across based on N determined weekly for 7 weeks. based on: Nitrification grouping of substances (category NOEC (7 wk): 50 mg/kg soilapproach) dw element (nominal) based on: Nitrification Test material (IUPAC name): zinc sulfate (See endpoint summary for justification of read- across) Species/Inoculum: soil NOEC (12 wk): 200 mg/kg 2 (reliable with Hemida et al. soil dw element (nominal) restrictions) (1997) Effects of two concentrations (200 and based on: Amidase activity 2000 µg/g soil) of Zn applied to clay or key study sandy soil for 12 weeks on the total NOEC (12 wk): 200 mg/kg counts of fungi, bacteria and soil dw element (nominal) read-across based on actinomycetes were studied. Activities based on: Amidase activity grouping of of 3 soil enzymes were also substances (category investigated. approach) Test material (IUPAC name): zinc sulfate (See endpoint summary for justification of read- across) Species/Inoculum: soil NOEC (30 min): 820 mg/kg 2 (reliable with Al-Khafaji and soil dw element (nominal) restrictions) Tabatabai (1979) Concentrations of 2.5 and 25 µmol based on: Arylsulphatase Zn/g soil were tested in 4 different key study soils on the effect of arylsulfatase. NOEC (30 min): 140 mg/kg soil dw element (nominal) read-across based on based on: Arylsulphatase grouping of substances (category NOEC (30 min): 164 mg/kg approach)

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 267 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

soil dw element (nominal) Test material based on: Arylsulphatase (IUPAC name): zinc sulfate (See endpoint NOEC (30 min): 820 mg/kg summary for soil dw element (nominal) justification of read- based on: Arylsulphatase across) Species/Inoculum: soil NOEC (12 wk): 63 mg/kg 2 (reliable with Hemida et al. soil dw element (nominal) restrictions) (1997) Effects of two concentrations (200 and based on: nitrate reductase 2000 µg/g soil) of Zn applied to clay or activity key study sandy soil for 12 weeks on the total counts of fungi, bacteria and read-across based on actinomycetes were studied. Activities grouping of of 3 soil enzymes were also substances (category investigated. approach) Test material (IUPAC name): zinc sulfate (See endpoint summary for justification of read- across) Species/Inoculum: soil EC10 (1 h): 508 mg/kg soil 2 (reliable with Svenson A. (1986) dw element (nominal) based restrictions) Phosphatase activity was tested with on: Phosphatase different concentrations of ZnSO4 in key study soil. read-across based on grouping of substances (category approach)

Test material (IUPAC name): zinc sulfate (See endpoint summary for justification of read- across) Species/Inoculum: soil NOEC (1 h): 590 mg/kg soil 2 (reliable with Svenson A. (1986) dw element (nominal) based restrictions) Phytase activity was tested with on: Phytase different concentrations of ZnSO4 in key study soil. read-across based on grouping of substances (category approach)

Test material (IUPAC name): zinc sulfate (See endpoint summary for justification of read- across) Species/Inoculum: soil NOEC (30 min): 164 mg/kg 2 (reliable with Juma N.G. & soil dw element (nominal) restrictions) Tabatabai M.A. Studie to evaluate the effects on based on: Phosphatase (1977) activity of acid and alkaline (acid-) key study

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 268 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 phosphatases of 20 trace elements on NOEC (30 min): 164 mg/kg read-across based on inhibition. Concentrations of 2.5 and soil dw element (nominal) grouping of 25 µmole/g were used. based on: Phosphatase substances (category (alkaline-) approach)

Test material (IUPAC name): zinc sulfate (See endpoint summary for justification of read- across) Species/Inoculum: soil NOEC (30 min): 1640 2 (reliable with Stott et al (1985) mg/kg soil dw element restrictions) Pyrophosphatase activity was (nominal) based on: measured 30 minutes after the addition Pyrophosphatase key study of zinc to the soil at concentrations of 2.5 and 25 µmol/g NOEC (30 min): 1640 read-across based on mg/kg soil dw element grouping of (nominal) based on: substances (category Pyrophosphatase approach)

NOEC (30 min): 1640 Test material mg/kg soil dw element (IUPAC name): zinc (nominal) based on: sulfate (See endpoint Pyrophosphatase summary for justification of read- across) Species/Inoculum: soil NOEC (30 min): 109 mg/kg 2 (reliable with Tabatabai M.A. soil dw element (nominal) restrictions) (1977) Urease activity was measured 30 based on: Urease minutes after the addition of zinc to the key study soil. NOEC (30 min): 52 mg/kg soil dw element (nominal) read-across based on based on: Urease grouping of substances (category NOEC (30 min): 64 mg/kg approach) soil dw element (nominal) based on: Urease Test material (IUPAC name): zinc sulfate (See endpoint summary for justification of read- across) Species/Inoculum: soil NOEC (26 wk): 500 mg/kg 2 (reliable with Maliszewska W, soil dw element (nominal) restrictions) Dec S, Wierzbicka Influence of various concentrations of based on: dehydrogenase H and zinc on dehydrogenase activity. activity key study Wozniakowska A (1985) EC10 (26 wk): 76 mg/kg soilread-across based on dw element (nominal) based grouping of on: dehydrogenase activity substances (category approach)

Test material (EC name): zinc sulphate (See endpoint summary for justification of read- across)

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 269 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Species/Inoculum: soil EC10 (24 h): 145 mg/kg soil 2 (reliable with Rogers J.E. & Li dw element (nominal) based restrictions) S.W. (1985) Soil dehydrogenase activity was on: Dehydrogenase assayed by the method of Klein et al. key study (1971) as modified by the authors. EC10 (24 h): 48 mg/kg soil dw element (nominal) based read-across based on on: Dehydrogenase grouping of substances (category approach)

Test material (IUPAC name): zinc sulfate (See endpoint summary for justification of read- across) Species/Inoculum: soil NOEC (7 wk): 233 mg/kg 2 (reliable with Necker & Kunze soil dw element (nominal) restrictions) (1986) “short-term” test in which the effect of based on: nitrate formation zinc (one concentration: 700 mg/kg) on rate (N mineralization) key study N-mineralization was measured during a 7-w period that started immediately read-across based on after the treatment of the soil samples grouping of with zinc in the laboratory. substances (category approach)

Test material (IUPAC name): zinc sulfate (See endpoint summary for justification of read- across) Species/Inoculum: soil NOEC (9 wk): 80 mg/kg soil2 (reliable with Stadelmann, F.X. dw element (nominal) based restrictions) and E. Santschi- Glucose added to soil: 5 g/kg d.w. on: glucose mineralization Fuhrimann (1987) Glucose mineralization (CO2 key study production in 24 h) was measured after 1, 31, and 63 days of exposure read-across based on grouping of substances (category approach)

Test material (IUPAC name): zinc sulfate (See endpoint summary for justification of read- across) Species/Inoculum: soil NOEC (52 d): 162 mg/kg 2 (reliable with Lahr J. et al (2008) soil dw element (meas. (not restrictions) Microcosm experiment to study effect specified)) based on: of zinc and earthworm density on respiration rate key study microbial processes. read-across based on grouping of substances (category approach)

Test material (EC name): zinc sulphate

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 270 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

(See endpoint summary for justification of read- across) Species/Inoculum: soil NOEC (21 d): 100 mg/kg 2 (reliable with Bollag J-M and soil dw element (nominal) restrictions) Barabasz W (1979) equivalent or similar to ISO 14238 based on: denitrifcation key study

read-across based on grouping of substances (category approach)

Test material (EC name): zinc nitrate (See endpoint summary for justification of read- across) Species/Inoculum: soil NOEC (28 d): 50 mg/kg soil 2 (reliable with Saviozzi A, Levi- dw element (nominal) based restrictions) Minzi R, Cardelli R In a laboratory study the effects on soil on: respiration rate (CO2 and Riffaldi R respiration of trace metals added at production) key study (1995) loading rates ranging from 0 to 1000 µg/g were determined. read-across based on grouping of substances (category approach)

Test material (EC name): zinc dichloride (See endpoint summary for justification of read-across) Species/Inoculum: soil NOEC (96 h): 300 mg/kg 2 (reliable with Ohya et al. (1985) soil dw element (nominal) restrictions) The effects of zinc added to a diluvial based on: glucose sandy clay loam soil were investigated. mineralization key study Changes in the soil microflora were followed by counting the microbes and read-across based on measuring their contribution to soil grouping of respiration. substances (category approach)

Test material (IUPAC name): zinc dichloride (See endpoint summary for justification of read-across) Species/Inoculum: soil EC10 (6 wk): 105 mg/kg soil2 (reliable with Haanstra L and dw element (nominal) based restrictions) Doelman P (1991) Effect of added Zn on arylsulphatase on: arylsulphatase activity activity key study EC10 (6 wk): 728 mg/kg soil dw element (nominal) based read-across based on grouping of

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 271 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

on: arylsulphatase activity substances (category approach) EC10 (6 wk): 151 mg/kg soil dw element (nominal) based Test material (EC on: arylsulphatase activity name): zinc chloride (See endpoint EC10 (6 wk): 2353 mg/kg summary for soil dw element (nominal) justification of read- based on: arylsulphatase across) activity Species/Inoculum: soil EC10 (6 wk): 1341 mg/kg 2 (reliable with Doelman P. & soil dw element (nominal) restrictions) Haanstra L. (1989) Short- and long-term effects of heavy based on: Phosphatase metal pollution on phosphatase activity key study was studied in five different soil types. EC10 (6 wk): 2623 mg/kg soil dw element (nominal) read-across based on based on: Phosphatase grouping of substances (category EC10 (6 wk): 160 mg/kg soilapproach) dw element (nominal) based on: Phosphatase Test material (IUPAC name): zinc dichloride (See endpoint summary for justification of read-across) Species/Inoculum: soil EC10 (6 wk): 70 mg/kg soil 2 (reliable with Doelman P and dw element (nominal) based restrictions) Haanstra L (1986) Effect of added Zn on urease activity on: urease activity key study EC10 (6 wk): 30 mg/kg soil dw element (nominal) based read-across based on on: urease activity grouping of substances (category EC10 (6 wk): 30 mg/kg soil approach) dw element (nominal) based on: urease activity Test material (EC name): zinc chloride EC10 (6 wk): 460 mg/kg soil(See endpoint dw element (nominal) based summary for on: urease activity justification of read- across) Species/Inoculum: soil NOEC (1 d): 240 mg/kg soil 2 (reliable with Smolders et al. dw element (nominal) based restrictions) (2003) The authors selected 24h respiration as on: Glucose mineralization an endpoint starting 2 days after key study equilibration of the soil with Zn salts. NOEC (1 d): 30 mg/kg soil The aims were to identify Zn toxicity dw element (nominal) based read-across based on thresholds (EC10, EC50 and NOEC on: Glucose mineralization grouping of values) in the soils examined, and to substances (category compare threshold values determined NOEC (1 d): 800 mg/kg soil approach) in field and laboratory-spikes soils. dw element (nominal) based on: Glucose mineralization Test material (IUPAC name): zinc NOEC (1 d): 100 mg/kg soil dichloride (See dw element (nominal) based endpoint summary on: Glucose mineralization for justification of read-across) NOEC (1 d): 400 mg/kg soil

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 272 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

dw element (nominal) based on: Glucose mineralization

NOEC (1 d): 1300 mg/kg soil dw element (nominal) based on: Glucose mineralization

NOEC (1 d): 600 mg/kg soil dw element (nominal) based on: Glucose mineralization

NOEC (1 d): 1400 mg/kg soil dw element (nominal) based on: Glucose mineralization

NOEC (1 d): 300 mg/kg soil dw element (nominal) based on: Glucose mineralization

NOEC (1 d): 50 mg/kg soil dw element (nominal) based on: Glucose mineralization

NOEC (1 d): 100 mg/kg soil dw element (nominal) based on: Glucose mineralization

NOEC (1 d): 100 mg/kg soil dw element (nominal) based on: Glucose mineralization Species/Inoculum: soil NOEC (28 d): 120 mg/kg 2 (reliable with Smolders et al. soil dw element (nominal) restrictions) (2003) equivalent or similar to OECD based on: Maize residue Guideline 217 (Soil Microorganisms: mineralization key study Carbon Transformation Test) NOEC (28 d): 200 mg/kg read-across based on soil dw element (nominal) grouping of based on: Maize residue substances (category mineralization approach)

NOEC (28 d): 469 mg/kg Test material soil dw element (nominal) (IUPAC name): zinc based on: Maize residue dichloride (See mineralization endpoint summary for justification of NOEC (28 d): 50 mg/kg soil read-across) dw element (nominal) based on: Maize residue mineralization

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 273 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

NOEC (28 d): 1300 mg/kg soil dw element (nominal) based on: Maize residue mineralization

NOEC (28 d): 1400 mg/kg soil dw element (nominal) based on: Maize residue mineralization

NOEC (28 d): 38 mg/kg soil dw element (nominal) based on: Maize residue mineralization

NOEC (28 d): 150 mg/kg soil dw element (nominal) based on: Maize residue mineralization

NOEC (28 d): 600 mg/kg soil dw element (nominal) based on: Maize residue mineralization

NOEC (28 d): 150 mg/kg soil dw element (nominal) based on: Maize residue mineralization

NOEC (28 d): 300 mg/kg soil dw element (nominal) based on: Maize residue mineralization Species/Inoculum: soil NOEC (7 d): 400 mg/kg soil 2 (reliable with Smolders et al. dw element (nominal) based restrictions) (2003) equivalent or similar to ISO 14238 on: Nitrification key study NOEC (7 d): 257 mg/kg soil dw element (nominal) based read-across based on on: Nitrification grouping of substances (category NOEC (28 d): 50 mg/kg soil approach) dw element (nominal) based on: Nitrification Test material (IUPAC name): zinc NOEC (14 d): 50 mg/kg soil dichloride (See dw element (nominal) based endpoint summary on: Nitrification for justification of read-across) NOEC (4 d): 424 mg/kg soil dw element (nominal) based on: Nitrification

NOEC (14 d): 38 mg/kg soil dw element (nominal) based on: Nitrification

NOEC (7 d): 206 mg/kg soil dw element (nominal) based

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 274 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

on: Nitrification

NOEC (4 d): 75 mg/kg soil dw element (nominal) based on: Nitrification

NOEC (4 d): 150 mg/kg soil dw element (nominal) based on: Nitrification

NOEC (4 d): 300 mg/kg soil dw element (nominal) based on: Nitrification

NOEC (23 d): 150 mg/kg soil dw element (nominal) based on: Nitrification

NOEC (4 d): 300 mg/kg soil dw element (nominal) based on: Nitrification

NOEC (10 d): 75 mg/kg soil dw element (nominal) based on: Nitrification Species/Inoculum: soil NOEC (2 d): 100 mg/kg soil 2 (reliable with Notenboom & dw element (nominal) based restrictions) Posthuma (1994) By adding sodiumglutamate to soil and on: glutamic acid measuring the release of CO2 we can mineralization key study Posthuma et al. see the response of soil (1998) microorganisms. This response varies NOEC (2 d): 100 mg/kg soil read-across based on incase of toxic products in the soil. dw element (nominal) based grouping of (Method from Haanstra & Doelman on: glutamic acid substances (category 1984) mineralization approach)

NOEC (2 d): 30 mg/kg soil Test material dw element (nominal) based (IUPAC name): zinc on: glutamic acid dichloride (See mineralization endpoint summary for justification of NOEC (2 d): 55 mg/kg soil read-across) dw element (nominal) based on: glutamic acid mineralization Species/Inoculum: soil NOEC (18 h): 303 mg/kg 2 (reliable with van Beelen et al. soil dw element (nominal) restrictions) (1994) The efffects of zinc on the based on: acetate mineralization of 1µg/L 14C acetate to mineralization key study 14CO2 was studied in an acid surface soil, an acid subsoil, an calcerous read-across based on surface soil and a calcereous sub-soil. grouping of substances (category approach)

Test material (IUPAC name): zinc dichloride (See endpoint summary for justification of

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 275 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

read-across) Species/Inoculum: soil EC10 (11 d): 39 mg/kg soil 2 (reliable with De Brouwere K. et dw element (nominal) based restrictions) al (2007) Net production of N2O and N2 was on: denitrification monitored during anaerobic key study incubations (25 °C, He atmosphere) of soils freshly spiked with ZnCl2 and of read-across based on corresponding soils that were gradually grouping of enriched with metals (mainly Zn) in substances (category the ﬁ eld by previous sludge approach) amendments or by corrosion of galvanized structures. Test material (EC name): zinc chloride (See endpoint summary for justification of read- across) Species/Inoculum: soil NOEC (55 d): 50 mg/kg soil 2 (reliable with Merckx R., Brans dw element (nominal) based restrictions) K., Smolders E. Effect of Zn on C-mineralisation was on: respiration rate (2001) assessed in 4 soils as measured by key study degradation of dissolved organic NOEC (55 d): 50 mg/kg soil carbon released after dry-wet cycles. dw element (nominal) based read-across based on on: respiration rate grouping of substances (category NOEC (55 d): 150 mg/kg approach) soil dw element (nominal) based on: respiration rate Test material (EC name): zinc chloride NOEC (55 d): 150 mg/kg (See endpoint soil dw element (nominal) summary for based on: respiration rate justification of read- across) Species/Inoculum: soil EC10 (8 d): 49 mg/kg soil 2 (reliable with Smolders E. et al dw element (meas. (not restrictions) (2003) The soil nitriﬁ cation potential was specified)) based on: measured from the increase in soil Potential nitrification rate key study nitrate (NO3-) in soils amended with ammonium (NH4+) salts. EC10 (28 d): 446 mg/kg soil read-across based on grouping of Basal respiration was measured by dw element (meas. (not substances (category trapping CO2 evolved from the soil specified)) based on: N- approach) sample in 1 M NaOH and titrating mineralisation against standardized HCl. EC10 (3 d): 140 mg/kg soil Test material (EC Substrate-induced respiration was dw element (meas. (not name): zinc chloride measured from the 24-h CO2 specified)) based on: (See endpoint production in glucose-amended soils. Potential nitrification rate summary for Nitrogen mineralization was measured justification of read- based on the Organisation for EC10 (28 d): 204 mg/kg soil across) Economic Cooperation and dw element (meas. (not Development-216 test protocol [14] specified)) based on: Basal using lucerne as the plant meal. respiration EC10 (28 d): 133 mg/kg soil dw element (meas. (not specified)) based on: Basal respiration

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 276 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Discussion

For microbial assays, in total 108 individual high quality NOEC/EC10s are selected for the PNEC derivation. These values represent 4 nitrogen transformation processes, 5 carbon transformation processes and 8 enzymatic processes and range from 17 mg Zn/kg dw for respiration (Chang and Broadbent, 1981 and Lighthart et al., 1983) to 2623 mg Zn/kg dw for phosphatase (Doelman and Haanstra, 1989).

All toxicity data are expressed as added Zn concentration in soil, based on either the nominal dose added or on measured, background corrected soil Zn concentrations.

The following information is taken into account for toxicity on soil micro-organisms for the derivation of PNEC:

For microbial assays, in total 108 individual high quality NOEC/EC10s represent 4 nitrogen transformation processes, 5 carbon transformation processes and 8 enzymatic processes and range from 17 mg Zn/kg dw for respiration to 2623 mg Zn/kg dw for phosphatase.

7.2.1.4. Toxicity to other terrestrial organisms

No data

7.2.2. Calculation of Predicted No Effect Concentration (PNEC soil)

The PNEC soil is derived from by statistical extrapolation from the species sensitivity distruibutions, described above. To set the PNEC from the HC5, an uncertainty analysis is made.

Uncertainty analysis

Based on uncertainty considerations an additional assessment factor between 1 and 5 must be applied on the 50% confidence value of the 5th percentile value (thus PNEC = HC5-50/AF). The choice of the assessment factor has to be judged on a case-by-case basis. Based on the available data, the following points have to be considered when determining the size of the assessment factor:

1. The overall quality of the database and the end-points covered, e.g., if all the data are generated from “true” chronic studies  The Zn-database coveres ecologically relevant endpoints. The selected endpoints were all relevant for potential effects at the population level: root, shoot or grain yield and blooming for the terrestrial plants; reproduction for the invertebrates; N- and C-transformation processes and enzyme activity for micro-organisms.  The reliable EC10/NOEC data were extracted from tests performed in natural and artificial soils, covering the wide range of soil characteristics in Europe (pH, organic matter, clay, CEC and background Zn, Table 82). It can be concluded that the soils covered by the toxicity data properly reflect the variability in physico-chemical conditions encountered in European soils.

Table 82. Soil characteristics of the selected toxicity studies. Parameter Plants Invertebrates Microbial tests pH 4.0-8.3 3.0-7.5 3.0-8.2 (3.0-7.7)* Org C (%) 0.3-5.8 0.4-23.0 0.4-37.1 (0.4-23.0) Clay (%) 2-69 1-51 1-60

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 277 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

CEC (cmolc/kg) 1.0-61.0 1.9-38.1 1.9-41.3 Background Zn 2-80 2-191 4-226 (mg/kg) *: range for data with information on abiotic factors allowing correction for differences in bioavailability among soils (if different from total range)

 All toxicity data are derived from chronic toxicity tests. For plants, exposure periods vary between 21 and 150 days. Endpoints studied cover all relevant life stages, including, emergence, growth and reproduction (grain yield, number of pods, blooming). Both Sheppard et al. (1993) and Warne et al. (2008b) observed that shoot yield (based on plant height or weight) is often a more sensitive endpoint compared to grain yield.  The exposure time for all invertebrate tests also varied between 21 and 150 days. Reproduction was for all organisms identified as the most sensitive endpoint.  Exposure times for microbial tests ranged between 30 minutes (enzyme tests in suspension) and 90 days.

Conclusion: True chronic data are available for multiple endpoints for plant species, invertebrate species and microbial processes, and for a representative range of soil types. The overall quality of the database can therefore be considered as optimal, there is no justification for an additional assessment factor.

2. The diversity and representativeness of the taxonomic groups covered by the database It is clear that the Zn-database largely fulfils the requirement of 10-15 different EC10/NOEC values (preferably more than 15) from chronic/long term studies for different species covering at least 8 different taxonomic groups from 3 trophic levels. In total, 168 individual NOEC/EC10 values are available, covering 9 plant species, 8 invertebrate species and 12 microbial processes.  Plants: 31 NOEC or EC10 values are available for normalization: monocotyle and dicotyle plants including agricultural species belonging to 9 different species and 4 different families: - Zea mays, Avena sativa, Hordeum vulgare and Triticum aestivum – family of the Poaceae; - Allium cepa – family of the Alliaceae; - Brassica rapa – family of the Brassicaceae; - Trigonella poenum, Trifolium pratense and Vicia sativa – family of the Fabaceae. Additionally, 14 extra NOEC or EC10 data for plants were available but did not have the required information on soil properties (CEC and pH). These data also include 9 species from 5 families (3 families not covered by normalization dataset): - Sorghum bicolor and Triticum vulgare – family of the Poaceae; - Lycopersicon esculentum – family of the Solanaceae; - Medicago sativa, Pisum sativum and Vigna mungo – family of the Fabaceae; - Lactuca sativa – family of the Asteraceae; - Spinacea oleracea and Beta vulgaris – family of the Amaranthaceae.

 Invertebrates: 61 NOEC or EC10 values are available for normalization, covering both hard- and soft-bodied organisms (arthropoda and oligochaeta, respectively) with different exposure routes and feeding strategies belonging to 8 different species and 4 different families: - Aporectadea caliginosa, Eisenia Andrei, Eisenia fetida, Lumbricus rubellus and Lumbricus terrestris – family of the Lumbricidae (oligochaeta); - Enchytraeus crypticus – family of the Enchytraeidae (oligochaeta); - Folsomia candida – family of the Isotomidae (Collembola, arthropoda); - Sinella curvesita – family of the Entomobrydae (Collembola, arthropoda). No data for Gastropoda and Crustacea exposed to soil are available. However, studies are available for snails fed with a Zn-spiked diet. As mentioned in the risk assessment, snails and crustaceae e.g. woodlice are living more on the soil than in the soil and are feeding especially on plants and litter/organic detritus, respectively. The lowest effect concentration in studies of snails (Helix aspersa, Gastropoda) exposed to such Zn-contaminated food was an EC20 value of 855 mg Zn/kg dry weight food (Laskowski and Hopkin, 1996; Gomot-De-Vaufleury, 2000). This is significantly above the normal plant tissue concentrations for Zn in unpolluted soils (15-150 mg Zn/kg, Chaney et al., 1983). Because Zn is an essential element, its uptake is regulated and Zn does not tend to accumulate in healthy plants. It is therefore concluded that a PNEC value protecting plants will also protect herbivorous organisms.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 278 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

 Microbial processes: 76 NOEC or EC10 values are available for normalization, covering 5 C- transformation reactions (respiration, glucose mineralization, acetate mineralization, glutamic acid mineralization and maize residue mineralization), 4 N-transformation reactions (nitrification, ammonification, N-mineralization and denitrification) and 4 enzymatic processes (arylsulphatase, dehydrogenase, phosphatase and urease). Additionally, 32 other NOEC or EC10 data for microbial processes were available but did not have the required information on soil properties (background Zn concentration). These data also include 4 additional enzymatic processes: phytase, nitrate reductase, amidase and pyrophosphatase. Conclusion: The database is composed of plant, invertebrate and microbial data and all major taxonomic groups are included. Therefore, the overall quality of the database is considered as optimal, there is no justification for an additional assessment factor.

3. Statistical uncertainties around the 5th percentile estimate, e.g., reflected in the goodness-of-fit or the size of confidence interval around the 5th percentile The log-normal distribution has been evaluated for different soil types or scenarios. Both statistical (e.g. Kolmogorov-Smirnov, Andersen-Darling tests) and visual (e.g. Q-Q plots) goodness-of-fit techniques were used in order to select the most appropriate distribution function for the compiled chronic data set. Most weight was given to the Anderson-Darling goodness-of-fit test as this test highlights differences between the tail of the distribution (lower tail is the region of interest) and the input data. For all distributions tested (generic scenario’s and 8 soil types), the log-normal distribution was accepted by the Anderson-Darling goodness-of-fit test at the P=0.1 level. Several observations suggest the combined use of single-species data for plants and invertebrates and multiple-species microbial data into one common species/process sensitivity distribution: - The distribution for microbial endpoint as presented in the RAR covers, in reality, the distribution of the responses of different soil communities to the same endpoints, and therefore, in the opinion of SCHER (2007), represents a distribution of “communities' sensitivities”. This is therefore comparable to the SSD concept as derived for single-species, and allows the combination of both distributions, once it has been demonstrated that the distributions are equivalent, for improving the statistical analysis. - The toxicity data clearly overlap among plants, invertebrates and microbial processes and there is no indication for differences in sensitivity among groups of organisms. - The abiotic availability factors for the various organisms (CEC and background Zn) are positively correlated indicating similar trend in toxicity with varying soil properties - The bioavailability factors for the common SSD agree with minimum BioF values derived for either plants and invertebrates or micro-organisms.

Conclusion: the log-normal distribution has been selected for derivation of the HC5-50 based on one species sensitivity distribution including plants, invertebrates and microbial processes, there is no justification for an additional assessment factor.

4. Evaluation of NOEC values below the HC5-50 A comparison of the normalized HC5-50 values with the normalized species/process mean NOEC/EC10 values for 8 soil scenarios (Tables 77 and 78) shows that only results for Hordeum vulgare consistently fall below the HC5-50 derived by the log-normal distribution. The normalised species mean value for Hordeum vulgare is only based on one datapoint (Luo and Rimmer, 1995), where the NOEC is estimated as LOEC/3 (23% effect at 100 mg Zn/kg) because of the high separation factor between doses (0-10-100 mg Zn/kg). This is a conservative approach. Indeed, two additional NOEC values of 100 and 215 mg Zn/kg are available for Hordeum vulgare (Aery and Jagetiya, 1997; Boawn and Rasmussen, 1971). These values are significantly higher than 33.3 mg Zn/kg, but unfortunately no soil properties were available for these extra data and therefore they could not be used in the normalisation approach. Conclusion: the low value for Hordeum vulgare is probably overly conservative and therefore the HC5-50 is considered protective for the terrestrial ecosystem, there is no justification for an additional assessment factor.

5. Comparisons between field/microcosm studies and the 5th percentile to evaluate the laboratory to field extrapolation. The method to derive the PNEC to terrestrial organisms typically relies on the use of single species tested in freshly laboratory spiked soils. Validation of the proposed HC5-50 could be carried out by comparing this HC5-50 value with NOEC values from field or microcosm studies. An extensive discussion was reported in

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 279 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

the RAR for Zn, concluding that “the HC5-50 value is generally in good agreement with actual no-effect levels in the field”. More recent observations confirm this conclusion: A study on Zn toxicity to wheat grown in 11 field-contaminated soils in Australia reports EC10added values between 91 and 4760 mg Zn/kg for grain yield at harvest and between 17.2 and 1210 mg Zn/kg for plant biomass after 8 weeks growth (Warne et al., 2008b). The normalised and lab-to-field corrected HC5-50 values are protective for both endpoints in all 11 soils, except for 8-week biomass in one soil. However that particular EC10 value (17.2 mg Zn/kg) has an extremely wide confidence interval (0.04 – 7580 mg Zn/kg) and the corresponding EC10 value for grain yield is two orders of magnitude higher (4760 mg Zn/kg). Therefore this EC10 for plant biomass is not considered reliable and the proposed HC5-50 values are considered protective for all soils. Chaudri et al., 2008 did not observe an effect of Zn concentrations up to 339-466 mg Zn/kg on the population of Rhizobium leguminosarum in 3 different agricultural sites 10 years after contamination with zinc carbonate or liquid sludges. These concentration are all well above the corresponding HC5-50 values for these soils. Only for soils amended with Zn-contaminated sludges, a significant effect was observed at Zn conentrations around 100 mg Zn/kg. This was attributed to the heterogeneous distribution of Zn in sludge-amended soils and the presence of local hot-spots with high Zn concentrations (>> 300 mg Zn/kg) in sludge particles. Conclusion: the available field studies indicate the normalised HC5-50 values to be protective under field conditions, there is no justification for an additional assessment factor.

Overall conclusion on PNECsoil:

Based on the above uncertainty analysis, and in particular a large toxicity database covering a representative range in plant and invertebrate species, microbial processes and soil conditions, an extensive field validation and the availability of normalisation models, it can be concluded that the available database and models allow for the derivation of an HC5-50 that is protective for the terrestrial environment. The application of an AF = 1 is therefore proposed on the HC5-50 derived with the statistical extrapolation method. This provides a robust and ecological relevant PNEC to be retained for the risk characterisation (derived by statistical extrapolation method with the log-normal distribution).

3 types of PNECadd soil can be considered:

- The generic PNECadd based on the entire ecotoxicity database is 35.6 mg Zn/kg.

-The generic PNECadd, ageing which is (in accordance with the EU risk assessment) resulting from multiplying the generic PNECadd with the default “lab-to-field” correction factor of 3 for taking into account differences of zinc bioavailability after ageing: generic PNECadd including ageing or generic PNECadd, ageing = 107mg/kg dw.

- If information on soil type and soil conditions is available, a soil-specific PNECadd,ageing can be calculated, by applying a further correction for bioavailability. A tool is available for this. For illustration, some examples were developed in the present analysis resulting in PNECadded values for soil types representative for the EU conditions between approx. 30 and 300mg Zn/kg (table 77).

Table 83. PNEC soil

PNEC Assessment Remarks/Justification factor PNEC soil: 35.6 1 Extrapolation method: statistical extrapolation mg/kg soil dw The given value is the generic PNECadd, i.e. it has to be added to the natural background concentration of zinc.

This generic PNECadd, should as a rule be multiplied with a factor 3 to take into account "lab-to-field" differences in toxicity. As such, the generic corrected PNECadd is 107 mg Zn/kg dw. Soil-specific PNEC values can further be calculated when the characteristics of the soil are documented.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 280 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

7.3. Atmospheric compartment

The EU risk assessment concluded on this compartment: “A quantitative risk characterisation for exposure of organisms to airborne zinc is not possible. This because there are no useful data on the effects of airborne zinc on environmental organisms and thus no PNEC for air could be derived. The PECs in air will be used for the risk assessment of man indirectly exposed via the environment”.

In accordance to the EU risk assessment, this compartment was not further assessed.

7.4. Microbiological activity in sewage treatment systems

Establishing the dataset

The EU risk assessment on zinc (ECB 2008) reviewed the available data for zinc toxicity to microorganisms, to set the PNEC for STP. The most recent reference mentioned in the risk assessment report is from 1997. A number of other results on micro-organisms were considered not relevant in the risk assessment process for reasons that are also applicable to the REACH analyses (REACH guidance chapter R.10, table R.10-6), e.g.: results on Pseudomonas fluorescens, Vibrio fischeri were considered less relevant. Results obtained on an industrial activated sludge were (and are for the present analysis) also not considered relevant for the setting of the PNEC-STP. In the EU risk assessment, a number of results obtained on protozoa were also reviewed, but for the setting of the PNEC-STP, preference was given to the activated sludge test.

According to the approach followed in the EU risk assessment, results on activated sludge respiration inhibition are also used for the present analysis. The available literature on this endpoint was checked for information becoming available after 1997.

In addition to the data from the RA, the following information has been reviewed and identified as relevant: a) zinc toxicity on activated sludge bacteria growth (Cabrero et al, 1998); b) zinc toxicity on activated sludge respiration and nitrification (Dalzell et al 2002).; c) ATP luminescence proved to be insensitive to zinc (Dalzell & Christophi 2002). Next to this, a nitrification inhibition test result was also available (Juliastuti et al. 2003). According to the guidance preference should be given to nitrification inhibition because respiration inhibition is a less sensitive endpoint. A study conducted by Juliastuti et al. (2003) reported a NOEC of 0.1 mg Zn/L in an ISO 9509 nitrification inhibition test. According to the guidance for PNEC setting (ECHA R.10, table R.10.6) this result is considered most relevant for setting the PNEC STP. Applying the rules of PNEC setting, this NOEC results in a PNEC STP of 100 µg Zn/L (AF of 1 ; ECHA R.10, table R.10.6).

Results relevant for PNEC-STP setting are summarised in table below. Details on these studies are given in table 84.

Table 84. Summary of study results for STP-PNEC study NOEC (mg Zn/l) E(I)C50 (mg Zn/l) reference Nitrification inhibition 0.1 0.35 Juliastuti et al. (2003) Respiration inhibition / 5.2 Dutka et al 1983 Respiration inhibition / 10 Cabrero et al 1998 Nitrification inhibition / >10 Sludge bacterial growth 5 / Dalzell et al 2002

7.4.1. Toxicity to aquatic micro-organisms

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 281 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

The results are summarised in the following table:

Table 85. Overview of effects on micro-organisms Method Results Remarks Reference activated sludge of a predominantly EC50 (3 h): 5.2 mg/L based 2 (reliable with Dutka BJ, Nyholm domestic sewage on: respiration rate restrictions) N and Petersen J. (1983) freshwater key study static read-across based on grouping of equivalent or similar to OECD substances (category Guideline 209 (Activated Sludge, approach) Respiration Inhibition Test) Test material (IUPAC name): zinc sulphate (See endpoint summary for justification of read-across) activated sludge of a predominantly NOEC (4 h): 0.1 mg/L 1 (reliable without Juliastuti, S.R.; domestic sewage element (nominal) based on: restriction) Baeyens J. ; nitrification rate Creemers C. (2003) freshwater key study IC50 (4 h): 0.35 mg/L static element (nominal) based on: read-across based on nitrification rate grouping of ISO DIS 9509 (Method for Assessing substances (category the Inhibition of Nitrification of approach) Activated Sludge Microorganisms by Chemicals and Waste Waters) Test material (EC name): Zinc Sulphate (See endpoint summary for justification of read-across) activated sludge NOEC (3 d): 5 mg/L 2 (reliable with Cabrero A, (nominal) based on: growth restrictions) Fernandez S, freshwater inhibition Mirada F and supporting study Garcia J. (1998) static read-across based on Lab-designed batch growth system for grouping of measurement of inhibition kinetics of substances (category activated sludge process. approach)

Test material (IUPAC name): zinc sulfate (See endpoint summary for justification of read- across) activated sludge, domestic EC50 (30 min): 1618 mg/L 2 (reliable with BASF AG (1988) test mat. (nominal) based on:restrictions) freshwater respiration rate supporting study static experimental result equivalent or similar to OECD Guideline 209 (Activated Sludge, Test material (CAS

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 282 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Method Results Remarks Reference

Respiration Inhibition Test) name): Ammonium chloride activated sludge IC50 (3 h): > 10 mg/L 2 (reliable with Dalzell DJB, Alte (nominal) based on: restrictions) S, Aspichueta E, de freshwater respiration rate la Sota A, supporting study Etxebarria J, static IC50 (3 h): 10 mg/L Guttierrez (2002) (nominal) based on: read-across based on ISO 8192 (Test for Inhibition of nitrification rate grouping of Oxygen Consumption by Activated substances (category Sludge) (for the respiration approach) measurements) Test material equivalent or similar to Swedish EPA (IUPAC name): zinc report Nr 4424 (1995) for the sulfate (See endpoint inhibition of nitrification summary for justification of read- across)

Discussion

Several data are available for this endpoint. Formerly (ECB 2008), and in the REACH-registrations of a number of zinc substances of November 2010, a PNEC of 52 µg/L for STP was derived, based on the lowest EC50 of 5.2 mg Zn/L observed in a sludge respiration inhibition test (Dutka et al. 1983). According to the guidance (ECHA 7.8.17.1) preference should be given to nitrification inhibition because respiration inhibition is a less sensitive endpoint. A study conducted by Juliastuti et al. (2003) reported a NOEC of 0.1 mg Zn/L in an ISO 9509 nitrification inhibition test. Applying the rules for PNEC setting (ECHA R.10, table R.10.6) this result yields a PNEC STP of 100 µg Zn/L (AF of 1).

The following information is taken into account for effects on aquatic micro-organisms for the derivation of PNEC:

A nitrification inhibition test with Zn sulfate on activated slugde originating from a municipal wastewater treatment plant was performed leading to a NOEC of 100 µg Zn/L.

Value used for CSA:

EC10/LC10 or NOEC for aquatic micro-organisms: 100 µg/L

7.4.2. PNEC for sewage treatment plant

According to the guidance for PNEC setting (ECHA R.10, table R.10.6) the nitrification inhibition test is considered most relevant for setting the PNEC STP. The PNEC derived from the NOEC value of 100 µg Zn/l of Juliastuti et al. (2003) is set, applying an assessment factor of 1: PNEC-STP of 100µg Zn/l.

Table 86. PNEC sewage treatment plant Value Assessment Remarks/Justification factor PNEC STP: 100 1 Extrapolation method: assessment factor µg/L Considering that the nitrification inhibition test is most relevant of the data available, the PNEC is derived from the NOEC (100 µg Zn/l ; Juliastuti et al. 2003) divided by AF 1 to give the PNEC-STP of 100µg Zn/l.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 283 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

7.5. Non compartment specific effects relevant for the food chain (secondary poisoning)

The EU risk assessment concluded the following related to this issue: “Based on the ICDZ data (Cleven et al., 1993) on bioaccumulation of zinc in animals and on biomagnification (i.e. accumulation and transfer through the food chain), it is concluded that secondary poisoning is considered to be not relevant in the effect assessment of zinc. Major decision points for this conclusion are the following. The accumulation of zinc, an essential element, is regulated in animals of several taxonomic groups, for example in molluscs, crustaceans, fish and mammals. In mammals, one of the two target species for secondary poisoning, both the absorption of zinc from the diet and the excretion of zinc, are regulated. This allows mammals, within certain limits, to maintain their total body zinc level (whole body homeostasis) and to maintain physiologically required levels of zinc in their various tissues, both at low and high dietary zinc intakes. The results of field studies, in which relatively small differences were found in the zinc levels of small mammals from control and polluted sites, are in accordance with the homeostatic mechanism. These data indicate that the bioaccumulation potential of zinc in both herbivorous and carnivorous mammals will be low. Based on the above data, secondary poisoning and the related issues bioaccumulation and biomagnification are not further discussed in this report” (EU risk assessment, ECB 2008).

7.5.1. Toxicity to birds

Data waiving

Information requirement: Toxicity to birds Reason: other justification Justification: zinc is an essential element which is regulated throughout the food chain. It does not bioaccumulate/biomagnify. Therefore testing of long-term toxicity to birds is considered not relevant.

Discussion

The aim of avian toxicity tests is to provide data for secondary poisoning, if the chemical safety assessment demonstrates the need for such a study (notably relevant for substances with a potential to bioaccumulate and high mammalian toxicity). Zinc is an essential element that is regulated throughout the food chain and does not bioaccumulate/biomagnify. For this reason, the potential for secondary poisoning is not considered relevant (EU risk assessment, ECB 2008), and testing of long-term toxicity to birds is considered not relevant.

The following information is taken into account for effects on birds for the derivation of PNEC:

Zinc is an essential element which is regulated throughout the food chain. It does not bioaccumulate/biomagnify. Therefore testing of long-term toxicity to birds is considered not relevant.

7.5.2. Toxicity to mammals

See 7.5.1.

7.5.3. Calculation of PNECoral (secondary poisoning)

Table 87. PNEC oral

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 284 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

PNEC Assessment Remarks/Justification factor No potential for Zinc is an essential element that is actively regulated within the body of bioaccumulation all organisms. Due to the general lack of increased whole body and tissue concentrations at higher exposure levels, the zinc BCF data show generally an inverse relationship to exposure concentrations (McGeer et al 2003). The physiological basis for the inverse relationship of BCF to zinc exposure concentration arises from Zn uptake and control mechanisms. At low environmental zinc levels, organisms are able to sequester and retain Zn in tissues for essential functions. When Zn exposure is higher, aquatic organisms are able to control uptake. There is clear evidence that many species actively regulate their body Zn concentrations, including crustaceans, oligochaetes, mussels, gastropods, fish, amphipods, chironomids by different mechanisms (McGeer et al 2003). The bioaccumulation potential in mammals is also considered low. Based on this, the EU risk assessment concludes that secondary poisoning is considered to be not relevant in the effect assessment for zinc.

7.6. Conclusion on the environmental hazard assessment and on classification and labelling

Environmental classification justification 7.6.1. Classification under Annex I dangerous substances directive 67/548/EEC

Diammonium tetrachlorozincate (2 -) was not classified under Directive 67/548/EEC, and does not figure in.Annex 1 of Directive 67/548/EEC (ECB 2008). 7.6.2. Classification under 2nd Adaptation to Technical Progress (ATP) to the CLP Regulation (2nd ATP CLP)

The ecotoxicity of Zn compounds is attributed to that of the Zn++ cation and thus depends on the solubility of the compound (i.e. its capacity to release the Zn++ cation). The following justification is given for classifying Diammonium tetrachlorozincate (2 -):

-classification for acute aquatic effects:

Considering a) the lowest acute aquatic ecotoxicity values of 136 µg Zn/l and 413 µg Zn/l for the zinc ion at pH 8 and 6 respectively, and b) the molecular weight ratio of Diammonium tetrachlorozincate (2 -) versus the Zn+ +ion (136 x MW (Cl4Zn.2H4N) / MW (Zn) = 136 x 243/65 = 509µg substance /l at pH 8 as worst case), Diammonium tetrachlorozincate (2 -) is classified acute 1 (H400; very toxic to aquatic life), with M-Factor 1.

-classification for chronic effect

The lowest chronic ecotoxicity value observed on a wide variety of species of different taxonomic groups is 19 µg Zn/l (section 7.1.1.2.), The reference value for the substance is calculated by factoring in the MW ratio between Zn and the substance (19µg/l x MW (Cl4Zn.2H4N) / MW (Zn) = 19 x 243/65 = 71µg of substance /l). For determination of the chronic aquatic effects classification according to the 2nd ATP CLP criteria, it has to be considered further if the substance is rapidly degradable or not.

The concept of “Degradability” was developed for organic substances and is not applicable as such to inorganic substances like zinc.As a surrogate approach for assessing “degradability”, the concept of “removal from the water column” was developed to assess whether or not a given metal ion would remain present in the water column upon addition (and thus be able to excert a chronic effect) or would be rapidly removed from the water column. In this concept, “rapid removal from the water column” (defined as >70% removal within 28 days) is considered as equivalent to “rapidly degradable”. Under section 4.6., the rapid removal of zinc from the water

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 285 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 column is documented. Consequently, zinc and zinc compounds are considered as equivalent to being ‘rapidly degradable” in the context of classification for chronic aquatic effects.

Considering the chronic ecotoxicity reference value for Diammonium tetrachlorozincate (2 -) of 71 µg /l, and considering zinc and its compounds as equivalent to being rapidly degradable, the classification of the substance for chronic aquatic effect is “chronic 2” (H411: Toxic to aquatic life with long lasting effects).

General discussion

A basic assumption made in this hazard assessment and throughout this CSR, (in accordance to the same assumption made in the EU RA process) is that the ecotoxicity of zinc and zinc compounds is due to the Zn+ +ion. As a consequence, all aquatic, sediment and terrestrial toxicity data in this report are expressed as “zinc”, not as the test compound as such, because ionic zinc is considered to be the causative factor for toxicity. A further consequence of this is that all ecotoxicity data obtained on different zinc compounds, are mutually relevant for each other. For that reason, the available ecotoxicity databases related to zinc and the different zinc compounds are combined before calculating the PNECs. The only way zinc compounds can differ in this respect is in their capacity to release zinc ions into (environmental) solution. That effect is checked eventually in the transformation/dissolution tests and may result in different classifications.

8. PBT AND VPVB ASSESSMENT 8.1. Assessment of PBT/vPvB Properties

According to regulation (EC) 1907/2006 (REACH) a PBT and vPvB assessment shall usually be conducted as foreseen in Article 14 (3) (d) in conjunction with Annex I Section 4 and according to the criteria laid down in Annex XIII. However, according to Annex XIII a PBT and vPvB assessment shall not be conducted for inorganic substances. Diammonium tetrachlorozincate (2 -) is an inorganic substance, thus a PBT and vPvB assessment is not required.

Still, the points below are raised:

8.1.1. Summary and overall conclusions on PBT or vPvB properties

Zinc is a natural, essential element, which is needed for the optimal growth and development of all living organisms, including man. All living organisms have homeostasis mechanisms that actively regulate zinc uptake and absorption/excretion from the body; due to this regulation, zinc and zinc compounds do not bioaccumulate or biomagnify.

Zinc is an element, and as such the criterion “persistence” is not relevant for the metal and its inorganic compounds in a way as it is applied to organic substances. The removal of inorganic substances from the water column has been discussed as a surrogate for persistence. In section 4.6., the rapid removal of zinc from the water column is documented. So, zinc does not meet this criterion, neither.

Considering the above, zinc and zinc compounds are not PBT or vPvB.

9. EXPOSURE ASSESSMENT (with local risk characterisation) Introduction

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 286 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

In the EU risk assessment on zinc metal and 5 zinc compounds (ZnO, ZnCl2, ZnSO4, Zn3(PO4)2, Zn- distearate), made in the framework of the Regulation 793/93/EEC, an extensive analysis was made for the local and regional exposures to man and environment. This extensive analysis is used as a starting basis for the present assessments, too.

However, some extensions and updates were made in the present analysis, where appropriate. These are shortly discussed below.

Local assessments for workers and environment. For assessment of exposures at local scale, several generic exposure scenarios (GES) were developed for each zinc substance. This was necessary because of the significant number of uses that was identified for each substance. The multitude of identified uses was assigned to the respective GES based on similarity of process, and , consequently, similarity in exposure and risk management measures. So, GES are relevant for the different identified uses that they group at the same time.

