Annex 9 Guidance on Hazards to the Aquatic Environment
Copyright@United Nations, 2017. All rights reserved
e erence
rch. nd. Hyg. ccup. ed undamental and pplied To icology. undamental and pplied To icology
Copyright@United Nations, 2017. All rights reserved
Annex 9
GUIDANCE ON HAZARDS TO THE AQUATIC ENVIRONMENT
Contents Page
A9.1 Introduction ...... 449 A9.2 The harmonized classification scheme ...... 452 A9.2.1 Scope ...... 452 A9.2.2 Classification categories and criteria ...... 452 A9.2.3 Rationale ...... 452 A9.2.4 Application ...... 453 A9.2.5 Data availability ...... 454 A9.2.6 Data quality ...... 454 A9.3 Aquatic toxicity ...... 455 A9.3.1 Introduction ...... 455 A9.3.2 Description of tests ...... 455 A9.3.3 Aquatic toxicity concepts ...... 457 A9.3.4 Weight of evidence ...... 459 A9.3.5 Difficult to test substances ...... 459 A9.3.6 Interpreting data quality ...... 465 A9.4 Degradation ...... 465 A9.4.1 Introduction ...... 465 A9.4.2 Interpretation of degradability data ...... 466 A9.4.3 General interpretation problems ...... 470 A9.4.4 Decision scheme ...... 472 A9.5 Bioaccumulation ...... 473 A9.5.1 Introduction ...... 473 A9.5.2 Interpretation of bioconcentration data ...... 473
A9.5.3 Chemical classes that need special attention with respect to BCF and Kow values ...... 476 A9.5.4 Conflicting data and lack of data ...... 478 A9.5.5 Decision scheme ...... 478 A9.6 Use of QSAR ...... 479 A9.6.1 History ...... 479 A9.6.2 Experimental artifacts causing underestimation of hazard ...... 479 A9.6.3 QSAR modelling issues ...... 480 A9.6.4 Use of QSARs in aquatic classification ...... 481 A9.7 Classification of metals and metal compounds ...... 483 A9.7.1 Introduction ...... 483 A9.7.2 Application of aquatic toxicity data and solubility data for classification ...... 485 A9.7.3 Assessment of environmental transformation ...... 486 A9.7.4 Bioaccumulation ...... 487 A9.7.5 Application of classification criteria to metals and metal compounds ...... 487 Appendix I Determination of degradability of organic substances ...... 491 Appendix II Factors influencing degradability in the aquatic environment ...... 497 Appendix III Basic principles of the experimental and estimation methods for determination of BCF and Kow of organic substances ...... 501 Appendix IV Influence of external and internal factors on the bioconcentration potential of organic substances ...... 505 Appendix V Test guidelines ...... 507 Appendix VI References ...... 511
- 447 - - 447 -
Copyright@United Nations, 2017. All rights reserved
ntro ction
this category are frequently referred to as “complex mixtures”. In most cases they can be characterized as a homologous the purpose of this guidance document, these materials will be referred to as “complex substances” or “multi component substances”.
n ironment Health and Safety ublications Series on Testing and ssessment No n ironment irectorate rgani ation for economic o operation and e elopment pril . - 449 - Copyright@United Nations, 2017. All rights reserved
reason, it has been felt necessary to de elop a transparent guidance document that would see to expand and explain the d comple or multi component substances: such substances, for e ample, hydrocarbon mi tures, criteria in such a way that a common understanding of their rationale and a common approach to data interpretation may fre uently cannot be dissolved into a homogeneous solution, and the multiple components ma e be achie ed. This is of particular importance since any harmonized system applied to the “universe of chemicals” will monitoring impossible. onsideration therefore needs to be given to using the data derived from rely hea ily on self classification by manufacturers and suppliers, classifications that must be accepted across national the testing of water accommodated fractions s for a uatic to icity, and the utilization of boundaries without always recei ing regulatory scrutiny. his guidance document, therefore, see s to inform the reader, such data in the classification scheme. iodegradation, bioaccumulation, partitioning behaviour in a number of ey areas, and as a result lead to classification in a consistent manner, thus ensuring a truly harmonized and water solubility all present problems of interpretation, where each component of the mi ture and self operating system. may behave differently
. . irstly, it will pro ide a detailed description of the criteria, a rationale for the criteria selected, and an e polymers: such substances fre uently have a wide range of molecular masses, with only a fraction o er iew of how the scheme will wor in practice ection . . his section will address the common sources of being water soluble. pecial methods are available to determine the water soluble fraction and data, the need to apply quality criteria, how to classify when the data set is incomplete or when a large data set leads to these data will need to be used in interpreting the test data against the classification criteria an ambiguous classification, and other commonly encountered classification problems. f inorganic compounds and metals: such substances, which can interact with the media, can produce . . econdly, the guidance will pro ide detailed expert ad ice on the interpretation of data deri ed from a range of a uatic to icities dependant on such factors as p , water hardness etc. ifficult the a ailable databases, including how to use non standard data, and specific quality criteria that may apply for interpretational problems also arise from the testing of essential elements that are beneficial at individual properties. The problems of data interpretation for “difficult substances”, those substances for which standard certain levels. or metals and inorganic metal compounds, the concept of degradability as applied testing methods either do not apply or gi e difficult interpretational problems, will be described and ad ice pro ided on to organic compounds has limited or no meaning. ually the use of bioaccumulation data should suitable solutions. he emphasis will be on data interpretation rather than testing since the system will, as far as be treated with care possible, rely on the best a ailable existing data and data required for regulatory purposes. he four core properties, acute and chronic aquatic toxicity ection . , degradability ection . and bioaccumulation