Structure of the local assessments Because they are so numerous, and for reasons of clarity and consistency, the local exposure scenarios, the local exposure assessments and the local risk characterisations related to each of the GES are presented together under chapter 9.

Approaches for local exposure assessment For the local exposure assessments, the data reported in the EU risk assessment are used as the starting point. Where appropriate (data gaps, scenarios showing risk), the exposure data from the RA were completed and updated, to better reflect the local situations today. The detail of the data reported by the companies is available for consultation at IZA. In general, measured data are used by preference for the exposure assessment. For a number of the scenarios, no measured exposure data are available; in that case exposure was modelled with established exposure models.  Assessment of workers exposure is related to the place /process the worker is involved in. The GES group different processes; exposure assessment is done using the worst case approach by considering full shift exposure at the workplace with highest potential for exposure. Risk management measures are specified accordingly.  Environmental emissions (notably to water) are usually integrating the totality of emissions from a given site, and cannot be distinguished for each process. Therefore assessments in the GES are done for the site as a whole.  A specific scenario was added on zinc in municipal waste water treatment plants

Consumer assessment. Conform to the approach followed in the EU risk assessment, consumer exposure has been assessed in an integrated way, by combining the exposures following from the use/consumption of different articles containing zinc from different zinc substances all together. This approach is reflecting reality. Since the consumption pattern of zinc containing articles has not significantly changed since the closure of the risk assessment, the analysis made in the RA is used for the present scenario, too. Because of the integrated approach, the consumer scenario is relevant for all zinc substances.

Indirect exposure via the environment Conform to the approach followed in the RA, a substance-specific assessment is presented for general and local indirect exposure via the environment. Considering that industrial emissions have further decreased since the closure of the RA, the analysis of the RA is used as a realistic worst case for this scenario.

Regional scenario Conform to the RA, a regional assessment of environmental exposure as a result of the use/consumption of zinc containing articles has been made. Since the emissions related to the use of the different substances all combine in the environment, no substance-specific assessment was made, but an overall assessment, combining all emissions from zinc containing products at the same time in one assessment. This approach was also followed in the EU RA. The environmental release factors for the different zinc articles were extensively discussed in the RA, and serve as a basis for the present analysis. The exposure assessment was however extended from the EU- 15 to the EU-27. As compared to the RA database, some important updates of emissions and, notably, of the environmental monitoring data were made. Like for consumers, this regional assessment is relevant for all zinc substances.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 287 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

9.1. Local scenarios

In table below, the generic exposure scenarios (GES) developed for diammonium tetrachlorozincate are summarised. Table 88. Generic exposure scenarios for diammonium tetrachlorozincate

Number Sector Uses Code

0 diammonium Manufacture GESZn(NH4)Clx 0 tetrachlorozincate production Substance

1 Formulation step Formulation GESZn(NH4)Clx 1 general

2 First tier applications Manufacturing of GESZn(NH4)Clx 2 other zinc compounds

3 Laboratory GESZn(NH4)Clx 3 reagent

4 As component GESZn(NH4)Clx 4 for solid blends & matrices

5 As component GESZn(NH4)Clx 5 for production of dispersions, pastes and other viscous matrices

6 Second tier applications DU of GESZn(NH4)Clx 6 Zn(NH4)Clx- containing solid preparations

7 DU of GESZn(NH4)Clx 7 Zn(NH4)Clx- containing liquid & pasty preparations

A specific consumer STP scenario (wide dispersive use) on evaluation of risks due to the presence of Zinc in European Sewage treatment plants was added (Ges Zn(NH4)Clx 8) (see Annex 2)

Numerous uses were identified for diammonium tetrachlorozincate . These are listed in table below, with the indication of the Generic Exposure Scenario (GES) that is relevant to these identified uses.

Table 89. Identified uses for Zn(NH4)Clx and corresponding Generic Exposure Scenario (GES) IU number Identified Use (IU) name GES code 1 diammonium tetrachlorozincate production and refining GESZn(NH4)Clx 0 5 Production of inorganic zinc compounds GESZn(NH4)Clx 2 6 Electroplating GESZn(NH4)Clx 2 7 Production of Zn(NH4)Clx-based fluxing agents GESZn(NH4)Clx 2

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 288 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

8 steel surface treatment prior to hot-dip galvanizing GESZn(NH4)Clx 4, GESZn(NH4)Clx 5 9 Use of Zn(NH4)Clx-based fluxing agents before welding/soldering GESZn(NH4)Clx 6, Generic processes consumer/environment* 10 Laboratory reagent GESZn(NH4)Clx 3 11 Production of organic zinc compounds GESZn(NH4)Clx 2 12 Production of coatings, paints, inks, enamels, varnishes GESZn(NH4)Clx 1, GESZn(NH4)Clx 4 13 Component for paper coating or treatment for paper products GESZn(NH4)Clx 1, GESZn(NH4)Clx 5 14 Use of Zn(NH4)Clx-containing paper coatings GESZn(NH4)Clx 6, Generic consumer/environment* 15 Textile and leather coating treatment GESZn(NH4)Clx 1, GESZn(NH4)Clx 5 16 Use of Zn(NH4)Clx-containing coatings for textile and leather GESZn(NH4)Clx 6, Generic consumer/environment* 17 Batteries /fuel cells GESZnCl 1, GESZn(NH4)Clx 4, GESZn(NH4)Clx 5 18 Component for production of rubber, resins and related preparations GESZn(NH4)Clx 1, GESZn(NH4)Clx 5 19 Production of polymer-matrices, plastics and related preparations GESZn(NH4)Clx 1, GESZn(NH4)Clx 5 20 Additive / component for the production of Sealants / Adhesives / GESZn(NH4)Clx 1, GESZn(NH4)Clx Mastics 5 21 Use of Zn(NH4)Clx-containing Sealants / Adhesives / Mastics GESZn(NH4)Clx 7, Generic consumer/environment* 22 Additive / component for the production of Lubricants / Grease / GESZn(NH4)Clx 1, GESZn(NH4)Clx Metal working fluids 5 23 Use of Zn(NH4)Clx-containing Lubricants / Grease / Metal workingGESZn(NH4)Clx 7, Generic fluids consumer/environment* 24 Additive / component for the production of Polishes / wax blends GESZn(NH4)Clx 1, GESZn(NH4)Clx 5 25 Use of Zn(NH4)Clx-containing Polishes / wax blends GESZn(NH4)Clx 7, Generic consumer/environment* 26 Use of Zn(NH4)Clx-containing catalysts GESZn(NH4)Clx 1, GESZn(NH4)Clx 5 27 Additive component for production of de-icing products GESZn(NH4)Clx 1, GESZn(NH4)Clx 5 28 Use of Zn(NH4)Clx-containing de-icing products GESZn(NH4)Clx 7, Generic consumer/environment* 29 Additive for the formulation of fertilizers GESZn(NH4)Clx 1, GESZn(NH4)Clx 4, GESZn(NH4)Clx 5 30 Use of Zn(NH4)Clx-containing fertilizer's formulations Generic consumer/environment* 31 Additive for the formulation of biocidal products GESZn(NH4)Clx 1, GESZn(NH4)Clx 4, GESZn(NH4)Clx 5 32 Additive for the formulation of cleaning products GESZn(NH4)Clx 1, GESZn(NH4)Clx 4, GESZn(NH4)Clx 5 33 Use of Zn(NH4)Clx-containing cleaning products GESZn(NH4)Clx 6, GESZn(NH4)Clx 7, Generic consumer/environment*

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 289 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

34 Additive in the formulation of cosmetics GESZn(NH4)Clx 1, GESZn(NH4)Clx 4, GESZn(NH4)Clx 5 35 Use of cosmetics GESZn(NH4)Clx 6, GESZn(NH4)Clx 7, Generic consumer/environment* 36 Additive in the formulation of pharma / veterinary products GESZn(NH4)Clx 1, GESZn(NH4)Clx 4, GESZn(NH4)Clx 5 37 Use of Pharma/veterinary products GESZn(NH4)Clx 6, GESZn(NH4)Clx 7, Generic consumer/environment*

* corresponds to “GES 8” in IUCLID

9.1.1. GES Zn(NH4)Clx-0: Industrial use of primary or secondary zinc bearing material in the manufacture of Zn(NH4)Clx in several process steps, collection of the substance produced and packaging.

Table 90. GES Zn(NH4)Clx-0 Exposure Scenario Format (1) addressing uses carried out by workers 9.1.1. Title of Exposure Scenario number GES Zn(NH4)Clx-0: Industrial use of primary or secondary zinc bearing material in the manufacture of Zn(NH4)Clx in several process steps, collection of the substance produced and packaging. List of all use descriptors related to the life cycle stage and all the uses under it; include market sector (by PC), if relevant; SU: 3, 8, 9 PROC: 2, 3, 5, 8b, 9, 26 PC: 12, 14, 19, 20, 21 AC: not applicable ERC: 1

9.1.1 Exposure Scenario 9.1.1.1 Contributing scenario (1) controlling environmental exposure for the Industrial use of primary or secondary zinc bearing material in the manufacture of Zn(NH4)Clx in several process steps, collection of the substance produced and packaging.

The manufacturing process includes:  Reception of zinc-bearing materials, if applicable, and transfer to the reaction tank (chloride and ammonia media)

 Reception of the Intermediate Ammonium zinc chloride solution in the reaction tank, if applicable

 Sequential addition of reagents for purification steps and filtration on press filter, when needed. Ventilation is adapted.

 Concentration by water evaporation, under exhaust hood.

 Pouring on a cooling belt

 Exposure to dust can occur during packing of the powder. Solutions are packed in intermediate bulk containers (ca. 1 m3 capacity); solids are packed in bags or drums.

 Maintenance activities

 Product characteristics

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 290 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Product related conditions:

 Zn(NH4)Clx is produced in minimum 80¨purity; higher grades (>95%) are usual.

Amounts used Daily and annual amount per site:

 maximum 12500 T/y;

Frequency and duration of use

 Continuous production

Environment factors not influenced by risk management Flow rate of receiving surface water:

 Default is used unless specified otherwise

Other given operational conditions affecting environmental exposure Other given operational conditions: e.g. technology or process techniques determining the initial release of substance from process (via air and waste water); dry or water based processes; conditions related to temperature and pressure; indoor or outdoor use of products; work in confined area or open air;

 Most of the operations are in wet phase.  Even when no process waters some non-process water can be generated containing zinc (e.g. from cleaning)  All processes are performed indoor in a confined area. All residues containing zinc are recycled.

Technical conditions and measures at process level (source) to prevent release Process design aiming to prevent releases and hence exposure to the environment; this includes in particular conditions ensuring rigorous containment; performance of the containment to be specified (e.g. by quantification of a release factor in section 9.x.2 of the CSR);  Process enclosures and closed circuits where relevant and possible.  Local exhaust ventilation on work areas with potential dust generation, dust capturing and removal techniques  Containment of liquid volumes in sumps to collect/prevent accidental spillage, acid solutions are treated with alkali.  Higher temperatures (~= 100°C) in the surroundings of the drying units are possible.

Technical onsite conditions and measures to reduce or limit discharges, air emissions and releases to soil Technical measures, e.g. on-site waste water and waste treatment techniques, scrubbers, filters and other technical measures aiming at reducing releases to air, sewage system, surface water or soil; this includes strictly controlled conditions (procedural and control technology) to minimise emissions; specify effectiveness of measures; specify the size of industrial sewage treatment plant (m3/d), degradation effectiveness and sludge treatment (if applicable);

 On-site waste water treatment techniques can be applied to prevent releases to water (if applicable) e.g.: chemical precipitation, sedimentation and filtration (efficiency 90-99.98%).  Careful use of chlorhydric acid, ammonic solutions and corrosive chloride solutions  Containment of liquid volumes in sumps to collect/prevent accidental spillage  Air emissions are controlled by use of bag-house filters and/or other air emission abatement devices e.g. fabric (or bag) filters (up to 99% efficiency), wet scrubbers (50-99% efficiency). This may create a general negative pressure in the building.

Organizational measures to prevent/limit release from site Specific organisational measures or measures needed to support the functioning of particular technical measures. Those measures need to be reported in particular for demonstrating strictly controlled conditions.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 291 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

. regular cleaning of equipment and floors, . procedures for process control and maintenance,...  Treatment and monitoring of releases to outside air, and exhaust gas streams (process & hygiene), according to national regulation.  SEVESO 2 compliance, if applicable

Conditions and measures related to municipal sewage treatment plant Size of municipal sewage system/treatment plant (m3/d); specify degradation effectiveness; sludge treatment technique (disposal or recovery); measures to limit air emissions from sewage treatment (if applicable); please note: the default size of the municipal STP (2000 m3/d) will be rarely changeable for downstream uses.

 In cases where applicable: default size, unless specified otherwise.

Conditions and measures related to external treatment of waste for disposal Fraction of used amount transferred to external waste treatment for disposal; type of suitable treatment for waste generated by work-ers uses, e.g. hazardous waste incineration, chemical-physical treatment for emulsions, chemical oxidation of aqueous waste; specify effectiveness of treatment;

Hazardous wastes from onsite risk management measures and solid or liquid wastes from production, use and cleaning processes should be disposed of separately to hazardous waste incineration plants or hazardous waste landfills as hazardous waste. Releases to the floor, water and soil are to be prevented. If the zinc content of the waste is elevated enough, internal or external recovery/recycling might be considered.

Fraction of daily/annual use expected in waste: zinc producers = 3.1 % zinc compound producers = 0.056 % downstream users = 0.30 %

Appropriate waste codes: 02 01 10*, 06 03 13*, 06 03 14, 06 03 15*, 06 04 04*, 06 04 05*, 06 05 02*, 08 01 11*, 10 05 01, 10 05 03*, 10 05 05*, 10 05 06*, 10 05 11, 10 05 99, 10 10 03, 10 10 05*, 10 10 07*, 10 10 09*, 10 10 10, 10 10 11*, 11 01 09*, 11 02 02*, 11 02 03, 11 02 07*, 12 01 03*, 12 01 04, 12 01 12*, 15 01 4*, 15 01 10*, 15 02 02*, 16 01 04*, 16 01 06*, 16 01 18*, 16 06 02*, 16 08 02*, 16 08 03*, 16 11 02, 16 11 03*, 16 11 04, 16 11 06, 17 04 07*, 17 04 09*, 17 09 04*, 19 02 05*, 19 10 02*, 19 12 03*

Suitable disposal: Keep separate and dispose of to either Hazardous waste incineration operated according to Council Directive 2008/98/EC on waste, Directive 2000/76/EC on the incineration of waste and the Reference Document on the Best Available Techniques for Waste Incineration of August 2006. Hazardous landfill operated under Directive 1999/31/EC.

A detailed assessment has been performed and is reported in the Waste report (ARCHE, 2012) (See Annex 1) Conditions and measures related to external recovery of waste Fraction of used amount transferred to external waste treatment for recovery: specify type of suitable recovery operations for waste generated by workers uses, e.g. re-distillation of solvents, refinery process for lubricant waste, recovery of slags, heat recovery out-side waste incinerators; specify effectiveness of measure;

 All residues from the wet process are recycled.  By-products (ashes) from the dry process that are formed in the reactor, are recovered and either recycled in the system or handled further according the waste legislation.  Users of Zn and Zn-compounds have to favour the recycling channels of the end-of-life products  Users of Zn and Zn-compounds have to minimize Zn-containing waste, promote recycling routes and,

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 292 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

for the remaining, dispose the waste streams according the Waste regulation.

9.1.1.2 Contributing scenario (2) controlling worker exposure for the industrial use of primary or secondary zinc bearing material in the manufacture of Zn(NH4)Clx in several process steps, collection of the substance produced and packaging. Product characteristic Product related conditions, e.g. the concentration of the substance in a mixture, the physical state of that mixture (solid, liquid; if solid: level of dustiness), package design affecting exposure)

Zn(NH4)Clx is hygroscopic in nature and is produced in a dust-free crystalline form (0.5 mm).

Amounts used Amounts used at a workplace (per task or per shift); note: sometimes this information is not needed for assessment of worker’s expo-sure

Maximum 60 T/day, 20T/shift

Frequency and duration of use/exposure Duration per task/activity (e.g. hours per shift) and frequency (e.g. single events or repeated) of exposure

8hrs shift

Human factors not influenced by risk management Particular conditions of use, e.g. body parts potentially exposed as a result of the nature of the activity

Uncovered body parts: (potentially) face

Other given operational conditions affecting workers exposure Other given operational conditions: e.g. technology or process techniques determining the initial release of substance from process into workers environment; room volume, whether the work is carried out outdoors/indoors, process conditions related to temperature and pressure.

 All processes are carried out indoor in confined areas.

Technical conditions and measures at process level (source) to prevent release Process design aiming to prevent releases and hence exposure of workers; this in particular includes conditions ensuring rigorous containment; performance of containment to be specified (e.g. by quantification of residual losses or exposure)

 Local exhaust ventilation on furnaces and other work areas with potential dust generation, dust capturing and removal techniques  Process enclosures closed circuits or semi-enclosures where appropriate.  Careful use of chlorhydric acid, ammonic solutions and corrosive chloride solutions  Containment of liquid volumes in sumps to collect/prevent accidental spillage  Local exhaust ventilation on furnaces and other work areas with potential dust and fumes generation, dust capturing and removal techniques.

Technical conditions and measures to control dispersion from source towards the worker Engineering controls, e.g. exhaust ventilation, general ventilation; specify effectiveness of measure

 Local exhaust ventilation systems (generic LEV (84%) as worst case; higher efficiencies (90-95%) are usual  Cyclones/filters (for minimizing dust emissions) : efficiency: 70-90% (cyclones), 50-80% (dust filters), 85-95% (double stage, cassette filters)  Process enclosure, especially in potentially dusty units  Dust control: dust and Zn in dust needs to be measured in the workplace air (static or individual) according to national regulations.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 293 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

 Special care for the general establishment and maintenance of a clean working environment by e.g.:

 Cleaning of process equipment and workshop

 Storage of packaged Zn product in dedicated zones

Organisational measures to prevent /limit releases, dispersion and exposure Specific organisational measures or measures needed to support the functioning of particular technical measures (e.g. training and supervision). Those measures need to be reported in particular for demonstrating strictly controlled conditions (to justify exposure based waiving).

In general integrated management systems are implemented at the workplace e.g. ISO 9000, ISO-ICS 13100, or alike, and are, when appropriate, IPPC-compliant.

Such management system would include general industrial hygiene practice e.g.: o information and training of workers on prevention of exposure/accidents, o procedures for control of personal exposure (hygiene measures) o regular cleaning of equipment and floors, extended workers instruction-manuals o procedures for process control and maintenance,... o personal protection measures (see below)

Conditions and measures related to personal protection, hygiene and health evaluation Personal protection, e.g. wearing of gloves, face protection, full body dermal protection, goggles, respirator; specify effectiveness of measure; specify the suitable material for the PPE (where relevant) and advise how long the protective equipment can be used before replacement (if relevant) Wearing of gloves and protective clothing is compulsory (efficiency >=90%). With normal handling, no respiratory personal protection (breathing apparatus) is necessary. If risk for exceedance of OEL/DNEL, use e.g.: -dust filter-half mask P1 (efficiency 75%) -dust filter-half mask P2 (efficiency 90%) -dust filter-half mask P3 (efficiency 95%) -dust filter-full mask P1 (efficiency 75%) -dust filter-full mask P2 (efficiency 90 %) -dust filter-full mask P3 (efficiency 97.5%)

Eyes: safety glasses are optional

Exposure estimation and risk characterisation

1. Environment

The risk assessment (RA) on ZnCl2 (ECB 2008) assessed the risks manufacture of ZnCl2 based on reported data that are summarised in table below. This dataset on ZnCl2 was updated with more recent data. Since the processes and uses related to Zn(NH4)Clx are very similar to ZnCl2 processes and uses, the exposures and exposure controls as described for ZnCl2 are crossread to Zn(NH4)Clx.

Table 91. Environmental risk characterisation for the manufacture of Zn(NH4)Clx, crossread fromZnCl2. Data from the EU tonnage Cadd*/PNEC** Cadd/PNEC** PEC/PNEC PEC/PNEC RA water sediment soil STP§ ZnCl2 manufacture (RA:

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 294 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

5 companies, cfr table 3.4.10) 1*** 6100 2.2 20 (8.3) 0.02 Not applicable 2 5700 0.001 0.002 0.02 0.55 3 3700 0.14 2.6 (1) 0.02 1.9 4 12500 0.02 0.3 0.02 0.01 5 588 0.56 10 (4.2) 0.02 1.3

Recent data**** PEC/PNEC PEC/PNEC water sediment Company A 6500 WWTP 0.18 0.3 0.39 NA cooling 0.62 4.5***** 0.39 NA

Company B 15000 0.17 0.23 0.39 NA

*In the RAs, “Cadd” the “added concentration by the emissions at a given site was used for the risk characterisation. As such, the risks related to the local emissions at the site were assessed only, without the added exposure from the regional background. **the risk ratios are those cited in the RA, i.e. with the PNECs derived in the RA; between brackets: risk ratio with updated PNEC. *** no on site WWTP or STP **** for the recent data, the regional exposure was taken into account (PEC/PNEC). *****internal recycling of cooling waters followed by WWTP treatment is planned. § PEC/PNEC ratio’s were recalculated with new PNEC for STP, leading to a RCR different than derived previously in RA

Table 92. Exposure assessment for the industrial manufacture Zn(NH4)Clx, crossread fromZnCl2. (recently reported data) ZnCl2 manufacture PEC water (µg PEC sediment mg PEC soil (mg PEC STP Zn/l) Zn/kgDW) Zn/kgDW) (mg Zn/l) Company A WWTP 3.6 70 42 NA Cooling 12.7 1053 42 NA

Company B 3.5 54.4 41 NA

Conclusion: based on the available data from the risk assessment and recently measured data on ZnCl2 GES-0, the risk ratios demonstrate no risk for the water and soil, but risk for the sediment in a few cases. In the sediment, the bioavailability of zinc will be determined by the content of acid volatile sulphide (AVS). It has been documented that there is covariance between zinc in sediment and AVS. In cases where AVS and sediment-Zn were measured near industrial sites, there was indeed a surplus on AVS observed, rendering the zinc present at that site non-bioavailable. Whether or not this phenomenon is also present at the sites mentioned in table above, can only be confirmed by local measurements. In any case, the data show that there are production sites for Zn(NH4)Clx (ZnCl2) for which no risk for the environment is calculated. It is concluded that, when the risk management measures described in this scenario are applied, no risk is predicted for environment, (including sediment) for the manufacture of Zn(NH4)Clx (e.g., it is noted that at site A reporting recent data, the emission from cooling waters will be replaced by internal recycling of these waters, with connection to the on-site WWTP. The emissions from the latter process don’t result in risk).

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 295 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

parameters that are needed for the calculations, are documented.  For water assessment, bioavailability model correction can be applied when the following water parameters are documented for the receiving water: Dissolved organic carbon (DOC), pH, hardness or Ca- concentration. For the calculations, the “zinc BLM-calculator” excel tool is used to this end (see “tools” on http://www.reach-zinc.eu/). When the local values of these parameters are unknown, regional data can be used as an alternative. Use of regional instead of local values should always be handled with caution.

 For sediment, a generic bioavailability factor of 2 is already integrated in the PNEC, based on AVS/SEM levels and according to the risk assessment (ECB 2008). A further refinement of local bioavailability can be made when local AVS/SEM concentrations are documented. The bioavailable fraction of zinc is given by subtracting local AVS from local SEM-Zn (SEM-Zn - AVS).

 For soil, a worst case bioavailability correction (corresponding to sandy soils) is already integrated. Further refinement for zinc bioavailability in other soil types is possible, when the local soil type is documented, together with pH, CEC (see “tools” on http://www.reach-zinc.eu/)

2. Workers Since the hazard profile Of ZnCl2 and Zn(NH4)Clx is the same, the GES-0 ZnCl2 is crossread in this section. Occupational exposure to Zn(NH4)Clx is possible due to emissions from parts of the process when solid, dusty ammonium zinc chloride is already formed. The packaging and repackaging of the produced ammonium zinc chloride in bags, big bags or bulk tankers may lead to contamination of the facility and to exposure (direct or indirect) of workers, by inhalation and dermal contact. Pulmonary absorption may occur but most of the material that is deposited in the head and the tracheobronchial region is rapidly translocated to the GI tract and part of it will be absorbed in the GI tract. The particle size of Zn(NH4)Clx is however rather large (99.66% larger than 15.8 μm). In the RA on ZnCl2, an exposure assessment was made based on reported data from 3 companies. The general conclusion was that no risks were observed (see table below);

Table 93. Occupational exposure data and risk characterisation for manufacture of zinc compounds RA data (RA Zn in Risk ratio Systemic Risk ratio Systemic Risk ratio ZnCl2 (table workplace air inhalation*** inhalation systemic dermal systemic total 4.1.3.2A) (mg/m3) exposure inhalation **(mg/d) total inhalable (mg/d)* 3 companies 0.2 0.2 0.8 0.2 0.4 0.12

* assuming a respiratory absorption of 40% for ZnCl2/ Zn(NH4)Clx /ZnSO4 and 20% for ZnO and other zinc compounds, and an inhalation volume of 10m3 ** assuming a dermal absorption of 0.2% for dust, no wearing of gloves assumed ***DNEL inhalation for Zn(NH4)Clx is 1.0 mg/m3

Conclusion: based on the data from the risk assessment, and in accordance to the conclusion drawn in the RA ZnCl2, there is no risk predicted for workers for this scenario if the risk management measures as described in the scenario are applied.

9.1.2. GES Zn(NH4)Clx -1: Industrial use of Zn(NH4)Clx in the formulation of preparations by mixing thoroughly, dry or in a solvent, the starting materials with potentially pressing, pelletising, sintering, possibly followed by packing

Table 94. GES Zn(NH4)Clx -1

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 296 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Exposure Scenario Format (1) addressing uses carried out by workers 9.1.2. Title of Exposure Scenario number Zn(NH4)Clx GES-1: Industrial use of Zn(NH4)Clx in the formulation of preparations by mixing thoroughly, dry or in a solvent, the starting materials with potentially pressing, pelletising, sintering, possibly followed by packing . List of all use descriptors related to the life cycle stage and all the uses under it; include market sector (by PC), if relevant; SU: 3,10 PROC: 1,2,3,4,5, 8b,9,13, 14, 15, 22, 26 PC: Not applicable (all) AC: not applicable ERC: 1,2 Further explanations (if needed)

Zn(NH4)Clx is used in the manufacture of preparations by mixing thoroughly the starting materials, followed by direct use of packaging of the preparation. Many different industrial uses are characterised by this process. Therefore these industrial uses are all covered by this generic exposure scenario.

9.1.2 Exposure Scenario 9.1.2.1. Contributing scenario (1) controlling environmental exposure for the Industrial use of Zn(NH4)Clx in the formulation of preparations by mixing thoroughly, dry or in a solvent, the starting materials with potentially pressing, pelletising, sintering, possibly followed by packing .

Further specification:

In the described process, the Zn(NH4)Clx is:  Removed from the packaging and stored in silos after delivery.  Extracted from the silo, dosed and fed with the other reagents to the mixing tank. Mixing occurs batch- wise or continuously, according the process receipt. The mixing occurs in a closed tank/chamber.  The preparation (dry or wet (solvent/paste) matrix) is further used as such or packed for further treatment/use.

Product characteristics Product related conditions:

Zn(NH4)Clx is used in minimum 80% purity; higher grades (>95%) are usual

Amounts used Daily and annual amount per site: maximum 5000 T/y;

Frequency and duration of use

Continuous production is assumed as a worst case. It is possible that use is not continuous; this has to be considered when estimating exposure.

Environment factors not influenced by risk management Flow rate of receiving surface water: default for generic scenario: 18,000 m3/d, unless specified otherwise

 All processes are performed indoor in a confined area. All residues containing zinc are recycled.  Even when no process waters (e.g. when dry process throughout), some non-process water can be generated containing zinc(e.g. from cleaning)

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 297 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Technical conditions and measures at process level (source) to prevent release Process design aiming to prevent releases and hence exposure to the environment; this includes in particular conditions ensuring rigorous containment; performance of the containment to be specified (e.g. by quantification of a release factor in section 9.x.2 of the CSR);  Process enclosures and closed circuits where relevant and possible.  Dust capturing and removal techniques are applied on local exhaust ventilation on furnaces and other work areas with potential dust generation.  Containment of liquid volumes in sumps to collect/prevent accidental spillage

 On-site waste water treatment techniques can be applied to prevent releases to water (if applicable) e.g.: chemical precipitation, sedimentation and filtration (efficiency 90-99.98%).  Air emissions are controlled by use of bag-house filters and/or other air emission abatement devices e.g. fabric (or bag) filters (up to 99% efficiency), wet scrubbers (50-99% efficiency). This may create a general negative pressure in the building.

 In general emissions are controlled and prevented by implementing an integrated management system e.g. ISO 9000, ISO 1400X series, or alike, and, when applicable, by being IPPC-compliant. o Such management system should include general industrial hygiene practice e.g.: . information and training of workers, . regular cleaning of equipment and floors, . procedures for process control and maintenance,...  Treatment and monitoring of releases to outside air, and exhaust gas streams (process & hygiene), according to national regulation.  SEVESO 2 compliance, if applicable

 In cases where applicable: default size, unless specified otherwise.

Fraction of daily/annual use expected in waste: zinc producers = 3.1 % zinc compound producers = 0.056 % downstream users = 0.30 %

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 298 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

 All residues are recycled or handled and conveyed according to waste legislation. .

9.1.2.2. Contributing scenario (2) controlling worker exposure for the Industrial use of Zn(NH4)Clx in the formulation of preparations by mixing thoroughly, dry or in a solvent, the starting materials with potentially pressing, pelletising, sintering, possibly followed by packing .

Further specification Zn(NH4)Clx is used in the manufacture of preparations by mixing thoroughly the starting materials, followed by direct use of packaging of the preparation. Many different industrial uses are characterised by this process. Therefore these industrial uses are all covered by this generic exposure scenario.

Product characteristic Product related conditions, e.g. the concentration of the substance in a mixture, the physical state of that mixture (solid, liquid; if solid: level of dustiness), package design affecting exposure)

 The concentration of Zn(NH4)Clx in the mixtures can cover a broad range (<= 5% up to >25%) depending on the application.  The preparation can be solid or liquid.  When the preparation is in solid state, it can be in a) powdery, b) glassy or c) pelletized form. In the powder form, it can be characterised by high dustiness in a worst case situation.

Amounts used Amounts used at a workplace (per task or per shift); note: sometimes this information is not needed for assessment of worker’s expo-sure

Max 5000T/y = 14T/d = 5T/shift depending on the application.

Frequency and duration of use/exposure Duration per task/activity (e.g. hours per shift) and frequency (e.g. single events or repeated) of exposure

8 hour shifts (default worst case) are assumed as starting point; it is emphasised that the real duration of exposure could be less. This has to be considered when estimating exposure.

Human factors not influenced by risk management

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 299 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Particular conditions of use, e.g. body parts potentially exposed as a result of the nature of the activity

Uncovered body parts: (potentially) face

 elevated temperature steps (~=100°C) can occur  all indoor processes in confined area.

 Process enclosures and closed circuits where relevant and possible.  Local exhaust ventilation on furnaces and other work areas with potential dust generation, dust capturing and removal techniques.  Containment of liquid volumes in sumps to collect/prevent accidental spillage

Technical conditions and measures to control dispersion from source towards the worker Engineering controls, e.g. exhaust ventilation, general ventilation; specify effectiveness of measure

 Local exhaust ventilation systems (high efficiency 90-95%)  Cyclones/filters (for minimizing dust emissions) : efficiency: 70-90% (cyclones), 50-80% (dust filters), 85-95% (double stage, cassette filters)  Process enclosure, especially in the drying /calcination / packaging (potentially dusty) units  Dust control: dust and Zn in dust needs to be measured in the workplace air (static or individual) according to national regulations.  Special care for the general establishment and maintenance of a clean working environment by e.g.:

 Cleaning of process equipment and workshop

 Storage of packaged Zn product in dedicated zones

Organisational measures to prevent /limit releases, dispersion and exposure

In general integrated management systems are implemented at the workplace e.g. ISO 9000, ISO-ICS 13100, or alike, and are, when appropriate, IPPC-compliant.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 300 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

-dust filter-half mask P2 (efficiency 90%) -dust filter-half mask P3 (efficiency 95%) -dust filter-full mask P1 (efficiency 75%) -dust filter-full mask P2 (efficiency 90 %) -dust filter-full mask P3 (efficiency 97.5%)

Eyes: safety glasses are optional

Exposure estimation and risk characterisation

1. Environment

The processes involved in this scenario can be dry or wet. Even when no process waters are involved, occasional non-process-waters can occur having some zinc content, due to e.g. dust cleaning. Therefore, all formulation processes with Zn(NH4)Clx and other zinc compounds should have some form of water treatment, on site or off-site, according to national legislation and permits.

The risk assessments on zinc and zinc compounds reported measured exposure data on a number of sectors falling under this scenario. In most cases, a) the formulation of the substance into the dry or wet preparation/mixture and b) the further industrial use of the preparation/mixture are integrated at the same industrial site. For this reason, environmental emissions data are integrating both process steps, and encompass the GES-1 and GES- 4/GES-5. Exposure related to the formulation of the pure Zn(NH4)Clx is considered to be the most critical, because the substance is used in its pure state.

The risk assessments (RAs) on several zinc compounds (ECB 2008) assessed the risks related to the industrial use of Zn(NH4)Clx (and other zinc compounds) for the formulation of Zn(NH4)Clx- (and other Zn-compound)- containing preparations, based on reported data. The resulting risk characterisations are summarized in table below. Distinction is being made between assessments based on measured data, and assessments based on modelling, using default release factors. Table below also summarizes the risk characterisation based on more recent data on manufacture of other Zn- compounds. The exposure estimates based on these more recent data are summarized in the second table below.

Table 95. Environmental risk characterisation for the Industrial use of ZnCl2 (crossread to Zn(NH4)Clx) as component for the manufacture of preparations for further downstream use. assessments from the EU RA by PEC/PNEC PEC/PNEC PEC/PNEC soil PEC/PNEC sector of use* water sediment (**) STP§ ZnCl2 (table3.4.10., RA ZnCl2, ECB 2008) Assessment based on measured data Agrochemical industry processing (1 0.03 0.51 0.02 0.19 single EU production site) Battery industry (1 company) 0 0 0.02 0

Assessments based on modelling Chemical industry: processing 0.19 1.7 (0.7) 19 23.5 Battery industry: processing 0.16 1.4 (0.6) 0.2 0.07 Dyes and inks industry: formulation 5.3 48 (20) 2.0 2.45

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 301 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Dyes and inks industry: formulation 150 1343 (560) 56 79

Additional recent data*** Fertiliser manufacture Company A 0.16 0.19 0.39 0 *PNECs from the RA are applied, integrating for sediment the generic bioavailability factor 0.5 and for soil the generic bioavailability factor 0.33 (RA, ECB 2008); Risk ratios for water and sediment are Cadd/PNEC; for STP and soil risk ratios are PEC/PNEC. **PEC/PNEC ratios for sediment between brackets apply the updated PNEC and generic bioavailability factor of the RA ***all risk ratios are PEC/PNECs § PEC/PNEC ratio’s were recalculated with new PNEC for STP, leading to a RCR different than derived previously in RA

Table 96. Exposure assessment for the industrial use of ZnCl2 (crossread to Zn(NH4)Clx) for the manufacture of wet or dry preparations, based on recently reported exposure data. PEC water (µg PEC sediment mg PEC soil (mg PEC STP Zn/l) Zn/kgDW) Zn/kgDW) (mg Zn/l)

Fertiliser manufacture Company A 3.4 45 41 0

Conclusion When local risks are assessed using measured emissions data, no risk for the environment is generally described for the formulation processes using Zn(NH4)Clx. Also recent data on fertiliser manufacture show no risk. Only when default release factors are applied (assessment based on modelling), risks are calculated. The measured data however overrule these modelled results, so it is concluded based on the measured data that there is no risk for the environment from this scenario, when risk management measures, as described, are applied. The “no risk” conclusion on the environmental assessment of formulation of Zn(NH4)Clx is confirmed by data on formulation with other zinc substances, see table below.

Table 97. Environmental risk characterisation for the industrial use of Zn compounds (e.g. Zn(NH4)Clx) as component for the manufacture of preparations for further downstream use. assessments from the EU RA by PEC/PNEC PEC/PNEC PEC/PNEC soil PEC/PNEC sector of use* water sediment (**) STP§ ZnS04 (table3.4.10., RA ZnSO4, ECB 2008) Assessment based on measured data Agricultural feed industry 0 0 0.02 0

Assessments based on modelling Agricultural pesticide industry 0.11 1 11 13 Agricultural fertiliser industry 19 175 7.3 9 Agricultural feed industry 1.0 9 0.4 0.47 Chemical industry: processing 0.19 1.7 19 23.5

ZnO (table3.4.33. RA ZnO, ECB 2008) Assessment based on measured data Tyre industry: processing 0 0 0.15 0

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 302 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

General rubber industry: processing 0 0 0.08 0 Ceramic industry processing typical 0 0 0.14 plant average 0 Ceramic industry processing typical 0 0 0.06-0.38 plant range 0 Ferrites industry (average of 4 (out of 0.27 2.5 (1.0) 0.4 5) plants 0.125 Varistors (average of 2 (out of 4) 0.06 1.2 (0.5) 0.09 plants*** 0.03 Catalysts processing**** <4.9 <45 0.02 <2.25 Feedstuff additive: formulation (site 0 0 0.02 specific) 0 Feedstuff additive: formulation 0 0 0.03 (generic average use) 0 Feedstuff additive: formulation 0 0 0.05 (generic largest use use) 0 Paints: formulation 0 0 0.02 0 Paints: processing (industry data) 0 0 0.02 0

Assessments based on modelling Glass industry: processing (average 2.5 23 0.93 use) 1.15 Glass industry: processing (largest 6.3 57 2.4 use) 2.9 Lubricants: formulation (average 7.5 67 2.7 use) 3.45 Lubricants: formulation (largest use) 13 118 5 6 Paints/ processing: generic data 1.6 14 0.6 0.75 Cosmetics pharmaceuticals: 2.5 23 0.93 formulation (average use) 1.15 Cosmetics pharmaceuticals: 21 188 8 formulation (largest use) 9.5

Zn phosphate (table3.4.9., RA Zn phosphate, ECB 2008) Assessment based on measured data Paint industry (average from 3 of 5 0.19 1.7 (0.35) Not calculated sites reported)***** 0.175

Assessments based on modelling Paint industry: formulation 8.3 75 3.1 3.85 Paint industry: processing, solvent 0.23 2.1 0.28 borne 0.105 Paint industry: processing, water 1.2 11 0.43 borne 0.55 *PNECs from the RA are applied, integrating for sediment the generic bioavailability factor 0.5 and for soil the generic bioavailability factor 0.33 (RA, ECB 2008); Risk ratios for water and sediment are Cadd/PNEC; for STP and soil risk ratios are PEC/PNEC. **PEC/PNEC ratios for sediment between brackets apply the updated PNEC and generic bioavailability factor of the RA ***data from site 3 (showing as only risk ratios>1), not considered, because it was explicitly mentioned that no WWTP or STP was present (RA ZnO). ****calculations from concentration in waste water reported as “<1mg Zn/l” (value of 1mg/l taken as maximum) For the one case with risk based on measured data observed in the RA, the catalysts producing sector, extensive additional data were generated; they demonstrate the absence of risks (see GES 1 ZnO).

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 303 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

*****Only reliable data are used, where the truly measured emission and/or effluent concentration was reported § PEC/PNEC ratio’s were recalculated with new PNEC for STP, leading to a RCR different than derived previously in RA

Calculation of local exposure- Bioavailability correction The local exposure at a given site can be calculated specifically using the excel sheet prepared by Arche (see “tools” on http://www.reach-zinc.eu/) In addition, bioavailability corrections can be integrated in the exposure assessment, if the environmental parameters that are needed for the calculations, are documented.  For water assessment, bioavailability model correction can be applied when the following water parameters are documented for the receiving water: Dissolved organic carbon (DOC), pH, hardness or Ca- concentration. For the calculations, the “zinc BLM-calculator” excel tool is used to this end (see “tools” on http://www.reach-zinc.eu/). When the local values of these parameters are unknown, regional data can be used as an alternative. Use of regional instead of local values should always be handled with caution.

2. Workers When blending/mixing Zn(NH4)Clx or other zinc compounds in a wet or dry preparation, occupational exposure is possible due to dust generation at several steps of the process. The highest potential for dust generation/exposure is at the unpacking of the dry Zn(NH4)Clx -powder and its mixing into the preparation matrix. At this stage mainly, dusts may lead to contamination of the facility and to exposure (direct or indirect) of workers, by inhalation and dermal contact. Pulmonary absorption may occur but most of the material that is deposited in the head and the tracheobronchial region is rapidly translocated to the GI tract and part of it will be absorbed in the GI tract. Zn(NH4)Clx has however not so small particle size (99.66% of the particles larger than 15.8 μ m).The size of the dry preparations is dependent on the application.

For assessing of worker exposure to zinc in formulation processes, exposure to high dustiness pure Zn(NH4)Clx at the unpacking stage is assumed as a worst case. The RA ZnSO4 (ECB 2008) mentions some measured data on ZnO related to this specific activity. These data can be used as worst case scenario for the coarser, hygroscopic Zn(NH4)Clx, and are given below. In addition, the workers exposure during Zn(NH4)Clx production (Zn(NH4)Clx GES-0) is also given as worst case similar scenario (second table below). Dermal exposure is modelled, assuming high dustiness as a worst case, but implementing the wearing of gloves.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 304 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Data from activity 8-hrs Risk ratio Inhalation Dermal Risk ratio ZnSO4 Inhalation inhalation** exposure exposure systemic RA:Sector exposure systemic (modelled) (mg Zn/m3) (mg Zn/d) systemic (mg/d) Paint Emptying of ZnO 0.17-0.28 0.03-0.06 0.34-0.56 0.2 0.05-0.07 industry* from big bags into dispensers Loading powders 0.1-0.5 0.02-0.1 0.2-1.0 0.2 0.04-0.12 from 25kg big bags Average: Average: 0.58 0.08 into dispensers 0.29 0.06 Loading powders 0.01-1.34 0.002-0.3 0.02-2.68 0.2 0.02-0.3 from big bags into Average 0.27 Average 0.54 0.07 dispensers 0.0.05 Ceramics (1 ZnO loaded from 0.1-0.98 0.02-0.2 0.2-2.0 0.2 0.04-0.2 company) bulk transport to bulk storage

ZnO Manufacture: 50P: 0.33 50P: 0.06 50P: 0.66 0.05 50P: 0.07 production, contributing scenario 90P: 2.0 90P: 0.4 90P: 4.0 90P: 0.4 dry 1 (ref ZnO GES-0)

Recent data: sector Catalyst Emptying of Mean: 0.37 Mean: 0.07 0.74 0.2 0.09 production containers Range: Range: <0.001-1.07 <0.0002-0.2 *values are for total dust; exposure to dust for short duration; data extrapolated to 8hrs exposure **DNEL inhalation for Zn(NH4)Clx /ZnSO4 and other soluble zinc substances is 1.0 mg/m3; for ZnO and other slightly soluble/ insoluble zinc substances: 5mg/m3

Table 99. Occupational exposure data and risk characterisation for the scenario “the industrial use of ZnO for the manufacture of ZnCl2” (ZnCl2 GES-0) RA data Zn in workplace air Risk ratio Systemic Risk ratio Systemic Risk ratio (RA ZnCl2, (mg/m3) inhalation*** inhalation inhalation dermal systemic total table total inhalable exposure systemic **(mg/d) 4.1.3.2A) (mg/d)* 3 0.2 0.2 0.8 0.08 0.4 0.12 companies * assuming a respiratory absorption of 40% for ZnCl2/ Zn(NH4)Clx /ZnSO4 and 20% for ZnO and other zinc compounds, and an inhalation volume of 10m3 ** assuming a dermal absorption of 0.2% for dust, no wearing of gloves assumed ***DNEL inhalation for ZnCl2/ Zn(NH4)Clx /ZnSO4 and other soluble zinc substances is 1.0 mg/m3; for ZnO and other slightly soluble/ insoluble zinc substances: 5mg/m3

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 305 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Conclusion: based on measured data from the RA, data from similar worst case scenario (dry ZnO manufacture), and additional recent measured data, no risk is demonstrated/predicted following the risk management measures indicated.

9.1.3. GES Zn(NH4)Clx -2: industrial use of ammonium zinc chloride or Zn(NH4)Clx -formulations in the manufacturing of other inorganic or organic zinc substances in a solvent-based matrix with potentially filtering and packaging.

Table 100. GES Zn(NH4)Clx -2 Exposure Scenario Format (1) addressing uses carried out by workers 9.1.3. Title of Exposure Scenario number GES Zn(NH4)Clx -2: industrial use of ammonium zinc chloride or Zn(NH4)Clx -formulations in the manufacturing of other inorganic or organic zinc substances in a solvent-based matrix with potentially filtering and packaging. List of all use descriptors related to the life cycle stage and all the uses under it; include market sector (by PC), if relevant;

 SU: 3, 8, 9, 10, 15, 17, 0 (Nace C25.6.1.)  PROC: 1, 2, 3, 4, 5, 8b, 9, 15, 21  PC : 7, 14, 19, 20, 21, 24, 29, 39  AC : 2, 7, 12  ERC : 1, 2, 5, 6a

Further explanations (if needed)

Zn(NH4)Clx is used as a starting material for the manufacturing of several other inorganic and organic zinc compounds. All the manufacturing processes are covered by the present scenario.

9.1.3. Exposure Scenario 9.1.3.1. Contributing scenario (1) controlling environmental exposure for the industrial use of ammonium zinc chloride or Zn(NH4)Clx -formulations in the manufacturing of other inorganic or organic zinc substances in a solvent-based matrix with potentially filtering and packaging. Further specification

Description of activities/process(es) covered in the Exposure Scenario

 Reception of the Zn(NH4)Clx or Zn(NH4)Clx -containing formulation, or Zn(NH4)Clx -bearing raw material in the reaction tank

 Sequential addition of reagents for purification steps and filtration on press filter, when needed (ventilation is adapted).

 Concentration by water evaporation, under exhaust hood, is optional.

 Possible pouring on a cooling belt, is optional as well

 Discharge and packaging of produced zinc compounds. Workers have to place and adjust the bag or drum under the discharge pipe and to set the process in motion. Filled bags or drums are subsequently closed and carried to the storage area.

 Exposure to dust can occur during packing of the powder. Solutions are packed in intermediate bulk containers (ca. 1 m3 capacity), solid products are packed in bags or drums.  Maintenance activities

Product characteristics Product related conditions, e.g. the concentration of the substance in a mixture; viscosity of product; package design affecting expo-sure

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 306 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Zn-compounds are produced in their pure form e.g.: >99%, or in solution.

Amounts used Daily and annual amount per site (for uses in industrial setting) or daily and annual amount for wide disperse uses;

Up to 75 T/d of Zn(NH4)Clx is transformed to equivalent Zn compound

Frequency and duration of use Intermittent ( used < 12 times per year for not more than 24 h) or continuous use/release

Continuous production is assumed as a worst case. It is possible that use is not continuous; this has to be considered when estimating exposure.

Environment factors not influenced by risk management Flow rate of receiving surface water (m3/d, usually 18,000 m3/d for the standard town by default; please note: the default flow rate will be rarely changeable for downstream uses.

Default for generic scenario: 18,000 m3/d, unless specified otherwise

 Wet processes (leaching, filtering, purification) followed by drying (possible grinding), and packaging;  All indoor processes, in confined area.

 Careful use of acids and corrosive solutions, if used  Sump containment is provided under the tanks and the filters i.o. to collect any accidental spillage  When applicable, process waters need to be specifically treated before release  Dosing and packaging operations occur under a special ventilation hood  Process air is filtered before release outside the building

 On-site waste water treatment techniques are (if applicable) e.g.: chemical precipitation, sedimentation, filtration (efficiency 90-99.98%).  Containment of liquid volumes in sumps to collect/prevent accidental spillage  Air emissions are controlled by use of bag-house filters and/or other air emission abatement devices e.g. fabric (or bag) filters (up to 99% efficiency), wet scrubbers (50-99% efficiency). This may create a general negative pressure in the building. Air emissions are continuously monitored.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 307 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

 In cases where applicable: default size, unless specified otherwise.

Fraction of daily/annual use expected in waste: zinc producers = 3.1 % zinc compound producers = 0.056 % downstream users = 0.30 %

A detailed assessment has been performed and is reported in the Waste report (ARCHE, 2012) (See Annex 1) Conditions and measures related to external recovery of waste Fraction of used amount transferred to external waste treatment for recovery: specify type of suitable recovery operations for waste generated by workers uses, e.g. re-destillation of solvents, refinery process for lubricant waste, recovery of slags, heat recovery out-side waste incinerators; specify effectiveness of measure; By-products formed during the process are either recycled, internally or externally, or handled further as waste , according the waste legislation

9.1.3.2. Contributing scenario (2) controlling worker exposure for the industrial use of ammonium zinc chloride or Zn(NH4)Clx -formulations in the manufacturing of other inorganic or organic zinc substances in a solvent-based matrix with potentially filtering and packaging.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 308 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

 Ammonium zinc chloride is transformed to equivalent pure zinc compound.

 The formed zinc compound can be produced as a powder with varying particle size (worst case scenario) or can be in solution.

Amounts used Amounts used at a workplace (per task or per shift); note: sometimes this information is not needed for assessment of worker’s exposure

Up to maximum 25T/shift

Frequency and duration of use/exposure Duration per task/activity (e.g. hours per shift) and frequency (e.g. single events or repeated) of exposure

8hrs shift (worst case)

Human factors not influenced by risk management Particular conditions of use, e.g. body parts potentially exposed as a result of the nature of the activity

Uncovered body parts: (potentially) face

All processes are carried out indoor in confined areas.

 Process enclosures or semi-enclosures where appropriate.  Local exhaust ventilation work areas with potential dust and fumes generation, dust capturing and removal techniques  Containment of liquid volumes in sumps to collect/prevent accidental spillage

Technical conditions and measures to control dispersion from source towards the worker Engineering controls, e.g. exhaust ventilation, general ventilation; specify effectiveness of measure

 Cleaning of process equipment and workshop

 Storage of packaged Zn product in dedicated zones

Organisational measures to prevent /limit releases, dispersion and exposure

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 309 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Specific organisational measures or measures needed to support the functioning of particular technical measures (e.g. training and supervision). Those measures need to be reported in particular for demonstrating strictly controlled conditions (to justify exposure based waiving).

In general integrated management systems are implemented at the workplace e.g. ISO 9000, ISO-ICS 13100, or alike, and are, when appropriate, IPPC-compliant.

Eyes: safety glasses are optional

Exposure estimation and risk characterisation

1. Environment

Like ZnCl2, Zn(NH4)Clx can be used for the manufacture of other zinc compounds e.g.:, Zn3(PO4)2, Zn(OH)CO3, ZnS... The risk assessments (RAs) (ECB 2008) assessed the risks related to the use of ZnCl2 in this respect for production of e.g. Zn3(PO4)2 based on reported data. Given the similar hazard profile of ZnCl2 and Zn(NH4)Clx, and the similarity of processes involved, the assessment for ZnCl2 (GES-2 ZnCl2) is crossread here for Zn(NH4)Clx. The risk characterisation is summarised in table below. Table below also summarises the risk characterisation based on more recent data on manufacture of other compounds. The exposure estimates based on these more recent data are summarised in the subsequent table below.

Table 101. Environmental risk characterisation for the scenario “industrial use of ZnCl2 for the manufacture of other zinc compounds” (crossread for Zn(NH4)Clx) Data from the EU RA Tonnage Cadd*/PNEC Cadd**/PNEC Cadd/PNEC PEC/PNEC (T/y) water sediment soil STP§ Manufacture of Zn3(PO4)2 (RA: 5 companies; cfr table 3.4.10 of RA) A 6000 0.06 0.5*** 0.03 NA

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 310 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

B 6000 0.01 0.3 0.02 NA C 3000 0.02 0.4 0.02 0.01 D 6000 0.05 0.4*** 0.02 NA E <1000 0.001 0.01 0.02 0.0005

Recent data ***

Zn3(PO4)2 manufacture Company F (recent 8000 (PEC/PNEC) (PEC/PNEC) (PEC/PNEC) (PEC/PNEC) data) 2005 0.18 0.32 0.39 NA 2006 0.18 0.37 0.39 NA 2007 0.18 0.38 0.39 NA 2008 0.18 0.31 0.39 NA 2009 0.18 0.31 0.39 NA

Zn(OH)CO3 manufacture Company A (direct) 5000 0.32 1.6 0.39 NA (2009) (indirect) 0.26 1.0 0.39 0.46

ZnBr2 manufacture Company A 72 0.17 0.19 0.39 0 *In the RAs, “Cadd” the “added concentration by the emissions at a given site was used for the risk characterisation. As such, the risks related to the local emissions at the site were assessed only, without the added exposure from the regional background. The risk ratios are those cited in the RA, i.e. with the PNECs derived in the RA; **after application of generic bioavailability factor 0.5 (RA 2008). ***for the recent data, the regional exposure was taken into account (PEC/PNEC). § PEC/PNEC ratio’s were recalculated with new PNEC for STP, leading to a RCR different than derived previously in RA

Table 102. Exposure assessment for the industrial use of ZnCl2 for the manufacture of other zinc compounds, based on recently reported exposure data (crossread for Zn(NH4)Clx).

Zn3(PO4)2 PEC water (µg PEC sediment mg PEC soil (mg PEC STP manufacture Zn/l) Zn/kgDW) Zn/kgDW) (mg Zn/l) Company F 2005 3.7 75 42 NA 2006 3.8 86 42 NA 2007 3.8 88 42 NA 2008 3.7 72 42 NA 2009 3.7 72 42 NA

Zn(OH)CO3 manufacture Company A (direct) 6.5 382 42 NA (2009) (indirect) 5.2 242 42 48

ZnBr2 manufacture Company A 0 0 42 0

Conclusion: based on the information of the risk assessment, there is no environmental risk at the sites covered by this scenario, when e.g. on site treatment of waste waters is present. The general absence of risk is confirmed by recently measured data covering also manufacture of other zinc compounds except for 1 company, where risk for sediment is predicted. It is noted that for this company a very small dilution (1.4) was reported. In the sediment, the bioavailability of zinc will be determined by the content

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 311 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

of acid volatile sulphide (AVS). It has been documented that there is covariance between zinc in sediment and AVS. In cases where AVS and sediment-Zn were measured near industrial sites, there was indeed a surplus on AVS observed, rendering the zinc present at that site non-bioavailable. Whether or not this phenomenon is also present at the sites mentioned in table above, can only be confirmed by local measurements.

It can be concluded that there is no risk for the environment from the manufacture of zinc compounds from Zn(NH4)Clx, if risk management measures as described in this scenario are applied.

2. Workers Occupational exposure (inhalation and dermal) to ammonium zinc chloride and the manufactured zinc compounds is possible due to manipulation of powdery materials, and from parts of the process when the zinc compound is already formed. The packaging and repackaging of the produced zinc compound in bags, big bags or bulk tankers may lead to contamination of the facility and to exposure (direct or indirect) of workers.  Pulmonary absorption may occur but most of the material that is deposited in the head and the tracheobronchial region is rapidly translocated to the GI tract and part of it will be absorbed in the GI tract.

 Zn(NH4)Clx has however rather coarse particle size (99.66% of the particles larger than 15.8 μ m).  Dermal contact with hands is prevented by mandatory wearing of gloves.  With normal handling, no respiratory personal protection (breathing apparatus) is necessary. Dust filter can be used (e.g. P2, efficiency 90%) if there is risk for exceedance of OEL/NOAEL.

Data reported on exposure in the risk assessment and more recently are summarised together with the risk characterisation in table below. Given the similar hazard profile between ZnCl2 and Zn(NH4)Clx, the assessment for ZnCl2 (GES-2) is crossread to Zn(NH4)Clx.

Table 103. Workplace exposure data and risk characterisation for the scenario “industrial use of ZnCl2 for the manufacture of other zinc compounds” (crossread to Zn(NH4)Clx). RA data Zn in workplace Risk ratio Systemic Risk ratio Systemic Risk ratio air (mg/m3) inhalation**** inhalation systemic dermal** systemic total inhalable exposure inhalation (mg/d) dermal (mg/d)*

Zn3(PO4)2 Rwc: 0.7 0.14 Rwc: 2.8 0.3 0.8 0.08

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 312 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Recent data

Zn3(PO4)2 0.5-1 0.1-0.2 1-2 0.1-0.2 No data, 0.05 (2007) assumed as Data reported modelled: by 1 company, 0.5*** at “most contaminated area”

Zn(OH)CO3 0.83 0.2 1.6 0.2 No data, 0.05 production assumed as (1 company) modelled: 0.5*** * assuming a respiratory absorption of 40% for Zn(NH4)Clx and 20% for ZnO, and an inhalation volume of 10m3. **dermal absorption of 0.2% for dust; no wearing of gloves assumed in RA ***24mg/d (MEASE) *0.2 (dermal absorption factor for dust) ; wearing of gloves obligatory; (PROC 4-5-8b; dispersive) **** DNEL inhalation for ZnCl2/ Zn(NH4)Clx /ZnSO4 and other soluble zinc substances is 1.0 mg/m3; for ZnO and other slightly soluble/ insoluble zinc substances: 5mg/m3

Conclusion: based on the measured data reported in the EU risk assessment, and confirmed by some more recent data, there is no risk for workers for this scenario.

9.1.4. GES Zn(NH4)Clx -3: Industrial and professional use of Zn(NH4)Clx as active laboratory reagent in aqueous or organic media, for analysis or synthesis.

Table 104. GES- Zn(NH4)Clx-3 Exposure Scenario Format (1) addressing uses carried out by workers 9.1.4. Title of Exposure Scenario number Zn(NH4)Clx GES-3: Industrial and professional use of Zn(NH4)Clx as active laboratory reagent in aqeous or organic media, for analysis or synthesis. List of all use descriptors related to the life cycle stage and all the uses under it; include market sector (by PC), if relevant; SU: 3,10, 22, 24 PROC: 1,2,3,4,5, 8b,9, 15 PC: 19, 21, 28, 39 AC: not applicable ERC: 1,2, 4, 6a, 6b, 8a, 8d

9.1.4. Exposure Scenario 9.1.4.1. Contributing scenario (1) controlling environmental exposure for the Industrial and professional use of Zn(NH4)Clx as active laboratory reagent in aqueous or organic media, for analysis or synthesis.

Further specification:

The ammonium zinc chloride is used for  Analysis: sample (solid or liquid) treatment or preparation: the substance is in the sample or in the reagens  or synthesis: manipulations are usually under ventilation (e.g. laminar flow, ventilation hood)  The substance is used o at the industrial scale, in industrial installations for air control and water treatment

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 313 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

o at the professional scale by laboratories

Product characteristics

Product related conditions:

Zn(NH4)Clx is used in minimum 80% purity; higher grades (>95%) are usual

Amounts used Daily and annual amount per site: maximum 5 T/y (industrial scale) maximum 0.5 T/y (professional scale)

Frequency and duration of use

Use is usually intermittent but continuous use is assumed as a worst case. It is possible that use is not continuous; this has to be considered when estimating exposure.

Environment factors not influenced by risk management Flow rate of receiving surface water:

If applicable: default for generic scenario: 18,000 m3/d, unless specified otherwise

 All processes are performed indoor in a confined area, with dedicated laboratory equipment. All solid residues containing zinc are recovered for recycling.

Technical conditions and measures at process level (source) to prevent release Process design aiming to prevent releases and hence exposure to the environment; this includes in particular conditions ensuring rigorous containment; performance of the containment to be specified (e.g. by quantification of a release factor in section 9.x.2 of the CSR);  Process enclosures and closed circuits where relevant.  If relevant, dust capturing and removal techniques are applied on local exhaust ventilation (centralised treatment, scrubbers, filters,...)  Containment of liquid volumes to collect waste streams

 At industrial scale, the waste waters will be treated in the on-site waste water treatment techniques that can be applied to prevent releases to water (if applicable) e.g.: chemical precipitation, sedimentation and filtration (efficiency 90-99.98%).  At professional scale, the emissions are treated usually by STP. Professional services will be used for treating waste streams e.g. for the recovery of metallic solids (for recycling), and for the recovery of e.g. acid solutions containing the substance.  Air emissions are controlled by use filters and/or other air emission abatement devices e.g. fabric (or bag) filters (up to 99% efficiency), wet scrubbers (50-99% efficiency). This may create a general negative pressure in the laboratory.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 314 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

 In general emissions are controlled and prevented by implementing an integrated management system e.g. ISO 9000/9001, ISO 1400X series, or alike, and, when applicable, by being IPPC-compliant. o Such management system should include general industrial hygiene practice e.g.: . information and training of laboratory personnel, . regular cleaning of equipment and floors, . procedures for process control and maintenance,...  Treatment and monitoring of releases to outside air, and exhaust gas streams according to national regulation.

 In cases where applicable: default size, unless specified otherwise.

 At industrial scale: Hazardous wastes from onsite risk management measures and solid or liquid wastes from production, use and cleaning processes should be disposed of separately to hazardous waste incineration plants or hazardous waste landfills as hazardous waste. Releases to the floor, water and soil are to be prevented. If the zinc content of the waste is elevated enough, internal or external recovery/recycling might be considered.

Fraction of daily/annual use expected in waste: zinc producers = 3.1 % zinc compound producers = 0.056 % downstream users = 0.30 %

A detailed assessment has been performed and is reported in the Waste report (ARCHE, 2012) (See Annex 1)

 At professional scale:

Fraction of daily/annual use expected in waste: 42% of all articles, 58% of the zinc used is recycled.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 315 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Appropriate waste codes: 20 01 34, 20 01 40, 20 03 01, 20 03 07

Suitable Disposal: Waste from end-of-life articles can be disposed of as municipal waste, except when they are separately regulated, like electronic devices, batteries, vehicles, etc. Disposal of wastes is possible via incineration (operated according to Directive 2000/76/EC on the incineration of waste) or landfilling (operated according to Reference Document on the Best available Techniques for Waste Industries of August 2006 and Council Directive 1999/31/EC and Council Decision 19 December 2002).

A detailed assessment has been performed and is reported in the Waste report (ARCHE, 2012) (See Annex 1)

Conditions and measures related to external recovery of waste Fraction of used amount transferred to external waste treatment for recovery: specify type of suitable recovery operations for waste generated by workers uses, e.g. re-distillation of solvents, refinery process for lubricant waste, recovery of slags, heat recovery out-side waste incinerators; specify effectiveness of measure;

 All residues are recycled or handled and conveyed according to waste legislation. .

9.1.4.2. Contributing scenario (2) controlling worker exposure for the Industrial use of Zn(NH4)Clx as active laboratory reagent in aqeous or organic media, for analysis or synthesis. Product characteristic Product related conditions, e.g. the concentration of the substance in a mixture, the physical state of that mixture (solid, liquid; if solid: level of dustiness), package design affecting exposure)

 Zn(NH4)Clx is used in minimum 80% purity; higher grades (>95%) are usual  The sample can be solid or liquid.  When the preparation is in solid state, it can be in a) powdery, b) glassy or c) pelletized form. In the powder form, it can be characterised by high dustiness in a worst case situation.

Amounts used Amounts used at a workplace (per task or per shift); note: sometimes this information is not needed for assessment of worker’s expo-sure maximum 5 T/y (industrial scale) maximum 0.5 T/y (professional scale) . Frequency and duration of use/exposure Duration per task/activity (e.g. hours per shift) and frequency (e.g. single events or repeated) of exposure

Use is usually intermittent but continuous use is assumed as a worst case. It is possible that use is not continuous; this has to be considered when estimating exposure.

Human factors not influenced by risk management Particular conditions of use, e.g. body parts potentially exposed as a result of the nature of the activity

Uncovered body parts: (potentially) face

 high temperature steps can occur in protected zones (fume cupboards);  all indoor processes in confined area, including hazardous substances cabinets.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 316 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

 Process enclosures and closed circuits where relevant and possible.  Local exhaust ventilation on work areas with potential generation of dust or fumes, dust capturing and removal techniques (fume cupboards).  Containment of liquid volumes and collection in special circuits

Technical conditions and measures to control dispersion from source towards the worker Engineering controls, e.g. exhaust ventilation, general ventilation; specify effectiveness of measure

 Local exhaust ventilation systems are provided where needed on the benches and in the fume cupboards.  Process enclosures if relevant  Dust control: dust to be measured in the workplace air according to national regulations.  Special care for the general establishment and maintenance of a clean working environment by e.g.:

 Cleaning of process equipment and laboratory

 Storage of Zn products in dedicated zones, e.g.: hazardous substances cabinets

Organisational measures to prevent /limit releases, dispersion and exposure

In general integrated management systems are implemented at the workplace e.g. ISO 9000/9001, ISO-ICS 13100, or alike, and are, when appropriate, IPPC-compliant.

Such management system would include general industrial hygiene practice e.g.: o information and training of personnel on prevention of exposure/accidents, o procedures for control of personal exposure (hygiene measures) o regular cleaning of equipment and floors, extended workers instruction-manuals o procedures for process control and maintenance,... o personal protection measures (see below)

Conditions and measures related to personal protection, hygiene and health evaluation Personal protection, e.g. wearing of gloves, face protection, full body dermal protection, goggles, respirator; specify effectiveness of measure; specify the suitable material for the PPE (where relevant) and advise how long the protective equipment can be used before replacement (if relevant) Wearing of protective clothing is compulsory (efficiency >=90%). Gloves can be used occasionally if risk for direct contact with the substance With normal handling, no respiratory personal protection (breathing apparatus) is necessary. If risk for exceedance of OEL/DNEL, use e.g.: -dust filter-half mask P1 (efficiency 75%) -dust filter-half mask P2 (efficiency 90%) -dust filter-half mask P3 (efficiency 95%) -dust filter-full mask P1 (efficiency 75%) -dust filter-full mask P2 (efficiency 90 %) -dust filter-full mask P3 (efficiency 97.5%)

Eyes: safety glasses are optional but usually taken as “normal laboratory practice”

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 317 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Exposure estimation and risk characterisation

1. Environment

In laboratory use of the substance, 2 situations can be distinguished related to environmental management/exposure:  At industrial scale, the waste waters from the laboratory will be connected to the on-site waste water treatment system and will be treated like the industrial process and other waters. The techniques that can be applied to prevent releases to water (if applicable) are thus the same as for the industrial waste waters e.g.: chemical precipitation, sedimentation and filtration (efficiency 90-99.98%).  At professional scale, the emissions to water are treated usually by STP. Still, professional services will be used for treating waste streams e.g. for the recovery of metallic solids (for recycling), and for the recovery of acid solutions containing the substance. These controlled waste streams will contain the bulk of the emissions.  Air emissions in general are controlled by use of filters and/or other air emission abatement devices e.g. fabric (or bag) filters (up to 99% efficiency), wet scrubbers (50-99% efficiency). This may create a general negative pressure in the laboratory For estimating the industrial laboratory emissions, the release factors are used, because they can be considered a worst case situation, because the zinc compounds are in their pure form, and the laboratories will be connected to the waste water treatment system. The release factors are summarised in table below:

Table 105. Environmental release factors for the manufacture of different zinc compounds, to be used for industrial laboratories using zinc compounds. Zn compound manufacture Release factor to air Release factor to water (g/g) Zinc Carbonate (1 company) 0.00012 0.000009 Ammonium zinc chloride (with / 0.0000063 WWTP) (1 company) Zinc oxide(1 company) 0.00046 0.000006 (1 company) 0.000012 0.0000044 (3 companies) 0.000017-0.00009 0 Zinc phosphate (1 company) 0.00003-0.0005 0.000016-0.00001

For the industrial use of zinc compounds, the highest of the measured release factors are used: 0.0005 for release to air, and 0.000016 for release to water. The exposure estimates and risk characterisation based on the modeling of emissions with these release factors are summarized in the table below.

For professional laboratory, the STP treatment is applied. The Eurometaux SPERC for “use of metal compounds” is 0.1% for air, 3% for water. The exposure estimates and risk characterisation based on the modeling of emissions with these release factors are also summarized in the table below.

Table 106. Exposure assessment and risk characterisation for the industrial and professional use of Zn(NH4)Clx in laboratory. Maximum PEC PEC/P PEC PEC/PNEC PEC soil PEC/PNEC PEC PEC/PNEC tonnage used water NEC sedime sediment* (mg/kgDW) soil* STP STP§ (T/y) (µg/l) water* nt (mg/l) mg/kg DW)

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 318 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Industr 5 3.4 0.17 46 0.2 41 0.39 0.2 0 ial Profes 0.5 4.7 0.23 184 0.79 41 0.39 0.034 0.33 sional *PECs include the regional PEC § PEC/PNEC ratio’s were recalculated with new PNEC for STP, leading to a RCR different than derived previously in RA

Conclusion: The risk ratios for the industrial uses of zinc compounds in laboratory are very low. For the professional uses, even when the very conservative release factor to water (Eurometaux SPERC) is applied, still PEC/PNECs < 1 are predicted. In conclusion, the use of zinc compounds in industrial and professional laboratory does not lead to a risk for the environment, when the risk management measures as described in this scenario are applied.

2) Workers Laboratory personnel applies general risk management measures to prevent exposure to zinc fumes/dust. Given the small quantities that are used in industrial, as well as in professional use, the human exposure is very limited.

Table below summarises the MEASE predictions of human exposure.

Table 107. Occupational exposure data and risk characterisation for the Industrial and professional use of Zn(NH4)Clx and other zinc compounds in the laboratory. Laboratory use 8-hrs Risk ratio Inhalation Dermal Risk ratio Inhalation inhalation**** exposure exposure systemic exposure (mg systemic (mg (modelled) Zn/m3) Zn/d)** systemic (mg/d) *** Industrial/professional ≤0.023 ≤0.023 0.04 0.005 0.004 PROC 1, 2, 3, 4, 5, 8b, 9, 15* *MEASE parameters:

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 319 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Aqueous solution >25% content Professional use Wide dispersive use Direct handling Intermittent exposure LEV generic No RPE gloves **assuming a respiratory absorption of 40% for Zn(NH4)Clx /ZnSO4 as a worst case for all zinc compounds (20% for ZnO and other zinc compounds), and an inhalation volume of 10m3 *** assuming a dermal absorption of 0.2% for dust, wearing of gloves assumed ****DNEL inhalation for Zn(NH4)Clx /ZnSO4 and other soluble zinc substances is 1.0 mg/m3; for ZnO and other slightly soluble/ insoluble zinc substances: 5mg/m3

Conclusion: based on modelling of exposure with MEASE, no risk is predicted for workers in the laboratory using Zn(NH4)Clx and other zinc compounds, following the risk management measures as described.

9.1.5. GES Zn(NH4)Clx -4 : Industrial use of Zn(NH4)Clx or Zn(NH4)Clx -formulations as component for the manufacture of solid blends and matrices for further downstream use.

Table 108.GES Zn(NH4)Clx -4 Exposure Scenario Format (1) addressing uses carried out by workers 9.1.5. Title of Exposure Scenario number GES Zn(NH4)Clx - 4 : Industrial use of Zn(NH4)Clx or Zn(NH4)Clx -formulations as component for the manufacture of solid blends and matrices for further downstream use. List of all use descriptors related to the life cycle stage and all the uses under it; include market sector (by PC), if relevant; SU: 1, 3, 4, 5, 8, 9, 10, 11, 12, 14, 15, 16, 18, 19, 0 (Nace C21.1., 23.9.9., 27.2.) PROC: 1, 2, 3, 4, 5 ,8b, 9, 13, 14, 15, 25 PC: 1, 8, 9a, 9b, 9c, 12, 14, 15, 18, 19, 20, 21, 26, 28, 29, 32, 35, 37, 38 AC:1, 2, 3, 7 ERC: 1, 2, 3, 4, 5, 7, 8a, 8b, 8d, 10a, 10b, 11a Further explanations (if needed)

Zn(NH4)Clx or Zn(NH4)Clx -containing preparations are used in the manufacture of dry preparations by mixing thoroughly the starting materials, possibly followed by pressing or pelletizing, and finally packaging of the preparation.

9.1.5. Exposure Scenario 9.1.5.1. Contributing scenario (1) controlling environmental exposure for the Industrial use of Zn(NH4)Clx or Zn(NH4)Clx -formulations as component for the manufacture of solid blends and matrices for further downstream use.

Further specification:

In the described process, the Zn(NH4)Clx (or Zn compound) containing preparation/mixture is optionally:  Pressed at high temperature (>1000°C), grinded and re-pressed or fritted at high temperature  Molten at high temperature (>500°C) and further cast as glassy material  Pressed and pelletized at low temperature And subsequently packed, or used as such, in further treatment/use

Product characteristics

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 320 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Product related conditions:

Zn(NH4)Clx (or Zn compound) in the preparation can be > 25%, usually <5%

Amounts used Daily and annual amount per site: maximum 5000 T/y;

Frequency and duration of use

Continuous production is assumed as a worst case. It is possible that use is not continuous; this has to be considered when estimating exposure.

Environment factors not influenced by risk management Flow rate of receiving surface water: default for generic scenario: 18,000 m3/d, unless specified otherwise

 All dry processes throughout, no process waters. Even when no process waters occur (with dry process throughout), some non-process water can be generated containing zinc (e.g. from cleaning)  High temperature steps are possible.  All processes are performed indoor in a confined area. All residues containing zinc are recycled.

Technical conditions and measures at process level (source) to prevent release Process design aiming to prevent releases and hence exposure to the environment; this includes in particular conditions ensuring rigorous containment; performance of the containment to be specified (e.g. by quantification of a release factor in section 9.x.2 of the CSR);  Local exhaust ventilation on furnaces and other work areas with potential dust generation.  Dust capturing and removal techniques are applied.  Process enclosures where relevant and possible.

 No process waters, so possible emissions to water are limited and non-process related.  On-site waste water treatment techniques can be applied to prevent releases to water (if applicable) e.g.: chemical precipitation, sedimentation and filtration (efficiency 90-99.98%).  Air emissions are controlled by use of bag-house filters and/or other air emission abatement devices e.g. fabric or bag filters, wet scrubbers. This may create a general negative pressure in the building.

In general emissions are controlled and prevented by implementing an integrated management system e.g. ISO 9000, ISO 1400X series, or alike, and, when appropriate, by being IPPC-compliant. o information and training of workers, o regular cleaning of equipment and floors, o procedures for process control and maintenance,...  Treatment and monitoring of releases to outside air, and exhaust gas streams (process & hygiene), according to national regulation.  SEVESO 2 compliance, if applicable.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 321 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

In cases where applicable: default size, unless specified otherwise.

Fraction of daily/annual use expected in waste: zinc producers = 3.1 % zinc compound producers = 0.056 % downstream users = 0.30 %

A detailed assessment has been performed and is reported in the Waste report (ARCHE, 2012) (See Annex 1)

 All residues are recycled or handled and conveyed according to waste legislation. .

9.1.5.2. Contributing scenario (2) controlling worker exposure for the Industrial use of Zn(NH4)Clx or Zn(NH4)Clx -formulations as component for the manufacture of solid blends and matrices for further downstream use. Name of contributing scenario 2:

Industrial formulation of dry preparations/mixtures by mixing thoroughly the Zn(NH4)Clx (or other zinc compounds) with the other starting materials, with possible pressing, pelletising, sintering and packaging of the

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 322 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 preparations/mixtures

 The concentration of Zn(NH4)Clx in the mixtures can be up to >25% but is usually of the order of <= 5%, depending on the application.  The preparation is in the solid state, usually with a low level of dustiness; however, powder forms can occur, the high dustiness is therefore applied as a worst case.  Zn(NH4)Clx particles are coarser then e.g. ZnO; 99.66% of the particles is larger than 15.8 μ m

Amounts used Amounts used at a workplace (per task or per shift); note: sometimes this information is not needed for assessment of worker’s expo-sure

Max 5000T/y = 15T/d = 5T/shift depending of application.

Frequency and duration of use/exposure Duration per task/activity (e.g. hours per shift) and frequency (e.g. single events or repeated) of exposure

8 hour shifts (default worst case) are assumed as starting point; it is emphasised that the real duration of exposure could be less. This has to be considered when estimating exposure.

Human factors not influenced by risk management Particular conditions of use, e.g. body parts potentially exposed as a result of the nature of the activity

Uncovered body parts: (potentially) face

 Dry processes: dry operational conditions throughout the process; no process waters;  high temperature steps can occur;  indoor processes in confined area.

 Local exhaust ventilation on furnaces and other work areas with potential dust generation, dust capturing and removal techniques  Process enclosures where appropriate

Technical conditions and measures to control dispersion from source towards the worker Engineering controls, e.g. exhaust ventilation, general ventilation; specify effectiveness of measure

 Local exhaust ventilation systems and process enclosures are generally applied  Cyclones/filters (for minimizing dust emissions): efficiency 70%-90% (cyclones); dust filters (50-80%)  LEV in work area: efficiency 84% (generic LEV)

In general integrated management systems are implemented at the workplace e.g. ISO 9000, ISO-ICS 13100, or

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 323 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 alike, and are, when appropriate, IPPC-compliant.

Exposure estimation and risk characterisation

1. Environment

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 324 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

The processes involved in this scenario are all dry, so there are no process waters. Even when no process waters are involved, occasional non-process-waters can occur having some zinc content, due to e.g. from dust cleaning. Therefore, all formulation processes with Zn(NH4)Clx and other zinc compounds should have some form of water treatment, on site or off-site, according to national legislation and permits. The physical form of the preparations is usually solid; the dustiness is then much lower than with the original substance Zn(NH4)Clx. However, since powder forms can occur, high dustiness (cfr ZnO) is applied as worst case.

The risk assessments on zinc and zinc compounds reported measured exposure data on a number of sectors falling under this scenario. In most cases, two sequential process steps are integrated at the same industrial site: a) the formulation of the substance into the dry preparation/mixture and b) the further industrial use of the preparation/mixture.

For this reason, environmental emissions data are in most cases integrating both process steps, and encompass both the generic scenarios GES-1 (formulation of Zn(NH4)Clx Zn compound into mixture) and GES-4. In the integrated process, exposure related to the formulation of the pure Zn(NH4)Clx is considered to be the most critical, because the substance is used in powdery form in its pure state. Therefore the data reported in the risk assessments for the formulation of ZnCl2 and other Zn compounds in mixtures (cfr GES 1) are used as worst case for the present scenario. The resulting risk characterisations are summarized in table below. Distinction is being made between assessments based on measured data, and assessments based on modelling, using default release factors. Preference is given to the measured data. Table below also summarizes the risk characterisation based on more recent data on manufacture of other compounds. The exposure estimates based on these more recent data are summarized in the second table below.

Table 109. Environmental risk characterisation for the Industrial use of Zn(NH4)Clx as component for the manufacture of solid blends and matrices for further downstream use crossread from other zinc compounds). assessments from the EU RA by PEC/PNEC PEC/PNEC PEC/PNEC soil PEC/PNEC sector of use* water sediment (**) STP§ ZnCl2 (table3.4.10., RA ZnCl2, ECB 2008) Assessment based on measured data Agrochemical industry processing (1 0.03 0.51 0.02 0.19 single EU production site)

Assessments based on modelling Chemical industry: processing 0.19 1.7 (0.71) 19 23.5

Additional recent data*****

Fertiliser manufacture Company A 0.16 0.19 0.39 0 *PNECs from the RA are applied, integrating for sediment the generic bioavailability factor 0.5 and for soil the generic bioavailability factor 0.33 (RA, ECB 2008); Risk ratios for water and sediment are Cadd/PNEC; for STP and soil risk ratios are PEC/PNEC. **PEC/PNEC ratios for sediment between brackets apply the updated PNEC and generic bioavailability factor of the RA ***all risk ratios are PEC/PNECs § PEC/PNEC ratio’s were recalculated with new PNEC for STP, leading to a RCR different than derived

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 325 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

previously in RA

Fertiliser manufacture Company A 3.4 45 41 0

Conclusion When local risks are assessed using measured emissions data, no risk is described for the formulation processes using ZnCl2. Also recent data on an additional sector (fertiliser) show no risk. Only when default release factors are applied (assessment based on modelling), risks are calculated. The measured data however overrule these modelled results, so it is concluded based on th measured data that there is no risk for the environment from this scenario, when risk management measures, as described, are applied. The conclusion on the environmental assessment on the use of ZnCl2 for the manufacture of solid blends and matrices for further downstream use is confirmed by data on formulation with other zinc substances, see table below.

Table 111. Environmental risk characterisation for the Industrial use of Zn compounds as component for the manufacture of solid blends and matrices for further downstream use. assessments from the EU RA by PEC/PNEC PEC/PNEC PEC/PNEC soil PEC/PNEC sector of use* water sediment (**) STP§ ZnS04 (table3.4.10., RA ZnSO4, ECB 2008) Assessment based on measured data Agricultural feed industry 0 0 0.02 0

RA ZnO (table 3.4.33., ECB 2008) Assessment based on measured data Ceramic industry processing typical 0 0 0.14 plant average 0 Ceramic industry processing typical 0 0 0.06-0.38 plant range 0 Ferrites industry (average of 4 (out 0.27 2.5 (1.0) 0.4 of 5) plants 0.125 Varistors (average of 2 (out of 4) 0.06 1.2 (0.5) 0.09 plants*** 0.03 Catalysts processing**** <4.9 <45 0.02 <2.25 Feedstuff additive: formulation (site 0 0 0.02 specific) 0 Feedstuff additive: formulation 0 0 0.03 (generic average use) 0 Feedstuff additive: formulation 0 0 0.05 (generic largest use use) 0

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 326 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Assessments based on modelling Glass industry: processing (average 2.5 23 0.93 use) 1.15 Glass industry: processing (largest 6.3 57 2.4 use) 2.9

*PNECs from the RA are applied, integrating for sediment the generic bioavailability factor 0.5 and for soil the generic bioavailability factor 0.33 (RA, ECB 2008); Risk ratios for water and sediment are Cadd/PNEC; for STP and soil risk ratios are PEC/PNEC. **PEC/PNEC ratios for sediment between brackets apply the updated PNEC and generic bioavailability factor of the RA ***data from site 3 (showing as only risk ratios>1), not considered, because it was explicitly mentioned that no WWTP or STP was present (RA ZnO). ****calculations from reported maximum concentration in waste water (<1mg Zn/l); For the one case with risk based on measured data observed in the RA, the catalysts producing sector, extensive additional data were generated; they demonstrate the absence of risks (see GES 1 ZnO). § PEC/PNEC ratio’s were recalculated with new PNEC for STP, leading to a RCR different than derived previously in RA

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 327 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

2. Workers Occupational exposure to Zn(NH4)Clx when dry mixing/blending Zn(NH4)Clx / Zn(NH4)Clx -preparations into dry solid matrices for further downstream use is possible due to dust emissions at several steps of the process. These dusts may lead to contamination of the facility and to exposure (direct or indirect) of workers, by inhalation and dermal contact. However, most of the products formed (pellets, fluxes, ..) are compacted solids, so dustiness is limited. Pulmonary absorption may occur but most of the material that is deposited in the head and the tracheobronchial region is rapidly translocated to the GI tract and part of it will be absorbed in the GI tract. It is noted that Zn(NH4)Clx particles are coarser then e.g. ZnO; 99.66% of the particles is larger than 15.8 μ m. The particle size distribution of the preparations is dependent on the application.

For assessing worker exposure, different lines of evidence can be used:  As a worst case, data reported in the risk assessment for the integrated process of mixing of the ZnO/Zn compound into the dry preparation and the subsequent further processing (table 112 below).  As a worst case, recent data on this step in e.g. catalyst production, pigment production (table 112 below).  Data for the formulation of pure Zn(NH4)Clx /Zn compounds can also be applied as a worst case. In this scenario, the main possibility for exposure is at the unpacking step of the pure Zn compound, and its mixing into the other components of the mix. In further processes, the concentration of the Zn compound is lower, and, consequently, exposure is also predicted to be lower. The risk assessments of e.g. ZnSO4 (ECB 2008) mentioned data for this step (table 113 below).  the manufacture of Zn(NH4)Clx can be considered as an additional worst case (highest concentration, pure Zn(NH4)Clx) (table 114).

Table 112. Occupational exposure data for the Industrial use of Zn(NH4)Clx and other Zn compounds as component for the manufacture of solid blends and matrices for further downstream use. Risk assessment Inhalation Risk ratio Inhalation Dermal exposure Total Risk ratio data (RA ZnCl2, exposure inhalation*** systemic RA systemic systemic (based on ECB 2008) (mg (mg/d) (MEASE- exposur measured Zn/m3) modelled) e (mg/d) data- (total mg/d** systemic) scenario inhalable) Production of animal Typical: / Rwc : 0.5 Rwc : 2* 2.3 4.3 0.43 feedstuff * Rwc: 0.5 Production of Rwc: 0.2 Rwc:0.2 0.8 0.2 1 0.1 fertilisers*

Recent data (on ZnO) Sector: activity Catalyst production: Mean: Mean: 0.07 0.74 0.2 0.94 0.09 0.37 Emptying of Range: Up to Up to Up to 0.2 containers Range: <0.0002-0.21 2.14 2.34 <0.001- 1.07 Catalyst production: Mean: Mean: 0.07 0.74 (0.2) 0.94 0.09 drying 0.37 Range: 0.01- Up to 1.7 Up to Up to 0.19 Range: 0.2 1.9 0.07-0.84 Catalyst production: Mean: Mean: 0.04 0.38 (0.2) 0.58 0.06 mixing 0.19 Range: Up to Up to Up to 0.1

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 328 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Range: <0.002-0.09 0.88 1.1 <0.01- 0.44 Catalyst production: Mean: 0.2 Mean: 0.04 0.4 (0.2) 0.6 0.06 forming Range: Range: Up to 2.8 Up to 3 Up to 0.3 0.004- 0.0008-0.3 1.42 Catalyst production: Mean: Mean: 0.15 1.5 (0.2) 1.7 0.17 precipitation/filtration 0.73 Range: Up to 2.7 Up to Up to 0.3 Range: 0.012-0.3 2.9 0.06-1.37 Catalyst production: Mean : Mean: 0.08 0.82 (0.2) 1.0 0.1 screening 0.41 Range: Up to 3.9 Up to Up to 0.4 Range: <0.002-0.4 4.1 <0.01- 1.96 Catalyst production: Mean: Mean: 0.12 1.2 (0.2) 1.4 0.14 filling 0.61 Range: Up to 3.3 Up to Up to 0.4 Range: 0.0008-0.33 3.5 0.004- 1.66 Catalyst production: Mean: Mean: 0.1 1.0 (0.2) 1.2 0.12 binning off 0.52 Range: Up to 2.6 Up to Up to 0.28 Range: <0.002-0.03 2.8 <0.01- 1.32 Catalyst production: Mean: Mean: 0.07 0.74 (0.2) 0.94 0.09 maintenance 0.37 Range: 0.03- Up to 1.2 Up to Up to 0.14 Range: 0.12 1.4 0.16-0.59 Recent data on ZnO and other Zn compounds Pigment production: 0.83 0.17 1.6 (0.2) 1.8 0.18 dosing and mixing (2005) Pigment production: 0.29 0.06 0.6 (0.2) 0.8 0.08 dosing and mixing (2006) Pigment production: 0.14 0.03 0.3 (0.2) 0.5 0.05 dosing and mixing (2009) Pigment production: 0.33 0.07 0.66 (0.2) 0.9 0.09 calcinations (charge) (2004) Pigment production: 0.055 0.01 0.11 (0.2) 0.3 0.03 calcinations (charge) (2005) Pigment production: 0.43 0.009 0.9 (0.2) 1.1 0.1 calcinations (charge) (2009)

Unspecified (“ZnO 0.1 0.02 0.2 (0.2) 0.4 0.04 8b”): preparing granulates Unspecified (“ZnO 0.1 0.02 0.2 (0.2) 0.4 0.04 8b”): pressing measuring 2005

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 329 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

*RA ZnSO4. 40% respiratory absorption was assumed for ZnSO4, and 10m3 respiratory volume/d **It is noted that in the RA, dermal exposures are estimated higher than in the present MEASE modelling, because the use of specialised working gloves is mandatory. ***DNEL inhalation for Zn(NH4)Clx /ZnSO4 and other soluble zinc substances is 1.0 mg/m3; for ZnO and other slightly soluble/ insoluble zinc substances: 5mg/m3

Table 113. Additional occupational data and risk characterisations also relevant for the industrial use of Zn(NH4)Clx and other Zn compounds as component for the manufacture of solid blends and matrices for further downstream use. Data activity 8-hrs Risk ratio Inhalation Dermal Systemic Risk from Inhalation inhalation** exposure exposure exposure ratio ZnSO4 exposure systemic (modelled) total (mg systemic RA by (mg (mg Zn/d) systemic Zn/d) Sector Zn/m3) (mg/d) Paint Emptying of 0.17-0.28 0.17-0.28 0.34-0.56 0.2 0.54- 0.05-0.08 industry* ZnO from big 0.76 bags into dispensers Loading 0.1-0.5 0.1-0.5 0.2-1.0 0.2 0.4-1.2 0.04-0.12 powders from Average: Average: 0.58 0.78 0.08 25kg big bags 0.29 0.29 into dispensers Loading 0.01-1.34 0.01-1.34 0.02-2.68 0.2 0.22- 0.02-0.3 powders from 2.88 Average Average 0.54 0.07 big bags into 0.27 0.27 0.74 dispensers Ceramics ZnO loaded 0.1-0.98 0.1-0.98 0.2-2.0 0.2 0.4-2.2 0.04-0.2 (1 from bulk company) transport to bulk storage *values are for total dust; exposure to dust for short duration; data extrapolated to 8hrs exposure ** DNEL inhalation for Zn(NH4)Clx /ZnSO4 and other soluble zinc substances is 1.0 mg/m3; for ZnO and other slightly soluble/ insoluble zinc substances: 5mg/m3

Table 114. Occupational exposure data and risk characterisation for the scenario “ZnCl2 manufacture” (crossread to Zn(NH4)Clx) RA data (RA Zn in workplace Risk ratio Systemic Risk ratio Systemic Risk ZnCl2, table air (mg/m3) inhalation*** inhalation systemic dermal ratio 4.1.3.2A) total inhalable exposure inhalation **(mg/d) systemic (mg/d)* dermal 3 companies 0.2 0.2 0.8 0.08 0.4 0.04

* assuming a respiratory absorption of 40% for Zn(NH4)Clx /ZnSO4 and 20% for ZnO and other zinc compounds, and an inhalation volume of 10m3 ** assuming a dermal absorption of 0.2% for dust, no wearing of gloves assumed ***DNEL inhalation for AZ Zn(NH4)Clx /ZnSO4 and other soluble zinc substances is 1.0 mg/m3; for ZnO and other slightly soluble/ insoluble zinc substances: 5mg/m3

Conclusion: based on measured data from the risk assessments and data from similar worst case scenarios, no risk is predicted for workers, following the risk management measures indicated in this scenario.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 330 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

9.1.6. GES Zn(NH4)Clx -5: Industrial use of Zn(NH4)Clx or Zn(NH4)Clx -formulations as component for the manufacture of dispersions, pastes or other viscous or polymerized matrices.

Table 115. GES Zn(NH4)Clx -5 Exposure Scenario Format (1) addressing uses carried out by workers 9.1.5. Title of Exposure Scenario number GES Zn(NH4)Clx -5 : Industrial use of Zn(NH4)Clx or Zn(NH4)Clx -formulations as component for the manufacture of dispersions, pastes or other viscous or polymerized matrices. List of all use descriptors related to the life cycle stage and all the uses under it; include market sector (by PC), if relevant; SU: 1, 3, 5, 6b, 7, 8, 9, 10, 11, 12, 14, 15, 16, 18, 19, 20, 0 (Nace C21.1., 23.9.9., 27.2) PROC: 1, 2,3,4,5,6,7, 8b,9,10, 11, 13, 14, 19, 20, 21, 24, 25, 26 PC: 4, 8, 9a, 9b, 9c, 12, 14, 15, 18, 19, 20, 21, 23, 24, 25, 28, 29, 31, 32, 33, 34, 35, 37, 38, 39, 40 AC:1, 2, 3, 5, 6, 7, 10, 11, 13 ERC:1, 2, 3, 4, 5, 6a, 6b, 6d, 8a, 8b, 8c, 8f, 9a, 9b, 10a, 10b,11a Further explanations (if needed)

9.1.5. Exposure Scenario 9.1.5.1. Contributing scenario (1) controlling environmental exposure for the industrial use of Zn(NH4)Clx or Zn(NH4)Clx -formulations as component for the manufacture of dispersions, pastes or other viscous or polymerized matrices. Name of contributing scenario

Further specification:

In the described process, the ammonium zinc chloride containing preparation/mixture is:  unpacked and stored in silos  Extracted from the silo, dosed and fed with the other reagents and/or solvents to the mixing tank, batch- wise or continuously, according the process receipt.  The resulting zinc salt containing mixture (solution, dispersion, paste) is directly further processed, or packed, for further treatment/use.

Product characteristics Product related conditions:

Zn(NH4)Clx in preparation can be > 25%

Amounts used Daily and annual amount per site: maximum 5000 T/y;

Frequency and duration of use

Continuous production is assumed as a worst case. It is possible that use is not continuous; this has to be considered when estimating exposure.

Environment factors not influenced by risk management Flow rate of receiving surface water: default for generic scenario: 18,000 m3/d, unless specified otherwise

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 331 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

 In parallel, non-process water can be generated containing zinc (e.g. from cleaning)  All processes are performed indoor in a confined area.  All residues containing zinc are recycled.

Technical conditions and measures at process level (source) to prevent release Process design aiming to prevent releases and hence exposure to the environment; this includes in particular conditions ensuring rigorous containment; performance of the containment to be specified (e.g. by quantification of a release factor in section 9.x.2 of the CSR);  Local exhaust ventilation on mixing tanks and other work areas with potential dust generation.  Dust capturing and removal techniques are applied.  Process enclosures where relevant and possible.

 Most of the operations imply wet process-steps  Sump containment is provided under the tanks and the filters i.o. to collect any accidental spillage  On-site waste water treatment techniques can be applied to prevent releases to water (if applicable) e.g.: chemical precipitation, sedimentation and filtration (efficiency 90-99.98%).  Air emissions are controlled by use of bag-house filters and/or other air emission abatement devices e.g. fabric or bag filters, wet scrubbers. This may create a general negative pressure in the building.

 In general emissions are controlled and prevented by implementing an integrated management system e.g. ISO 9000, ISO 1400X series, or alike, and, when applicable, by being IPPC-compliant.

o Such management system should include general industrial hygiene practice e.g.: . information and training of workers, . regular cleaning of equipment and floors, . procedures for process control and maintenance,...  Treatment and monitoring of releases to outside air, and exhaust gas streams (process & hygiene), according to national regulation.  SEVESO 2 compliance, if applicable.

In cases where applicable: default size, unless specified otherwise.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 332 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Fraction of daily/annual use expected in waste: zinc producers = 3.1 % zinc compound producers = 0.056 % downstream users = 0.30 %

All residues are recycled or handled and conveyed according to waste legislation. .

9.1.5.2. Contributing scenario (2) controlling worker exposure for the industrial use of Zn(NH4)Clx or Zn(NH4)Clx -formulations as component for the manufacture of dispersions, pastes or other viscous or polymerized matrices. Product characteristic Product related conditions, e.g. the concentration of the substance in a mixture, the physical state of that mixture (solid, liquid; if solid: level of dustiness), package design affecting exposure)

 The concentration of Zn(NH4)Clx in the mixtures can be >25%, depending on the application.  The preparation is in the liquid state, as a paste or dispersion or other viscous or polymerized matrix, with a low level of dustiness; however, powder forms can occur, medium dustiness is therefore applied as a worst case

Amounts used Amounts used at a workplace (per task or per shift); note: sometimes this information is not needed for assessment of worker’s expo-sure

Max 5000T/y = 20 T/d = 7 T/shift depending of application.

Frequency and duration of use/exposure Duration per task/activity (e.g. hours per shift) and frequency (e.g. single events or repeated) of exposure

8 hour shifts (default worst case) are assumed as starting point; it is emphasised that the real duration of exposure could be less. This has to be considered when estimating exposure.

Human factors not influenced by risk management Particular conditions of use, e.g. body parts potentially exposed as a result of the nature of the activity

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 333 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Uncovered body parts: (potentially) face

 Wet processes  All indoor processes in confined area.

 Local exhaust ventilation on mixing tanks, furnaces and other work areas with potential dust generation, dust capturing and removal techniques  Process enclosures where appropriate

Technical conditions and measures to control dispersion from source towards the worker Engineering controls, e.g. exhaust ventilation, general ventilation; specify effectiveness of measure

In general integrated management systems are implemented at the workplace e.g. ISO 9000, ISO-ICS 13100, or alike, and are, when appropriate, IPPC-compliant.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 334 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Eyes: safety glasses are optional

Exposure estimation and risk characterisation

1. Environment

The processes involved in this scenario are all wet. Even when no process waters are involved, occasional non- process-waters can occur having some zinc content, due to e.g. dust cleaning. Therefore, all formulation processes with Zn(NH4)Clx and other zinc compounds should have some form of water treatment, on site or off-s- ite, according to national legislation and permits.

The physical form of the preparations is a wet matrix, so the dustiness is usually very low than with the original substance Zn(NH4)Clx. However, since powder forms can occur, medium dustiness is applied as worst case.

The risk assessments on zinc and zinc compounds reported measured exposure data on a number of sectors falling under this scenario. In most cases, two sequential process steps are integrated at the same industrial site:

a) the formulation of the substance into the wet preparation/mixture and

b) the further industrial use of the preparation/mixture.

For this reason, environmental emissions data are in most cases integrating both process steps, and encompass both the generic scenarios GES-1 (formulation of Zn(NH4)Clx /Zn compound into mixture) and GES-5. In the integrated process, exposure related to the formulation of the pure Zn(NH4)Clx is considered to be the most critical, because the substance is used in powdery form in its pure state, which gives highest potential for environmental exposure (even when no process waters, by non-process emissions, e.g. by cleaning). Therefore the data reported in the risk assessments for the formulation of ZnCl2 /Zn compounds in mixtures (cfr GES 1) are used as worst case for the present scenario.

The resulting risk characterisations are summarized in table below. Distinction is being made between assessments based on measured data, and assessments based on modelling, using default release factors. Preference is given to the measured data.

Table below also summarizes the risk characterisation based on more recent data on manufacture of other compounds. The exposure estimates based on these more recent data are summarized in the second table below.

Table 116. Environmental risk characterisation for the Industrial use of Zn(NH4)Clx as component for the manufacture of liquid blends and matrices for further downstream use. assessments from the EU RA by PEC/PNEC PEC/PNEC PEC/PNEC soil PEC/PNEC sector of use* water sediment (**) STP§ ZnCl2 (table3.4.10., RA ZnCl2, ECB 2008) Assessment based on measured data

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 335 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Agrochemical industry processing (1 0.03 0.51 0.02 single EU production site) 0.195 Battery industry (1 company) 0 0 0.02 0

Assessments based on modelling Chemical industry: processing 0.19 1.7 (0.71) 19 23.5 Battery industry: processing 0.16 1.4 0.2 0.075 Dyes and inks industry: formulation 5.3 48 2.0 2.45 Dyes and inks industry: formulation 150 1343 56 69

Additional recent data***

Table 117. Exposure assessment for the industrial use of ZnCl2 as component for the manufacture of liquid blends and matrices for further downstream use (crossread to Zn(NH4)Clx). PEC water (µg PEC sediment mg PEC soil (mg PEC STP Zn/l) Zn/kgDW) Zn/kgDW) (mg Zn/l) Fertiliser manufacture Company A 3.4 45 41 0

Conclusion When local risks are assessed using measured emissions data, no risk is generally described for the formulation processes using ZnCl2. Recent data on an additional sector (fertiliser) show also no risk. Only when default release factors are applied (assessment based on modelling), risks are calculated. The measured data however overrule these modelled results, so it is concluded based on the measured data that there is no risk for the environment from this scenario, when risk management measures, as described, are applied. The conclusion on the environmental assessment of formulation of ZnCl2 is confirmed by data on formulation with other zinc substances, most of the RA, with recent data added, see table below. By crossreading , this conclusion holds also for Zn(NH4)Clx.

Table 118. Environmental risk characterisation for the Industrial use of Zn compounds as component for the manufacture of liquid blends and matrices for further downstream use. assessments from the EU RA by PEC/PNEC PEC/PNEC PEC/PNEC soil PEC/PNEC sector of use* water sediment (**) STP§ ZnS04 (table3.4.10., RA ZnSO4, ECB 2008) Assessment based on measured data Agricultural feed industry 0 0 0.02 0

Assessments based on modelling Agricultural pesticide industry 0.11 1 11 13

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 336 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Agricultural fertiliser industry 19 175 7.3 9 Agricultural feed industry 1.0 9 0.4 0.47 Chemical industry: processing 0.19 1.7 19 23.5

ZnO (table3.4.33., ECB 2008) Assessment based on measured data Tyre industry: processing 0 0 0.15 0 General rubber industry: processing 0 0 0.08 0 Feedstuff additive: formulation (site 0 0 0.02 specific) 0 Feedstuff additive: formulation 0 0 0.03 (generic average use) 0 Feedstuff additive: formulation 0 0 0.05 (generic largest use use) 0 Paints: formulation 0 0 0.02 0 Paints: processing (industry data) 0 0 0.02 0

Assessments based on modelling Lubricants: formulation (average 7.5 67 2.7 use) 3.45 Lubricants: formulation (largest use) 13 118 5 6 Paints/ processing: generic data 1.6 14 0.6 0.75 Cosmetics pharmaceuticals: 2.5 23 0.93 formulation (average use) 1.15 Cosmetics pharmaceuticals: 21 188 8 formulation (largest use) 9.5

Zn phosphate (table3.4.9., RA Zn phosphate, ECB 2008) Assessment based on measured data Paint industry (average from 3 of 5 0.19 1.7 (0.35) Not calculated sites reported)*** 0.175

Assessments based on modelling Paint industry: formulation 8.3 75 3.1 3.85 Paint industry: processing, solvent 0.23 2.1 0.28 borne 0.105 Paint industry: processing, water 1.2 11 0.43 borne 0.55

Additional recent data**** Use of Zn(H3PO4)2 in liquid blend 0.23 0.79 0.39 / *PNECs from the RA are applied, integrating for sediment the generic bioavailability factor 0.5 and for soil the generic bioavailability factor 0.33 (RA, ECB 2008); Risk ratios for water and sediment are Cadd/PNEC; for STP and soil risk ratios are PEC/PNEC. **PEC/PNEC ratios for sediment between brackets apply the updated PNEC and generic bioavailability factor of the RA ***Only reliable data are used, where the truly measured emission and/or effluent concentration was reported **** all risk ratios are PEC/PNECs § PEC/PNEC ratio’s were recalculated with new PNEC for STP, leading to a RCR different than derived previously in RA

Table 119. Exposure assessment for the industrial use of Zn compounds as component for the manufacture of liquid blends and matrices for further downstream use.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 337 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Making liquid PEC water (µg PEC sediment mg PEC soil (mg PEC STP blends of Zn/l) Zn/kgDW) Zn/kgDW) (mg Zn/l) Zn(H3PO4)2 Company A 4.7 187 41 /

A specific use of Zn(NH4)Clx in liquid blend is as a flux solution in batch (general) hot dip galvanising. The EU RA on ZnCl2 assessed in detail the exposure from this use. In the EU RA, exposure from hot dip galvanising (HDG) was extensively analysed (ECB 2008). For general (batch) HDG, extensive data were available from galvanising plants in FR, UK, DE, NL. Emissions to water, including emissions from run-off of stockpiled galvanised steel articles, were assessed in detail. A complete and detailed measured dataset on 20 NL galvanising plants was used for risk characterisation. The NL dataset was indeed, together with the data on the other countries mentioned, considered to be representative for the EU, “because the process itself does not result in zinc emissions (holds for EU in general) and the releases from non-process sources are expected not to be significantly different between EU countries” (EU RA, ECB 2008). These data are related to combined uses of Zn metal, ZnCl2 and Zn(NH4)Clx , and therefore considered relevant for the use of Zn(NH4)Clx in this process and summarised in table below.

Table 120. Exposure assessment and risk characterisation for the Industrial use of Zinc, alloyed or not, ZnCl2 and Zn(NH4)Clx for metal surface treatment (hot dip galvanising) or for pyrometallurgical extraction processes. Data reported in the PEC/PNEC water PEC/PNEC PEC/PNEC soil PEC/PNEC STP§ EU risk assessment on sediment zinc* General (batch) hot dip 0.0018-0.037 0.03-0.66 0 0.0008-0.017 galvanising *data from RA zinc, table 3.4.67. Risk ratios with PNECs from RA, and as reported in the RA: for water/sediment: Cadd/PNEC; for STP/soil: PEC/PNEC. § PEC/PNEC ratio’s were recalculated with new PNEC for STP, leading to a RCR different than derived previously in RA

Conclusion: The data from the risk assessment demonstrate that also for general hot dip galvanising, there is no risk for environment.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 338 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

2. Workers Occupational exposure to zinc when wet mixing/blending Zn(NH4)Clx / Zn(NH4)Clx preparations into wet matrices for further downstream use is possible due to dust emissions at the initial step of the process (mixing of the dry Zn(NH4)Clx into the other components). These dusts may lead to contamination of the facility and to exposure (direct or indirect) of workers, by inhalation and dermal contact. However, the mixtures that are formed (pastes, dispersions or other viscous or polymerized matrices) are wet, so dustiness is very limited. Pulmonary absorption may occur but most of the material that is deposited in the head and the tracheobronchial region is rapidly translocated to the GI tract and part of it will be absorbed in the GI tract. It is noted that Zn(NH4)Clx particles are coarser then e.g. ZnO; 99.66% of the particles is larger than 15.8 μm.

For assessing worker exposure, different lines of evidence can be used:  data reported in the risk assessment for the integrated process of mixing of the ZnCl2 or other Zn compounds into the wet preparation and the subsequent further processing. Some data on dry mixing are included here as worst case (table 121 below).  recent data on this step in e.g. catalyst production, pigment production (dry formulation: worst case) are also included as worst case (table 121).  Data for the formulation of high dustiness pure ZnO/Zn compounds can also be applied as a worst case. In this scenario, the main possibility for exposure is at the unpacking step of the ZnO/Zn compound, and its mixing into the other components of the mix. The risk assessments of e.g. ZnSO4 (ECB 2008) mentioned data for this step (table 122 below).  the manufacture of ZnCl2 can be considered as an additional worst case (high dustiness) (table 114).

Table 121. Occupational exposure data for the Industrial use of formulations containing ZnO and/or other zinc compounds as component for the manufacture of mixtures for further downstream use.

Risk assessment data Inhalation Risk ratio Inhalation Dermal Total Risk ratio (ECB 2008) exposure inhalation***** systemic exposure RA systemic (systemic) (mg (mg/d) systemic exposure total Zn/m3) (MEASE- (mg/d) scenario (total modelled) inhalable) mg/d**** Production of animal Typical: / Rwc : 0.5 Rwc : 2.3 4.3 0.43 feedstuff * Rwc: 0.5 2** (ZnO) Production of Rwc: 0.2 Rwc:0.2 0.8 0.2 1 0.1 fertilisers* Production of paints 2 4 4.8 (0.2) 8.8 (4.2) 0.88 containing ZnO** (0.42) Production of rubber 2 4 4.4 (0.2) 8.4 (4.2) 0.84 products containg (0.42) ZnO** Production of 0.4 0.8 0.9 (0.2) 1.7 (1.0) 0.17 (0.1) paint***

Recent data Sector: activity

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 339 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Catalyst production : Mean: Mean: 0.07 0.74 0.2 0.94 0.09 emptying of 0.37 Range: containers Range: <0.0002-0.21 (dry formulation: <0.001- worst case) 1.07 Catalyst production: Mean: Mean: 0.07 0.74 (0.2) 0.94 0.09 drying 0.37 Range: 0.01-0.2 Up to 1.7 1.9 0.19 (dry process: worst Range: case) 0.07-0.84 Catalyst production: Mean: Mean: 0.04 0.38 (0.2) 0.58 0.06 mixing 0.19 Range: <0.002- Up to Up to Up to 0.1 (dry process: worst Range: 0.09 0.88 1.1 case) <0.01-0.44 Catalyst production: Mean: 0.2 Mean: 0.04 0.4 (0.2) 0.6 0.06 forming Range: Range: 0.0008- Up to 2.8 Up to 3 Up to 0.3 (dry process: worst 0.004-1.42 0.3 case) Catalyst production: Mean: Mean: 0.15 1.5 (0.2) 1.7 0.17 precipitation/filtration 0.73 Range: 0.012- Up to 2.7 Up to Up to 0.3 (dry process: worst Range: 0.3 2.9 case) 0.06-1.37 Catalyst production: Mean : Mean: 0.08 0.82 (0.2) 1.0 0.1 screening 0.41 Range: <0.002- Up to 3.9 Up to Up to 0.4 (dry process: worst Range: 0.4 4.1 case) <0.01-1.96 Catalyst production: Mean: Mean: 0.12 1.2 (0.2) 1.4 0.14 filling 0.61 Range: 0.0008- Up to 3.3 Up to Up to 0.4 (dry process: worst Range: 0.33 3.5 case) 0.004-1.66 Catalyst production: Mean: Mean: 0.1 1.0 (0.2) 1.2 0.12 binning off 0.52 Range: <0.002- Up to 2.6 Up to Up to (dry process: worst Range: 0.03 2.8 0.28 case) <0.01-1.32

Catalyst production: Mean: Mean: 0.07 0.74 (0.2) 0.94 0.09 maintenance 0.37 Range: 0.03- Up to 1.2 Up to Up to (dry process: worst Range: 0.12 1.4 0.14 case) 0.16-0.59

Pigment production: 0.83 0.17 1.6 (0.2) 1.8 0.18 dosing and mixing (2005) Pigment production: 0.29 0.06 0.6 (0.2) 0.8 0.08 dosing and mixing (2006) Pigment production: 0.14 0.03 0.3 (0.2) 0.5 0.05 dosing and mixing (2009) Pigment production: 0.33 0.07 0.66 (0.2) 0.9 0.09 calcinations (charge) (2004) Pigment production: 0.055 0.01 0.11 (0.2) 0.3 0.03 calcinations (charge) (2005) Pigment production: 0.43 0.009 0.9 (0.2) 1.1 0.1 calcinations (charge) (2009)

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 340 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Unspecified (“ZnO 0.1 0.02 0.2 (0.2) 0.4 0.04 8b”): preparing granulates Unspecified (“ZnO 0.1 0.02 0.2 (0.2) 0.4 0.04 8b”): pressing measuring 2005

*RA ZnSO4. 40% respiratory absorption was assumed for ZnSO4, and 10m3 respiratory volume/d **RA ZnO ***RA Zn3(PO4)2 ****It is noted that in the RA, dermal exposures are estimated higher than in the present MEASE modelling, because the use of specialised working gloves is mandatory. ***** DNEL inhalation for Zn(NH4)Clx /ZnSO4 and other soluble zinc substances is 1.0 mg/m3; for ZnO and other slightly soluble/ insoluble zinc substances: 5mg/m3

Table 122. Occupational exposure data also relevant for the industrial formulation of wet preparations/mixtures by mixing thoroughly zinc compounds with other materials Data activity 8-hrs Risk ratio Inhalation Dermal Systemic Risk ratio from Inhalation inhalation exposure exposure exposure Systemic ZnSO4 exposure systemic (mg (modelled) total (mg total RA by (mg Zn/d) systemic Zn/d) Sector Zn/m3) (mg/d) Paint Emptying of 0.17-0.28 0.03-0.06 0.34-0.56 0.2 0.54-0.76 0.05-0.08 industry* ZnO from big bags into dispensers Loading 0.1-0.5 0.02-0.1 0.2-1.0 0.2 0.4-1.2 0.04-0.12 powders Average: Average: 0.58 0.78 0.08 from 25kg 0.29 0.06 big bags into dispensers Loading 0.01-1.34 0.002-0.3 0.02-2.68 0.2 0.22-2.88 0.02-0.3 powders Average Average 0.54 0.74 0.07 from big 0.27 0.06 bags into dispensers Ceramics ZnO loaded 0.1-0.98 0.02-0.2 0.2-2.0 0.2 0.4-2.2 0.04-0.2 (1 from bulk company) transport to bulk storage

*values are for total dust; exposure to dust for short duration; data extrapolated to 8hrs exposure

Table 123. Occupational exposure data and risk characterisation for the scenario “ZnCl2 manufacture” RA data (RA Zn in workplace Risk ratio Systemic Systemic Risk ratio ZnCl2, table air (mg/m3) inhalation inhalation dermal systemic total 4.1.3.2A) total inhalable exposure **(mg/d) (mg/d)* 3 companies 0.2 0.2 0.8 0.4 0.12

* assuming a respiratory absorption of 40% for Zn(NH4)Clx ZnCl2//ZnSO4 and 20% for ZnO and other zinc

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 341 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

compounds, and an inhalation volume of 10m3 ** assuming a dermal absorption of 0.2% for dust, no wearing of gloves assumed

A specific use of Zn(NH4)Clx in liquid blend is as a flux solution in batch (general) hot dip galvanising. The exposure of workers (via inhalation or the dermal route) comes from the Zn(NH4)Clx that is used as a flux on the bath. Since it is not possible to make distinction between exposure from the molten zinc (which is very low at the melting temperature of zinc in the bath) and from the Zn(NH4)Clx flux, both exposures are combined in the exposure estimation below, according to the EU risk assessment on Zn (ECB 2008). In continuous hot-dip galvanizing Zn(NH4)Clx is not used as a fluxing or pre-treatment agent.

Table 124. Occupational exposure data for the industrial use of Zinc, alloyed or not, or Zn(NH4)Clx or metal surface treatment (batch hot dip galvanising) or for pyrometallurgical extraction processes. Estimations of worker 8-hrs Inhalation Inhalation Dermal exposure Risk ratio exposure exposure (mg exposure systemic systemic (mg/d) Systemic total Zn/m3) (mg Zn/d) Exposure to ZnO and 0.2 + 0.2 (0.8 + 0.4) = 1.2** 0.3*** 0.15 Zn(NH4)Clx (simultaneous exposure) (Data from the RA on ZnCl2*) *Data reported for batch hot-dip galvanising (RA ZnCl2, table 4.1.3.2.A.).

**assuming 40% respiratory adsorption for ammonium zinc chloride, 20% for zinc oxide

***data from RA ZnCl2 taken as worst case (no wearing of gloves assumed)

Conclusion: based on measured data from the risk assessments and data from similar worst case scenarios (e.g. dry ZnO manufacture), and data for Zn/ Zn(NH4)Clx use in batch hot dip galvanising, no risk is predicted for workers, following the risk management measures indicated in this scenario.

9.1.7. GES Zn(NH4)Clx - 6 : Industrial and professional use of solid substrates containing less than 25%w/w of Zn(NH4)Clx.

Table 125. GES Zn(NH4)Clx -6 Exposure Scenario Format (1) addressing uses carried out by workers 9.1.7. Title of Exposure Scenario number GES Zn(NH4)Clx - 6 : Industrial and professional use of solid substrates containing less than 25%w/w of Zn(NH4)Clx. List of all use descriptors related to the life cycle stage and all the uses under it; include market sector (by PC), if relevant; SU: 3, 5, 6b, 9, 10, 16, 17, 18, 20, 22, 0 (Nace 23.9.9) PROC: 2, 4, 5 ,6, 8b, 9,10, 11, 13, 19 PC: 1, 7, 8, 9a, 9b, 9c,14,15, 18 , 20, 21, 23, 25, 28, 29, 34, 35, 38, 39, AC: 1, 2, 3, 5, 6, 7, 0 (coatings for art and creative items) ERC: 3, 5, 8a, 8d, 10a, 11a

9.1.7. Exposure Scenario 9.1.7.1. Contributing scenario (1) controlling environmental exposure for the Industrial and professional use of solid substrates containing less than 25%w/w of Zn(NH4)Clx

Further specification:

This scenario covers both the industrial scale processes and professional use. In the described process, the Zn(NH4)Clx containing preparation/mixture is further processed, involving potentially the following steps:  Reception/unpacking of material

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 342 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

 Final application, embedding, or shaping to produce the end product or article.

Product characteristics Product related conditions:

Zn(NH4)Clx (or Zn compound) in the article is < 25%

Amounts used Daily and annual amount per site:

 The quantities involved in this scenario are 10-50 times smaller than in blending (GES 4-GES 5); the concentration of the zinc substance is also lower (<25%).  Typical quantities for both Industrial and professional are 50T/y (typical), maximum 500T/y (in industrial setting).

Frequency and duration of use

Continuous production is assumed as a worst case. Usually, use is not continuous; this has to be considered when estimating exposure.

Environment factors not influenced by risk management Flow rate of receiving surface water: default for generic scenario: 18,000 m3/d, unless specified otherwise

 Solid, so in principle all dry processes throughout, no process waters. Even when no process waters occur (with dry process throughout), some non-process water can be generated containing zinc (e.g. from cleaning)  In industrial and professional setting, all processes are performed indoor in a confined area. All residues containing zinc are recycled.

Technical conditions and measures at process level (source) to prevent release Process design aiming to prevent releases and hence exposure to the environment; this includes in particular conditions ensuring rigorous containment; performance of the containment to be specified (e.g. by quantification of a release factor in section 9.x.2 of the CSR);  In industrial and professional setting the following applies: o Local exhaust ventilation on furnaces and other work areas with potential dust generation. o Dust capturing and removal techniques are applied. o Process enclosures where relevant and possible.

 In industrial and professional setting, the following applies: o No process waters, so possible emissions to water are limited and non-process related. o If zinc emissions to water, on-site waste water treatment techniques can be applied to prevent releases to water (if applicable) e.g.: chemical precipitation, sedimentation and filtration (efficiency 90-99.98%). . By exposure modelling it is predicted that at use quantities of >100T/y, refinement of the exposure assessment to water and sediment needs to be made (exposure assessment based on real measured data and local parameters). Treatment of the emissions to water may be needed under such conditions (see “exposure estimation

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 343 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

and risk characterisation”). o Air emissions are controlled by use of bag-house filters and/or other air emission abatement devices e.g. fabric or bag filters, wet scrubbers. This may create a general negative pressure in the building.

In general, emissions are controlled and prevented by implementing an appropriate management system. This would involve:  information and training of workers,  regular cleaning of equipment and floors,  procedures for process control and maintenance,...  Treatment and monitoring of releases to outside air, and exhaust gas streams, according to national regulation.  SEVESO 2 compliance, if applicable.

In cases where applicable: default size, unless specified otherwise.

Fraction of daily/annual use expected in waste: zinc producers = 3.1 % zinc compound producers = 0.056 % downstream users = 0.30 %

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 344 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Hazardous landfill operated under Directive 1999/31/EC.

A detailed assessment has been performed and is reported in the Waste report (ARCHE, 2012) (See Annex 1)

 At professional scale:

Fraction of daily/annual use expected in waste: 42% of all articles, 58% of the zinc used is recycled.

Appropriate waste codes: 20 01 34, 20 01 40, 20 03 01, 20 03 07

A detailed assessment has been performed and is reported in the Waste report (ARCHE, 2012) (See Annex 1)

 All residues are recycled or handled and conveyed according to waste legislation.

9.1.7.2. Contributing scenario (2) controlling worker exposure for the Industrial and professional use of solid substrates containing less than 25%w/w of Zn(NH4)Clx. Product characteristic Product related conditions, e.g. the concentration of the substance in a mixture, the physical state of that mixture (solid, liquid; if solid: level of dustiness), package design affecting exposure)

The concentration of Zn(NH4)Clx (or Zn compound) in the mixture is < 25%  The mixture is in the solid state, with a low level of dustiness; however, powder forms can occur, the medium dustiness is therefore applied as a worst case.

Amounts used Amounts used at a workplace (per task or per shift); note: sometimes this information is not needed for assessment of worker’s expo-sure

 The quantities involved in this scenario are 10-50 times smaller than in blending (GES 4-GES 5); the concentration of the zinc substance is also lower (<25%).  Typical quantities for both Industrial and professional are 50 T/y (typical), or 0.15 T/day, 0.05 T/shift  maximum use quantity is 500T/y (1.5T/d, 0.5T/shift) in industrial setting.

Frequency and duration of use/exposure Duration per task/activity (e.g. hours per shift) and frequency (e.g. single events or repeated) of exposure

8 hour shifts (default worst case) are assumed as starting point; it is emphasised that the real duration of exposure could be less. This has to be considered when estimating exposure.

Human factors not influenced by risk management

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 345 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Particular conditions of use, e.g. body parts potentially exposed as a result of the nature of the activity

Uncovered body parts: (potentially) face

 Industrial / Professional: o Dry processes: dry operational conditions throughout the process; no process waters; o indoor processes in confined area.

 Industrial /professional o Local exhaust ventilation on work areas with potential dust generation, dust capturing and removal techniques o Process enclosures where appropriate  Technical conditions and measures to control dispersion from source towards the worker Engineering controls, e.g. exhaust ventilation, general ventilation; specify effectiveness of measure

 Industrial /professional: o Local exhaust ventilation systems and process enclosures are generally applied o Cyclones/filters (for minimizing dust emissions): efficiency 70%-90% (cyclones); dust filters (50-80%) o LEV in work area: efficiency 84% (generic LEV)  Organisational measures to prevent /limit releases, dispersion and exposure Specific organisational measures or measures needed to support the functioning of particular technical measures (e.g. training and supervision). Those measures need to be reported in particular for demonstrating strictly controlled conditions (to justify exposure based waiving).

In general, management systems are implemented; They include general industrial hygiene practice e.g.: o information and training of workers on prevention of exposure/accidents, o procedures for control of personal exposure (hygiene measures) o regular cleaning of equipment and floors, extended workers instruction-manuals o procedures for process control and maintenance,... o personal protection measures (see below)

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 346 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

-dust filter-full mask P3 (efficiency 97.5%) Eyes: safety glasses are optional

Exposure estimation and risk characterisation

1. Environment

The processes involved in this scenario are all dry, so there are no process waters. Even when no process waters are involved, occasional non-process-waters can occur having some zinc content, due to e.g. from dust cleaning. The physical form of the preparations is usually solid; the dustiness is then much lower than with the original substance Zn(NH4)Clx. However, since powder forms can occur, medium dustiness is applied as worst case.

The risk assessments on zinc and zinc compounds reported no measured exposure data on these scenarios. Therefore exposure has to be modelled. Based on the modelling, recommendations can be made (see discussion below).

The resulting exposure estimates and risk characterisations are summarized in table below. The specific environmental release factors are (Verdonck et al., 2010):

-to air: 0.03% - 0.0003 g/g

-to water: 0.02% - 0.0002 g/g

Table 126. Environmental risk characterisation for the Industrial and professional use of solid substrates containing less than 25% w/w of Zn(NH4)Clx. tonnage PEC PEC/PNEC PEC PEC/PNEC PEC soil PEC/PNEC PEC PEC/PNEC used (T/y) water water* sedimen sediment* (mg/kgD soil* STP STP§ (µg/l) t W) (mg/l Mg/kgD ) W) Industrial 50 3.9 0.19 101 0.43 41 0.39 0.014 0.13 (typical) Professional 100 5.1 0.25 231 0.98 41 0.39 0.046 0.435 (trigger for refined assessment *PECs include the regional PEC § PEC/PNEC ratio’s were recalculated with new PNEC for STP, leading to a RCR different than derived previously in RA

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 347 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Discussion: The model predictions indicate that up to a use of 100T/y, no risks are predicted for this scenario. At uses > 100T/y, a more refined assessment of the possible emissions to water should be made (exposure assessment based on real measured data and local parameters). If needed, some form of water treatment, on site or off-site, according to national legislation and permits, should be applied.

Conclusion When considering typical use quantities, model calculations predict no risk for the environment for the downstream use processes using Zn(NH4)Clx. When use quantities exceed a critical level of 100T/y, more refined assessment should be done and risk management measures should be applied to ensure safe use.

2. Workers Occupational exposure to Zn(NH4)Clx when using Zn(NH4)Clx -containing preparations in solid form is possible due to possible dust formation. These dusts may lead to contamination of the facility and to exposure (direct or indirect) of workers, by inhalation and dermal contact. However, most of the products formed (pellets, fluxes ..) are solid, so dustiness is limited. The mixture is in the solid state, with a low level of dustiness; however, powder forms can occur, the medium dustiness is therefore applied as a worst case. Pulmonary absorption may occur but most of the material that is deposited in the head and the tracheobronchial region is rapidly translocated to the GI tract and part of it will be absorbed in the GI tract. Because of lack of any measured data, worker exposure is assessed using the MEASE model. As a worst case, professional situation is taken for the model calculations. Distinction is being made between indoor and outdoor use. No LEV is assumed present in the latter situation.

Table 127: Occupational exposure data for the Industrial and professional use of solid substrates containing less than 25%w/w of Zn(NH4)Clx. Inhalation Risk ratio Inhalation Dermal Total Risk ratio exposure inhalation** systemic exposure systemic (systemic) (mg Zn/m3) (mg/d)** systemic exposure total (total (mg/d) (mg/d) inhalable)* ****

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 348 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

MEASE modelling: ≤0.675 ≤0.675 ≤4.0 ≤0.12 ≤4.1 ≤0.4 professional (= worst case for industrial) indoor full shift, PROC 2, 4, 5, 6, 8b, 9,10, 13 MEASE modelling: professional (= worst case for industrial) indoor full shift, PROC 11, 19 ≤3 ≤3 ≤12 ≤0.12 ≤12 ≤1.2 with respiratory protection if > 1 hr ≤ 0.75 ≤ 0.75 ≤3.0 ≤0.12 ≤ 3.1 ≤0.3 (e.g.: P1 mask (=MEASE: AFP4- mask) MEASE modelling: professional (= worst case for industrial) outdoor full shift, PROC 2, 4, 5, ≤3 ≤3 ≤6 ≤0.12 ≤6 ≤0.6 6, 8b, 9,10, 13, 19 with respiratory protection if > 1 hr (e.g.: P1 mask ≤ 0.75 ≤ 0.75 ≤ 3.0 ≤0.12 ≤3.1 ≤0.3 (=MEASE: AFP4- mask) MEASE modelling: professional (= worst case for industrial) outdoor full shift, PROC 11 12 12 48 ≤0.12 48 4.8 with respiratory protection if > 1 hr (e.g.: MEASE: AFP4- mask) 0.6 0.6 1.2 ≤0.12 1.3 0.13

*MEASE parameters were set as follows:

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 349 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

 Solid, medium dustiness  Content in preparation: 5-25%  Process category (PROC): as indicated  Process T: 25°C  Scale of operation: professional  Duration of exposure: >240 minutes  Pattern of use: wide dispersive  Patter of exposure control: direct handling  Contact level: extensive  Implemented RMMs: LEV generic (indoor); no RMM (outdoors)  Efficiency based on: median estimate  No RPE  Use of gloves: properly designed

** DNEL inhalation for Zn(NH4)Clx is 1.0 mg/m3, for ZnO 5mg/m3 *** 40% respiratory absorption was assumed for Zn(NH4)Clx/ZnCl2 and ZnSO4, 20% for ZnO; 10m3 respiratory volume/d ****dermal absorption of 0.2% for dust

Conclusion: Based on modeling of exposure, no risk is predicted indoor for workers/professional users, for the majority of the PROC codes (2, 4, 5, 6, 8b, 9,10, 13) of this scenario, following the risk management measures indicated. For the PROCs 11, 19 however, respiratory protection (e.g. with P1 mask) is required when exposure exceeds 4 hrs.

Outdoors, risk is predicted for the PROCs 2, 4, 5, 6, 8b, 9,10, 13, 19. Wearing of a P1 mask is necessary when exposure exceeds 4 hrs. For PROC 11 higher level respiratory protection is recommended as a function of exposure time, e.g. mask with >95% efficiency when exposure exceeds 4 hrs, mask with efficiency > 90% when exposure exceeds 1 hr, but is less than 4hrs.

9.1.8. GES Zn(NH4)Clx -7 : Industrial and professional use of dispersions, pastes and polymerised substrates containing less than 25%w/w of Zn(NH4)Clx.

Table 128. GES Zn(NH4)Clx -7 Exposure Scenario Format (1) addressing uses carried out by workers 9.1.8.Title of Exposure Scenario number GES Zn(NH4)Clx - 7 : Industrial and professional use of dispersions, pastes and polymerised substrates containing less than 25%w/w of Zn(NH4)Clx. List of all use descriptors related to the life cycle stage and all the uses under it; include market sector (by PC), if relevant; SU: 5, 6, 9, 11, 12, 13, 15, 17, 18, 19, 20, 22 PROC: 7, 8b, 9, 10, 11, 13, 14, 17, 19, 21 PC:1, 4, 8, 9a, 9b, 9c, 14, 19, 20, 21, 24, 25, 28, 29, 31, 32, 35 , 39 AC: 1, 2, 7, 11 ERC: 8a, 8c, 8d, 8f, 10a

9.1.8. Exposure Scenario 9.1.8.1. Contributing scenario (1) controlling environmental exposure for the Industrial and professional use of dispersions, pastes and polymerised substrates containing less than 25%w/w of Zn(NH4)Clx.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 350 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Further specification:

Product characteristics Product related conditions:

Zn(NH4)Clx (or Zn compound) in the article is < 25%

Amounts used Daily and annual amount per site:

 The quantities involved in this scenario are 10-50 times smaller than in blending (GES 4-GES 5); the concentration of the zinc substance is also lower (<25%).  Typical quantities for both industrial and professional are 50T/y (typical), maximum 500T/y (in industrial setting).

Frequency and duration of use

Continuous production is assumed as a worst case. Usually, use is not continuous; this has to be considered when estimating exposure.

Environment factors not influenced by risk management Flow rate of receiving surface water: default for generic scenario: 18,000 m3/d, unless specified otherwise

 Wet processes. All process and non-process waters should be recycled internally to a maximal extent. Even when no process waters occur, some non-process water can be generated containing zinc (e.g. from cleaning)  In industrial and professional setting, all processes are performed in a confined area. All residues containing zinc are recycled.

Technical conditions and measures at process level (source) to prevent release Process design aiming to prevent releases and hence exposure to the environment; this includes in particular conditions ensuring rigorous containment; performance of the containment to be specified (e.g. by quantification of a release factor in section 9.x.2 of the CSR);  In industrial and professional setting the following applies:  Process enclosures where relevant and possible  Local exhaust ventilation on furnaces and other work areas with potential dust generation.  Dust capturing and removal techniques are applied.  Containment of liquid volumes in sumps to collect/prevent accidental spillage

 In industrial and professional setting, the following applies:

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 351 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

o If zinc emissions to water, on-site waste water treatment techniques can be applied to prevent releases to water (if applicable) e.g.: chemical precipitation, sedimentation and filtration (efficiency 90-99.98%). . By exposure modelling it is predicted that at use quantities of >100T/y, refinement of the exposure assessment to water and sediment needs to be made (exposure assessment based on real measured data and local parameters). Treatment of the emissions to water may be needed under such conditions (see “exposure estimation and risk characterisation”). o Air emissions are controlled by use of bag-house filters and/or other air emission abatement devices e.g. fabric or bag filters, wet scrubbers. This may create a general negative pressure in the building.

In cases where applicable: default size, unless specified otherwise.

Fraction of daily/annual use expected in waste: zinc producers = 3.1 % zinc compound producers = 0.056 % downstream users = 0.30 %

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 352 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

A detailed assessment has been performed and is reported in the Waste report (ARCHE, 2012) (See Annex 1)

 At professional scale:

Fraction of daily/annual use expected in waste: 42% of all articles, 58% of the zinc used is recycled.

Appropriate waste codes: 20 01 34, 20 01 40, 20 03 01, 20 03 07

A detailed assessment has been performed and is reported in the Waste report (ARCHE, 2012) (See Annex 1)

 All residues are recycled or handled and conveyed according to waste legislation.

9.1.8.2. Contributing scenario (2) controlling worker exposure for the Industrial and professional use of dispersions, pastes and polymerised substrates containing less than 25%w/w of Zn(NH4)Clx. Product characteristic Product related conditions, e.g. the concentration of the substance in a mixture, the physical state of that mixture (solid, liquid; if solid: level of dustiness), package design affecting exposure)

The concentration of Zn(NH4)Clx (or Zn compound) in the mixture is < 25%  Particles can occur sporadically, the low level of dustiness is basically applied.  Most of the processes imply the use of solutions or pastes; the “solution status” is therefore taken as the worst case.

Amounts used Amounts used at a workplace (per task or per shift); note: sometimes this information is not needed for assessment of worker’s expo-sure

 The quantities involved in this scenario are 10-50 times smaller than in blending (GES 5-GES 5); the concentration of the zinc substance is also lower (<25%).  Typical quantities for both Industrial and professional are 50 T/y (typical), or 0.15 T/day, 0.05 T/shift.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 353 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

 maximum use quantity is 500T/y (1.5T/d, 0.5T/shift) in industrial setting.

Frequency and duration of use/exposure Duration per task/activity (e.g. hours per shift) and frequency (e.g. single events or repeated) of exposure

8 hour shifts (default worst case) are assumed as starting point; it is emphasised that the real duration of exposure could be less. This has to be considered when estimating exposure.

Human factors not influenced by risk management Particular conditions of use, e.g. body parts potentially exposed as a result of the nature of the activity

Uncovered body parts: (potentially) face

 Industrial / Professional: o Wet processes, all indoor in confined area.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 354 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

-dust filter-half mask P1 (efficiency 75%) -dust filter-half mask P2 (efficiency 90%) -dust filter-half mask P3 (efficiency 95%) -dust filter-full mask P1 (efficiency 75%) -dust filter-full mask P2 (efficiency 90 %) -dust filter-full mask P3 (efficiency 97.5%) Eyes: safety glasses are optional

Exposure estimation and risk characterisation

1. Environment

The processes involved in this scenario are wet. Non-process-waters can occur having some zinc content, due to e.g. from dust cleaning. The physical form of the preparations is usually liquid; the dustiness is then much lower than with the original substance Zn(NH4)Clx. However, aerosols can occur occasionally, “solution status” is applied as worst case for dustiness in modelling. The risk assessments on zinc and zinc compounds reported no measured exposure data on these scenarios. Therefore exposure has to be modelled. Based on the modelling, recommendations can be made (see discussion below).

The resulting exposure estimates and risk characterisations are summarized in table below. The specific environmental release factors are (Verdonck et al 2010): -to air: 0.03% - 0.0003 g/g -to water: 0.02% - 0.0002 g/g

Table 129. Environmental risk characterisation for the Industrial and professional use of liquid substrates containing less than 25% w/w of Zn(NH4)Clx. tonnage PEC PEC/PN PEC PEC/PNEC PEC soil PEC/PNEC PEC PEC/P used (T/y) water EC sediment sediment* (mg/kgDW) soil* STP NEC (µg/l) water* Mg/kgDW) (mg/l) STP§ Industrial 50 3.9 0.19 101 0.43 41 0.39 0.014 0.13 (typical) Professiona 100 5.1 0.25 231 0.98 41 0.39 0.046 0.435 l (trigger for refined assessment *PECs include the regional PEC § PEC/PNEC ratio’s were recalculated with new PNEC for STP, leading to a RCR different than derived previously in RA

Conclusion When considering typical use quantities, model calculations predict no risk for the environment for the

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 355 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 downstream use processes using Zn(NH4)Clx. When use quantities exceed a critical level of 100T/y, more refined assessment should be done and risk management measures should be applied to ensure safe use.

Calculation of local exposure- Bioavailability correction The local exposure at a given site can be calculated specifically using the excel sheet prepared by Arche (see “tools” on http://www.reach-zinc.eu/) In addition, bioavailability corrections can be integrated in the exposure assessment, if the environmental parameters that are needed for the calculations, are documented.  For water assessment, bioavailability model correction can be applied when the following water parameters are documented for the receiving water: Dissolved organic carbon (DOC), pH, hardness or Ca-concentration. For the calculations, the “zinc BLM-calculator” excel tool is used to this end (see “tools” on http://www.reach-zinc.eu/). When the local values of these parameters are unknown, regional data can be used as an alternative. Use of regional instead of local values should always be handled with caution.

2. Workers Occupational exposure to diammonium tetrachlorozincate when using Zn(NH4)Clx -containing preparations in liquid form is possible due to aerosol formation. These aerosols may lead to exposure (direct or indirect) of workers, by inhalation and dermal contact. However, most of the products formed (dispersions, pastes and polymerised substrates, fluxes ..) are liquid, so dustiness is very limited, and “solution status” is applied as worst case for dustiness in modelling. Pulmonary absorption may occur but most of the material that is deposited in the head and the tracheobronchial region is rapidly translocated to the GI tract and part of it will be absorbed in the GI tract.

Because of lack of any measured data, worker exposure is assessed using the MEASE model. As a worst case, professional situation for “solutions” handling is taken for the model calculations. Distinction is made between indoor and outdoor professional use. In the latter situation, no LEV is assumed present.

Table 130. Occupational exposure data for the Industrial and professional use of solid substrates containing less than 25%w/w of Zn(NH4)Clx. Inhalation Risk ratio Inhalation Dermal Total Risk ratio exposure (mg inhalation** systemic exposure systemic (systemic) Zn/m3) (mg/d)*** systemic exposure (total (mg/d)**** (mg/d) inhalable)*

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 356 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

MEASE modelling*: ≤ 0.675 ≤0.675 ≤ 2.7 ≤ 0.06 ≤2.7 ≤ 0.27 professional (= worst case for industrial); indoor PROCs: 8b, 9, 10, 13, 14, 15, 17, 21 MEASE modelling: professional (= worst case for industrial) ; indoor full shift, PROC 7, 11  without ≤ 2.7 ≤ 2.7 ≤ 10.8 0.09 ≤10.9 ≤ 1.1 respiratory protection* ≤ 0.7 ≤ 0.7 ≤ 1.4 0.09 ≤1.4 ≤ 0.14  with respiratory protection (e.g.: P1 mask (=MEASE AFP 4 Mask) MEASE modelling: professional (= worst case for industrial) ; indoor full shift, PROC 19* 0.3 0.3 1.2 0.03 1.2 0.12  = respiratory protection already included MEASE modelling*: professional (= worst case for industrial); outdoor ≤ 0.6 ≤0.6 ≤0.06 0.3 ≤0.36 ≤0.04 PROCs: 8b, 9, 10, 13, 14, 17, 21 MEASE modelling*: professional (= worst case for industrial); outdoor (no LEV assumed) 12 12 48 0.3 48 4.8 PROC 11

PROC 11 with mask e.g. 0.6 0.6 2.4 0.3 2.7 0.3 AFP 20 (95% efficiency)

*MEASE parameters were set as follows:  aqueous solution  Content in preparation: 5-25%  Process category (PROC): as indicated

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 357 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

 Process T: 25°C  Scale of operation: professional  Duration of exposure: >240 minutes,  Pattern of use: wide dispersive  Patter of exposure control: direct handling  Contact level: intermittent  Implemented RMMs: LEV generic , for outdoor: none  Efficiency based on: median estimate  No RPE, unless indicated otherwise  Use of gloves: properly designed

** DNEL inhalation for Zn(NH4)Clx is 1.0 mg/m3 ***40% respiratory absorption was assumed for Zn(NH4)Clx /ZnCl2 and ZnSO4, and 10m3 respiratory volume/d ****dermal absorption of 0.2% for dust, 2% for liquid

Conclusion: For indoor processes, based on modelling of exposure, no risk is predicted for workers/professional users for most of the PROC codes of this scenario (5, 8b, 9, 10, 13, 14, 17, 21), following the risk management measures indicated. However, for PROCs 7,11 and 19, respiratory protection is required when exposure exceeds 1 hr. For outdoor professional use, for PROC 11 (non-industrial spraying), a mask should be worn, with an efficiency depending on the time of exposure, e.g.: when >240 minutes exposure: with 95% efficiency.

9.1.9. GES Zn(NH4)C1x-8: Generic wide dispersive use of Zn

A generic scenario on consumer STP (wide dispersive use) was developed.

Table 131. GES Zn(NH4)C1x -8: Generic wide dispersive use of Zn 9.1.9. Generic wide dispersive use of zinc Use descriptors 8a, 8b, 8c, 8d, 8e, 8f, 9a, 9b, 10a, 10b, 11a, 11b Additional information This generic exposure scenario has been created based on measured zinc concentrations in effluents from municipal STPs 9.1.9.1 Controlling environmental exposure Product characteristics Zinc is used in a variety of formulations or articles used by consumers Amounts used Total amounts used are not relevant since the assessment is done based on concentrations in STPs Frequency and duration of use Releases occur for 365 day/year, it’s a wide dispersive use and STPs are also operating 365 day/year. Environment factors not influenced by risk management Information type Dilution factor Remarks Selected for Exposure Scenario 10 Freshwater default Other given operational conditions affecting environmental exposure Indoor or outdoor use of products containing zinc is possible; zinc can be used in formulations that go down the drain but also in articles with non-intended releases. Conditions and measures related to municipal sewage treatment plant All releases are going directly into a municipal sewer. The releases are treated in an STP with removal efficiency for zinc of 80 %. The STP is dimensioned according to the defaults in EUSES. I.e. 10,000 inhabitants equivalents and 2,000 m3/day water treated per day. Zinc concentrations in effluents of municipal STPs have been collected in a separate report. (Evaluation of risks due to the presence of zinc in European Sewage Treatment Plants, 2013) The 90th percentile is 91.6 µg Zn/L and reflect the situation of a realistic worst case region, Flanders (Belgium), in terms of population density, and density of agricultural and industrial activities. The natural sources are subtracted from that 90th percentile resulting in a zinc concentration of 77.6 µg/L coming from wide dispersive use of zinc in consumer products and

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 358 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 articles. 9.1.9.2 Exposure estimation and reference to its source

Compartment Operational conditions Value Unit PNECadd RCR

PECtot stp 77.6 µg/L 100 0.78 E th PECtot freshwater 90 P of zinc conc. in effluents of STPs minus the 6.4 µg/L 34.3 0.19 S PECtot sediment natural sources: 77.6 µg Zn/L 73.4 mg/kg dw 117.8 0.62 1 PECtot soil 55.0 mg/kg dw 107 0.51 9.1.9.3 Guidance to DU to evaluate whether he works inside the boundaries set by the ES The DU works inside the boundaries set by the ES if either the proposed risk management measures as described above are met or the DU can demonstrate on his own that his implemented risk management measures are adequate. Detailed guidance for evaluation of ES can be acquired via your supplier or from the ECHA website (guidance R16). For environmental exposure, a DU-scaling tool (free download: http://www.arche-consulting.be/Metal-CSA-toolbox/du-scaling-tool) is available.

Exposure estimation and risk characterisation for environment

The trend (1990-2009) of zinc entering the aquatic environment from Sewage Treatment Plants (STP) is generally decreasing mainly due to a decrease of discharges from industrial settings (CBS, 2011). The potential risk of the presence of zinc in European STPs has been analysed in more detail in the present analysis, notably focusing on the following elements:

- Extension of the Zn STP database with recent data sets that came available on STPs in order to come to a complete and consistent picture of the Zn mass balance (influent/effluent/removal/river/soil) in domestic STPs.

- Revision of the PNEC STP based on a new nitrification inhibition study (Juliastuti, 2003).

- Incorporation of bioavailability for the sediment compartment as this compartment acts as the final repository for zinc present in the aquatic compartment.

These data were subsequently used to conduct a refined assessment of potential risks and to develop a local emission scenario for wide dispersive (consumer) use (Vangheluwe et al., 2013; Annex 2). In order to calculate zinc risk characterisation ratios from a wide dispersive consumer use exposure scenario for zinc, the effluent concentrations derived from the data set for Flanders (Belgium) is been used together with other input parameters that has been refined. The updated identified key parameters listed in Error: Reference source not found32 were entered into an EUSES-spread sheet and Zn concentrations in both the receiving aquatic and terrestrial compartments were calculated.

Table 132. Input parameters for EUSES REACH CSR Refined value used for the final calculations

Effluent Zn concentration (P90) 91.6 µg/L Removal efficiency 82% 79%

PNECadd STP 52 µg/L 100 µg/L Kd susp. solids 110,000

PNECadd Sediment 117.8 mg/kg dw BioF Sediment 0.5 0.2

PNECadd Freshwater 20.6 µg/L BioF Freshwater 0.6 PNEC Soil 106.8 mg/kg dw Effluent Zn concentration (P90) 91.6 µg/L Removal efficiency 82% 79%

The 90th percentile of the Flanders data set amounts to 92 µg/L. This value represents a total value and covers all sources. In order to come to an estimate for only the use of consumer products the human excretion and the natural input of zinc contributing to the zinc concentration in an STP i.e. 11 µg/L (human excretion) + 3 µg/L (leaf decay and rainwater) were subtracted. This yielded a final zinc concentration reflecting predominantly

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 359 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 domestic sources of approximately 78 µg/L. Using the parameters discussed in the above paragraphs safe use can be demonstrated for the STP scenario for the wide dispersive use of zinc in consumer products. RCR ratios are for all environmental compartments are below 1 (Table 131). The full report can be found back in Annex 2 (Vangheluwe et al., 2013)

9.2. Consumer exposure

Introduction

Zn(NH4)Clx can be used in several consumer products. Consumer exposure to zinc compounds was assessed in detail in the EU risk assessments (Part: human health). In this assessment, it was remarked that the total daily consumer exposure could be higher than from the substance alone, by the use of consumer products containing other zinc substances at the same time. Therefore, the RA made an integrated analysis of human consumer exposure from all main consumer products (containing different zinc substances) combined. Since this combined exposure is the reality of consumer exposure, this approach is also followed in the present analysis.

The risk assessment identified the main possible sources of consumer exposure. Since the pattern of consumption of consumer products containing zinc substances has not changed significantly after the closure of the risk assessment, the analysis made in the RA is considered still relevant for the consumer exposure at present and taken over in this CSR. Conform to the approach followed in the RA, the consumer exposure analysis is identical for all zinc substances. The RA analysis indeed included not only exposure from the products, containing the 6 zinc substances evaluated under 793/93/EEC, but also the exposure from products containing other, e.g. organic zinc substances. As such, it covered also the consumer exposure to Zn(NH4)Clx.

Related to the calculations of exposure, the main assumptions made in the RA were that uptake through inhalation was negligible and that the dermal absorption of the zinc compounds from any of the consumer products is 2% for solutions/suspensions, and 0.2% for dust/powder (same values as applied in the industrial environment).

Consumer exposure analysis of the RA (ECB 2008)

Remark: The section below is identical for all six zinc compounds evaluated under EU Regulation 793/93. Specific information is available for five of the six zinc compounds under evaluation (zinc phosphate, zinc distearate, zinc oxide, zinc chloride and zinc sulphate), as well as for some other zinc compounds not under evaluation. The latter information has also been included, because consumers (knowingly or unknowingly) at the same time can be exposed to several zinc-containing products, and irrespective of the original zinc compounds in these products, exposure will ultimately be to Zn2+. paint  Anti-corrosive primer containing 30% zinc phosphate. Assuming a frequency of 0.5 events/year with a dermal exposure of 2.7 g (paintbrush) or 10.8 g (spraying; roughly estimated as 4x paintbrush) primer/event, the maximum exposure will be 1.62 g zinc phosphate/year  2.25 mg Zn2+/day. With a dermal absorption of 2% the uptake is estimated to be 0.045 mg Zn2+ /day.

 Impregnating agent containing 40% zinc naphthenate. Assuming a frequency of 0.5 events/year with a dermal exposure of 2.7 g impregnating agent/event, the exposure will be 0.54 g zinc naphthenate/year  0.44 mg Zn2+/day (percentage of zinc in zinc naphthenate is estimated at 30%). With a dermal absorption of 2% the uptake is estimated to be 0.0088 mg Zn2+/day. cosmetics  Eye shadow containing 10% zinc distearate (it mainly concerns glossy, emulsion-like eye shadows). By an application of 10 mg/event for 3 times/day, the exposure to eye shadow is 30 mg/day, which contains 3 mg zinc distearate  0.31 mg Zn2+/day. Assuming a dermal absorption of 2% the uptake is estimated to be 0.0062 mg Zn2+ /day.

 Sunscreen containing 10% zinc oxide (refers to a protection factor 20-25!).

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 360 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

By an application of 9 g sunscreen/event, 3 events/day during 18 days/year the exposure will be 1332 mg sunscreen/day, being 107 mg Zn2+/day. Assuming a dermal absorption of 2% the uptake is estimated to be 2.14 mg Zn2+/ day.

 Deodorant contains 10-20% large organic zinc compounds, but apparently no ZnO. The dermal exposure is 3 g or 0.5g/event by using a spray or a roll-on, respectively. In both cases the use is once a day. Maximum dermal exposure to deodorant is 3000 mg/day  300 mg zinc compounds/day  30 mg Zn2+/day (percentage of zinc in these zinc compounds is estimated at 10%). Assuming a dermal absorption of 2% the uptake is estimated to be 0.6 mg Zn2+/day.

 Dandruff shampoo containing 5% zinc compounds such as zinc pyrithione and zinc omadine (5% is estimated based on other active components in dandruff shampoos). By a usage of 12 g shampoo/event for 4 times/week, the dermal exposure to shampoo will be 6800 mg/day with a content of 340 mg zinc compounds. Assuming that 10% of these compounds consist of zinc and that the dermal absorption is 2%, the uptake via the use of dandruff shampoo will be 0.68 mg Zn2+/day. drugstore products  ‘Baby care’ ointment containing 15% zinc oxide for the irritated skin (intensive ointment) or 5% zinc oxide for protective treatment when changing diapers. The assumption was made that the usage will be 50 g of the intensive ointment/year, leading to a dermal exposure of 7.5 g ZnO/year  16.5 mg Zn2+/day. Assuming a dermal absorption of 2% the uptake is estimated to be 0.33 mg Zn2+/day.

 Gargle containing 6.88 mg zinc chloride/ml. Assuming a use of 10 g gargle/event ( 10 ml/event), 4 times/day for 4 weeks/year, the exposure during these 4 weeks will be 1120 g gargle/year  3.1 g gargle/day, which is  10 mg Zn2+/day. Assuming that almost nothing will be swallowed, there is only buccal uptake via the mucous membranes. As the contact time is very short, the uptake is assumed to be very limited. Hence, with an arbitrary absorption value of 2% the uptake is estimated to be 0.2 mg Zn2+/day.

 Eye drops containing 0.25% zinc sulphate (2.5 mg/ml). The assumption was made that the usage will be 2 eye drops (0.025 ml/drop)/event, 6 times/day during 4 weeks/year, leading to an exposure of 8.4 ml eye drops/year  23 mg eye drops/day  0.058 mg zinc sulphate/day  0.023 mg Zn2+/day. Assuming an absorption of 2% the uptake is estimated to be 0.00046 mg Zn2+/day.

 Zinc oil containing 60% ZnO, which is merely used medically for the treatment of skin disorders. The assumption was made that the usage will be 100 g/year, leading to an exposure of 60 g ZnO/year  0.131 g Zn2+/day. Assuming a dermal absorption of 2% the uptake is estimated to be 2.62 mg Zn2+/day. Remark: it is noted that with skin disorders uptake might be higher than 2%. However, how much more is not known. Besides, it is not expected that the possible higher amount absorbed will disturb the homeostatic balance.

 Dietary supplements containing zinc. Results from a recent report on the food intake of the general population in the Netherlands (Hulshof et al., 1998) indicate that approximately 10% of the population uses dietary supplements, which amongst others can contain zinc. As it is not known how much zinc-containing dietary supplements are used and in what frequency, it is difficult to estimate the exposure to zinc from dietary supplements from this report. A dietary survey in the UK showed that <1-3% of the participants in different age groups took zinc supplements, providing median zinc intakes of 0.3-3.4 mg/day. However, the contribution of this supplemental zinc intake to the population average zinc intakes from food and supplements combined was negligible (EVM, 1999).

Conclusion The compound specific exposure estimates for the different zinc compounds are taken across to the risk characterisation. However, the total daily exposure to zinc can be higher since several zinc compounds are used in consumer products. Not all of these products are used regularly or at the same time (see above). It is assumed that dandruff shampoo, deodorant, eye shadow, and possibly baby care ointment will be used on a regular basis (more than once a week), resulting in a cumulative uptake of approximately 1.6 mg Zn2+/day. Therefore this

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 361 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 value will also be taken across to the risk characterisation, as this is a more realistic calculation of the daily consumer exposure to zinc.

9.3. Indirect exposure of humans via the environment

The EU risk assessment (ECB 2008) did not assess the risks for Zn(NH4)Clx. Considering that: -the production and use processes of Zn(NH4)Clx are similar to those of ZnCl2, -the production and use pattern of zinc and zinc compounds has not changed significantly since the closure of the RA, and -the RA calculated the exposure through air and water based on the reported emissions data for zinc towards the air and water environment. These emissions have further decreased since the closure of the RA, so the analysis of the RA can be considered as realistic, but conservative for the situation today. The analysis made in the framework of the EU RA for ZnCl2 is taken over in the present analysis as a conservative, realistic assessment of exposure of man through environment for Zn(NH4)Clx.

The related sections out of the zinc chloride RA are taken over below. It should be noted that in this section the zinc cation is discussed, not the salt from which it originates.

General exposure

The most important exposure to zinc for the general population is by the ingestion of foods. Especially meat and meat products, milk and milk products, bread and starchy foods contribute to the dietary zinc intake. The risk assessment summarised that average dietary intake of zinc by adults in nine European countries was 9.1-12.3 mg/day. Only for adult males in Germany and Italy a higher daily dietary intake of 14-15 mg/day was reported. Figures for the Netherlands reporting an average daily intake of zinc of 9.4 mg (minimum of 0.6 mg and a maximum of 39 mg) confirmed this range. A 95-percentile value of 15.4 mg (P5=4.7, P10=5.5, median=9.0, P90=13.8) was calculated. The Dutch intake figures were based on a random group of 6,250 persons. The differences in zinc intake vary due to source and variety of the food.

An epidemiological study has been carried out by Kreis (1992) in which the health effects of cadmium (and zinc) were investigated in a contaminated area in the southern part of the Netherlands (Kempenland). A population sample aged 30-69, with a residence of at least 15 years in a rural village in Kempenland (NL) was compared with a control population of an unpolluted area. About 75% of the inhabitants of both areas consumed at least half of their vegetables from local gardens. The plasma concentration of zinc did not differ between the exposed (n = 299) and the reference population (n = 295) after adjustment for age and gender. The author concluded that, in contrast to cadmium, zinc exposure probably did not differ between the two villages.

For zinc levels in groundwater, the RA mentioned data for the Netherlands. The National Soil Monitoring Network in the Netherlands reported maximum zinc concentrations in upper groundwater of 1.1 mg/l (cattle farms) and 3.1 mg/l (forest locations). Recent zinc concentrations in large surface water in the Netherlands are reported to be all below 0.1 mg/l. Recent atmospheric zinc concentrations in the Netherlands were reported to be below 0.1 µg/m3 (annual averages). Higher concentrations, up to 14 µg/m3, were reported for Belgium (older data). Under normal conditions, drinking water and ambient air are minor sources of zinc intake. Cleven et al. (1993) estimated the intake by drinking water and ambient air to be <0.01 mg/day and 0.0007 mg/day, respectively. The monitoring data above indicate somewhat higher intakes, but it is to be noted that nowadays in the EU upper groundwater and large surface water are not directly representative for drinking water. It was mentioned that in the Netherlands, monitoring of zinc in drinking water was ceased (at water companies) or about to be ceased (at pumpstations) (pers.comm. by RIVM-LWD, 1999). It was concluded that the recent average dietary intake of zinc is around 10 mg/day. This value is taken across to the risk characterisation. Compared to this intake via food, intake via drinking water and ambient air is considered negligible.

Local exposure diammonium tetrachlorozincate, crossreading from zinc chloride

Estimated local zinc concentrations in water and air around industrial facilities In the EU risk assessment, surface water maximum local zinc concentrations (PECadds) of 45.6 µg/l and 3154

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 362 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

µg/l (total zinc) were estimated for the production and processing of zinc chloride, respectively (RA ZnCl2, ECB 2008; see 3.2.1.2). 3 3 Maximum atmospheric zinc concentrations (PECadds) are 0.0525 µg/m and 3.2 µg/m , for production and processing, respectively (RA ZnCl2, ECB 2008; see 3.2.1.2).

The PECadds mentioned above were taken across to the risk characterisation.

9.4. Regional exposure concentrations

The regional (environmental) exposure assessment integrates the environmental exposure following from the use of zinc containing articles by consumers. All exposures to the zinc ion coming from the use of all zinc substances are combined to an “overall” zinc emission and exposure, so no distinction is being made between zinc substances. Consequently, this overall “regional” analysis is relevant for the environmental exposure following from consumer use of all zinc substances. The analysis presented below is based on the extensive regional exposure assessment conducted in the framework of the Zn risk assessment (ECB, 2008). The following updates were made in the present analysis:  Extension of the EU RAR diffuse data set developed for EU-15 towards the EU-27 and recalculation of the regional and continental exposure concentrations

 Update of the monitoring data

The EUSES calculations have been run using the new data and the EUSES 2.0 model. 9.4.1. Modelling approach: diffuse source analysis

9.4.1.1. Overview national emission data For the EU RA, emission data were reported for The Netherlands (1999), Belgium (1995), Sweden (1990-1995), Germany (1998) and UK (1999 and 2000). The diffuse sources analysis for the Netherlands (Table 133) revealed that for the water compartment STP effluents, industry, and traffic are important sources of zinc. For air, industry is the largest contributor. The emissions data reported in the RA zinc were the result of an in-depth discussion and are still considered to be relevant for the situation today, because the use pattern of zinc has not significantly changed since the closure of the RA. However, some updates have been done:

Table 133. Zinc emissions to water, soil and air in the Netherlands (data from the RA from 1999, with some updates) (in t/y). Waste water Surface water Soil Air

Agriculture 4 4 850 5) Industry 63 31 64 Waste treatment 4 - Traffic 140 541) 150 22 Consumers 212 8 4 5 Trade and Services 3) 37 2 Effluents STP - 95 Others 0.4 502) 2384) Atmospheric deposition - 8 90 Total 460 254 1332 91

1) Original CCDM figure of 84 t/y was corrected for new (preliminary) estimates for emissions from ship anodes (7 t/y instead of 23.9 t/y) and anodes on lock gates (14 t/y instead of 27.7 t/y) 2) Including emissions from a.o. overflows and separated (rainwater) sewer.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 363 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

3) Trade and Services (HDO in Dutch) comprises emissions from a.o. car trade, storage firms, educational institutes, medical care, government agencies, recreation and sports and catering industry 4) Emissions from ‘Others’ to soil mainly comprises emissions from composted/re-used or incinerated sewage treatment sludge . 5) Soil is the primary receiving compartment for zinc emissions from agricultural activities. It has to be noted, however, that owing to runoff etc. a significant part of this load will end up in surface water. Recent figures of 2008 showed a net input (corrected for harvest removal) of 850 t/y (CBS, 2008)

Zinc emissions from sacrificial anodes on e.g. lock gates and ships contribute to water emissions under Traffic. Other zinc emissions from traffic, other than corrosion of crash barriers, are related to wearing of tyres and brakes and emissions from fuel and oil. Within the target groups Consumers, Effluent STP and Others a significant part of the zinc emissions is due to corrosion of roofing and gutters of houses etc. Agricultural activities represent the largest source of zinc emission to soil, mainly caused by excretion from animals (manure). For the Netherlands, a total agricultural bruto zinc emission to soil of 2,220 t/a was given for 1999. The estimate was mainly based on total usage of animal feed, its zinc contents and absorption rates of zinc in animals. A large proportion of the feed given to the animals is not absorbed (20-50%). This fraction will pass straight into the manure. Corrected for harvest removal a net input of 1,620 t/y was obtained (ECB 2008). Recent information of the CBS (Netherlands, 2008) revealed that this emission is significantly reduced over the years. For the reference year 2008 a bruto emission to soil of 1,480 t/y and a net input of 850 t/y has been reported. These figures will be used for further calculations. Table 134 provides a comparison of the zinc emissions to the different environmental compartments (air, water and soil) from the identified diffuse sources between the Netherlands, Germany, Belgium, Sweden and the UK. In this overview those emissions are excluded which are not relevant to the circumstances in other countries (e.g. mining).

Table 134. Comparison of total emission rates (tonnes/year) for The Netherlands, Germany, Belgium and Sweden Compartment Netherland (1999) Germany Belgium (1995) Sweden (1990- 1995) Air 91 6,640 440 230 + diffuse emissions Water 254 5,200 527 260 + diffuse emissions Soil 2,720 8,670 Not available 1,2661 Total 3,065 20,510 967 1,786 + diffuse emissions 1) calculated from Table 3.2 in Landner and Lindeström report (1998). Total of 527 g/ha/y and area of 24,000 km2.

As discussed in the RA, it would be too speculative to draw sound conclusions on the differences between these four countries because of the imbalance in the data set, the different assessment methods etc. The available dataset of Belgium was e.g. incomplete and the Swedish dataset was rather dated and less complete compared to that of the Netherlands. The information for Germany seemed to be rather complete, although they were compiled from several sources and the reference period for the water emissions was unclear. Nevertheless data from Sweden and the Netherlands were, roughly taken, in the same order of magnitude (total volume of 3065 t/y (NL) versus 2966 (2301) t/y (S). The German data seemed to support this conclusion, as the size of the country and its number of inhabitants in comparison with the Netherlands was reflected in the total emission data for zinc. Generally, the UK total zinc emission input to soil also seemed to fit with the German data for soil,

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 364 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 regarding the size of both countries (ECB 2008). In the EU RA, the Netherlands was selected as EU-region because the most recent, extended and detailed information was available for this country. The area of the Netherlands also corresponds with the area of a regional system (40,000 km2). Finally, the zinc emissions of the Netherlands were assumed to be representative for an EU regional system, which was generally supported by the above-mentioned comparison with other EU countries. For reasons of consistency, the same approach, i.e. using the NL as regional model, was applied in the present analysis. It must of course be stated that for specific emission sources (e.g. agriculture) rather large differences may occur between EU regions. Given however the high population density of the Netherlands and the concentration of main diffuse sources e.g. agriculture, the NL data can be considered as realistic worst case for the EU.

9.4.1.2. Continental releases and PEC calculations The emissions used for the continental scale (foreign emissions) are defined according to the following TGD default equation:

Continental Emission  10 * Regional Emission  Regional Emission

However, for zinc most continental emissions to air, water and soil are initially not calculated with this equation, because more realistic extrapolation factors are available from other sources. These extrapolation factors are applied to the emission inventory of the Netherlands in order to extrapolate to the EU 27. For industrial emissions (water and air) a factor of 31 is used to extrapolate the NL data to the EU. This extrapolation factor of 31 is based on the ratio NL inhabitants (16 million) versus EU inhabitants (501 million). The assumption is that there is a relationship between the number of inhabitants and the industrial activities. This is an arbitrary choice, but the standard TGD factor of 10 is considered to be too low for zinc. This because it is known that there are a number of EU Member States with (much) higher zinc production and processing activities than the Netherlands. The EU atmospheric emission becomes about 2,711 t/a (Error: Reference source not found 24; Note: this value also includes traffic emissions for which another extrapolation factor is used; see below). This value is lower than the estimate available for Germany (7,190 t/a). The background of this value is unknown however, and, additionally, it is unknown to which period the German data refers. It is further known that the last decade a considerable number of emission reduction measures has been taken by industry. Some support for the current estimate of 1,984 t/y for EU extrapolated air emissions from industry comes from very recent and reliable US data. The TRI database gives a total zinc atmospheric emission figure of 6,500 t/a for the US industry in 1998. The TRI database gives a total US industrial emission value of 800 t/y which is very close to the total EU estimated surface water emission of 961 t/y for industry. For comparison, the EU ePRTR (2008) mentions the following emissions for the total EU industry: to air: 1077T; to water: 3070T/y. For agricultural soil emissions a factor of 10 is used on the recent Dutch estimate of 850 t/y (corrected for harvest removal). This is the TGD default factor, and was considered in the EU RA to reflect the extensive concentration of agriculture in the Netherlands, as compared to other countries. . The EU estimate amounts to 10 * 850 t/y = 8,500 t/y. For consumers, waste treatment, trade and services, STP effluent and others the NL/EU inhabitants correction factor of 31 is used as these sectors are all related to consumption aspects. Emissions from corrosion contribute significantly to the emissions from these target groups. The extrapolation factor of 31 is considered applicable to emissions from corrosion as well. It is noted that due to progressive reduction of atmospheric SO2 emissions in Europe during the last decade, the atmospheric corrosion of zinc will have decreased further over the last decade. Support for the extrapolation for consumers etc. on the basis of the inhabitants ratio is given by

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 365 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

the fact that more or less similar zinc levels are monitored in the communal sewage sludge of a number of EU countries (see section 9.3.2.1.4.). For traffic emissions to soil an extrapolation factor from the Netherlands to EU of 26 is used based on recent road transport statistics (EUROSTAT, 2009). The same factor of 26 is used for traffic emissions to waste water and surface water. However, a considerable part (about 20 tonnes) of the traffic emissions to surface water comes from emissions of zinc anodes. A lower factor (default) of 5 is used for emissions from anodes, as this usage is expected to be relatively high in the Netherlands. Details and results of the calculations for the conversion of NL data into the EU are presented in table below

Table 135. Conversion of the NL emission data to EU. Surface water Soil Air NL emission EU (t/y) NL emission EU (t/y) NL emission EU (t/y) (t/y) and (t/y) and (t/y) and relevant relevant relevant extrapolation extrapolation extrapolation factor factor factor Agriculture 4*10 40 850*10 8,500 Industry 31*31 961 64*31 1,984 Waste treatment - Traffic 34*26 884 150*26 3,900 22*26 572 20*7 140 consumers 8*31 248 4*31 124 5*31 155 Trade and 2*31 62 services Effluents STP 95*31 2,945 Others 50*31 1,550 238*31 7,378

EU total 6,830 19,902 2,711

Agricultural soil 8,500 Industrial soil 11,402

Calculation of PECadd

As mentioned, EUSES 2.0 has been used for calculating the regional PECadd values for each environmental compartment. The input for the regional assessment are the emissions to air, wastewater, surface water and agricultural soil. For modelling the behaviour of zinc in the environment, the octanol-water partition coefficient (Kow) and the aqueous solubility are not appropriate. Measured solids-water partition coefficients for sediment, suspended matter and soil (Kp values) are used instead (Chapter R.16 ECHA, 2010). The modelling for the freshwater compartment have been done using the average Kp SS value of 110,000 l/kg, that was reported for the Netherlands and used throughout the EU RA. The vapour pressure has been fixed at a low value of 1.10-10 Pa and the biotic and abiotic degradation rates have been minimised (Chapter R.16 ECHA, 2010 and Appendix R.7.13.2, ECHA 2008). With EUSES the regional environmental concentrations are directly calculated from the regional and continental emission input.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 366 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

The definition of the continental scale (EU27) in the ECHA guidelines no longer corresponds to the EUSES model characteristics (EU15), since the significant increase in EU surface (4,694x106 km² versus 3.56x106) is standard not taken forward to the EUSES model calculations. This would consequently lead to a significant additional loading of both the continental and the regional scale, since the total manufacturing/import and related emissions for the EU 27 will be applied to the former EU 15 surface. In order to avoid this bias the continental area parameters have been adapted to the EU 27 situation before performing the EUSES calculations (i.e continental area EU 27 cont. + region incl sea = 8.36 E+06 km2 instead of 7.04E+06 km2 for the EU 15) .

The used regional and continental emissions are presented in Table 135. The distribution of the diffuse zinc emissions over the various environmental compartments in Table 133 is based on two additional assumptions: 1) the soil emissions from traffic are all allocated to industrial/urban soil and 2) the soil emissions from consumers and others are allocated to industrial/urban soil except for the emissions from greenhouses. Emissions from greenhouses are added to the agricultural soil (negligible compared to emissions from manure etc.). The sludge application route is not taken into account in this regional assessment, because sewage sludge is not used in several countries and, additionally, it would result in an over conservative agricultural soil scenario in combination with the spread of manure over the soil.

The resulting regional PECadd values (NL-region) are listed in Table 136. A PECadd total of 9.31 μg/l and a PECadd dissolved of 3.51 μg/l (Kp of 110,000 l/kg) is obtained for the aquatic compartment. For the marine aquatic environment a PECadd total of 2.96 μg/l and a PECadd dissolved of 2.88 μg/l (Kp of 6,010 l/kg) is obtained. For sediments a PECadd of 168 mg/kg wwt (Kp of 73,000 l/kg) is obtained.

It is stated that the PECadd values are not corrected for the natural background concentrations in surface water, sediment and soil. In the Zn RAR the influence was mentioned of zinc emissions to agricultural soil on the surface water concentrations by leaching and run-off (CIW, 2003). This aspect can be further investigated quantitatively in the EUSES calculations by varying the various emission input routes (e.g. estimation of PEC water with zinc emissions to agricultural soil set at zero, etc.). The impact of agricultural zinc emissions on the regional PEC water is found to be significant (approximately 60%), which is within the same order of magnitude as the preliminary conclusions of the CIW (2003) report (40%). Emissions to industrial soil (mainly from traffic) have a smaller, but still substantial (app. 20%) impact on the PEC water.

Table 136. Input data and results of the regional exposure assessment (all data refer to NL-region). Input Regional: Amount released to air 91 t/y Aount released to surface water 254 t/y Amount released to agricultural soil 850 t/y Amount released to industrial/urban soil 392 t/y Input Continental: Amount released to air 2,711-91= 2,620 t/y Amount released to surface water 6,830-254 = 6,576 t/y Amount released to agricultural soil 8,500-850= 7,650 t/y Amount released to industrial/urban soil 11,402-392= 11,0102 t/y Results Regional: 3 PECadd air 0.0078 g/m 1) PECadd surface water (total) Kp 110,000 l/kg- 9.31 g/l freshwater 1) PECadd surface water (dissolved) Kp 110,000 3.51 g/l l/kg- freshwater

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 367 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

1) PECadd surface water (total) Kp 6,010 l/kg – 2.96 g/l marine water 1) PECadd surface water (dissolved) Kp 6,010 2.88 g/l l/kg-marine water

PECadd sediment (freshwater) Kp 73,000 159 mg/kgwwt (418 mg/kgdwt) l/kg- freshwater

PECadd sediment (marine) Kp 6,010 l/kg- 7.52 mg/kgwwt (19.8 mg/kgdwt) marine water

PECadd agricultural soil 14.2 mg/kgwwt (16.1 mg/kgdwt)

PECadd natural soil 0.87 mg/kgwwt (0.9 mg/kgdwt)

PECadd industrial/urban soil 38.8 mg/kgwwt (44. mg/kgdwt) 1) This value is calculated with a default suspended matter concentration of 15 mg/l.

9.4.2. Measured regional data in the environment

In this section measured zinc concentrations in various environmental compartments are presented.

Measured regional data aquatic compartment

Monitoring data before 2000 have been summarized and extensively discussed in the Zn RAR (ECB, 2008). In this section the regional analysis is discussed using data from 2000 onwards.

Water

Freshwater data In the framework of the revision of the list of priority substances under the EU Water Framework Directive, recent monitoring data on zinc were reported by the member states to the commission (INERIS and IOW, 2009). For zinc, information was reported by the national water authorities of 20 of 27 member states. The data are from the national water monitoring programmes, and contain substantial influence from local issues e.g. industrial point source monitoring, monitoring of local mining activities, etc. Still, these data are all from the period after the year 2000, so they represent an update on the monitoring data reported in the RA, and are more relevant for the present day water quality in the EU. Analysis of EU-P90 data The dataset on updated monitored data for zinc (total concentrations) in EU waters yields an overall EU-90P zinc concentration of 32 µg Zn/l. This value is based on 5,881 stations in 16 countries, and 118,827 data points (INERIS and IOW 2009). A number of countries also reported dissolved zinc concentrations. Surprisingly, the 90P of the dissolved zinc concentrations is higher than the 90P of the total concentrations. Yet, the dissolved fraction is only part of the total, and thus should be lower, logically. This anomaly can be explained by a number of reasons:  The country-specific analysis (see below) indicates that 85% of the data in the dissolved dataset were reported by only one country (Spain). A basic methodological problem was identified with this Spanish

dataset: the samples were at the spot directly stabilised with HNO 3 to pH 1-2, before sending them out for analysis. As such, zinc was extracted prior to analysis, and the results can not be considered as “dissolved” zinc concentration. The bulk of the “dissolved” dataset is thus not reflecting dissolved concentrations. For this reason, the calculated 90P dissolved concentration (INERIS and IOW 2009) cannot be used6.

 It is well documented that significant contamination of the sample can occur when the sample is filtered in order to separate the dissolved from the total fraction. As a result of this manipulation, dissolved concentrations higher than total have been measured (STOWA 2007). In the zinc data set, there are 192,051 analyses, of which 118,825 are on whole water and 62,082 on dissolved water (of which 47,818 were from Spain). In most cases however, the analyses are conducted on different stations and separate samples. Only 74 stations have 1,525 samples with analyses on dissolved and whole water at the same date, of which 20 stations corresponding to 36 samples have dissolved concentration above whole water concentration. Although this limited comparison shows that contamination of the samples is possible, the paired data are too limited to conclude on the importance of this effect.

6 Due to lack of the original data, a 90P for the remaining data (without the data for Spain) cannot be calculated.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 368 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

 The dissolved and total zinc concentration data were in most cases not monitored at the same stations nor even in the same countries/regions. The datasets on dissolved and on total are thus covering different areas.

The above may explain why the dissolved 90P is higher than the total 90P. Until there is more detailed information on the origin of the data, this remains unsolved. However, the fundamental uncertainty related to the relevancy of the dissolved concentrations for the current exercise remains. In this respect, it should be recognised that elevated zinc background due to local geological conditions can also be a reason for higher monitored values.

In conclusion, there is significant uncertainty regarding the relevancy of the measured dissolved zinc data for assessing the general water quality in the EU. Given that the total data are measured across the EU, that samples for total concentration analysis are not further manipulated, minimising the chance for contamination and other artefacts, and that there is fundamental uncertainty related to the quality and relevancy of the dissolved monitored data, the total measured zinc concentrations are considered more reliable and more relevant for conducting the regional exposure assessment for zinc.

Based on the general “EU-wide” 90P total concentration and recognizing that the PNEC used in this RA is an added value and is expressed on a dissolved basis, the methodology as set out in the EU RA, needs to be applied in order to convert the total concentrations into a bioavailable/dissolved) PEC add: a) Correction for natural background: The PNEC used in the RA is an added value, so has to be added to the background zinc concentration. According to the zinc RA, the natural background must be subtracted from the monitored PEC value. The average zinc background in the EU waters is set at 12 µg/l, with a lower estimate of 3µg/l (RA, EC 2008). So, the PEC add becomes: 32-12 = 20 µg Zn/l (average BG estimation), and 32-3 = 29 µg/l (lower BG estimation). b) Dissolved concentrations: The PNEC is expressed on a dissolved concentration basis. PEC and PNEC must be expressed on the same basis, so, the dissolved fraction of the PEC must be calculated from the total fraction. According to general RA methodology, the dissolved concentration can be calculated from the total, using the partition coefficient between zinc in water and suspended matter: [Me]diss = [Me]total / 1+(Kp*Cs), where -Kp: water-suspended matter partition coefficient for metal X (for zinc: 110000 l/kg (EU RA)) -Cs = suspended matter concentration (TGD default: 15 mg/l) Calculated for zinc: [Zn]diss = [Zn]total / 1 + (110000 l/kg * 0.000015g/l) = [Zn]total / 2.7 The dissolved zinc concentration, to be compared with the PNEC is thus: 20/2.7=7.4 µg/l (average BG correction), and 29/2.7=10.7 µg Zn/l (lower BG correction). c) Inclusion of bioavailability: Finally, a correction for bioavailability of zinc in the natural waters needs to be made (ECB 2008). Lacking evidence on the physicochemical conditions of the waters in the updated monitoring database, several lines of evidence on EU water conditions can be used for setting an overall, representative but at the same time conservative value for zinc bioavailability:  Using 50P values reported for dissolved organic carbon (DOC), pH and hardness in the EU, a typical bioavailability of zinc of 0.44 can be calculated (FOREGS 2006).

 In the RA, physicochemical conditions were reported for several EU waters. They allow to calculate typical bioavailability factors for zinc of 0.4-0.5. More recent data confirm this range (IZA 2010)

 In a detailed UK report (UK EA 2005), a general bioavailability factor of 0.6 was applied.

 The value of 0.6 is applied in the present analysis as an average, but conservative estimation of bioavailability of zinc in the EU waters.

The bioavailable fraction of zinc, to be compared with the PNEC according to the RA, is thus: 7.4 µg/l*0.6= 4.4 µg/l, and 10.7*0.6= 6.4 µg/l, resp. Table below gives a summary of the calculations.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 369 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Table 137. Assessment of the recent monitored data on zinc, reported by 20 of 27 EU member states, according to the methodology as applied in the EU RAR on Zinc (ECB, 2008) Description Zn concentration (µg/l) Measured 90 P [Zn] total 32 (20/27 countries ; “PEC2”)* PEC added 20 (29)** (correction for natural background**) PEC dissolved, calculated from total [Me]diss = [Me]total / 7.4 (10.7) 1+(Kp*Cs) PEC added, bioavailable (Bioavailability correction; 4.4 (6.4) typical case EU waters: 0.6***) *due to significant uncertainties about the quality of the bulk of the dissolved data, only the PEC total is considered to reflect the general water quality across the EU. Since all data need to be considered for making the assessment of water quality, including data below detection limit, the PEC2 (INERIS 2009) is used in the further analysis. **equilibrium partitioning coefficient (2.7) and regional background taken from the RA (ECB 2008). Two values are given: average background according to RA, and (between brackets) the calculation done with the lower estimate of the natural background in EU waters (RA) ***Conservative estimation of the average bioavailability of zinc in EU waters, based on data reported in the EU RA.

Analysis of country data In the same reporting exercise, data on total zinc concentration were reported by 16 member states individually. The country-PEC is, expressed as the 90P of the arithmetic means by sampling station. The PECs are given by country in Table 138. Values in black are obtained from calculating the 90P from > 30 sampling stations, and are considered reliable. Values indicated in black italic are calculated from less than 30 stations and are therefore statistically not reliable (pers. Com. EU Commission, 2010); they are given as supporting evidence, only.

Table 138. Monitored total and added zinc concentrations (µg Zn/l) in EU member states. 90th 90P add 90Padd 90P add percentile dissolved dissolved (all bio- Country Nr of stations Nr of analyses analyses) available Austria 383 13,330 5.94 2.9 1 0.6 Belgium 27 269 30.55 27.5-18.6 10.1-6.9 6.1-4.1 Cyprus 31 84 121.67 118.7-109.7 44.0-40.6 26.3-24.3 Czech republic 312 20,641 38.47 35.5-26.5 13.1-9.8 7.9-5.9 Denmark 16 317 21.41 18.4-9.4 6.8-3.5 4.1-2.1 Estonia 11 142 19.82 16.8-7.8 6.2-2.9 3.7-1.7 France 968 8,630 36.28 33.3-24.3 12.3-9.0 7.4-5.4 Germany 200 8,644 36.22 33.2-24.2 12.3-9.0 7.4-5.4 Greece 165 628 38.36 35.4-26.4 13.1-9.8 7.9-5.9 Lithuania 21 233 35.55 32.6-23.6 12.1-8.7 7.3-5.2 Luxembourg 7 180 31.15 28.2-19.2 10.4-7.1 6.3-4.3 Netherlands 32 799 29.82 26.8-17.8 9.9-6.6 6.0-4.0 Portugal 318 8,021 21.28 18.3-9.3 6.8-3.4 4.1-2.1 Romania 104 768 64.85 61.9-52.9 22.9-19.6 13.7-11.7 Slovakia 27 764 93.71 90.71-81.7 30.3-33.6 18.2-20.2 United Kingdom 3,254 52,022 26.78 23.8-14.8 8.8-5.5 5.3-3.3

Marine waters Recent monitoring data (2005-2009) on dissolved Zn concentrations are available for marine and coastal waters of the Netherlands (NL Ministry Rijkswaterstaat, 2010). Similar data (but less recent 1995-1998) are available for the Belgian part of the North Sea. Netherlands

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 370 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Dissolved Zn concentrations were reported for 8 marine stations, close to the coast or in open sea. Two stations (Walcheren and Schouwen) are located in Zeeland, South of The Netherlands; near the mouth of the Scheldt River. Goeree and Noordwijk are found in Holland (Goeree in the South of Holland close to Zeeland, and Noordwijk is situated North of Den Haag) and Terschelling and Rottumerplaat are two stations located in the “Wadden” sea. Dissolved concentrations are available for the years 2005-2009. The overall percentiles based on the whole dataset as well as station percentiles are presented in table below. Data represent the situation from 2007 to 2009. Given the position of Dutch coastal waters at the mouth of major rivers in EU (Rhine, Meuse, Scheldt,...), this area can be considered as a realistic worst case of coastal water influence in the EU.

Table 139. Monitored dissolved zinc concentrations (µg Zn/l) in coastal waters and open sea of the Netherlands (2007-2009) Station # Percentile Dissolved Zn Dissolved Zn samples concentrations concentrations (µg/l) (µg/l) add* Goeree (2 km off the coast) 34 90P 1.679 0.679 Goeree (6 km off the coast) 34 90P 1.597 0.597 Noordwijk (10 km off the 35 90P 1.606 0.606 coast) Noordwijk (2 km off the 39 90P 1.464 0.464 coast) Rottumerplaat (3 km off the 32 90P 1.628 0.628 coast) Schouwen (10 km off the 34 90P 1.025 0.025 coast) Terschelling (10 km off the 33 90P 1.562 0.562 coast) Walcheren (2 km off the 34 90P 1.195 0.195 coast) Overall 275 50P 0.3 Overall 275 90P 1.58 0.58 *A correction for BG was made on zinc as the proposed marine PNEC is an added value. The default concentration used for background zinc is of 1 µg/L, which is the value proposed for the North Sea in the Zn RA based on a study made by Cleven et al., 1993.

Belgium

Monitoring data for coastal waters were obtained from the North Sea data centre from the Management Unit of the North Sea Mathematical models (MUMM, 2010). They are less recent (going from 1995 to 1998) than the data found on the Dutch Ministry website. Dissolved zinc concentrations reported as 10, 50 and 90P values are

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 371 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

presented in table below.

Table 140. Monitored dissolved zinc concentrations (µg Zn/l) in coastal waters of Belgium (1995-1998) Area # samples Unit 10P 50P 90P 90Padd* Coastal 141 µg/l 0.41 0.98 1.7 0.7 *A correction for BG was made on zinc as the proposed marine PNEC is an added value. The default concentration used for background zinc is of 1 µg/L, which is the value proposed for the North Sea in the Zn RA based on a study made by Cleven et al., 1993.

Sediment Freshwater sediments

Similar to the water compartment data on zinc concentrations in sediments were collected in the framework of the revision of the list of priority substances under the WFD (INERIS and IOW 2009). These data are all from the period after the year 2000, so they represent update on monitoring data for zinc and more relevant than the data in the RA.

Analysis of EU-P90 data The dataset on updated monitored data for zinc in EU freshwater sediments (fraction < 2 mm) yields an overall EU-90P zinc concentration of 270 mg Zn/kg dwt. This value is based on 1,958 stations and 6,312 data points (INERIS and IOW 2009).

Marine sediments Monitoring data for marine sediments are available for the Netherlands (NL Ministry Rijkswaterstaat, 2010) and Belgium (MUMM, 2010). Netherlands Zinc concentrations based on the 63 µm for the reference years 2000, 2003 and 2006 are available from 35 sampling stations (table below).

Table 141. Monitored sediment concentrations (mg Zn/kg dry wt) in the Netherlands (2000, 2003 and 2006) Compartment # samples Unit 50P 90P 90P whole fraction add* Marine sediment 141 mg/kg 140 210 82.8 (fraction < 63 dwt. µm) *conversion to whole sediment fraction and background correction.

Since the PNEC is related to the whole fraction in sediment, a conversion factor was applied to the data based on the 63µm fraction. This factor was obtained from an analysis made by INERIS (2009) of extensive data all over the EU of zinc and cadmium concentrations in the whole (“2mm”) fraction and the 63µm fraction. The 90P for both fractions are compared as follows:

Zinc: 90P 2mm fraction: 270 mg.kg dry wt-1 / 90P 63µm fraction: 399 mg.kg dry wt-1 = 0.68

After conversion to the whole fraction and a background correction (Zn 50P value from CSR = 60 mg Zn/kg dry wt) a 90P value of 82.8 mg Zn/kg dwt. is obtained.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 372 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Belgium

Monitoring data on coastal areas could also be retrieved from the Belgian North Sea Datacentre (http://www.mumm.ac.be/datacentre/index.php), where recent data (2000-2008) can be downloaded using a password. Overall, the North Sea database contains mainly sediment data based on the 63 µm fraction (“fractionated sediments”). The 10,000 µm fraction ("unfractionated sediments") is reported to a much less extent, and as such, is not representative of the coastal area that we consider here for the regional assessment. The monitoring data based on the 63 µm fraction are presented in table below.

Table 142. Monitored sediment concentrations (mg Zn/kg dry wt) in Belgium Compartment # samples Unit 50P 90P 90P whole fraction add* Marine sediment 140 mg/kg 120 190.6 69.6 (fraction < 63 dwt. µm)- coastal area *conversion to whole sediment fraction and background correction.

Since the PNEC is related to the whole fraction in sediment, a conversion factor was applied to the data based on the 63µm fraction. The same factor as applied above for the NL coastal waters, was used: 0.68.

After conversion to the whole fraction and a background correction (Zn 50P value from CSR = 60 mg Zn/kg dry wt) a 90P value of 69.6 mg Zn/kg dwt. is obtained for coastal areas.

Measured regional data Soil

An extensive soil data set is available from the Eurogeosurveys geochemical mapping of agricultural and grazing land soils project (GEMAS) for agricultural (arable) and grassland soils (Reimann et al, 2009). Arable land The GEMAS data contained 2,211 agricultural (arable) soil samples spread over Europe (Table 143).

Table 143. Monitored total zinc concentrations (mg Zn/kg dwt.) in arable soils in Europe.

Country Arable soils 90 p 50 p 10 p # (mg Zn/kg dwt.) (mg Zn/kg dwt.) (mg Zn/kg dwt.) Austria 37 99.3 72.1 42.6 Belgium 14 147.4 53.2 41.4 Bosnia Herzegovina 16 97.0 76.6 49.1 Bulgaria 46 71.4 50.9 36.0 Croatia 31 84.0 65.8 39.5 Cyprius 6 68.4 54.4 36.4 Czech republic 34 114.1 62.9 45.4 Denmark 17 38.3 33.4 16.8

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 373 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Estonia 18 63.5 30.5 23.8 Finland 155 76.0 29.1 11.8 Macedonia 10 132.1 66.2 56.2 France 225 101.9 46.7 19.5 Germany 154 87.0 44.8 19.0 Greece 87 79.4 54.3 29.8 Hungary 41 76.8 45.8 21.3 Ireland 22 84.9 57.9 31.6 Italy 124 90.9 61.7 38.1 Latvia 27 49.6 24.7 17.3 Lithuania 27 36.8 24.8 18.2 Luxembourg 1 35.8 35.8 35.8 Montenegro 6 107.0 83.9 62.8 Netherlands 15 102.2 46.3 19.8 Norway 136 94.3 44.6 21.1 Poland 135 60.7 24.2 10.4 Portugal 38 90.9 43.1 5.1 Switzerland 18 110.3 62.0 46.1 Slovakia 21 75.3 56.4 37.3 Slovenia 10 96.5 69.3 60.9 Spain 213 70.5 32.7 13.1 Serbia 36 84.9 62.6 48.6 Sweden 182 85.7 46.7 21.9 Ukraine 165 50.0 26.9 9.4 United Kingdom 145 111.8 59.4 25.9

Zinc 90 th percentile concentrations for arable soils ranged between 35.8 and 147.4 mg Zn/kg dwt. with a median RWC PEC of 84.9 mg Zn/kg dwt. Grassland The GEMAS data contained 2,119 grassland soil samples spread over Europe (Table 144).

Table 144. Monitored total zinc concentrations (mg Zn/kg dwt.) in arable soils in Europe.

Country Grassland soils 90 p 50 p 10 p # (mg Zn/kg dwt.) (mg Zn/kg dwt.) (mg Zn/kg dwt.) Austria 37 110.8 76.9 52.9 Belgium 14 120.8 71.4 37.7 Bosnia Herzegovina 16 89.4 70.4 55.1 Bulgaria 46 74.8 53.4 35.8 Croatia 31 101.7 61.0 47.8 Cyprius 6 43.8 32.8 22.7 Czech republic 34 111.5 68.5 37.1 Denmark 17 38.6 26.7 13.8 Estonia 18 125.1 29.6 16.5 Finland 43 50.7 22.0 8.6 Macedonia 10 93.8 64.0 35.6

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 374 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

France 230 96.7 48.3 19.7 Germany 153 102.5 51.4 19.8 Greece 87 98.0 54.7 27.7 Hungary 41 79.1 50.1 25.0 Ireland 32 105.3 54.3 20.3 Italy 124 108.3 68.2 36.5 Latvia 27 46.5 23.7 11.1 Lithuania 27 35.5 27.0 17.5 Luxembourg 1 34.4 34.4 34.4 Montenegro 6 139.6 78.3 53.8 Netherlands 15 103.0 43.5 15.6 Norway 135 83.4 39.1 18.5 Poland 135 70.0 24.8 12.3 Portugal 38 95.1 32.2 4.0 Switzerland 18 115.4 85.4 54.6 Slovakia 21 75.9 56.2 35.3 Slovenia 10 110.6 75.3 61.7 Spain 213 70.0 34.1 11.9 Serbia 36 90.9 64.9 32.9 Sweden 187 86.6 43.2 21.6 Ukraine 166 50.0 26.5 8.4 United Kingdom 145 111.8 59.4 25.9

Zinc 90 th percentile concentrations for grassland soils ranged between 34.4 and 139.6 mg Zn/kg dwt. with a median RWC PEC of 93.8 mg Zn/kg dwt.

Measured regional data Sludge and STP effluent

In the RA, a lot of information was available on the quality of sludge from both communal and private (mostly industrial) STPs in the Netherlands (CBS, 1999). Mean zinc concentrations in communal and private STP sludge amounted to, respectively, 865 mg/kg dwt and 143 mg/kg dwt in 1997. Much higher levels were found in the early eighties: 1739 mg/kg dwt in communal STPs and 617 mg/kg dwt in private STPs (1981 data). Error: Reference source not found13 gives the distribution of the sludge in Dutch communal STPs into four zinc concentration classes in 1997 and 1981.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 375 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

>1500 mg/kg n o i

t >1000-1500 a r t mg/kg n 1997 e c

n 1981

o >500-1000 c mg/kg c n i z 0-500 mg/kg

0 20 40 60 80 percentage

Figure 15. Zinc concentrations (classes) in sludge from communal waste water treatment plants in the Netherlands in 1981 and 1997 (after CBS, 1999).

The EU RA mentioned also other data (all referred in the RA report): UK data (1996/7) were available on the quality of sludge used in agriculture: the median zinc content was 559 mg/kg dwt and the 90th percentile-value was 1,076 mg/kg dwt. Median sludge levels in 1982/3 and 1990/1 were, respectively, 1205 and 889 mg/kg dwt, indicating a decrease in zinc levels during the period 1982-1997. The decreasing trend seemed to be similar to the situation in the Netherlands.

The same held for Germany. In the years 1982 and 1983-85 the Zn contents in sewage sludge used for agriculture were 1480 and 1318 mg/kg/dwt, respectively. More recent German data amounted to 863 (1995), 831 (1996) and 809 (1997) mg/kg dwt, pointing to a clear decrease in zinc levels in German sewage sludge from 1982 to 1997.

For Denmark the calculated load of zinc as a result of normal sewage sludge application in 1997, as a worst case situation in 1997 and 2000, were 3040 g/ha, 16,000 g/ha and 12,000 g/ha respectively. All figures were calculated based on the latest sludge directive in Denmark of 1996. In 1997 the weighted mean for zinc in Danish sewage sludge was 760 mg/kg dw for all sludges and 678 mg/kg dwt for sludges used to amend soils. The 90 P values were 1068 and 1069 mg/kg dwt, respectively. In conclusion, the zinc sewage sludge concentration in various EU countries (the Netherlands, Germany, UK and Denmark, see table below) all decreased clearly during the 1980s-1990s and they were found to be at more or less the same levels in the RA. Reduced corrosion run-off rates, due to lower SO2 levels may be one of the explanations for the decreasing trend. The observation of approximately the same (absolute) levels in different countries may point to a more or less similar zinc consumption pattern (at least via the sewage sludge route) in these EU countries.

Table 145. Former and recent zinc sewage sludge concentrations in various EU countries. Sludge concentration (mg/kg dwt) Former data 1997 The Netherlands 1,739 (mean: early eighties) 865 (mean)

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 376 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Germany 1,480 (mean ?; 1982) 809 (mean ?) U.K. 1,205 (median; 1982/3) 559 (median) Denmark - 760 (mean)

For the Netherlands measured effluent concentrations were reported from a large number of (communal) sewage treatment plants in the range of 25-160 µg/l (respectively 5 and 95 percentile; RA zinc).

Measured regional data Air

Zinc concentrations in air reported in the RA are summarised in table below. These data were from the 1980s- 1990s. Given the general decrease in zinc emissions to air observed over the last decade, these data can be considered as conservative estimates for the situation today.

Table 146. Measured zinc concentrations in air (EU RA, for references, see ECB 2008). Location Concentration (g/m3) Source The Netherlands (1995) 0.037-0.054 (annual mean, 4 locations) Monitoring data LML (1995) The Netherlands (1992) 0.038-0.057 (annual mean, 4 locations) Aben et al. (1994) Bilthoven (NL), 1990/1992 0.08 / 0.043 (annual mean) CCRX, 1991/1994 0.160 (98%) Vlaardingen (NL), 1990/1992 0.08 / 0.057 (annual mean) CCRX, 1991/1994 0.210 (98%) Houtakker (NL), 1990/1992 0.07 / 0.054 (annual mean) CCRX, 1991/1994 0.210 (98%) Belgium (1989/1990) 0.03-42.0 / 0.03-1.56 (monthly averages) IDE (B), 1991 (A3) Flanders (B), 1992-1993 0.07-1.02 (mean) Vlaamse Milieumij, 1993 7.75-14.62 (maximum) (A4) The Netherlands 0.065 (calculated annual mean) Cleven et al, 1993 North Limburg (B) (industrial 1-2 (mean) Cleven et al, 1993 area) The Netherlands 1996-1998 0.05 (annual mean 1996) RIVM, 1999 0.04 (annual mean 1997) 0.04 (annual mean 1998) Beerse and Engis (B) 1985/1986 3 (annual mean) Cleven et al, 1993 (industrial area)

9.4.3. Comparison of measured and calculated regional zinc concentrations The risk characterisation should be based on the most realistic exposure information. Hence, it must be decided whether calculated regional concentrations or monitoring data are more useful for the exposure assessment. In this section a comparison is made between the measured concentrations of zinc in the various environmental

compartments (section 9.3.2) and the corresponding calculated PECadd values (section 9.3.1). It must be noted that measured concentrations can only directly be compared with calculated concentrations when the natural background concentration is added to the calculated values (Table 147).

Table 147. Regional concentrations in the environment

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 377 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Predicted (total) regional Measured (total) regional Exposure Concentrations exposure concentrations Source of measured data value Unit 90P value unit EU-90P Ineris 12.3-21.3a Total µg Zn /l 32 Total µg Zn /l Freshwater database (2009) dissolved µg 2.88 1.58 dissolved µg Zn/l NL 2010 Marine water Zn/l EU-90P Ineris 558b mg/kg dwt. 270 mg/kg dwt. Freshwater sediments database (2009) MUMM database 79.8c mg/kg dwt. 190.6-210 mg/kg dwt. Marine sediments (2010) Rijkswaterstaat 210 mg/kg dwt. (2010) PEC (Median of 90P) 64.1d mg/kg dwt. 84.9 mg/kg dwt. Gemas database Agricultural soil (2009) PEC (median of 90P) mg/kg dwt. 93.8 mg/kg dwt. Gemas database Grassland (2009) Netherlands, (1995- 0.0078 µg added/m3 0.04-0.05 µg/m3 1998) Air a natural background of 12 and 3 µg/l is included b natural background of 140 mg/kg dwt is included c natural background of 60 mg/kg dwt (50th percentile) is included d natural background of 48 mg/kg dwt (Foregs database 50th percentile, aqua regia) is included

Water

The calculated regional (NL region) concentrations (PECadd) of zinc in surface water is 9.31 g/l (Csusp. = 15 mg/l) expressed as total Zn (dissolved Zn = 3.51 µg/l). A meaningful comparison of measured and calculated data is possible because a large set of reliable monitoring data of zinc concentrations in surface water is available. Natural background values of 3 and 12 g/l are added to the calculated concentrations. From this comparison it can be concluded that the modelled data underestimate the measured data. For the risk characterisation the measured data will be used.

Sediment Freshwater sediment Also for sediment a meaningful comparison of measured and calculated data is possible. A natural background level of 140 mg/kg dwt can be added to the calculated value (418 mg/kg dwt) yielding a total sediment concentration of 558 mg/kg dwt. The 90th percentile sediment concentration from the recently collected EU data amounts to 270 mg/kg dwt (INERIS & IOW 2009) which is lower than the calculated values, including an added natural background estimate of 140 mg/kg dwt. Marine sediment For marine sediments a natural background value of 60 mg/kg dwt. is added. Measured data (190-210 mg/kg dwt) are higher than the modelled data (79.8 mg/kg dwt.) Both for freshwater sediments and marine sediments preference is given to the measured data and these data have been taken forward to the risk characterisation section.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 378 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Air 3 The calculated regional concentrations (PECadd) of zinc in air are 0.078 g/m . Recent monitoring data of the Netherlands (0.04-0.05g/m3 for 1995-1998) are found to be in the same order of magnitude. Available Belgian monitoring data are higher than the calculated PECadd for air. It is however noted that these data are older and were obtained partially in industrial areas. Soil

The calculated regional concentrations (PECadd NL region) of zinc in agricultural and natural soils are respectively 14.2 mg/kgwwt (16.1 mg/kgdwt) and 0.87 mg/kgwwt (0.9 mg/kgdwt). A comparison of the modelled agricultural data with monitoring data for arable land and grassland is performed by adding a natural background of 48 mg/kg dwt (FOREGS database, 50P, aqua regia). The calculated value from agricultural soil (64.1 mg/kg dry wt.) is slightly lower than the monitoring data for arable soil (84.1 mg/kg dwt) In the risk characterisation both calculated and measured data will be used for the regional scale, but the emphasis will be put on the large number of measured data from various EU regions.

The risk characterisation on the data presented in this section 9.3. is made under section 10.3.

10. RISK CHARACTERISATION

10.1. Local scenarios 10.1.1. Human health

10.1.1.1. Workers

See section 9.1. for the local risk characterisations at the workplace under the different exposure scenarios.

10.1.1.2. Consumers

Conform to the approach followed in the EU risk assessment, no separate assessments of consumer exposure are made for each exposure scenario, but all possible consumer exposures are combined in one integrated scenario, see sections 10.2.1. Human health (combined for all exposure routes), 10.2.1.1. Consumers

10.1.1.3. Indirect exposure of humans via the environment

Conform to the approach followed in the EU risk assessment, no separate assessments of indirect exposure of humans via the environment are made for each exposure scenario, but indirect exposures are combined in one integrated scenario, see sections 10.2.1. Human health (combined for all exposure routes), 10.2.1.2. Indirect exposure of humans via the environment.

10.1.2. Environment

See section 9.1. for the local risk characterisations of the environmental compartments under the different exposure scenarios.

10.2. Overall exposure (combined for all relevant emission/release sources)

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 379 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

10.2.1. Human health (combined for all exposure routes)

10.2.1.1. Consumers

Conform to the approach followed in the EU risk assessment, consumer exposure was assessed by combining the main possible sources of consumer products containing all zinc substances together. Table below (taken from the RA) summarises this combined exposure.

Table 148. Consumer exposure estimates (Table 4.1.3.3 of the RA) internal exposure internal exposure (not (compound specific) compound specific) zinc metal negligible zinc oxide 2.5 mg Zn2+/day (5.1 including medically used zinc oil) ammonium zinc chloride 0.2 mg Zn2+/day zinc sulphate 0.00046 mg Zn2+/day zinc phosphate 0.045 mg Zn2+/day zinc distearate 0.0062 mg Zn2+/day personal care products used 1.6 mg Zn2+/day regularly

Only data on the use of ammonium zinc chloride in gargle are available. For this use, a consumer exposure of 0.2 mg zinc/day was calculated.

Considering these data, the EU risk assessment concluded the following for zinc chloride:

Acute toxicity/Irritation/Corrosivity/Sensitisation Given the data available, the limited use of gargle and the resulting low exposure, it is concluded that zinc chloride is of no concern for consumers with respect to acute toxicity, skin, eye and respiratory tract irritation, corrosivity and skin sensitisation (conclusion ii).

Repeated dose toxicity Starting point for the risk characterisation for systemic effects is the human oral NOAEL of 50 mg zinc/day. Assuming 20% absorption, this NOAEL corresponds to an internal dose of 10 mg zinc/day. The risk ratio between this (internal) NOAEL and the internal exposure by the use of gargle (0.2 mg/day) is 0.02. However, consumer products containing zinc chloride are probably not used regularly. Besides, consumers can also be exposed to other zinc compounds in consumer products, some of which may be used on a regular basis (more than once a week). The use of regularly used products (dandruff shampoo, deodorant, eye shadow, and possibly baby care ointment) results in a cumulative (internal) exposure of approximately 1.6 mg zinc/day (see table 4.1.3.3 in the RA). Comparing the (internal) NOAEL with this more realistic exposure, a risk ratio of 0.16 can be calculated. These risk ratios are considered sufficient and it can be concluded that there is no concern for consumers (conclusion ii), neither for zinc chloride nor for regularly used zinc compounds taken together.

Mutagenicity/Carcinogenicity/Reproductive toxicity Given the results from the mutagenicity studies, it is concluded that zinc chloride is of no concern for consumers with regard to mutagenicity (conclusion ii). As there is no experimental or epidemiological evidence for carcinogenicity, there is no concern for carcinogenicity (conclusion ii). Given the data available, it is concluded that zinc chloride is of no concern for reproductive toxicity (conclusion ii).

Considering the similar hazard profile and similar production/use pattern for ZnCl2 and Zn(NH4)Clx, these conclusions for ZnCl2 are considered also relevant for Zn(NH4)Clx.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 380 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

10.2.1.2. Indirect exposure of humans via the environment.

Repeated dose toxicity

General exposure In the EU risk assessment, the ingestion of foods was considered to be the most important exposure route of zinc for the general population, compared to which the intake by drinking water and ambient air is negligible. Recently, the average dietary intake of zinc is reported to be around 10 mg/day with a minimum of 0.6 mg and a maximum 39 mg. Both the reported average intake and the maximum intake are well below the human oral NOAEL of 50 mg/day and also well below the upper limit of safe intake as recommended by WHO (45 mg/day; 1996). Hence, it can be concluded that there is no concern for the general population exposed indirectly to zinc via the environment.

Local assessment According to the EU RA, the starting point for the risk characterisation for systemic effects are the local PECadds in air and water and the human oral NOAEL of 50 mg zinc/day. Assuming 20% absorption, this NOAEL corresponds to an internal dose of 10 mg zinc/day. The local PECadds in air and water are converted to internal doses by correction for inhalatory and oral absorption (20% and 12%, respectively), and by assuming a breathing volume of 20 m3/day and a drinking water consumption of 2 l/day (see table below. Data for ZnCl2 crossread to Zn(NH4)Clx).

Table 149. Internal exposure levels via water and air at local scale (taken over from Table 4.1.3.4.A of the RA ZnCl2 (ECB 2008))

PECadd-water internal exposure PECadd-air internal exposure (in g/l) (in mg zinc/day) (in g/m3) (in mg zinc/day) production 45.6 0.018 0.0525 0.00042 processing 3154 1.3 3.2 0.026

Considering these data, the EU risk assessment concluded the following for zinc chloride: “Comparing the (internal) NOAEL with the internal exposures, MOSs are in the range 7.7-23810. These MOSs are considered sufficient, and it can be concluded that there is no concern for human health. Moreover, it must be noted that the internal exposures via water are overestimates. They are based on untreated surface water, which nowadays in the EU is not directly representative for drinking water”.

The EU risk assessment also made the following additional statements related to this scenario:

Mutagenicity/Carcinogenicity/Reproductive toxicity

General and Local exposure Given the results from the mutagenicity studies, it is concluded that zinc chloride is of no concern with regard to mutagenicity for the general population exposed indirectly to zinc via the environment (conclusion ii). As there is no experimental or epidemiological evidence for carcinogenicity, there is no concern for carcinogenicity (conclusion ii). Given the data available, it is concluded that zinc chloride is of no concern for reproductive toxicity (conclusion ii).

Considering the similar hazard profile and similar production/use pattern for ZnCl2 and Zn(NH4)Clx, these conclusions for ZnCl2 are considered also relevant for Zn(NH4)Clx.

10.2.2. Environment (combined for all emission sources)

“Regional” PEC add values, integrating the environmental exposure following from the use/consumption of articles containing zinc (from all zinc substances combined) by consumers have been derived by modelling (section 9.4.1.) and based on measured data (section 9.4.2.).

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 381 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

10.2.2.1. Risk characterisation based on modelled exposure In table below, the regional PECs derived by modelling are compared with the PNECs for the respective environmental compartments.

Table 150. Modelled PECadd values and risk characterisation for zinc in the regional analysis. compartment PECadd PNECadd Risk ratio Freshwater (µg 3.51 20.6 0.17 Zn/l) Marine water (µg 2.88 6.1 0.47 Zn/l) Freshwater 418 235.6* 1.8 sediment (mg Zn/kg DW) Marine sediment 19.8 113* 0.18 (mg Zn/kg DW) Agricultural soil 16.1 106.8** 0.15 (mg Zn/kg DW) Natural soil (mg 0.9 106.8 0.008 Zn/kg DW) Urban/industrial 44.0 106.8 0.41 soil (mg Zn/kg DW) * containing a generic bioavailability factor of 2 by default. ** containing a generic bioavailability /ageing factor of 3 by default.

10.2.2.2. Risk characterisation based on measured data

a) Freshwater

The risk characterisation based on the monitored data (EU average 90P and 90P values for EU member states) is summarised in table below:

Table 151. Monitored PEC add values and risk characterisation for the EU freshwater 90P add dissolved PEC/PNEC bio-available Country EU (90P of averages of all stations) 4.4-6.4 0.2-0.3

Austria 0.6 0.03 Belgium 6.1-4.1 0.2-0.3 Cyprus 26.3-24.3 1.2-1.3 Czech republic 7.9-5.9 0.3-0.4 Denmark 4.1-2.1 0.1-0.2 Estonia 3.7-1.7 0.08-0.2 France 7.4-5.4 0.3-0.4 Germany 7.4-5.4 0.3-0.4 Greece 7.9-5.9 0.3-0.4 Lithuania 7.3-5.2 0.3-0.4 Luxembourg 6.3-4.3 0.2-0.3 Netherlands 6.0-4.0 0.2-0.3 Portugal 4.1-2.1 0.1-0.2 Romania 13.7-11.7 0.6-0.7 Slovakia 18.2-20.2 0.9-1.0

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 382 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

United Kingdom 5.3-3.3 0.2-0.3

From the monitored data mentioned above, it is concluded that there is no regional risk for zinc in freshwater. This conclusion is confirmed by the analysis based on modelling. All but one country in the EU show risk ratios below 1; in the one exception (Cyprus) the 90P value is related to the monitoring of a mixed waste dump site which is in the process of being remediated (personal communication A. Kolios). Also in other countries, there is influence from specific local influences, e.g.: the Romanian dataset is influenced by specific geological conditions, and deliberate monitoring of local mines and direct industrial point source monitoring (personal communication C. Toader), and higher zinc levels in the Czeck republic are related to the monitoring of identified industrial point sources and possible influence of local geological conditions (personal communication P. Strzyž).

b) Marine waters

Monitored PECadd values (90P) for coastal waters of 2 EU countries are given in table below. The coastal waters that were monitored are in the surroundings of the mouth of major rivers flowing through major industrial areas and regions with very high density population. They can thus be considered as a realistic worst case for the situation in the EU.

Table 152. Monitored PEC add values and risk characterisation for EU marine waters country PECadd (µg Zn/l) PEC/PNEC The Netherlands 0.58 0.1 Belgium 0.70 0.11

From the monitored data mentioned above, it is concluded that there is no regional risk for zinc in marine water. This conclusion is confirmed by the analysis based on modelling.

c) Sediments: freshwater and marine

Monitored PECadd values (90P) for zinc in sediments (freshwater and marine) are given in table below.

Table 153. Monitored PEC add values and risk characterisation for EU sediment (freshwater and marine) dataset PECtotal (mg PECadd PEC/PNEC** Zn/kg DW) Freshwater: EU (90P of 270 130-210* 0.55-0.89 averages of all stations)

Marine 90P The 82.8 0.7 Netherlands 90P Belgium 69.6 0.6 *two background values were used for calculating the PECadd: 1) the value of 140 mg/kg DW, applied throughout the EU RA, and b) the 50P value for EU sediments (60mg/kg DW), reported by FOREGS (2006) **PNECs integrate default bioavailability factor 0.5

From the monitored data mentioned above, it is concluded that there is no regional risk for zinc in sediment, neither in freshwater, nor in marine waters. For marine waters, this conclusion is confirmed by the analysis based on modelling. For freshwater sediment, the modelled PECadd exceeds the PNECadd. Given the extent (1958 stations) and EU relevancy of the monitored dataset, the measured data are considered to reflect better the environmental reality of EU freshwater sediment, and it is concluded that there is no regional risk.

d) soils

Monitored PECadd values (90P) reported for zinc in agricultural soils are given in table below.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 383 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Table 154. Monitored PECvalues and risk characterisation for EU agricultural soils (arable land and grassland) soil PECtotal PECadd PEC/PNEC (mg/kg DW) Agricultural: 93.8 45.8 0.4 grassland (EU 90P) Agricultural: 84.9 36.9 0.3 arable land (EU 90P)

EU countries: arable soils Austria 110.8 62.8 0.6 Belgium 120.8 72.8 0.7 Bosnia 0.4 Herzegovina 89.4 41.4 Bulgaria 74.8 26.8 0.3 Croatia 101.7 53.7 0.5 Cyprius 43.8 /* / Czech republic 111.5 63.5 0.6 Denmark 38.6 / Estonia 125.1 73.5 0.7 Finland 50.7 2.7 0.02 Macedonia 93.8 45.8 0.4 France 96.7 48.7 0.5 Germany 102.5 54.5 0.5 Greece 98.0 50 0.5 Hungary 79.1 31.1 0.3 Ireland 105.3 57.3 0.5 Italy 108.3 60.3 0.6 Latvia 46.5 / / Lithuania 35.5 / / Luxembourg 34.4 / / Montenegro 139.6 91.6 0.9 Netherlands 103.0 55 0.5 Norway 83.4 35.4 0.3 Poland 70.0 22 0.2 Portugal 95.1 47 0.4 Switzerland 115.4 67 0.6 Slovakia 75.9 28 0.3 Slovenia 110.6 63 0.6 Spain 70.0 22 0.2 Serbia 90.9 43 0.4 Sweden 86.6 39 0.4 Ukraine 50.0 2 0.01 United Kingdom 111.8 64 0.6

EU countries: grassland soils Austria 99.3 51 0.5 Belgium 147.4 99 0.9 Bosnia 0.5 Herzegovina 97.0 49 Bulgaria 71.4 23.4 0.2 Croatia 84.0 36 0.3

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 384 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Cyprus 68.4 20 0.2 Czech republic 114.1 66 0.6 Denmark 38.3 / / Estonia 63.5 16 0.2 Finland 76.0 28 0.3 Macedonia 132.1 84 0.8 France 101.9 54 0.5 Germany 87.0 39 0.4 Greece 79.4 31.4 0.3 Hungary 76.8 29 0.3 Ireland 84.9 37 0.4 Italy 90.9 43 0.4 Latvia 49.6 2 0.02 Lithuania 36.8 / / Luxembourg 35.8 / / Montenegro 107.0 59 0.6 Netherlands 102.2 54.2 0.51 Norway 94.3 46 0.4 Poland 60.7 13 0.1 Portugal 90.9 43 0.4 Switzerland 110.3 62 0.6 Slovakia 75.3 27 0.3 Slovenia 96.5 49 0.5 Spain 70.5 23 0.2 Serbia 84.9 37 0.4 Sweden 85.7 38 0.4 Ukraine 50.0 2 0.02 United Kingdom 111.8 64 0.6 *the PECadd with 50P EU background is <0, so PEC/PNEC cannot be calculated. This is probably due to lower local background than the one considered for this analysis.

From the monitored data mentioned above, it is concluded that there is no regional risk for zinc in agricultural soils (arable land and grassland). This conclusion is confirmed by the analysis based on modelling. An extensive analysis of the possible risk of zinc in agricultural soils in the Netherlands was made in the framework of the EU risk assessment (De Vries et al 2004). The conclusion of this analysis, and of the EU risk assessment, was also that there was no risk for agricultural soils (ECB 2008).

A no risk conclusion can also be drawn for the natural soils and urban/industrial soils, based on the modelled PECadd.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 385 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

REFERENCES

Aamodt RL, Rumble WF, Babcock AK, Foster DM, and Henkin RI (1982). Effects of oral zinc loading on zinc metabolism in humans-I: experimental studies. Metabolism, 31(4):326-334. Testing laboratory: Clinical Center, NIH, Bethesda, Md.

ACGIH (1991). American Conference of Governmental Industrial Hygienists Inc., Documentation of the threshold limit values and biological exposure indices, 6th edition.

Adams K and Kirkpatrick D (1994). Zinc monoglycerolate: mammalian cell mutation assay. EU risk assessment report 2004. Testing laboratory: Huntington Research Centre, Huntington, England. Report no.: KM930593.

Admiraal, W., (1977). Tolerance of Estuarine Benthic Diatoms to High Concentrations of Ammonia, Nitrite Ion, Nitrate Ion and Orthophosphate. Mar. Biol. 43(4), 307-315, (1977).

Aery NC and Jagetiya BL (1997). Relative toxicity of Cadmium, Lead, and zinc on Barley. Commun. Soil Sci. Plant Anal., 28(11&12),949-960 (1997).

Agren M S (1991b). Influence of two vehicles for zinc oxide on zinc absorption through intact skin and wounds. Acta. Derm. Venereol. (Stockh). 71: 153-156.

Agren MS (1990). Percutaneous absorption of zinc from zinc oxide applied topically to intact skin in man. Dermatologica. 180: 36-39.

Agren MS, Krusell M and Franzen L (1991a). Release and absorption of zinc from zinc oxide and zinc sulfate in open wounds. Acta. Derm. Venereol. (Stockh). 71: 330-333.

Ahsanullah M and Arnott GH (1978). Acute toxicity of copper, cadmium, and zinc to larvae of the crab Paragrapsus quadridentatus (H. Milne Edwards), and implications for water quality criteria. Aust J Mar Freshwater Res 29(1):1-8.

Ahsanullah M and Williams AR (1991). Sublethal effects and bioaccumulation of cadmium, chromium, copper, and zinc in the marine amphipod Allorchestes compressa. Mar Biol 108:59-65.

Akhurst LC and Kitching JD (1994). Zinc monoglycerolate: metaphase chromosome analysis of human lymphocytes cultured in vitro. EU risk assessment report for zinc metal 2004. Testing laboratory: Huntington Research Centre, Huntington, England. Report no.: KC930592.

Al-Khafaji and Tabatabai (1979). Effects of trace elements on arylsulfatase activity in soils. Soil Science Vol. 127. No. 3.

Amacher DE and Paillet SC (1980). Induction of trifluorothymidine-resistant mutants by metal ions in L5178y/TK+/-cells. Mutat Res. 78:279-288.

Amdur M O, McCarthy J F and Gill M W (1982). Respiratory response of guinea pigs to zinc oxide fume. Am Ind Hyg Assoc J 43(12):887-889.

Ameille J, Brechot JM, Brochard P, Capron F and Dore MF (1992). Occupational hypersensitivity pneumonitis in a smelter exposed to zinc fumes. Chest. 101, 862-863. [Cited from EU 2004 a, b].

Anderson BS and Hunt JW (1988). Bioassay Methods for Evaluating the Toxicity of Heavy Metals, Biocides and Sewage Effluent Using Microscopic Stages of Giant Kelp Macrocystis pyrifera (Agardh): A Preliminary Report. Mar Environ Res 26(2):113-134.

Anderson BS, Hunt JW, Martin M, Turpen SL and Palmer FH (1988). Marine Bioassay Project. 3rd Report. Protocol Development: Reference Toxicant and Initial Complex Effluent Testing. Div. of Water Qual. Rep. No.88-7WQ, State Water Resour. Control Board, State of California, Sacramento, CA:154 p.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 386 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Arbejdstilsynet (1992). Grænseværdier for stoffer og materialer. Copenhagen, Denmark, Arbejdstilsynet

Arthur, J. W. et al., (1987). Seasonal Toxicity of Ammonia to Five Fish and Nine Invertebrate Species. Bull. Environ. Contam. Toxicol. |38(2),324-331, (1987).

Arts, MHE (1996). Acute (4-hour) inhalation toxicity study with Zinc powder in rats. EU risk assessment zinc metal, 2004. Testing laboratory: TNO, Zeist, The Netherlands. Report no.: TNO-report V96.734.

ATSDR (Agency for Toxic Substances and Disease Registry) (1994). Toxicological profile for zinc (update). U.S. Department of Health and Human Services, Public Health Service, Agency for Toxic Substances and Disease Registry, Atlanta, GA, USA.

ATSDR (Agency for Toxic Substances and Disease Registry) (2005). Toxicological profile for zinc. U.S. Department of Health and Human Services, Public Health Service, Agency for Toxic Substances and Disease Registry, Atlanta, GA, USA.

Attar EN., and Maly EJ. (1982). Acute toxicity of cadmium, zinc and cadmium-zinc mixtures to Daphnia magna. Arch. Environm. Contam. Toxicol. 11,291-296.

Aughey E, Grant L, Furman BL and Dryden WF (1977). The effects of oral zinc supplementation in the mouse. J. Comp. Path. 87: 1-14.

Aulerich RJ, Bursian SJ, Poppenga RH, Braselton WE and Mullaney TP (1991). Toleration of high concentrations of dietary zinc by mink. J. Vet. Diagn. Invest. 3: 232-237.

Azuara-Gracia R, Sarma SS., and Nandini S. (2006). the combined effects of zinc and alga on the life table demography of Anuraeopsis fissa and Brachiones rubens (Rotifera). Journal of Environm. science and health, part A, 41:559-572.

Babcock AK, Henkin RI, Aamodt RL, Foster DM and Berman M (1982). Effects of oral zinc loading on zinc metabolism in humans II: In Vivo Kinetics. Metabolism 31(4): 335-47.

Banu BS, Devi KD, Mahboob M and Jamil K (2001). In vivo genotoxic effect of zinc sulfate in mouse peripheral blood leukocytes using comet assay. Drug and Chem. Tox. 24(1), 63–73. Testing laboratory: Indian Institute of Chemical Technology, Biology and Biotechnology Division, Toxicology Unit, Hyderabad, A. P. India 500 007.

Barata C, Baird D and Markisch S. (1998). Influence of genetic and environmental factors on the tolerance of Daphnia magna Straus to essential and non-essential metals. Aquatic toxicology 42, 115-137.

Barceloux DG (1999). Zinc. J. Toxicol. Clin. Toxicol. 37, 279-292. [Cited from EU 2004 b].

Barnett YA and King CM (1995). An investigation of antioxidant status, DNA repair capacity and mutation as a function of age in humans. Mutat. Res. 338, 115-128. [Cited from EU 2004, a- f].

BASF AG (1968). Abt. Toxikologie, unveroeffentlichte|Untersuchungen, (XVIII/29), 05.04.

BASF AG (1988). Pruefbericht ueber eine Untersuchung der Wirkung einer Substanz (Ammonium chloride) auf die Atmungsaktivitaet von Belebtschlamm im Kurzzeitatmungstest. unpublished data. Testing laboratory: Department of ecology. Owner company: BASF SE. Study number: GLP3: KA3. Report date: 1988-08-22.

Bauchinger M, Schmid E, Einbrodt HJ and Dresp J (1976). Chromosome aberrations in lymphocytes after occupational exposure to lead and cadmium. Mutat. Res. 40:57-62.

Beiras R and Albentosa M (2004). Inhibition of embryo development of the commercial bivalves Ruditapes decussatus and Mytilus galloprovincialis by trace metals; implications for the implementation of seawater quality criteria. Aquaculture 230: 205-213.

Belanger SE, Farris JL, Cherry DS and J Cairns. (1986). Growth of Asiatic clams (Corbicula sp.) during and

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 387 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 after long-term zinc exposure in field-located and laboratory artificial streams. Archives of Environmental Contamination and Toxicology, 15:427-434.

Belanger SE and Cherry DS (1990). Interacting effects of pH acclimation, pH, and heavy metals on acute and chronic toxicity to ceriodphnia dubia (cladocera). Journal of crustacean biology 10(2): 225-235.

Bengtsson, BE (1974). Effect of zinc on growth of the minnow Phoxinus phoxinus. OIKOS 25, 370-373.

Benoit GA. and Holcombe GW. (1978). Toxic effects of zinc on fathead minnows (Pimephales promelas) in soft water. J. Fish. Biol. 13, 701-708.

Bentley PJ and Grubb BR (1991). Experimental dietary hyperzincemia tissue disposition of excess zinc in rabbits. Trace. Elem. Med. 8(4):202-207

Besser et al. (1998). Effect of zeolite on toxicity of ammonia in freshwater sediments: implications for toxicity identification evaluation procedures. Environmental Toxicology and Chemistry, Vol. 17, No. 11, pp. 2310– 2317, 1998. Testing laboratory: U. S. Geological Survey, Biological Resources Division, Environmental and Contaminants Research Center, 4200 New Haven Road, Columbia, Missouri 65201.

Biesinger et al (1986). Effects of metal salt mixtures on Daphnia magna reproduction. Ecotox. Environm. Safety 11,9-14.

Biesinger KE. and Christensen GM. (1972). Effects of various metals on survival, growth, reproduction, and metabolism of daphnia magna. Journal of the Fisheries research board of Canada, Vol 29, nr 12, 1691-1700.

Black MR, Medeiros DM, Brunett E and Welke R (1988). Zinc supplements and serum lipids in young adult white males. Am. J. Clin. Nutr. 47, 970-975.

Blanc P, Wong H, Bernstein MS and Boushey HA (1991). An experimental human model of metal fume fever. Ann. Intern. Med. 114(11):, 930-936.

Blanc PD, Boushey HA, Wong H, Wintermeyer SF and Bernstein MS (1993). Cytokines in metal fume fever. Am.Rev. Respir. Dis. 147(1): 134-138.

Bleavins MR, Aulerich RJ, Hochstein JR, Hornshaw TC and Napolitano AC (1983). Effects of excessive dietary zinc on the intrauterine and postnatal development of Mink. J. Nutr. 113: 2360-2367.

Blinova I, Ivask A Heinlaan M, Mortimer M and Kahru A. (2010). Ecotoxicity of nanoparticles of CuO and ZnO in natural water. Environmental pollution 158, 41-47.

Boawn LC and Rasmussen PE (1971). Crop Response to Excessive Zinc Fertilization of Alkaline Soil. Agronomy Journal., Vol. 63, November-December, 1971.

Bollag J-M and Barabasz W (1979). Effects of heavy metals on the denitrification process in soils. J. Env. Quality, vol 8 no 2 (1979). Testing laboratory: Agric Univ of Cracow (Poland).

Bonham M, O’Connor JM, Alexander HD, Coulter J, Walsh PM, McAnena LB, Downes CS, Hannigan BM and Strain JJ (2003). Zinc supplementation has no effect on circulating levels of peripheral blood leucocytes and lymphocyte subsets in healthy adult men. Br. J. Nutr. 89: 695–703.

Borgmann et al. (1993). Chronic toxicity of ammonia to the amphipod Hyalella azteca: Importance of ammonium ion and water hardness. Environmental Pollution 86 (1994) 329-335 1994 Elsevier Science Limited. Testing laboratory: Department of Fisheries and Oceans, Great Lakes Laboratory for Fisheries and Aquatic Sciences, Burlington, Ontario, Canada, L7R 4A6.

Brandao-Neto J, Vieira JG, Shuhama T, Russo EM, Piesco RV and Curi PR (1990). Interrelationships of zinc with glucose and insulin metabolism in humans. Biol. Trace Elem. Res. 24(1):73-82.

Brinkman S and Woodling J (2005). Zinc toxicity to the mottled sculpin (Cottus bairdi) in high-hardness water. Environm; Toxic; & Chemistry, vol 24, nr 6, 1515-1517.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 388 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Brown RFR, Marrs TC, Rice P and Masek LC (1990). The histopathology of rat lung following exposure to zinc oxide/hexachioroethane smoke or instillation with zinc chloride followed by treatment with 70% oxygen. Environmental Health Perspectives: 85: 81-87.

Buhl K. and Hamilton S. (1990). Comparative toxicity of inorganic contaminants released by placer mining to early life stage salmonids. Ecotoxicology and environmental safety 20, 325-342.

Burton AG., Nguyen LTH, Janssen C., Baudo R., McWilliam R., Bossuyt B., Beltrami M. and Green A (2005). Field validation of sediment zinc toxicity. Env. Tox. and Chem. Vol 24, No 3: 541-553.

Cabrero A, Fernandez S, Mirada F and Garcia J. (1998). Effects of copper and zinc on the activated suldge bacteria growth kinetics. Water Research Vol 32, nr 5, 1355-1362.

Cairns J, Buikema, AL., Heath AG. and Parker BC. (1978). Effects of temperature on aquatic organisms sensitivity to selected chemicals. Virginia water resources research center Bulletin 106. Testing laboratory: dept of Biology and centre for environmental studies Virginia polytechnic Instititute and state University. Report no.: report on OWRT project B-084-VA. Owner company: Virginia state university. Report date: 1978-02-01.

Cairns MA., Garton RR. and Tubb RA. (1982). Use of fish ventilation frequency to estimate chronically safe toxicant concentrations. Trans. Am. Fish. Soc. 111, 70-77.

Cameron TP (1991). Short-term test program sponsored by the division of cancer etiology, NCI. [[Cited from EU 2004, a-f].

Campbell JK and Mills CF (1979). The toxicity of zinc to pregnant sheep. Environmental Research 20: 1-13.

Carlson AR., Nelson H. and Hammermeister D. (1986). Evaluation of site specific criteria for zinc and copper: an integration of metal addition toxicity, effluent and receiving water toxicity and ecological survey data. technical report. Testing laboratory: US environmental protection agency. Report no.: EPA/600/3-86/026. Owner company: US EPA. Report date: 1986-04-01.

Carpenter JM and Ray JH (1969). The effect of zinc chloride on the production of mutations in Drosophila melanogaster. Am. Zool. 9: 1121.

Casas AM and Crecelius EA. 1994. Relationship between acid volatile sulfide and the toxicity of zinc, lead and copper in marine sediments. Environmental Toxicology and Chemistry 13:529-536.

CBS (Netherlands) 2008. http://statline.cbs.nl Tabellen per thema: Zware metalen op landbouwgrond. selectie netto belasting (aanvoer minus afvoer van zware metalen met gewassen) directe link: http://statline.cbs.nl/StatWeb/selection/?VW=P&DM=SLNL&PA=37878&D1=0-6,12,16,19&D2=a&D3= %28l-11%29-l&HDR=T&STB=G1,G2&P=T

CBS, PBL, Wageningen UR (2011). Emissies naar oppervlaktewater en riool, 1990-2009 (indicator 0549, versie 04, 27 oktober 2011). www.compendiumvoordeelomgeving.nl. CBS, Den Haag; Planbureau voor de leefomgeving, Den Haag/Bilthoven en Wageningen, UR, Wageningen.

Cesar A, Marín-Guirao L, Vita R and Marín A (2002). Sensitivity of Mediterranean amphipods and sea urchins to reference toxicants. Ciencias Marinas 28(4): 407–417. Chandra RK (1984). Excessive intake of zinc impairs immune responses. JAMA 252, 1443-1446. Chaney, RL. et al. 1983. Potential effects of waste constituents on the food chain. In: Parr, J.F., P.B. Marsh, and J.M. Kla, eds. Land treatment of hazardous wastes. Park Ridge, NJ: Noyes Data Corp. pp. 152-240.

Chang CH (1976). Modification of DNA, RNA and ATP synthesis in liver and spleen by ZnCl2, 1,10- Phenanthroline and the zinc complex of 1,10-Phenanthroline; Teratogenic effects of these agents in mice. Diss Abstr Int B: 6103- 6104.

Chang C-H, Mann Jr DE and Gautieri RF (1977) Teratogenicity of zinc chloride, 1,10-Phenanthroline, and a zinc-1,10-Phenanthroline complex in mice. J Pharm Sci 66: 1755—1758

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 389 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Chang F-H and Broadbent FE (1981). Influence of trace metals on carbon dioxide evolution from a yolo soil. Soil Science, vol 132 No 6, december 1981. Testing laboratory: Dpt. of Land, Air and Water resources, University of California, Davis. Report date: 1981-03-05.

Chang F-H and Broadbent F E (1982). Influence of trace metals on some soil nitrogen transformations. Journal of Environmental Quality, Vol 11, No1. Testing laboratory: Dpt. of Land, Air and Water resources, University of California, Davis.

Chapman G, Ota S and Recht F. (1980). Effects of water hardness on the toxicity of daphnia magana. study report. Testing laboratory: Corvallis Environmental research laboratory, office of research and develoment, US Environmental protection Agency, Corvallis, Oregon, 97330. Report no.: status report September 1980. Report date: 1980-09-01.

Chapman G., Ota, S and Recht, F. (1980). Effects of water hardness on the toxicity of metals to daphnia magna. Stydy report (september 1980). Testing laboratory: Corvallis Environmental research laboratory, office of research and develoment, US Environmental protection Agency, Corvallis, Oregon, 97330. Report no.: status report -september 1980.

Chapman PM and McPherson C (1993). Comparative zinc and lead toxicity tests with arctic marine invertebrates and implications for toxicant discharges. Polar record 29 (168): 45-54.

Chapman PM, Wang F, Adams W and Green A (1999). Appropriate Applications of Sediment Quality Values for Metals and Metalloids. Env. Sci. And Technology 33, 3937-3941.

Chaudri A., McGrath S., Gibbs P., Chambers B., Carlton-Smith C., Bacon J., Campbell C. and Aitken M. (2008). Population size of indigenous Rhizobium leguminosarum biovar trifolii in long-term field experiments with sewage sludge cake, metal-amended liquid sludge or metal salts: Effects of zinc, copper and cadmium. Soil Biology & Biochemistry 40, 1670-1680.

Chemical and Physiological Inter Relationships in Protein-Metal Interactions Friedman M (ed.). Plenum Press, New York, NY, 299-328. [Cited from EU 2004,a- f].

Chen C and Liao S (2003). Neurotrophic and neurotoxic effects of zinc on neonatal cortical neurons. Neurochem. Int. 42: 471–479.

Chen, J. C., Lin, C. Y., (1992). Effect of ammonia on growth of Penaeus penicillatus juveniles. Comp. Biochem. Physiol. Vol.101C(3),443-447, (1992).

Chobanian SJ (1981). Accidental ingestion of liquid zinc chloride: Local and systemic effects. Ann. Emerg. Med. 10: 91-93.

CIW 2003 Commissie Integraal Waterbeheer, 2003. Lelystad the Netherlands

Clements WH. (2004). Small-scale experiments support causal relationships between metal contamination and macroinvertebrate community responses. Ecol Appl, 14: 954-967.

Cleven RFMJ, Janus JA, Annema JA and Slooff W (1993). Integrated criteria document zinc. RIVM Report No.710401028, Bilthoven, The Netherlands. [Cited from EU 2004, a-f].

Clifford M and McGeer J. C. (2009). Development of a biotic ligand model for the acute toxicity os zinc to Daphnia pulex in soft waters. Aquatic toxicology 91, 232.

Conner MW, Flood WH and Rogers AE (1988). Lung injury in guinea pigs caused by multiple exposures to ultrafine zinc oxide: Changes in pulmonary lavage fluid. Toxicol Environ Health 25: 57-69.

Conrad GW (1988). Heavy metal effects on cellular shape changes, cleavage, and larval development of the marine gastropod mollusk, (Ilyanassa obsoleta say). Bulletin of Environmental Contamination and Toxicology 41: 79-85.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 390 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Conroy PT, Hunt JW and Anderson BS (1996). Validation of a short-term toxicity test endpoint by comparison with longer-term effects on larval Red abalone Haliotis rufescens. Environmental Toxicology and Chemistry 15 (7): 1245–1250.

Cooper NL., Bidwell JR., Kumar A. (2009). Toxicity of copper, lead and zinc mixtures to Ceriodaphnia dubia and Daphnia carinata. Ecotoxicology and Environmental Safety 72, 1523-1528.

Costello LC and Franklin RB.(2006) The clinical relevance of the metabolism of prostate cancer: zinc and tumor suppression: connecting the dots Mol Cancer.15;5:17. Review.

Costello LC and Franklin RB. (2007) Re: Silvano Gallus, Roberto Foschi, Eva Negri et al. Dietary zinc and prostate cancer risk: a case-control study from Italy Eur Urol 52: 1052-7; Eur Urol. 52(4):1262-3; author reply 1263-4.

Courtois Ph, Guillard O, Pouyollon M, Piriou A and Warnet J-M (1978). Comparison of the acute toxicity and the ulcer inducing power of zinc sulphate and pantothenate carried out in animals. Toxicol Eur Res 1: 371-373.

Cousins RJ (1985). Absorption, transport, and hepatic metabolism of copper and zinc, special reference to metallothionein and ceruplasmin. Physiol. Rev. 65, 238-309. [Cited from ATSDR, 1994].

Cousins RJ (1989). Theoretical and practical aspects of zinc uptake and absorption. Adv. Exp. Med. Biol. 249, 3-12[Cited from EU 2004, a-f].

Crane M, Kwok KWH, Wells C, Whitehouse P and Lui GCS. (2007). Use of field data to support European water framework directive quality standards for dissolved metals. Environm. Science and technology 41,5014- 5021.

Crebelli R, Paoletti A, Falcone E, Aquilina G, Fabri G and Carere A (1985). Mutagenicity studies in a tyre plant: in vitro activity of workers' urinary concentrates and raw materials. Br. J. Ind. Med. 42(7):481-487

Cullumbine H (1957). The toxicity of screening smokes. J. Roy. Army. Med. Corps 103, 109-122. [Cited from EU 2004 c].

Cunnane CS (1988). Zinc, clinical and biochemical significance. CRC Press, Boca Raton, Fl, 69-78. [Cited in EU 2004, a-f].

Cutler, D. W, (1933). Some aspects of the physiology of certain nitrate forming bacteria. Ann. appl. Biol, 20, 291-296.

Dalzell DJB, Alte S, Aspichueta E, de la Sota A, Etxebarria J, Guttierrez M, Hoffmann CC, Sales D, Obst U and Christophi N (2002). A comparison of five rapid direct toxicity assessment methods to determine toxicity of pollutants to activated sludge. Chemosphere 47, 535-545.

Dang et al. (1990). Effect of Cd, Ni, Pb and Zn on growth and chemical composition of onion and fenugreek. Commun. in Soil Sci. Plant Anal., 21(9&10), 717-735 (1990).

Davies AG and Sleep JA (1979). Inhibition of carbon fixation as a function of zinc uptake in natural phytoplankton assemblages. J. mar. biol. Ass. U. K. 59: 937-949.

Davis CD, Milne DB and Nielsen FH (2000). Changes in dietary zinc and copper affect zinc- status indicators of postmenopausal women, notably, extracellular superoxide dismutase and amyloid precursor proteins. Am. J. Clin.Nutr. 71, 781-788.

Dawson DA., Stebler EF., Burks SL. and Bantle JA. (1988). Evaluation of the developmental toxicity of metal contaminated sediment using short-term fathead minnow and frog embryo-larval assays. Environm. Tox. & Chem. Vol 7, 27-34.

De Brouwere K. et al (2007). Zinc toxicity on N2O reduction declines with time in laboratory spiked soils and is undetectable in ﬁeld contaminated soils. Soil Biology & Biochemistry 39 (2007) 3167–3176.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 391 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

De Haan et al. (1985). Acceptable levels of heavy metals (Cd, Cr, Cu, Ni, Pb, Zn) in soils, depending on their clay and humus content and cation-exchange capacity. Testing laboratory: Instituut voor bodemvruchtbaarheid haren-gr. Report no.: rapport 9-85.

De Schamphelaere KAC., Heijerick DG and Janssen CR. (2003a). Development and validation of biotic ligand models for predicting chronic tioxicity to fish, daphnids and algae. final report. Testing laboratory: laboratory of environmental toxicology and aquatic ecology, University of Gent, (B). Report no.: ZEH-WA-01. Owner company: International lead and zinc research orgainsation (ILZRO), now: IZA. Report date: 2003-02-28.

De Schamphelaere KAC., Heijerick DG., and Janssen CR. (2003b). Development and validation of biotic ligand models for predicting chronic tioxicity to fish, daphnids and algae. final report. Testing laboratory: laboratory of environmental toxicology and aquatic ecology, University of Gent, (B). Report no.: ZEH-WA-01. Owner company: International lead and zinc research orgainsation (ILZRO), now: IZA. Report date: 2003-02-28.

De Schamphelaere KAC., Lofts S. and Janssen CR (2005). Bioavailability models for predicting acute and chronic toxicity of zinc to algae, Daphnids and fish in natural surface waters. Environm. Toxicol. & Chemistry 24 nr 5, 1190-1197.

Deknudt G (1982). Clastogenic effects of zinc in mammals. CR Soc. Biol. 176, 563-567. [[Cited from EU 2004, a-f].

Deknudt G and Gerber GB (1979). Chromosomal aberrations in bone-marrow cells of mice given a normal or a calcium-deficient diet supplemented with various heavy metals. Mutat. Res. 68: 163-168.

Deknudt Gh, Deminatti M (1978). Chromosome Studies in Human Lymphocytes After In Vitro Exposure to Metal Salts. Toxicology, 10: 67-75.

Delistraty, D. A. et al. (1977). unknown. Proc. Annu. Meeting, WorldMariculture Soc. 8, 647-672, (1977).

Derry JE, McLean WM and Freeman JB (1983). A study of the percutaneous absorption from topically applied zinc oxide ointment. J. Parenter. Enteral. Nutr. 7(2): 131-135.

Deutsche Forschungsgemeinschaft (DFG): Senatskommission zur Prüfung gesundheitsschädlicher Arbeitsstoffe. MAKund BAT-Werte-Liste (1997). Maximale Arbeitsplatzkonzentrationen und biologische Arbeitsstofftoleranzwerte.Weinheim, FRG.

Dimitropoulou P, Nayee S, Liu JF, Demetriou L, Tongeren MV, Hepworth SJ and Muir KR (2008). Dietary zinc intake and brain cancer in adults: A case–control study. British Journal of Nutrition. 99: 667–673.

Dinslage-Schlünz A and Rosmanith J (1976). The zinc elimination from the rat lung after repeated zinc oxide inhalation. Beitr. Silikose-Forsch. (Pneumokon) 28, 80-89. [Cited from EU 2004, a-f].

DiPaoIo JA and Casto BC (1979). Quantitative studies of in vitro morphological transformation of Syrian hamster cells by inorganic metal salts. Canc. Res. 39: 1008-1013.

Doelman P and Haanstra L (1986). Short- and long-term effects of heavy metals on urease activity in soils. Biol Fertil Soils (1989) 2:213-218. Testing laboratory: Research institute for nature management, Arnhem, The Netherlands.

Doelman P. and Haanstra L. (1989). Short- and long-term effects of heavy metals on phosphatase activity in soils: An ecological dose-response model approach. Biol Fertil Soils (1989) 8:235-241.

Domingo J L, Llobet J M, Paternain J L and Corbella J (1988). Acute zinc intoxication: Comparison of the antidotal efficacy of several chelating agents. Vet Hum Toxicol 30(3):224-228.

Dorgelo J., Meester, H. and Van Velzen C. (1995). Effects of diet and heavy metals on growth rate and fertility in the deposit-feeding snail Potamopyrgus jenkisi (Smith) (Gastropoda: Hydrobiidae). Hydrobiologia 316:199- 210.

Dowden, B. F., Bennett, H. J. (1965). Toxicity of selected chemicals to certain animals. JWPCF 37(9), 1308-

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 392 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

1316, (1965).

Dutka BJ, Nyholm N and Petersen J. (1983). Comparison of several microbiological toxicity screening tests. Water research volume 17, nr10, 1363-1368.

Dwyer et al. (2005). Assessing Contaminant Sensitivity of Endangered and Threatened Aquatic Species: Part III. Effluent Toxicity Tests. Arch. Environ. Contam. Toxicol. 48, 174–183 (2005). Testing laboratory: Columbia Environmental Research Center.

E. I. Dupont de Nemours and Co (1992). Acute toxicity chemical/physical properties final results. EPA government. Testing laboratory: Report no.: 8EHQ-1092-11302A. Owner company: E. I. Dupont de Nemours and Co.

European Center for Ecotoxicology and Toxicology of Chemicals (ECETOC) 2003. Aquatic Hazard Assessment II. Technical Report No. 91.

ECHA Guidance on information requirements and chemical safety assessment Chapter R8: Characterisation of dose [concentration]-response for human health, May 2008)

ECHA Guidance on information requirements and chemical safety assessment Chapter R7:endpoint specific guidance, November 2012

ECHA Guidance on information requirements and chemical safety assessment Chapter R10:Characterisation of dose concentration-response for environment , May 2008

Edwards K and Buckley P (1995). Study report zinc monoglycerolate: 13 week feeding study in rats. EU risk assessment report for zinc metal. Testing laboratory: Environmental Safety Laboratory, Unilever Research, Bedford, England. Report no.: FT930588.

EHC (1996). Environmental Health Criteria for zinc (draft). IPCS-WHO, Geneva [Cited from EU 2004, a-f]

Eisler R (1977). Acute toxicities of selected heavy metals to the soft-shell clam, Mya arenaria. Bull. Environ. Contam. Toxicol. 17, 137–145.

Eklund B (2005). Development of a growth inhibition test with the marine and brackish water red alga Ceramium tenuicorne. Marine Pollution Bulletin 50: 921-930.

Elements in Human Health and Disease. Prasad AS (ed.) Vol. 1, Zinc and copper. New York, NY. Academic Press, 345-361. [Cited from EU 2004, a-f].

Elinder CG (1986). Zinc in handbook on the toxicology of metals. Friberg L, Nordberg GF, Vouk VB and Kessler E (eds.), Elsevier, Volume 2, 664-679.

Ellis TM, Masters HG and Mayberry C (1984). Examination of the susceptibility of different breeds of sheep to zinc intoxication. Aust. Vet. J. 61(9):296-298. Testing laboratory: Animal Health Laboratories, Department of Agriculture, Jarrah Road, South Perth, Western Australia 6151.

Enserink E. L., Maas-Diepeveen J. L. and Van Leeuwen C. J. (1991). Combined effects of metals: an ecotoxicological evaluation. Wat. Res. Vol 25, No 6, 679-687.

EPA (1997). Exposure Factors Handbook. Volume 1 – General factors. Update to Exposure Factors Handbook EPA/600/8-89/043 – May 1989. EPA, Office of Research and Development, National Center for Environmental Assessment, U.S. Environmental Protection Agency, Washington DC, EPA/600/P-95/002Fa – August 1997. . [Cited in EU 2004, a-f].

EPA/600/8-89/043 – May 1989. EPA, Office of Research and Development, National Center for Environmental Assessment, U.S. Environmental Protection Agency, Washington DC, EPA/600/P-95/002Fa – August 1997. [Cited in EU 2004,a-f]

EU (2004 a) Zinc metal. Risk assessment report, 2nd priority list, Volume 42, European Commission, Joint

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 393 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Research Center, Institute for Health and Consumer Protection, European Chemicals Bureau, Ispra, Italy

EU (2004 b) Zinc oxide. Risk assessment report, 2nd priority list, Volume 43, European Commission, Joint Research Center, Institute for Health and Consumer Protection, European Chemicals Bureau, Ispra, Italy

EU (2004 c) Zinc chloride. Risk assessment report, 2nd priority list, Volume 45, European Commission, Joint Research Center, Institute for Health and Consumer Protection, European Chemicals Bureau, Ispra, Italy

EU (2004 d) Trizinc bis(orthophosphate). Risk assessment report, 2nd priority list, Volume 47, European Commission, Joint Research Center, Institute for Health and Consumer Protection, European Chemicals Bureau, Ispra, Italy

EU (2004 e) Zinc sulphate. Risk assessment report, 2nd priority list, Volume 46, European Commission, Joint Research Center, Institute for Health and Consumer Protection, European Chemicals Bureau, Ispra, Italy

EU (2004 f) Zinc distearate. Risk assessment report, 2nd priority list, Volume 44, European Commission, Joint Research Center, Institute for Health and Consumer Protection, European Chemicals Bureau, Ispra, Italy

European Commission – Joint Research Centre, Institute for Health and Consumer Protection, European Chemicals Bureau (ECB). 2008. European Union Risk Assessment Report Zinc metal, Volume 42. Final report. (S.J. Munn et al. eds.) 812 pp.

European Commission Joint Research Centre Institute for health and consumer protection http://ecb.jrc.ec.europa.eu/esis/ accessed 2010

Eurostat 2009. Statistics in focus: Inland freight and passenger transport in the EU-27 up to 2007. ISSN 1977- 0316

Evans EH (1945). Casualties following exposure to zinc chloride smoke. Lancet ii, 368-370. [Cited from EU 2004 c].

Farley KJ, Rader K J and Miller BE 2008. Tableau Input Coupled Kinetics Equilibrium Transport (TICKET) model. Environ. Sci. Technol. 42, 838-844.

Farrar JD. and Bridges TS. (2003). Effetcs of zinc on Hyalella azteca, Chironomus tentans and Tubifex tubifex following chronic whole sediment exposures. US Army Engeneer Research and Development Center Vicksburg, Mississipi.

Farris JL, Belanger SE, Cherry DS and. Cairns Jr. J (1989). Cellulolytic activity as a novel approach to assess long-term zinc stress to Corbicula. Wat Res 23: 1275-1283.

Farris JL, Grudzien JL. Belanger SE, Cherry DS and Cairns Jr J. (1994). Molluscan cellulolytic activity responses to zinc exposure in laboratory and field stream comparisons. Hydrobiologia 287: 161-178.

Ferry JJ (1966). Communication to TLV Committte from the General Electric Co., Schenedectady, NY. [Cited from EU 2004 c].

Ferry JJ (1974). Letter to the National Institute for Occupational Safety and Health from the General Electric Co.,Schenedectady, NY. [Cited from EU 2004 c].

Fischer PWF, Giroux A and L’Abbé MR (1984). Effect of zinc supplementation on copper status in adult man. Am.J. Clin. Nutr. 40, 743-746.

Fischer PWF, L'Abbé MR and Giroux A (1990). Effects of age, smoking, drinking, exercise and estrogen use on indices of copper status in healthy adults. Nutr. Res. 10, 1081-1090. [Cited from EU 2004, a-f].

Fisher NS and Frood D (1980). Heavy Metals and Marine Diatoms: Influence of Dissolved Organic Compounds on Toxicity and Selection for Metal Tolerance Among Four Species. Marine Biology 59, 85-93.

Fisher NS, Jones GJ and Nelson DM (1981). Effects of Copper and Zinc on Growth, Morphology, and

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 394 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Metabolism of Asterionella japonica (Cleve). J Exper Marine Biol & Ecol 51/1: 37-56.

Fisher, T., et al (1956). Nitrite Production by Heterotrophic Bacteria. J. gen. Microbiol. 14, 238-247.

Flanagan PR, Haist J and Valberg LS (1983). Zinc absorption, intraluminal zinc and intestinal metallothionein levels in zinc-deficient and zinc-repleted rodents. J. Nutr. 113, 962-972. [Cited from EU 2004, a-f].

Foekema EM, Kramer KJM, Kaag NHBM, Sneekes AC, Bierman S, Hoornsman G, Koelemij E. (2012). Determination of the biological effects and fate of dissolved zinc in outdoor marine mesocosms. IMARES Wageningen UR report C108/12, 5 Oct 2012.

Food and Drug Research Labs., Inc (1973). Teratologic evaluation of FDA 71-49 (zinc sulfate). Report no.: PB- 221 805. [Cited from EU 2004, a-f].

Food and Drug Research Labs., Inc (1974a). Mutagenic evaluation of compound FDA 71-49. Zinc sulfate. Testing laboratory: Litton Bionetics Inc. Report no.: PB-245 451. [Cited from EU 2004, a-f].

Food and Drug Research Labs., Inc (1974b). Teratologic evaluation of compound FDA 71-49. Zinc sulfate in rabbits. Not reported. Testing laboratory: Food and Drug Research Labs., Inc. Report no.: PB-267 191. [Cited from EU 2004, a-f].

FOREGS Geochemical atlas of Europe. 2006.

Franklin RB, Milon B, Feng P and Costello LC.(2005) Zinc and zinc transporters in normal prostate and the pathogenesis of prostate cancer. Front Biosci. 1;10:2230-9. Review.

Franklin RB and Costello LC. (2007) Zinc as an anti-tumor agent in prostate cancer and in other cancers. Arch Biochem Biophys. 15;463(2):211-7. Review.

Frederickson CJ, Suh SW, Silva D, Frederickson CJ, Thompson RB. (2000) Importance of zinc in the central nervous system: the zinc-containing neuron J Nutr. 130(5S Suppl):1471S-83S. Review.

Freijer JI, Cassee FR, Subramaniam R, Asgharian B, Miller FJ, van Bree L and Rombout PJA (1999). Multiple Path Particle Deposition Model (MPPDep version 1.11) – A model for human and rat airway particle deposition. RIVM publication 650010019, Bilthoven, The Netherlands. [Cited from EU 2004, a-f].

Fulcher SM. (2002). DAP 4-WEEK GENERAL TOXICITY AND REPRODUCTIVE/DEVELOPMENTAL TOXICITY SCREENING TEST. Testing laboratory: Huntingdon Life Sciences Ltd, Woolley Road, Alconbury, Huntingdon, Cambridgeshire, PE284HS, ENGLAND. Report no.: 022948. Owner company: The Weinberg Group Inc., 1220 Nineteenth St., N. W., Suite 300, Washington, DC 20036, USA. Study number: WBG 001/022948. Report date: 2002-11-20.

Furchner JE and Richmond CR (1962). Effect of dietary zinc on the absorption of orally administered Zn65. Health Phys. 8:35-40.

Gächter R. (1976). Untersuchungen über die Beëinflussung der planktischen Photosynthese durch anorganische Metallsalze im eutrophen Alpnachersee und der mesotrophen Horwer Bucht. Hydrologie, 38, 97-119.

Gale SA, CK King and RV Hyne (2006). Chronic sublethal sediment toxicity testing using the estuarine amphipod, Melita plumulosa (Zeidler): evaluation using metal-spiked and field-contaminated sediments. Environmental Toxicology and Chemistry 25:1887-1898.

Gallus S, Foschi R, Negri E, Talamini R, Franceschi S, Montella M, Ramazzotti V, Tavani A, Dal Maso L and La Vecchia C (2007). Dietary zinc and prostate cancer risk: A case-control study from Italy. Eur. Urol. 52(4):1052-1057.

Galvez-Morros M, Garcia-Martinez O, Wright AJA and Southon S (1992). Bioavailability in the rat of zinc and iron from the basic salts Zn5(OH)8Cl2. H2O, Fe(OH) SO4 and Fe4(OH)11NO3.2H2O. Food Chem. 43(5):377- 381.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 395 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Gamer AO, Leibold E, and van Ravenzwaay B (2006). The in vitro absorption of microfine zinc oxide and titanium dioxide through porcine skin. Toxicology in Vitro 20 (2006) 301–307. Testing laboratory: BASF Aktiengesellschaft, Department of Experimental Toxicology and Ecology, GVT Z 470, Carl-Bosch-Str. 38, D- 67056 Ludwigshafen/Rhein, Germany.

Genter RB, Cherry DS, Smith EP and J Cairns. (1987). Algal periphyton population and community changes from zinc stress in stream mesocosms. Hydrobiologia, 153:261-275.

Gersich, F. M. and Hopkins, D. L., (1986). Site-specific acute and chronic toxicity of ammonia to. Daphnia magna Straus. Env. Tox. Chem. 5, 443-447 (1986).

Gocke E, King M-T, Eckhardt K and Wild D (1981). Mutagenicity of cosmetics ingredients licensed by the European Communities. Mutat. Res. 90: 91-109.

Goldman et al. (1964). Salicylate intoxication and congenital anomalies. Arch Environ Health 8:648-656.

Gomot-De Vaufleury, A., 2000. Standardized growth toxicity testing (Cu, Zn, Ph, and pentachlorophenol) with Helix aspersa. Ecotoxicology and Environmental Safety 46, 41-50.

Gordon T, Chen LC, Fine JM, Schlesinger RB, Su WY, Kimmel TA and Amdur MO (1992). Pulmonary effects of inhaled zinc oxide in human subjects, guinea pigs, rats, and rabbits. Am Ind Hyg Assoc J 53(8): 503-509.

Gordon EF, Gordon RC and Passal DB (1981). Zinc metabolism: Basic, clinical, and behavioral aspects. J Pediatr 99: 341-349.

Gorski J and Nugegoda D (2006). Sublethal toxicity of trace metals to larvae of the blacklip abalone, Haliotis rubra. Environmental Toxicology and Chemistry 25: 1360-1367.

Greaves MW and Skillen AW (1970). Effects of long-continued ingestion of zinc sulphate in patients with venous leg ulceration. Lancet.II, 889-891.

Groat S, Searl A, Kenny LC, Howe A and Chung K (1999). Preliminary investigation into the size distribution of zinc aerosol in the galvanising, brass casting and zinc oxide production industries. Research project report for ILZRO Program ZEH-4.

Grötsch (1999). Cutaneous permeation of zinc oxide and zinc sulphate through pig skin in vitro. EU risk assessment for zinc oxide 2004. Testing laboratory: Labor L+S AG, Bad Bocklet, Germany. Report no.: 02073979/02073989.

Guidance on information requirements and chemical safety assessment Chapter R8: characterisation of dose [concentration]-response for human health, May (2008)

Gunshin H, Noguchi T and Naito H (1991). Effect of calcium on the zinc uptake by brush-border membrane vesicles isolated form the rat small intestine. Agric. Biol. Chem. 35, 2813-2816. [Cited from EU 2004, a-f].

Gupta T, Talukder G and Sharma A (1990). Cytotoxicity of zinc chloride in mice in vivo. Biol. Trace. Elem. Res. 30(2): 95-101.

Haanstra L and Doelman P (1991). An ecological dose-response model approach to short- and long-term effects of heavy metals on arylsulphatase activity in soil. Biol Fertil Soils (1991) 11:18-23. Testing laboratory: Research institute for nature management, Arnhem, The Netherlands.

Hallbook T and Lanner E (1972). Serum-zinc and healing of venous leg ulcers. Lancet.II, 780-782. [Cited from EU 2004, a-f].

Hallmans G (1977). Treatment of burns with zinc-tape. A study of local absorption of zinc in humans. Scand. J. Plast. Reconstr. Surg. 11(2):155-161.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 396 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Hallmans G and Liden S (1979). Penetration of 65Zn through the Skin of Rats. Acta. Derm. Venereol. 59(2):105- 112.

Hamdi EA (1969). Chronic exposure to zinc of furnace operators in a brass foundry. Brit. J. Ind. Med. 26, 126- 134.

Han T and Choi G-W (2005). A novel marine algal toxicity bioassay based on sporulation inhibition in the green macroalga Ulva pertusa (Chlorophyta). Aquatic Toxicology 75:202–212.

Harmon VL and Langdon CJ (1996). A 7-d toxicity test for marine pollutants using the pacific mysid Mysidopsis intii. 2. Protocol evaluation. Environmental Toxicology and Chemistry, Vol. 15, No. 10, pp. 1824– 1830.

Hart BT, Currey NA, and Jones MJ (1992). Biogeochemsitry and effects of copper, manganese and zinc added to enclosures in Billabong, Magela creek northern Australia. Hydrobiologia 230, 93-134.

He LS, Yan XS and Wu DC (1991). Age-dependent variation of zinc-65 metabolism in LACA mice. Int. J. adiat. Biol. 60(6): 907-16.

Health Canada http://www.hc-sc.gc.ca/accessed 2010

Heijerick DG, De Schamphelaere KAC and Janssen CR (2003). Application of Biotic Ligand Models for Predicting Zn Toxicity in European surface Waters. Report prepared for the International Lead and Zinc Research Organisation (ILZRO). Final report, March 2003.

Heijerick DG, De Schamphelaere KAC. Van Sprang PA. and Janssen CR. (2005). Development of a chronic biotic zinc biotic ligand model for Daphnia magna. ecotoxicology and environmental safety 62, 1-10.

Helling CS., Chesters G and Corey RB (1964). Contribution of organic matter and clay to soil cation exchange capacity as affected by the pH of the saturating solution. Soil Sci. Soc. Am. J. 28, 517-520.

Hemida et al. (1997). Microbial populations and enzyme activity in soil treated with heavy metals. Water, Air, and Soil Pollution 95: 13-22, 1997.

Hempe JM and Cousins RJ (1992). Cysteine-rich intestinal protein and intestinal metallothionein. An inverse relationship as a conceptual model for zinc absorption in rats. J. Nutr. 122, 89-95. [Cited from EU 2004, a-f].

Henkin RI (1974). Metal-albumin, amino acid interactions: Chemical and physiological interrelationships. In:Chemical and Physiological Inter Relationships in Protein-Metal Interactions Friedman M (ed.). Plenum Press, New York, NY, 299-328 [Cited from EU 2004, a-f].

Henkin RI, Mueller CW and Wolf RO (1975). Estimation of zinc concentration of parotid saliva by flamelessatomic absorption spectrophotometry in normal subjects and in patients with idiopathic hypogeusia. J. Lab. Clin.Med. 86, 175-180. [Cited from EU 2004, a-f].

Heydon JL and Kagan AN (1990). Metal fume fever. N. Z. Med. J. 103, 52. [Cited from EU 2004 b].

Hickey, (1999). Chronic Toxicity of Ammonia to the Freshwater Bivalve Sphaerium novaezelandiae. Arch. Environ. Contam. Toxicol. 36, 38–46 (1999). Testing laboratory: National Institute ofWater and Atmosphere Research Ltd, PO Box 11-115, Hamilton, New Zealand.

Hikiba H, Watanabe E, Barrett JC & Tsutsui T (2005). Ability of Fourteen Chemical Agents Used in Dental Practice to Induce Chromosome Aberrations in Syrian Hamster Embryo Cells. J. Pharmacol. Sci. 97(1):146-152.

Hirano S, Higo S, Tsukamoto N, Kobayashi E and Suzuki KT (1989). Pulmonary clearance and toxicity of zinc oxide instilled into the rat lung. Arch. Toxicol. 63(4):336-342.

Hirano T, Murakami M, Fukada T, Nishida K, Yamasaki S, Suzuki T.(2008) Roles of zinc and zinc signaling in immunity: zinc as an intracellular signaling molecule Adv Immunol. 97:149-76. Review.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 397 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Hirohito Hikiba; Eiko Watanabe, J; Carl Barrett and Takeki Tsutsui1, (2005). Ability of fourteen chemical agents used in dental practice to induce chromosome aberrations in Syrian hamster embryo cells. J Pharmacol Sci 97: 146-152.

Hitomi Someya, Yukari Higo, Maki Ohno, Takeo W. Tsutsui, Takeki Tsutsui (2008). Clastogenic activity of seven endodontic medications used in dental practice in human dental pulp cells. Mutation Research 650:39–47.

Hjortsø E, Qvist J, Bud MI, Thomsen JL, Andersen JB, Wiberg-Jørgensen F, Jensen NK, Jones R, Reid LM and Zapol WM (1988). ARDS after accidental inhalation of zinc chloride smoke. Intensive Care Med. 14, 17-24. [Cited from EU 2004, a-f].

Holcombe GW., Benoit DA. and Leonard EN (1979). Long-term effects of zinc exposures on brook trout (salvenilus fontinalis). Transactions of the american fisheries society 108, 76-87.

Homma S, Jones R, Qvist J, Zapol WM and Reid L (1992). Pulmonary vascular lesions in the adult respiratory distress syndrome caused by inhalation of zinc chloride smoke: a morphometric study. Hum. Pathol. 23, 45-50. [Cited from EU 2004, a-f].

Hooftman, RN. and Henzen R (1996). De effekten van zink op de groei van de plantensoort Trifolium pratense. Testing laboratory: TNO Voeding, afdeling milieutoxicologie. Report no.: V96.357.

Hooper PL, Visconti L, Garry PJ and Johnson GE (1980). Zinc lowers high-density lipoprotein-cholesterol levels.JAMA 244, 1960-1961.

Houle RE and Grant WM (1973). Zinc chloride keratopathy and cataracts. Am. J. Ophthalmol. 75, 992-996. [Cited from EU 2004, a-f].

HSE (1998). Health and Safety Executive. Occupational exposure limits 1998. Sudbury, England: HSE Books.

Huber, M. C. et al (1997). Ammonia tolerance of the bivalve Milinia lateralis sublethal toxicity test. Bull. Environ. Contam. Toxicol. 59,292-297 (1997).

Huff et al. (2009). Draft 2009 Update Aquatic Life Ambient Water Quality Criteria For Ammonia – Freshwater. Report no.: EPA-822-D-09-001.

Hunt IF, Murphy NJ, Cleaver AE, Faraji B, Swendseid ME, Coulson AH, Clark VA, Browdy BL, Cabalum MT, & Smith JCJ (1984). Zinc supplementation during pregnancy: effects on selected blood constituents and on program and outcome of pregnancy in low income women of Mexican descent. Am J Clin Nutr, 40: 508–521.

Hunt JR, Lykken GI and Mullen LK (1991). Moderate and high amounts of protein from casein enhance human absorption of zinc from whole wheat or white rolls. Nutr. Res. 11, 413-418. [Cited from EU 2004, a-f].

Hunt JW and Anderson BS (1989). Sublethal effects of zinc and municipal effluents on larvae of the red abalone Haliotis rufescens. Marine Biology 101, 545 552.

Hunt JW, Anderson BS, Turpen SL, Englund MA and Piekarski W (1997). Precision and Sensitivity of a Seven- Day Growth and Survival Toxicity Test Using the West Coast Marine Mysid Crustacean Holmesimysis costata. Environ. Toxicol. Chem. 16(4) (4):824-834.

Hyne RV., Pablo F, Moreno J, Markisch SJ. (2005). Influence of water chemistry on the acute toxicity of copper and zinc to the cladoceran Ceriodaphnia dubia. Environm. Toxic. & Chemistry 24,1667-1675.

Ishidate et al. (1983). Primary mutagenicity screening of food additives currently used in Japan. Food and Chemical Toxicology Volume 22, Issue 8, August 1984, Pages 623-636.

ICRP (1994). Annals of the ICRP (International Commission on Radiological Protection). Human respiratory tract model for radiological protection. ICRP Publication 66. Pergamon/Elsevier Science, UK, USA, Japan. [Cited from EU 2004, a-f].

Ikarashi Y, Tsuchiya T and Nakamura A (1992). Detection of contact sensitivity of metal salts using the murine

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 398 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 local lymph node assay. Toxicol. Lett. 62: 53-61.

INERIS and IOW (International Office of Water) (2009). Implementation of requirements on priority substances within the context of the water framework directive. Prioritisation process: monitored-based ranking. Annex VI.2. report made for the EU Commission, DG ENV., contract nr 07010401/2008/508122/ADA/D2.

IZA (2010). Zinc in European waters: key information of relevance to prioritisation under the water framework directive. International Zinc Association publ. Brussels, Belgium

Janssen, RPT et al. (1997). Equilibrium partitioning of heavy metals in Dutch field soils. I. Relationship between metal partition coefficients and soil characteristics. Environ. Toxicol. Chem. 16, 2470-2478.

Johnson FA and Stonehill RB (1961). Chemical pneumonitis from inhalation of zinc chloride. Dis. Chest. 40: 619-624. Testing laboratory: The Pulmonary Disease Service, Department of Medicine, United States Air Force Hospital, Lackland, United States Air Force Aerospace Medical Center, Lackland Air Force Base, Texas.

Johnson HL, Stauber JL, Adams MS & Jolley DF (2007). Copper and Zinc Tolerance of Two Tropical Microalgae After Copper Acclimation. Environmental Toxicology 22 (3) 234 – 244.

Johnson et al. (1984). Ambient water criteria for ammonia. EPA#: 440/5-85-001. Testing laboratory: n. a. Report no.: n. a. Owner company: n. a.

Jones E and Gant RA (1994). Zinc monoglycerolate: bacterial mutation assay. EU risk assessment report for zinc metal 2004. Testing laboratory: Huntington Research Centre, Huntington, England. Report no.: KA930591.

Juliastuti, S. R.; Baeyens J.; Creemers C. (2003). Inhibition of Nitrification by Heavy Metals and Organic Compounds: The ISO 9509 Test. Environmental Engineering science: 20(2):79-90

Juma NG and Tabatabai MA. (1977). Effects of Trace Elements on Phosphatase Activity in Soils. Soil. Sci. Soc. AM. J., Vol. 41, 1977.

Kada T, Hirano K and Shirasu Y (1980). Screening of environmental chemical mutagens by the Rec-assay system with Bacillus subtilis. Chapter 6, Contribution No. 1209 of the National Institute of Genetics, Misima, Sizuoka 411, Japan.

Källqvist T, Rosseland B, Hytterod S and Kristiansen T (2003). Effect of zinc on the early life stages of brown trout (Salmo trutta) at different levels of water hardness. Report. Testing laboratory: Norwegian Institute for Water research. Report no.: Nr 4687-2003. Owner company: International lead and Zinc research organisation ILZRO (now: IZA). Report date: 2003-05-09.

Kalyanaraman SB. and Sivagurunathan P (1993). Effect of cadmium, copper, and zinc on the growth of blackgram. Journal of plant nutrition, 16(10), 2029-2042.

Kapur SP, Bhussry BR, Rao S and Harmuth-Hoene E (1974). Percutaneous uptake of zinc in rabbit Skin. Proc. Soc. Exp. Biol. Med. 145(3):932-937.

Karbe L (1972). Marine Hydroiden als Test Organismen zur Priifung der Toxizit/it yon Abwasserstoffen. Die Wirkung von Schwermetallen auf Kolonien yon Eirene viridula. Marine Biology 12, 316-328.

Karlsson N, Cassel G, Fangmark I and Bergman F (1986). A comparative study of the acute inhalation toxicity of smoke from TiO2-hexachloroethane and Zn-hexachloroethane pyrotechnic mixtures. Arch. Toxicol. 59(3): 160-166.

Keen CL, Hurley BS and Hurley LS (1977). Zinc absorption through skin: Correction of zinc deficiency in the Rat. Am. J. Clin. Nutr. 30: 528-530.

Kelleher P, Pacheco K and Newman LS (2000). Inorganic dust pneumonias, the metal-related parenchymal disorders. Environ. Health Perspect 108 (suppl), 685-695. [Cited from EU 2004, a-f].

Kessler E (eds.), Elsevier, Volume 2, 664-679. [Cited from EU 2004, a-f].

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 399 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Ketcheson MR, Barron GP and Cox DH. (1969). Relationship of maternal dietary zinc during gestation and lactation to development and zinc, iron and copper content of the postnatal rat. J Nutr. 98(3): 303-311.

Khalil M. A. et al. (1996). Effects of metals and metal mixtures on survival and cocoon production of the earthworm Aporrectodea caliginosa. Pedobiologia 40, 548556.

Khan AT, Atkinson A, Graham TC, Green M, Ali S, Thompson SJ and Shireen KF (2001) Effects of low levels of zinc on reproductive performance of rats. Environ Sci 8: 367—381

Khan AT, Atkinson A, Graham TC, Thompson SJ, Ali S and Shireen KF (2003) Effects of low levels of zinc on reproductive performance of mice. Environ Sci 10: 279—290

Khan AT, Graham TC, Ogden L, Salwa AS, Thompson SJ, Shireen KF, and Mahboob M (2007). A two- generational reproductive toxicity study of zinc in rats. J. Environ. Sci. Health Part B 42: 403–415.

Kimber I and Weisberger C (1989). A murine local lymph node assay for the identification of contact allergens. Arch Toxicol 63: 274-282.

Kimber I, Hilton J and Botham PA (1990). Identification of contact allergens using the murine local lymph node assay: comparisons with the Buehler occluded patch test in guinea pigs. J Appl Toxicol 10: 173-180.

King CK & Riddle MJ (2001). Effects of metal contaminants on the development of the common antarctic sea urchin Sterechinus neumayeri and comparisons of sensitivity with tropical and temperate echinoids. Marine Ecology-Progress Series 215: 143-154.

Klein W and Glaser U (1989). Acute Toxicity of Zinc Phosphate. Study by Fraunhofer-Institut für Umweltchemie und Ökotoxikologie, Schmallenberg, Germany. [Cited in EU risk assessment report for zinc phosphate 2004.

Klimisch H-J, Hildebrand B and Freisberg KO (1982). Acute inhalation toxicity study (LC50, 4 hours, rat) with zinc oxide containing manganese II. EU risk assessment for zinc oxide. Testing laboratory: BASF Aktiengesellschaft, Abteilung Toxikologie, Ludwigshafen.

Kohn et al. (1994). Acute Toxicity of Ammonia to Four Species of Marine Amphipod. Marine Environmental Research 38 (1994) 1-15.

Kolonel LN, Yoshizawa CN and Hankin JH (1988). Diet and prostatic cancer: a case-control study in Hawaii. Am. J. Epidemiol. 127(5): 999-1012.

Konoha K, Sadakane Y and Kawahara M (2006). Zinc neurotoxicity and its role in neurodegenerative diseases. Journal of Health Science, 52 (1) 1-8.

Kossakowski S and Grosicki A (1983). Effect of mercuric chloride upon zinc distribution in the rat. Bull. vet. Inst. Pulawy. 26 (1-4): 67-76. Testing laboratory: Department of Radiobiology, The Veterinary Research Institute, Pulawy.

Kozik MB, Gramza G and Pietrzak M (1981). Neurosecretion of the hypothalamo-hypophyseal system after intragastric administration of zinc oxide. Folia Histochem. Cytochem. 19, 115-122. [Cited from EU 2004, a-f].

Kozik MB, Maziarz L and Godlewski A (1980). Morphological and histochemical changes occurring in the brain of rats fed large doses of zinc oxide. Folia. Histochem. Cytochem. (Krakow) 18(3): 201-206.

Kraak MHS et al (1994). Chronic ecotoxicity of zinc and lead to the Zebra mussel Dreissena polymorpha. Aquatic toxicology 30, 77-89.

Kreis IA (1992). Health effects of cadmium contamination in Kempenland. PhD thesis. Kristal AR, Stanford JL, Cohen JH, Wicklund K and Patterson RE (1999). Vitamin and mineral supplement use is associated with reduced risk of prostate cancer. Cancer Epidemiology, Biomarkers and Prevention. 8: 887– 892.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 400 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Kumar S (1976). Effect of zinc supplementation on rats during pregnancy. Nutrition Reports International. 13(1): 33-36. Testing laboratory: National Institute of Nutrition, Indian Council of Medical Research, Hyderabad, India.

Kuschner WG, D’Alessandro A, Wintermeyer SF, Wong H, Boushey HA and Blanc PD (1995). Pulmonary responses to purified zinc oxide fume. J. Investig. Med .43, 371-378.

Kwok KWH, Leung KMY, LUI GSG, Chu VKH, Lam PKS, Morritt D, Maltby L, Brock TCM, Vanden Brink PJ, Warne MStJ and Crane M. 2007.Comparison of tropical and temperate freshwater animal species’ acute sensitivities to chemiicals: implications for deriving safe extrapolation factors. Integrated Environmental assessment and management Vol 3, nr 1, pp 49-67.

Kynast G and Saling E (1986) Effect of oral zinc application during pregnancy. Gynecol Obstet Invest, 21: 117– 123.

Laane RWPM. (Ed.). (1992). Background concentrations of natural compounds in rivers, sea water, atmosphere and mussels. Tidal Waters Division, DGW report 92.033, The Hague, The Netherlands. 84 pp

Lahermo et al. (1996). Geochemical atlas of Finland. Part 3: Environmental geochemistry – stream waters and sediments. Espoo: Geologian tutkimuskeskus. 1996. 149 pp. ISBN 951-690-678-8 (part 3) 951-690-374-6 (serie).

Lahr J. et al (2008). Combined effects of zinc and earthworm density on soil ecosystem functioning. Soil Biology & Biochemistry 40 (2008) 334–341.

Lam HF, Chen LC, Ainsworth D, Peoples S and Amdur MO (1988). Pulmonary function of guinea pigs exposed to freshly generated ultrafine zinc oxide with and without spike concentrations. Am. Ind. Hyg. Assoc. J. 49(7): 333-341.

Lam HF, Conner MW, Rogers AE, Fitzgerald S and Amdur MO (1985). Functional and morphologic changes in the lungs of guinea pigs exposed to freshly generated ultrafine zinc oxide. Toxicol. Appl. Pharmacol. 78(1):29- 38.

Lam HF, Peisch R and Amdur MO (1982). Changes in lung volumes and diffusing capacity in guinea pigs exposed to a combination of sulfur dioxide and submicron zinc oxide mixed in a humidified furnace. Toxicol Appl Pharmacol 66:427-433.

Landner L. and Lindeström L. (1998). Zinc in society and in the environment. An account of the facts on fluxes, amounts and effects of zinc in Sweden. Swedish Environmental Research Group (MFG). 160 pp.

Langham Brown JJ (1988). Zinc fume fever. Br. J. Radiol. 61, 327-329. [Cited from EU 2004 a, b].

Lansdown ABG (1991). Interspecies variations in response to topical application of selected zinc compounds. Fd Chem Toxic 29 (1): 57-64. Testing laboratory: Charing Cross and Westminster Medical School, Department of Comparative Biology, London, UK.

Laskowski, R. and Hopkin, SP. 1996. Effect of Zn, Cu, Pb, and Cd on fitness in snails (Helix aspersa). Ecotoxicology and Environmental Safety 34, 59-69.

Le Dean L and Devineau J (1987). In search of standardisation: A comparison of toxicity bioassays on two marine crustaceans (Palaemon serratus and Tigriopus brevicornis). Rev. Trav. Inst. Peches marit. 49 (3 et 4):187-198.

Lee CH, Ryu TK, Chang M and Choi JW (2004). Effect of Silver, Cadmium, Chromium, Copper, and Zinc on the Fertilization of the Northern Pacific Asteroid, Asterias amurensis. Bull. Environ. Contam. Toxicol. 73:613– 619.

Lee HH, Prasad AS, Brewer GJ and Owyang C (1989). Zinc absorption in human small intestine. Am. J. Physiol.256, G87-G91. [Cited from EU 2004, a- f].

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 401 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Leitzmann MF, Stampfe MJ, Wu K, Colditz GA, Willett WC and Giovannucci EL (2003). Zinc supplement use and risk of prostate cancer. J. Nat. Can. Inst. 95(13): 1004-1007.

Léonard A, Gerber GB and Léonard F (1986). Mutagenicity, carcinogenicity and teratogenicity of zinc. EU risk assessment report for zinc chloride 2004.

Lewis RJ, ed. (1992). Sax’s dangerous properties of industrial materials. 8th ed. Van Nostrand Reinhold, New York, NY, 3538-3539. [Cited from EU 2004 c].

Liang and Tabatabai (1977). Effects of trace elements on nitrogen mineralisation in soils. Environ. Pollut. (12) (1977).

Liang and Tabatabai (1978). Effects of trace elements on nitrification in soils. J. Environ. Qual., Vol. 7, no. 2, 1978.

Liber K., Call D., Markee TP, Schmude KL, Balcer MD, Whiteman FW and Ankley G (1996). Effects of acid- volatile sulfide on zinc bioavailability and toxicity to benthic macroinvertebrates: a spiked sediment field experiment. Env. Tox. and Chem. Vol. 15 No. 12: 2113-2125.

Lighthart et al. (1983). Microbial Respiration and Chemical Speciation in Metal-Amended Soils. J. Environ. Qual., Vol. 12, no. 4, 1983.

Liipo J, Karhu J, Metsärinta M-L, Virta K, Wiik E, Santala M, Pihlasalo J, Salminen J, Tiihonen J and Harle S. (2010). Characterization of Pure Zinc Compounds – Ammonium Zinc Chloride. Outotec report. Testing laboratory: Outotec Oy, Pori, Finland. Report no.: 10049-ORC-T. Report date: 2010-03-17.

Litton bionetics (1974). EU risk assessment report for zinc sulphate 2004. Testing laboratory: Litton Bionetics. Report no.: FDA 75-14.001314-13-2.

Litton Bionetics (1976). Mutagenic evaluation of compound Zinc oxide USP. EU risk assessment for zinc oxide 2004. Testing laboratory: Litton Bionetics. Report no.: FDA 75-14.001314-13-2.

Llobet JM, Domingo JL, Colomina MT, Mayayo E and Corbella J (1988). Subchronic oral toxicity of zinc in rats. Bull. Environ. Contam. Toxicol. 41(1):36-43.

Lock et al. (2003). Laboratory zinc ecotoxicity testing for soil invertebrates. Testing laboratory: Ghent University Laboratory of Environmental Toxicology and Aquatic Ecology Jozef Plateaustraat 22, B-9000 Gent Belgium. Report no.: ZEH-SO-3.

Lock K. and Janssen C. R. (2001). Modeling zinc toxicity for terrestrial invertebrates. Environmental Toxicology and Chemistr y, Vol. 20, No. 9, pp. 1901–1908, 2001.

Lock K. and Janssen C. R. (2002). Multi-generation toxicity of zinc, cadmium, copper and lead to the potworm Enchytraeus albidus. Environmental Pollution 117: 89–92.

Logue JN, Koontz MD and Hattwick MA (1982). A historical prospective mortality study of workers in copper and zinc refineries. J. Occup. Med. 24(5):398-408.

Long ER, Wolfe DA, Scott KJ, Thursby GB, Stern EA, Peven C, Schwartz T. (1995). Magnitude and Extent of Sediment Toxicity in the Hudson-Raritan Estuary. NOAA Technical Memorandum NOS ORCA 88.

Lorber SA, Gold FM, Maglione AA and Rubenfeld S (1970). 69m Zn-chloride - a new scanning agent, a study of its dosimetry and biological fate. J. Nucl. Med. 11, 699-703. [Cited from EU 2004, a-f].

Lorke D (1983). A new approach to practical acute toxicity testing. Arch. Toxicol. 54: 275-287.

Löser E (1972). Acute toxicity of anorganic pigments. EU risk assessment for zinc oxide 2004. Testing laboratory: Bayer Institut für Toxikologie, Wuppertal-Elberfeld.

Löser E (1977). Acute oral toxicity and skin and eye irritation studies. EU risk assessment for zinc oxide 2004.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 402 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Testing laboratory: Bayer Institut für Toxikologie, Wuppertal-Elberfeld.

Luo Y and Rimmer DL. (1995). Zinc-copper interaction affecting plant growth on a metal-contaminated soil. Environmental Pollution 88 (1995) 79-83.

Macaulay MB and Mant AK (1964). Smoke-bomb poisoning. A fatal case following the inhalation of zinc chloride smoke. J. R. Army Med. Corps 110, 27-32. [Cited from EU 2004 c].

MacDonald DD, Ingersoll CG and Berger TA (2000) Development and Evaluation of Consensus-Based Sediment Quality Guidelines for Freshwater Ecosystems. Arch. Environ. Contam. Toxicol. 39, 20–31

MacFarlane GR. and Burchett MD (2002). Toxicity, growth and accumulation relationships of copper, lead and zinc in the grey mangrove Avicennia marina (Forsk.) Vier. Mar. Env. Res. 54: 65-84.

MacLean AJ. (1974). Effects of soil properties and amendments on the availability of zinc in soils. Can. J. Soil Sci. 54: 369-378 (Nov. 1974).

Magliette RJ. (1995). Need for environmental quality guidelines based on ambient freshwater quality criteria in natural waters -case study "zinc". Bull. Environm. Contam. Toxicol. 54, 626-632. Testing laboratory: Merck Research laboratories, P. O. Box 2000, Rahway, New Jersey 07065, USA.

Mahomed K, James DK, Golding J and McCabe R (1989). Zinc supplementation during pregnancy: A double blind randomised controlled trial. Br. Med. J. 299: 826-30.

Maita K, Hirano M, Mitsumori K, Takashi K and Shirasu Y (1981). Subacute toxicity studies with zinc sulfate in mice and rats. J. Pesticide Sci. 6:327-336.

MAK-Maximale Arbeitsplatzkonzentrationen (2009). Draft Zink_090716.doc

Maliszewska W, Dec S, Wierzbicka H and Wozniakowska A (1985). The influence of various heavy metal compounds on the development and activity of soil micro-organisms. Environmental Pollution (series A) 37 (1985) 195-215. Testing laboratory: Dpt. of Microbiology, Institute of soil science and plant cultivation, Pulaway, Poland.

Malo JL, Malo J, Cartier A and Dolovich J (1990). Acute lung reaction due to zinc inhalation. Eur. Respir. J. 3(1) :111-114

Malten KE and Kuiper JP (1974). Allergie cutanée de contact dans 100 cas d’ulcères varieux. Phlébologie 27, 417-420. [Cited from EU 2004, a-f].

Marquart H, Smid T, Heederik D and Visschers M (1989). Lung function of welders of zinc-coated mild steel: cross-sectional analysis and changes over five consecutive work shifts. Am. J. Ind. Med. 16(3): 289-96.

Marshall JS, Parker JI, Mellinger DL and C Lei. (1983). Bioaccumulation and effects of cadmium and zinc in a Lake Michigan plankton community. Can. J. Fish Aquatic Science, 40:1469-1479.

Martin M, Hunt JW, Anderson BS, Turpen SL & Palmer FH (1989). Experimental Evaluation of the Mysid Holmesimysis costata as a Test Organism for Effluent Toxicity Testing. Environ Toxicol Chem. 8(11):1003- 1012.

Marzin DR and Phi HV (1985). Study of the mutagenicity of metal derivatives with Salmonella typhimurium TA102. Mutat. Res. 155: 49-51.

Masters JA., Lewis MA., Davidson DH. and Bruce RD. (1991). Validation of a four-day Ceriodaphnia test and statistical considerations in data analysis. Environ. Toxicol. Chem. 10, 47-55.

Matarese SL and Matthews JI (1986). Zinc chloride (smoke bomb) inhalational lung injury. Chest 89, 308-309. [Cited from EU 2004 c].

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 403 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Mayes M. A. et al (1986). Acute and chronic toxicity of ammonia to freshwater fish: A site-specific study. Env. Tox. Chem. 5, 437-442 (1986).

McCormick, J. H., (1984). Toxicity of ammonia to early life stages of the green sunfish Lepomis cyanellus. Env. Pollut. (Series A), 36, 147-163 (1984).

McGeer J, Brix KV, Skeaff JM, De Forest DK, Bingham SI, Adams WJ and Green A (2003). Inverse relationship between bioconcentration factors and exposure concentration of metals: implications for hazard assessment of metals in the aquatic environment. Env. Tox. & Chem. 22, nr 5, 1017-1037.

McKinney PE, Brent J and Kulig K (1994). Acute zinc chloride ingestion in a child: local and systemic effects. Ann. Emerg. Med. 23(6):1383-1387. Testing laboratory: Rocky Mountain Poison and Drug Center, Denver General Hospital, University of Colorado Health Sciences Center, Colorado.

McKinney PE, Brent J and Kulig K (1995). Zinc chloride ingestion in a child: exocrine pancreatic insufficiency. Ann. Emerg. Med. 25(4):562.

Ménache MG, Miller FJ and Raabe OG (1995). Particle inhalability curves for humans and small laboratory animals. Ann. Occup. Hyg. 39, 317-328. [Cited from EU 2004, a-f].

Merckx R, Brans K., Smolders E. (2001). Decomposition of dissolved organic carbon after soil drying and rewetting as an indicator of metal toxicity in soils. Soil Biology & Biochemistry 33, 235-240.

Microbiological and Biochemical Assay Laboratories (1985). ReportNo. J-0833. Testing laboratory: Microbiological and Biochemical Assay Laboratories. Report no.: J-0833.

Miller, D. C. et al. (1990). The acute and chronic toxicity of ammonia to marine fish and mysid. Arch. Environ. Contam. Toxicol. 19, 40-48, (1990).

Milliken JA, Waugh D and Kadish ME (1963). Acute interstitial pulmonary fibrosis caused by a smoke bomb. Can. Med. Ass. J. 88, 36-39. [Cited from EU 2004 c].

Milne DB, Davis CD and Nielsen FH (2001). Low dietary zinc alters indices of copper function and status in postmenopausal women. Nutr. 17, 701-708.

Mirbeau T, Guillaumat PPO and Pelcot C (1999). Acute eye irritation in rabbits (phosphate de zinc PZ20). EU risk assessment for zinc phosphate. Testing laboratory: Centre International de Toxicologie. Report no.: 17755 TAL.

Moore R (1978). Bleeding gastric erosion after oral zinc sulphate. Br. Med. J. I: 754. Testing laboratory: Department of Medicine, Addenbrooke's Hospital, Cambridge.

Morgan SL, Strong DC, Kitchin EM, Johnston KE, Disilvestro RA and Tamura T (2005). Zinc status of women with low bone mineral density who receive calcium supplements. Biol Trace Elem Res. 110(3): 211-22.

Mount DI and Norberg TJ (1984). A seven-day life cycle cladoceran toxicity test. Environm. Toxicol. & Chemistry Vol 3, 425-434.

Mueller (1995). The dynamics of ammonia in the 10-day sediment test using the amphipod, Ampelisca abdita. Second SETAC World Congress, p. 265 Pensacola, FL, USA: SETAC.

Mueller EJ and Seger DL (1985). Metal fume fever - a review. J. Emerg. Med. 2, 271-274. [Cited from EU 2004 b].

Mukherjee MD, Sandstead HH, Ratnaparkhi MV, Johnson LK, Milne DB and Stelling HP (1984). Maternal zinc, iron, folic acid, and protein nutriture and outcome of human pregnancy. Am. J. Clin. Nutr. 40(3):496-507.

MUMM (2010). Belgian North Sea datacentre. 2010. Direct link: http://www.mumm.ac.be/datacentre/index.php

Münzinger A and Monicelli F (1991). A comparison of the sensitivity of three Dapnia magna populations under

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 404 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 chronic heavy metal stress. Ecotoxicology and Environmental safety 22,24-31.

Murphy JV (1970). Intoxication following ingestion of elemental zinc. JAMA 212, 2119-2120. [Cited from EU 2004 a].

Mutch Associates (2010a). Validation of time-variable solutions in the unit world model for metals in lakes: pahse 1. Mutch Associates, Ramsey, New Jersey, US. Report nr IZAS.001., February 2010.

Mutch Associates (2010b). Assessing zinc removal from surface waters using the unit world model for metals in lakes and the EUSES model lake parameters. Mutch Associates, Ramsey, New Jersey, US. Report nr IZAS.002., March 2010.

Muysen BTA., Bossuyt BTA. and Janssen CR (2005). Inter- and intra-species variation in acute zinc tolerance of field-collected cladoceran populations. Chemosphere 61, 1159-1167.

Muyssen BTA and Janssen CR. (2002). Tolerance and acclimation to zinc of Ceriodaphnia dubia. Environmental pollution 117, 301-306

Muyssen B., Bossuyt B. and Janssen C. R. (2003a). ecotoxicity of zinc to algae and daphnids tested in natural soft waters. final report. Testing laboratory: laboratory of environmental toxicology and aquatic ecology, Gent University (B). Report no.: final report. Owner company: international lead and zinc research organisation (ILZRO), now: IZA. Report date: 2003-04-01.

Muyssen B., Bossuyt B. and Janssen C. R. (2003b). ecotoxicity of zinc to algae and daphnids tested in natural soft surface waters. study report. Testing laboratory: laboratory of environmental toxicology and aquatic ecology, Gent University (B). Report no.: final report. Owner company: International lead and zinc research organisation (ILZRO), now: IZA. Report date: 2003-04-01.

National Board of Occupational Safety and Health (1993). Occupational exposure limit values. Solna, Sweden.

Necker and Kunze (1986). Stickstoffmineralisation durch Pilze und Bakterien in schwermetallbelastetem Boden in Brutversuch. Angew. Botanik 60, 81-93.

Neuberger JS and Hollowell JG (1982). Lung cancer excess in an abandoned lead-zinc mining and smelting area. Thee Science of the Total Environment 25:287-294.

Nève J, Hanocq M, Peretz A, Abi Khalil F, Pelen F, Famaey JP and Fontaine J (1991). Pharmacokinetic study of orally administered zinc in humans: evidence for an enteral recirculation. Eur. J. Drug Metab. Pharmacokinet. 16(4): 315-23.

Nguyen LTH, Roman YE, Vandegehuchte M and Janssen CR (2005a). Ecotoxicity of zinc to Lumbriculus variegatus and Gammarus pulex tested in natural freshwater sediment. Final report submitted to International Lead Zinc Research Organisation (August 2005).

Nguyen LTH, Roman YE, Vandegehuchte M and Janssen CR (2005b). Ecotoxicity of zinc to the amphipod Hyalella azteca tested in natural freshwater sediment. Final report submitted to International Lead Zinc Research Organisation (July 2005).

Nguyen LTH, Roman YE, Vandegehuchte M and Janssen CR (2005c). Ecotoxicity of zinc to the mayfly Ephoron virgo tested in natural freshwater sediment. Final report submitted to International Lead Zinc Research Organisation (August 2005).

Niederlehner BR, Cairns J. (1993). Effects of previous zinc exposure on pH tolerance of periphyton communities. Environ Toxicol Chem, 12: 743-753.

NIOSH (1975). Criteria for a recommended standard. Occupational exposure to zinc oxide. US Department of Health, Education and Welfare, Public Health Service, Center for Disease Control, National Institute for Occupational Safety and Health, Washington

NL Ministry verkeer en Waterstaat. 2010. Direct link: http://www.rijkswaterstaat.nl/water/scheepvaartberichten_waterdata/historische_waterdata/waterbase/index.aspx

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 405 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 n. n. (2003). SIDS Initial Assessment Report For SIAM 17 - Ammonium chloride. SIDS Initial Assessment Report For SIAM 17.

Norberg TJ. and Mount DI. (1985). A new fathead minnow (Pimephales promelas) subchronic toxicity test. Environm. Tox & Chem. Vol 4, 711-718.

Norberg-King T. (1989). An evaluation of the fathead minnow seven-day subchronic test for estimating chronic toxicity. Environm. Tox. & Chem. vol 8, 1075-1089.

Norwood WPT. Watson-Leun and Milani D (2009). Impacts of Zn-Spiked Sediments on Four Invertebrates: Implications for the Canadian Sediment Quality Guideline. Environment Canada, 2009.

Notenboom and Posthuma (1994). Validatie toxiciteitsgegevens en risicogrnzen bodem: voortgangsrapportage 1993. Testing laboratory: Rijksinstituut voor Volksgezondheid en Milieuhygiëne, Laboratorium voor Ecotoxicologie. Report no.: RIVM-719102029.

NOTOX B.V., ‘s-Hertogenbosch, The Netherlands. Guinea Pig (Maximisation-Test). Project 263429. [Cited from EU 2004 b ].

NOTOX B.V., ‘s-Hertogenbosch, The Netherlands. Albino Guinea Pig (Maximisation-Test). Project 254328. [Cited from EU 2004 e ].

Novelli AA, Losso C, Ghetti PF and Volpi Ghirardini A (2003). Toxicity of heavy metals using sperm cell and embryo toxicity bioassays with Paracentrotus lividus (Echinodermata: Echinoidea): comparisons with exposure concentrations in the lagoon of Venice, Italy. Environmental Toxicology and Chemistry, Vol. 22, No. 6, pp. 1295–1301.

Oberdörster G, Hochrainer D and Ma RH (1980). Zinc oxide aerosols: generation, lung clearance and effects on lung clearance. J Aerosol Sci Med Fed Biomed Influence Aerosol Conf 7th: 132-137.

Occupational Safety and Health Administration, OSHA (1989). U.S. Department of Labor.

Occupational Safety and Health, Washington. [Cited from EU 2004 b].

OECD OECD TG 209. Report date: 2010-07-20

Ohya et al. (1985). Zinc effects on soil microflora and glucose metabolites in soil amended with 14C-glucose. Biol. Fert. Soils 1: 117-122.

Owojori OJ., Reinecke AJ. and Rozanov AB. (2008). Effects of salinity on partitioning, uptake and toxicity of zinc in the earthworm Eisenia fetida. Soil Biology & Biochemistry 40 (2008) 2385–2393.

Pack S. (1993). A review of statistical data analysis and experimental design in OECD aquatic toxicology test guidelines. Shell Research Ltd., Sittingbourne Research. Centre, Sittingbourne, Kent, U.K. 42 pp

Painter, H. A., (1970). A review of literature on inorganic nitrogen metabolism in microorganisms. Water Reseach Pergamon Press, 4, 393-450

Pal N and Pal B (1987). Zinc feeding and conception in the rats. Internat. J. Vit. Nutr. Res. 57: 437-440. Testing laboratory: Department of Biochemistry, School of Tropical Medicine, Calcutta, India.

Palauskis JD. and Winner RW. (1988). Effects of water hardness and humic acid on zinc toxicity to Daphnia magna Straus. Aquatic Toxicologhy 12,273-290.

Pare CMB and Sandler M (1954). Smoke-bomb pneumonitis: description of a case. J. R. Med. Corps 100, 320- 322. [Cited from EU 2004 c].

Pascoe GA, Blanchet RJ and Linder G (1996). Food chain analysis of exposures and risks to wildlife at a metal contaminated wetland. Arch. Environ. Contam. and Toxicol. 30,306-318.

Paulsson M, Nyström B and Blanck H (2000). Long-term toxicity of zinc to bacteria and algae in periphyton communities from the river Göta älv, based on a microcosm study. Aquatic toxicology; 47:243-257.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 406 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Pavicic J, Skreblin M, Kregar I, Tusekznidaric M & Stegnar P (1994). Embryolarval tolerance of Mytilus galloprovincialis, exposed to the elevated sea-water metal concentrations.1. Toxic effects of Cd, Zn and Hg in relation to the metallothionein level. Comparative Biochemistry and Physiology C Pharmacology Toxicology & Endocrinology 107: 249-257.

Payton KB, Flanagan PR, Stinson EA, Chodirker DP, Chamberlain MJ and Valberg LS (1982). Technique for determination of human zinc absorption from measurement of radioactivity in a fecal sample or the body. Gastroenterology. 83(6):1264-1270.

Pennington JAT, Young BE and Wilson DB (1989). Nutritional elements in U.S. diets, results from the Total diet study, 1982 to 1986. J. Am. Diet Assoc. 89, 659-664.[[Cited from EU 2004, a-f].

Pirot F, Millet J, Kalia YN and Humbert P (1996a). In vitro study of percutaneous absorption, cuneous bioavailability and bioequivalence of zinc and copper from five topical formulations. Skin Pharmacol 9: 259- 269.

Pirot F, Panisset F, Agache P, Humbert P (1996b). Simultaneous absorption of copper and zinc through human skin in vitro. Skin Pharmacol 9: 43-52.

Pistorius D, Rosmanith J and Breining H (1976). Intake and distribution of zinc in rat organisms after zinc oxide inhalation in male and female animals. Beitr Silikose Forsch (Pneumokon) 28: 92-101.

Plodikova, P., Rostislav, C. (2010). Ammonium chloride Assessment of toxicokinetic behaviour. Testing laboratory: Research Institute for Organic Syntheses Inc., CETA, Rybitvi 296, 53354 Rybitvi, Czech Republic. Report no.: 10-172. Owner company: Agrofert Holding a. s. Report date: 2010-07-12.

Posthuma et al. (1998). Validation of toxicity data and risk limits for soils: final report. Testing laboratory: National Institute of Public Health and the Environment. Report no.: RIVM-607505004.

Prasad AS, Schulert AR, Sandstead HH, Miale A Jr and Farid Z (1963). Zinc, iron, and nitrogen content of sweat in normal and deficient subjects. J Lab Clin Med 62: 84-89.

Prasad AS, Beck FWJ and Nowak J (1993). Comparison of absorption of five zinc preparations in humans using oral zinc tolerance test. The Journal of Trace Elements in Experimental Medicine. 6: 109-115.

Prasad AS. (2008) Zinc in human health: effect of zinc on immune cells Mol Med.14(5-6):353-7. Review.

Pratt FR, Niederlehner BR, Bowers NJ, Cairns J. (1987). Effects of zinc on freshwater microbial communities. In: Lindberg, S.E. and Hutchinson, T.C. (Eds). Proceedings of the international conference on heavy metals in the environment, Vol 2. September 10-13, 1987, New Orleans, LA, USA, pp. 324-326

Premi and Cornfield (1969). Effects of addition of copper, manganese, zinc and chromium compounds on ammonification and nitrification during incubation of soil. Plant and Soil XXXI, no. 2.

Prinsen MK (1996). Acute oral toxicity study (limit study) with zinc powder in rats. EU Risk Assessment Zinc metal 2004. Testing laboratory: TNO, Zeist, The Netherlands. Report no.: TNO-report V96.515.

Przytocka-Jusiak, M. et al, (1977). Properties of Chlorella vulgaris strain adapted to high concentration of ammonium nitrogen. ACTA MICROBIOLOGICA POLONICA,26(2), 185-197 (1977).

Pullen RGL, Franklin PA and Hall GH (1990). 65Zinc uptake from blood into brain and other tissues in the rat. EU risk assessment report for zinc chloride

Puscas I, Baican M, Coltãu M, Puscas C and Domuta G (1999). Erythrocyte superoxide dismutase activity in patients with digest cancer, adjuvant diagnosis test. Cancer Lett. 143, 95-98. [Cited from EU 2004, a-f].

Radenac G, Fichet D & Miramand P (2001). Bioaccumulation and toxicity of four dissolved metals in Paracentrotus lividus sea-urchin embryo. Marine Environmental Research 51: 151-166.

Rainbow PS and White SL. (1989). Comparative strategies of heavy metal accumulation by crustaceans: zinc,

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 407 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 copper and cadmium in a decapod, an amphipod and a barnacle. Hydrobiologia 174,245-262.

Rand GM, Hoang TC and Brausch JM (2010). Effects of zinc on freshwater microcosms: Preliminary report. Submitted to the International Zinc Association (13 August 2010).

Rand GM, Hoang TC and Brausch JM (2012). Effects of zinc on freshwater microcosms: Final report. Submitted to the International Zinc Association (20 February 2012).

Research Institute for Organic Synthesis Inc., CETA, Taublova E (2010) Zinc bis(dihydrogen phosphate): bacterial reverse mutation test Report no 10-137

Reimann C and the EuroGeoSurveys Geochemistry expert group. (2009). Geochemical mapping of agricultural and grazing land soils project (GEMAS) - database.

Reinhold JG, Faradji B, Abadi P and Ismail-Beigi F (1991). Decreased absorption of calcium, magnesium, and phosphorous by humans due to increased fiber and phosphorous consumption as wheat bread. Nutr. Rev. 49, 204-206. [Cited from EU 2004, a-f].

Reish DJ and Carr RS (1978). The effect of heavy metals on the survival, reproduction, development and life cycles of two species of polychaetous annelids. Mar Pollut Bull 9:24-29.

Reish DJ and Gerlinger TV (1984). The Effects of Cadmium, Lead, and Zinc on Survival and Reproduction in the Polychaetous annelid Neanthes arenaceodentata (F. Nereididae). In: Hutchings PA (Ed.) Proc. of the First Int. Polychaete Conf., Sydney, Aust., July 1983, The Linnean Society of New South Wales, Australia:383-389.

Reish DJ, Gerlinger TV, Phillips CA & Schmidtbauer PD (1977). Toxicity of Formulated Mine Tailings on Marine Polychaete. Marine Biological Consultants, Costa Mesa, CA:133.

Richards MP and Cousins RJ (1975). Mammalian zinc homeostasis: requirement for RNA and metallothionein synthesis. Biochem. Biophys. Res. Comm. 64, 1215-1223. [Cited from EU 2004, a- f].

Richards RJ, Atkins J, Marrs TC, Brown RFR and Masek L (1989). The biochemical and pathological changes produced by the intratracheal instillation of certain components of zinc-hexachloroethane smoke. Toxicol 54: 79-88.

Richelle-Maurer E. and Van de Vyver G. (2001). Chronic toxicity of zinc to freeshwater sponges. final report. Testing laboratory: Insitut de Biologie et de médecine moleculaires, Université libre de Bruxelles, Campus Charleroi, srvice de physiologie cellulaire. Report no.: ILZRO ZEH-00. Owner company: International lead and zinc reserach organisation (ILZRO), now: IZA. Report date: 2001-05-01.

Rivlin RS (1983). Misuse of hair analysis for nutritional assessment. Am. J. Med. 75, 489-493. [Cited from EU 2004, a-f].

Rogers JE. and Li SW (1985). Effect of Metals and Other Inorganic Ions on Soil Microbial Activity: Soil Dehydrogenase Assay as a Simple Toxicity Test. Bull. Environ. Contam. Toxicol. (1985) 34:858-865. Testing laboratory: Pacific Northwest Laboratories, P. O. Box 999, Richland, WA 99352.

Rognerud et al. (1999). Landsomfattande undersokelser av metaller i innsjosedimenter. State Pollution Authority, Report 759/99. 74 pp.

Rossman TG, Molina M and Meyer LW (1984). The genetic toxicology of metal compounds: I. Induction of λ prophage in E coli WP2s (λ). Environmental Mutagenesis. 6: 59-69.

Rossowka MJ and Nakamoto T (1992). Caffeine decreases zinc and metallothionein levels in heart of newborn and adult rats. Pediatr. Res. 32, 330-332. [Cited from EU 2004, a-f].

RTECS (1991). Zinc chloride. Risk assessment report, 2nd priority list, Volume 45, European Commission, Joint Research Center, Institute for Health and Consumer Protection, European Chemicals Bureau, Ispra, Italy

Sachtleben Chemie GmbH (2000a). Acute oral toxicity in rat. IUCLID 2000 ECB.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 408 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Sachtleben Chemie GmbH (2000b). Skin irritation in humans. IUCLID 2000 ECB.

Sachtleben Chemie GmbH (2000c). Eye irritation in humans. IUCLID 2000 ECB.

Sackner M A, Dougherty R L, Chapman G A, Cipley J, Perez D, Kwoka M, Reinhart M, Brito M and Schreck R (1981). Effects of brief and intermediate exposures to sulfate submicron aerosols and sulfate injections on cardiopulmonary function of dogs and tracheal mucous velocity of sheep. J Toxicol Environ Health 7:951-972.

Samanta K and Pal B (1986). Zinc feeding and fertility of male rats. Internat. J. Vit. Nutr. Res. 56: 105-107. Testing laboratory: Department of Biochemistry, School of Tropical Medicine, Calcutta, India.

Samman S and Roberts DCK (1987). The effect of zinc supplements on plasma zinc and copper levels and the reported symptoms in healthy volunteers. Med. J. Australia 146, 246-249.

Samman S and Roberts DCK (1988). The effect of zinc supplements on lipoproteins and copper status Atherosclerosis 70, 247-252.

Sanders A (2001a). Zinc sulphate hexahydrate tech. Grillowflow" (CAS no 13986-24-8) acute oral toxicity in the rat- acute toxic class method. EU risk assessment report zinc sulphate, 2004. Testing laboratory: Safepharm Laboratories Ltd, Derby U. K. Report no.: SPL project no 1353/030.

Sanders A (2001b). Zinc sulphate heptahydrate USP. " (CAS no 7446-20-0) Acute oral toxicity in the rat- acute toxic class method. EU risk assessment report for zinc sulphate 2004. Testing laboratory: Safepharm Laboratories Ltd, Derby U. K. Report no.: SPL project no 1353/031.

Sandifer RD. and Hopkin SP. (1996). Effects of pH on the toxicity of cadmium, copper, lead and zinc to Folsomia candida Willem, 1902 (Collembola) in a standard laboratory test system. Chemosphere, vol 33, No 12, pp 2475-2486, 1996. Testing laboratory: Ecotxicology Group, School of Animal and Microbial Sciences, Uni. of Reading, UK.

Sandifer RD. and Hopkin SP. (1997). Effects of Temperature on the relative toxicities of cadmium, copper, lead and zinc to Folsomia candida (Collembola). Ecotoxicology and environmental safety 37, 125-130 (1997). Testing laboratory: Ecotxicology Group, School of Animal and Microbial Sciences, Uni. of Reading, UK.

Sandstrom B and Sandberg AS (1992). Inhibitory effects of isolated inositol phosphates on zinc absorption in humans. J. Trace Elem. Electrolytes Health Dis. 6, 99-103. [Cited from EU 2004, a-f].

Saviozzi A, Levi-Minzi R, Cardelli R and Riffaldi R (1995). Influence of heavy metals on CO2 evolution from a typic xerochrept soil. Water, Air, & Soil Pollution, Volume 93, Numbers 1-4 / January, 1997. Testing laboratory: Institute of Agricultural Chemistry, University of Pisa, via S. Michele degli Scalzi 2, 56124 Pisa, Italy.

SCF 2003 - SCF (2003) Opinion of the Scientific Committee on Food on the tolerable upper intake level of zinc. Scientific Committee on Food SCF/CS/NUT/UPPLEV/62. Final, 19 March 2003 (expressed on 5 March 2003), Brüssel, Belgien

Schenker MB, Speizer FE and Taylor JO (1981). Acute upper respiratory symptoms resulting from exposure to zinc chloride aerosol. Environ. Res. 25, 317-324. [Cited from EU 2004 c].

SCHER (2007). Scientific Committee on Health and Environmental Risks. SCHER opinionon the Risk Assessment Report on Zinc-Environmental Part. EU Commission publ. Brussels.

Schlicker SA and Cox DH (1968). Maternal dietary zinc, and development and zinc, iron, and copper content of the rat fetus. J. Nutr. 95(2): 287-294.

Schroeder HA, Nason AP, Tipton IH and Balassa JJ (1967). Essential trace metals in man: zinc. Relation to environmental cadmium. J. Chronic Dis. 20(4): 179-210.

Schubauer-Berrigan Mk., Dierkes J R., Monson PD. and Ankley GT. (1993). pH-dependent toxicity of Cd, Cu, Ni, Pb and Zn to Ceriodaphnia dubia, Pimephales promelas, Hyalella azteca and Lumbricus variegatus. Environm. Toxicol. & Chemistry 12, 1261-1266.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 409 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Sheppard SC, Evenden WG, Abboud SA and Stephenson M (1993). A plant life-cycle bioassay for contaminated soil, with comparison to other bioassays: mercury and zinc. Arch. Environ. Contam. Toxicol. 25. 27-35.

Shiller AM and Boyle E (1985). Dissolved zinc in rivers. Nature Vol. 317, nr 6032, 49-52.

Shumskaya NI, Mel’nikova VV, Zhilenko VN and Berezhnova LI (1986). Hygienic assessment of zinc ions in rubber extracts in contact with food products. Gig Sanit 4: 89-90.

Sibley PK., Ankley GT., Cotter AM. and Leonard EN. (1996). Predicting chronic toxicity of sediment spiked with zinc: an evaluation of the acid-volatile sulphide model using a life cycle test with the midge Chironomus tentans. Environm. Tox; and Chem. Vol 15, Nr 12, 2102-2112.

Siebert D, Zimmermann FK and Lemperle E (1970). Genetic Effects of Fungicides. Mutat. Res. 10: 533-543.

Simmer K, Lort-Phillips L, James C, et al. 1991. A double-blind trial of zinc supplementation in pregnancy. Eur J Clin Nutr 45:139-144.

Sinawat et al. (2003). Fetal abnormalities produced after preimplantation exposure of mouse embryos to ammonium chloride. Human Reproduction Vol.18, No.10 pp. 2157±2165, 2003.

Singh I (1983). Induction of reverse mutation and mitotic gene conversion by some metal compounds in Saccharomyces cerevisiae. Mut. Res. 117: 149-152.

Sinley JR., Goettl JP. Jr. and Davies PH. (1974). the effects of zinc on rainbow trout (salmo gairdneri) in hard and soft water. Bull. of Env. Contam. & Toxicol. Vol 12, Nr 2, 193-201.

Skog E and Wahlberg JE (1964). A comparative investigation of the percutaneous absorption of metal compounds in the guinea pig by means of the radioactive isotopes: 51Cr, 58Co, 65Zn, 110mAg, 115mCd, 203Hg. EU risk assessment report for zinc chloride 2004.

Skornik WA and Brain JD (1983). Relative toxicity of inhaled metal sulfate salts for pulmonary macrophages. Am. Rev. Respir. Dis. 128: 297-303.

Smit CE. and Van Gestel CAM (1997). Influence of Temperature on the Regulation and Toxicity of Zinc in Folsomia candida (Collembola). Ecology and environmental safety 37, 213–222 (1997).

Smit EC. and Van Gestel CAM. (1998). Effects of soil type, prepercolation, and ageing on bioaccumulation and toxicity of zinc for the springtail Folsomia candida. Environmental Toxicology and Chemistry 17, 1132-1141.

Smith BL and Embling PP (1993). Sequential changes in the development of the pancreatic lesion of zinc toxicosis in sheep. Vet. Pathol. 30: 242-247.

Smolders E. et al (2003). Comparison of toxicity of zinc for soil microbial processes between laboratory- contamined and polluted field soils. Environmental Toxicology and Chemistry 22, 2592-2598.

Smolders et al. (2003). Effects of field and laboratory Zn contamination on soil microbial processes and plant growth. Testing laboratory: Laboratory of Soil and Water Management, KUL, Kardinaal Mercierlaan 92, 3001 Heverlee, Belgium.

Soderburg TA, Elemros T, Gref R and Hallmans G (1990). Inhibitory effect of zinc oxide on contact allergy due to colophony. Contact Dermatitis: 23: 346-351. Somasundaram B, King PE & Shackley S (1985). The Effect of Zinc on the ultrastructure of the posterior gut and pronephric ducts of the larva of Clupea harengus L. Comp. Biochem. Physiol. 81C (1) 29-37.

Somasundaram B, King PE and Shackley SE (1984). Some morphological effects of zinc upon the yolk- sac larvae of Clupea harengus L. J. Fish Biol 25:333-343.

Someya H, Higo Y, Ohno M, Tsutsui TW and Tsutsui T (2007). Clastogenic activity of seven endodontic medications used in dental practice in human dental pulp cells. Mutat Res. 650(1): 39-47.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 410 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Spehar RL. (1976). Cadmium an zinc toxicity to flagfish, Jordanella floridae. J. fish. res. board Canada, vol 33, 1939-1945.

Spencer H, Osis D, Kramer L and Norris C (1976). Intake, excretion, and retention of zinc in man. In: Trace Elements in Human Health and Disease. Prasad AS (ed.) Vol. 1, Zinc and copper. New York, NY. Academic Press, 345-361. [Cited from EU 2004, a-f].

Spurgeon DJ, Hopkin SP and Jones DT (1994). Effects of Cd, Cu, Pb and Zn on growth, reproduction and survival of the earthworm Eisena fetida (Sav.): assessing the environmental impact of point-source metal contamination in terrestrial ecosystems. Environmental pollution 84 (1994) 123-130. Testing laboratory: Dpt. of pure and applied zoology, University of Reading, PO box 228, Reading, UK.

Spurgeon DJ. and Hopkin SP (1996). Effects of variations of the organic matter content and pH of soils on the availability and toxicity of zinc to the earthworm Eisenia fetida. Pedobiologia 40,80-96 (1996).

Spurgeon DJ. and Hopkin SP. (1995). Extrapolation of the laboratory-based OECD earthworm toxicity test to metal-contaminated field studies. Ecotoxicology, 4, 190-205 (1995). Testing laboratory: Ecotoxicology Group, School of Animal and Microbial Science, University of Reading, PO box 228, Reading, UK.

Spurgeon DJ. et al (2000). Relative sensitivity of life-cycle and biomarker responses in four earthworm species exposed to zinc. Environmental Toxicology and Chemistr y, Vol. 19, No. 7, pp. 1800 –1808, 2000.

Spurgeon DJ., Tomlin MA. and Hopkin SP. (1997). Influence of Temperature on the Toxicity of Zinc to the Earthworm Eisenia fetida. Bull. Environ. Contam. Toxicol. (1997) 58:283-290.

Stadelmann FX. and Santschi-Fuhrimann E (1987). Beitrag zur Abstuetzung von Schwermetall-Richtwerten im boden mit Hilfe von Bodenatmungsmessungen. Testing laboratory: FAC. Liebefeld, Switserland.

Stott et al (1985). Inhibition of pyrophosphatase activity in soils by trace elements. Soil Science Vol. 139, No2.

STOWA (2007) Biologische beschikbaarheid en actuele risko’s van zware metalen in oppervlaktewater. STOWA report nr 2007-12. STOWA, Utrecht, The Netherlands, Publ.

Straube EF, Schuster NH and Sinclair AJ (1980). Zinc toxicity in the ferret. J. Comp. Path. 90: 355-361. Testing laboratory: Not reported. Report no.: Not applicable. Owner company: Not reported. Study number: Not applicable.

Straube EF, Schuster NH and Sinclair AJ (1980). Zinc toxicity in the ferret. J. Comp. Path. 90: 355-361.

Stromgren T (1979). The effects of zinc on the increase in length of five species of intertidal fucales. J Exp Mar Biol Ecol 40:95-102.

Sturniolo GC, Montino MC, Rossetto L, Martin A, D’Inca R, D’Odorico A and Naccarato R (1991). Inhibition of gastric acid secretion reduces zinc absorption in man. J. Am. Coll. Nutr. 10, 372-375. [Cited from EU 2004, a- f].

Suzuki H (1987). Assessment of the carcinogenic hazard of 6 substances used in dental practices. (II) Morphological transformation, DNA damage, and sister chromatid exchanges in cultured Syrian hamster embryo cells induced by formocresol, iodoform, zinc oxide, chloroform, chloramphenicol, tetracycline hydrochloride. Shigaku 74: 1385-1403.

Svenson A (1986). Effect of copper, zinc, and cadmium ions on the production of phosphate from phytic acid by the phytase system in spruce forest soil. Plant and Soil 94, 227-234 (1986).

SZW (1997). Ministerie van Sociale Zaken en Werkgelegenheid. Nationale MAC-lijst 1997-1998. The Hague, The Netherlands.

Tabatabai MA (1977). Effects of trace elements on urease activity in soils. Soil. Biol. Biochem. Vol. 9 pp. 9 to 13 Pergamon Press 1977.

Tacnet F, Watkins DW and Ripoche P (1990). Studies of zinc transport into brush-border membrane vesicles isolated from pig small intestine. Biochem. Biophys. Acta 1024, 323-330. [Cited from EU 2004, a-f].

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 411 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Taublova E (1985). Zinc bis(dihydrogen phosphate): bacterial reverse mutation test. Testing laboratory: Research Institute for Organic Synthesis Inc., CETA. Report no.: 10-137. Owner company: Chemetall GmbH. Study number: 7/10/2. Report date: 2010-06-08.

Thijssen J (1978). Eye irritation study with zinc oxide. EU risk assessment for zinc oxide, 2004. Testing laboratory: Bayer Institut für Toxikologie, Wuppertal-Elberfeld.

Thompson ED, McDermott JA, Zerkle TB, Skare JA, Evans BL, Cody DB (1989) Genotoxicity of zinc in 4 short-term mutagenicity assays. Mutat Res. 223(3):267-72.

Thurston, R. V. and Russo, R. C.: (1978). Acute Toxicity of Ammonia and Nitrite to Cutthroat Trout Fry. Trans. Am. Fish. Soc., 107, 361-368, (1978).

Trevisan A, Buzzo A and Gori GP (1982). Biological indicators in occupational exposure to low concentrations of zinc. Med. Lavoro 6, 614-618. [Cited from EU 2004, a-f].

UK Environment Agency report 6848 (2005)

Uriu-Hare JY, Stern JS, Keen CL (1989) Influence of maternal dietary Zn intake on expression of diabetes- induced teratogenicity in rats Diabetes 38(10):1282-90.

U.S. Environmental Protection Agency. 1996. EMAP-Estuaries Virginian Province Data 1990-1993. Available from: EMAP Home Page (http://www.epa.goc/emap). Accessed 14 July 2010.

US EPA, 2005- US-EPA (US Environmental Protection Agency) (2005) Toxicological review of zinc and compounds. Washington, DC, USA van Beelen et al. (1994). Stimulatory and Toxic Effects of Acid, Pentachlorophenol or Zinc on the Mineralization of Acetate in Acid or Calcareous Soils and Subsoils. J. Environ. Sci. Health, A29(7), 1391-1408 (1994).

Van de Meent. (1990). Desire for levels. Background study for the policy document ‘Setting Environmental Quality Standards for Water and Soil’. RIVM report 670101002, Bilthoven, The Netherlands.

Van der Hoeven N. and Henzen L (1994a). Combinatietoxicologie van de metalen zink en cadmium en de plantensoort Trifolium pratense op kunstmatige grond samengesteld volgens OECD Guideline no. 207 (wormengrond). Testing laboratory: Netherlands Organisation for Applied Scientific Research (TNO), The Netherlands. Report no.: MW-R94/297. Van der Hoeven N. and Henzen L (1994b) Groei van de plantensoorten Lolium perenne, Vicia sativa en Trifolium pratense op grond uit Budel en effecten van zink en cadmium op de groei, TNO rapport MW- R94/004, , Netherlands Organisation for Applied Scientific Research (TNO), The Netherlands

Van der Hoeven N. and Henzen L (1994c). De effecten van zink op de groei van de plantensoort Trifolium pratense (onder laboratorium- en veldomstandigheden) op grond uit Panheel. Testing laboratory: Netherlands Organisation for Applied Scientific Research (TNO), The Netherlands. Report no.: MW-R94/304.

Van der Werff M and Pruyt MJ. (1982). Long-term effects of heavy metals on aquatic plants. Chemosphere Vol 11, Nr 8, 727-739.

Van Eck et al. 1991. Water nr. 60, p. 164-181

Van Gestel CAM and Hensbergen P. J. (1997). Interaction of Cd and Zn toxicity for Folsomia candida willem (collembola: isotomidae) in relation to bioavailability in soil. Environmental Toxicology and Chemistry, Vol 16, No. 6. pp 1177-1186, 1997.

Van Gestel CAM. et al (1993). Accumulation and elimination of cadmium, chromium and zinc and effects on growth and reproduction in Eisenia andrei (oligochaeta, Annelida). The science of the Total Environment, Supplement 1993.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 412 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Van Ginneken I. (1994). The effect of zinc oxide on the growth of the unicellular green algae Selenastrum capricornutum. draft report. Testing laboratory: Janssen Pharmaceutica Beerse, B. Report no.: AASc/0022. Owner company: International lead and zinc research organisation (ILZRO) now: IZA. Report date: 1994-08- 16.

Van Huygevoort AHBM (1999 a). Assessment of acute dermal toxicity with zinc sulphate heptahydrate in the rat. Project 254385. NOTOX B.V., ‘s-Hertogenbosch, The Netherlands. [Cited from EU 2004 e].

Van Huygevoort AHBM (1999b). Primary skin irritation/corrosion study with zinc sulphate heptahydrate in the rabbit (4-hour semi-occlusive application). Project 254374. NOTOX B.V., ‘s-Hertogenbosch, The Netherlands. [Cited from EU 2004 e)].

Van Huygevoort AHBM (1999c). Acute eye irritation/corrosion study with zinc dust in the rabbit. Project 254363. NOTOX B.V., ‘s-Hertogenbosch, The Netherlands. [Cited from EU 2004 a, b].

Van Huygevoort AHBM (1999d). Acute eye irritation/corrosion study with zinc powder in the rabbit. Project 255072. NOTOX B.V., ‘s-Hertogenbosch, The Netherlands. [Cited from EU 2004 (a, b].

Van Huygevoort AHBM (1999 e). Acute eye irritation/corrosion study with zinc oxide in the rabbit. Project 254352. NOTOX B.V., ‘s-Hertogenbosch, The Netherlands. [Cited from EU 2004 b,].

Van Huygevoort AHBM (1999 f ). Acute eye irritation/corrosion study with zinc sulphate heptahydrate in the Rabbit. Project 254341. NOTOX B.V., ‘s-Hertogenbosch, The Netherlands. [Cited from EU 2004 e].

Van Huygevoort AHBM (1999 g). Assessment of contact hypersensitivity to Zincweiß Pharma A in the albino guinea pig (maximisation-test). Project 263429. NOTOX B.V., ‘s-Hertogenbosch, The Netherlands. [Cited from EU 2004 b ].

Van Huygevoort AHBM (1999 h1). Assessment of contact hypersensitivity to zinc oxide in the albino guinea pig (maximisation-test). Project 254339. NOTOX B.V., ‘s-Hertogenbosch, The Netherlands. [Cited from EU 2004 b].

Van Huygevoort AHBM (1999 h2). Assessment of contact hypersensitivity to zinc oxide in the albino guinea pig (maximisation-test). (An extension of NOTOX Project 254339). Project 261214. NOTOX B.V., ‘s- Hertogenbosch,The Netherlands. [Cited from EU 2004 b].

Van Huygevoort AHBM (1999 i). Assessment of contact hypersensitivity to zinc sulphate heptahydrate in the albino guinea pig (maximisation-test). Project 254328. NOTOX B.V., ‘s-Hertogenbosch, The Netherlands. [Cited from EU 2004 e].

Van Huygevoort AHBM (2007). Assessment of acute oral toxicity with SAT 060267 in the rat (acute toxic class method). Project 481106; NOTOX B.V., ‘s-Hertogenbosch, The Netherlands.

Van Leeuwen K, Emans H-J, Vande Plassche E and Canton H. (1994). The role of field tests in hazard assessment of chemicals. In: “Freshwater field tests for hazard assessment of chemicals”, Hill IR et al , eds., Lewis Publ. London.chapter 27, p425-437.

Van Sprang PA, Verdonck FAM, Van Assche F, Regoli L and De Schamphelaere KAC (2009). Environmental risk assessment of zinc in European freshwater: a critical appraisal. The Science of the Total Environment 407, 5373-5391.

Van Vlaardingen PLA., Traas T, Wintersen AM and Aldenberg T. (2004). ETX 2.0 A program to calculate hazardous concentrations and fractions affected, based on normally distributed toxicity data. RIVM report 601501028/2004

Van Woensel M. (1994). The effect of zinc powder on the growth of the unicellular algae Selenastrum capricornutum. final report. Testing laboratory: Janssen Pharmaceutica, Beerse, B. Report no.: AASc/0021. Owner company: International lead and zinc research organisation (ILZRO) now: IZA. Report date: 1994-12- 13.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 413 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Vangheluwe M.L., Eliat-Eliat M., Verdonck F, Van Sprang P, Vercaigne I, De Wilde T. 2013. Evaluation of risks due to the presence of Zinc in European Sewage Treatment Plants. Final report May 2013.

Vangheluwe ML, Heijerick D and Van Sprang P 2003. Probabilistic assessment of zinc bioavailability in sediments. Final report February 2003. Commissioned by the International Lead and Zinc Research Organization

Venugopal B and Lucky TD (1978). Metal toxicity in mammals 2, Chemical Toxicity of Metals and Metalloids. Plenum Press, New York and London, 68-75. [Cited from EU 2004, a-f].

Verdonck F, Verougstraete V, Van Assche F and the Euromeatux ES task force (2010). Specific environmental release categories (SPERCS) for metals and metal compounds. Eurometaux publication, Brussels.

Verhagen H, Rauma AL, Törrönen, De Vogel N, Bruintjes-Rozier GCDM, Dreve MA, Bogaards JJP and Mykkänen (1996). Effect of a vegan diet on biomarkers of chemoprevention in females. Hum. Exp. Toxicol. 15,821-825. [Cited from EU 2004, a-f].

Vilkina GA, Pomerantseva MD and Ramaila LK (1978) Absence of a mutagenic effect of cadmium and zinc salts in mouse somatic and sex cells Genetika 14(12):2212-4 [Cited from EU 2004, a-f].

VMM (2004) Luchtkwaliteit in het Vlaams Gewest 2003. VMM, Erembodegem, 2004.

Vogelmeier C, König G, Bencze K and Fruhmann G (1987). Pulmonary involvement in zinc fume fever. Chest. 92, 946-948. [Cited from EU 2004 a, b].

Voroshilin SI, Plotko EG, Fink TV and Nikiforova V Ya (1978). Cytogenetic action of inorganic compounds of tungsten, zinc, cadmium, and cobalt on human and animal somatic cells. Tsitol. Genet. 12: 241-243.

Vranken G, Vanderhaeghen R and Heip C (1991). Effects of Pollutants on Life-History Parameters of the Marine Nematode Monhystera disjuncta. ICES J. Mar. Sci. 48:325-334.

Wallenborn JG, Evansky P, Shannahan JH, Vallanat B, Ledbetter AD, Schladweiler MC, Richards JH, Gottipolu RR, Nyska A and Kodavanti UP (2008). Subchronic inhalation of zinc sulfate induces cardiac changes in healthy rats. Toxicology and Applied Pharmacology 232 (2008) 69–77.

Walters M and Roe FJC (1965). A study of the effects of zinc and tin administered orally to mice over a prolonged period. Fd. Cosmet. Toxicol. 3: 271-276.

Wang et al (2007). Contaminant sensitivity of freshwater mussels: Chronic toxicity of copper and ammonia to juvenile freshwater mussels. Environmental Toxicology and Chemistry, Vol. 26, No. 10, pp. 2048–2056, 2007. Testing laboratory: Columbia Environmental Research Center, U. S. Geological Survey, Columbia, Missouri. Report date: 2007-04-09.

Warne M. et al (2008). Modeling the toxicity of copper and zinc salts to wheat in 14 soils. Environmental Toxicology and Chemistr y, Vol. 27, No. 4, pp. 786–792, 2008. Warne, M.S.J., Heemsbergen, D., McLaughlin, M., Bell, M., Broos, K., Whatmuff, M., Barry, G., Nash, D., Pritchard, D., Penney, N., 2008b. Models for the field-based toxicity of copper and zinc salts to wheat in 11 Australian soils and comparison to laboratory-based models. Environmental Pollution 156, 707-714.

Wastney ME, Aamodt RL, Rumble WF and Henkin RI (1986). Kinetic analysis of zinc metabolism and its regulation in normal humans. Am. J. Physiol. 251 (Regulatory Integrative Comp. Physiol. 20): R398-R408.

Watling HR (1982). Comparative study of the effects of zinc, cadmium and copper on the larval growth of three oyster species. Bull Environm Contam Toxicol 28:195-201.

Whiteman (1995). Evaluation of interstitial water as a route of exposure for ammonia in sediment tests with Benthic macroinvertebrates. Environmental Toxicology and Chemistry, Vol. 15, No. 5, pp. 794–801, 1996. Testing laboratory: Lake Superior Research Institute, University of Wisconsin–Superior, 1800 Grand Avenue, Superior, Wisconsin 54880, USA.

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 414 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Whitton B. A. (1967). Studies on the growth of riverain Cladophora in culture. Arch. Microbiol.58, 21-29.

WHO (2001). Environmental Health Criteria 221 Zinc. http://www.inchem.org/documents/ehc/ehc/ehc221.htm#1.0

Wilde KL., Stauber JL., Markich SJ., Franklin NM. and Brown PL. (2006). The effect of pH on the uptake and toxicity of copper and zinc in a tropical freshwater alga (Chlorella sp.). Arch. Environm. Contam. Toxic. 51, 174-185.

Wilson (1977). Nitrification in three soils amended with zinc sulfate. Soil. Biol. Biochem. Vol. 9. pp. 277 to 280. Pergamon Press 1977.

Windebank S, Fedyk J and Henderson L (1995). Study report zinc monoglycerolate: micronucleus study in rat bone marrow. EU risk assessment report for zinc metal 2004. Testing laboratory: Environmental Safety Laboratory, Unilever Research, Bedford, England. Report no.: MN940183.

Windholz M, Budavari S, Blumetti RF and Otterbein ES (1983). The Merck Index, an encyclopedia of chemicals, drugs and biologicals, 10th edition. – [Cited from EU 2004 (a-f)].

Wolfe DA, Bricker SB, Long ER, Scott KJ, Thursby GB. 1994. Biological Effects of Toxic Contaminants in Sediments from Long Island Sound and Environs. NOAA Technical Memorandum NOS ORCA 80.

Wolter K, Rabsch U, Krischker P and Davies AG (1984). Influence of low concentrations of cadmium, copper and zinc on phytoplankton of natural water samples. Mar Ecol Prog Ser 19: 167-173.

Xu J. et al (2009). Evaluation of growth and reproduction as indicators of soil metal toxicity to the Collembolan, Sinella curviseta. Insect Science (2009) 16, 57-63.

Yadrick MK, Kenney MA and Winterfeldt EA (1989). Iron, copper, and zinc status, response to supplementation with zinc or zinc and iron in adult females. Am. J. Clin. Nutr. 49, 145-150.

Yanagisawa H, Miyakoshi Y, Kobayashi K, Sakae K, Kawasaki I, Suzuki Y and Tamura J (2009). Long-term intake of a high zinc diet causes iron deficiency anemia accompanied by reticulocytosis and extra-medullary erythropoiesis. Toxicol. Lett. 191: 15–19.

Yeardley, R. B. et al, (1995). Evaluation of alternative reference toxicants for use in the earthworth toxicity test. Env. Tox. Chem. 14(7), 1189-1194 (1995).

Zapol WM (1988). ARDS after accidental inhalation of zinc chloride smoke. Intensive Care Med. 14, 17-24. [Cited from EU 2004 c].

Zaporowska H and Wasilewski W (1992). Combined effect of vanadium and zinc on certain selected haematological indices in rats. Comp Biochem Physiol 103(1):143-147.

Zuurdeeg BW., van Enk RJ and Vriend SP (1992). Natuurlijke achtergrondgehalten van zware metalen en enkele andere sporenelementen in Nederlands oppervlaktewater. Ministerie van VROM, Dir.-Gen. Milieubeheer, VRPM/DGM/AMS 361346

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 415 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

ANNEX 1: Exposure scenario building and environmental release estimation for the waste life stage of the manufacture and the use of zinc and zinc compounds

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 416 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Double click on this page for complete report (pdf)

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 417 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5 ANNEX 2: Evaluation of risks due to the presence of Zinc in European sewage treatment plants

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 418 EC number: diammonium tetrachlorozincate(2-) CAS number: 238-687-6 14639-97-5

Double click on this page for complete report (pdf)

2013-05-27 CSR-PI-5.2.1 CHEMICAL SAFETY REPORT 419