ection . are g surface active substances: such substances can form emulsions in which the bioavailablity is treated separately. difficult to ascertain, even with careful solution preparation. icelle formation can result in an overestimation of the bioavailable fraction even when “solutions” are apparently formed. This . . he range of interpretational problems can be extensi e and as a result such interpretation will always presents significant problems of interpretation in each of the water solubility, partition coefficient, rely on the ability and expertise of the indi iduals responsible for classification. owe er, it is possible to identify some bioaccumulation and a uatic to icity studies commonly occurring difficulties and pro ide guidance that distils accepted expert udgement that can act as an aid to achie ing a reliable and consistent result. uch difficulties can fall into a number of o erlapping issues h ionizable substances: such substances can change the e tent of ionization according to the level of counter ions in the media. cids and bases, for e ample, will show radically different partitioning a he difficulty in applying the current test procedures to a number of types of substance behaviour depending on the p
b The difficulty in interpreting the data derived both from these “difficult to test” substances and i coloured substances: such substance can cause problems in the algal a uatic plant testing because from other substances of the bloc ing of incident light
c he difficulty in interpretation of di erse data sets deri ed from a wide ariety of sources. impurities: some substances can contain impurities that can change in and in chemical nature between production batches. Interpretational problems can arise where either or both the to icity . . or many organic substances, the testing and interpretation of data present no problems when and water solubility of the impurities are greater than the parent substance, thus potentially applying both the rele ant uideline and the classification criteria. here are a number of typical interpretational influencing the to icity data in a significant way. problems, howe er, that can be characterized by the type of substance being studied. hese are commonly called “difficult substances”: . . These represent some of the problems encountered in establishing the ade uacy of data, interpreting the data and applying that data to the classification scheme. etailed guidance on how to deal with these problems, as a poorly soluble substances these substances are difficult to test because they present problems in well as other issues related will be presented in the following sections. The interpretation of data on acute and on solution preparation, and in concentration maintenance and erification during aquatic toxicity chronic a uatic to icity will be covered in ection . . This section will deal with the specific interpretational testing. In addition, many available data for such substances have been produced using “solutions” problems encountered for the above “difficult substances”, including providing some advice on when and how such in excess of the water solubility resulting in ma or interpretational problems in defining the true data can be used within the classification scheme. lso covered will be a general description of the test data used and or for the purposes of classification. Interpretation of the partitioning beha iour the testing methodologies suitable for producing such data. can also be problematic where the poor solubility in water and octanol may be compounded by insufficient sensiti ity in the analytical method. ater solubility may be difficult to determine and . . wide range of degradation data are available that must be interpreted according to the criteria for is frequently recorded as simply being less than the detection limit, creating problems in rapid degradability. uidance is thus needed on how to use these data obtained by employing non standard test interpreting both aquatic toxicity and bioaccumulation studies. In biodegradation studies, poor methods, including the use of half lives where these are available, of primary degradation, of soil degradation rates and solubility may result in low bioa ailability and thus lower than expected biodegradation rates. he their suitability for e trapolation to a uatic degradation and of environmental degradation rates. short description of specific test method or the choice of procedures used can thus be of ey importance estimation techni ues for evaluating degradability in relation to the classification criteria is also included. This guidance will be provided in ection . . b unstable substances such substances that degrade or react rapidly in the test system present both testing and interpretational problems. It will be necessary to determine whether the correct . . ethods by which the potential to bioaccumulate can be determined will be described in ection methodology has been used, whether it is the substance or the degradation reaction product that . . This section will describe the relationship between the partition coefficient criteria and the bioconcentration factor has been tested, and whether the data produced is rele ant to the classification of the parent , provide guidance on the interpretation of e isting data, how to estimate the partition coefficient by the use of substance s when no e perimental data are available and in particular deal with the specific problems identified above for difficult substances. The problems encountered when dealing with substances of high molecular mass will also be c olatile substances such substances that can clearly present testing problems when used in open covered. systems should be e aluated to ensure adequate maintenance of exposure concentrations. oss of test material during biodegradation testing is ine itable in certain methods and will lead to . . section is also included which covers general issues concerning the use of s within the misinterpretation of the results system, when and how they may be used, for each of the three properties of concern. s a general approach, it is widely accepted that e perimental data should be used rather than data when such data are available. The use of s
- 450 - Copyright@United Nations, 2017. All rights reserved
reason, it has been felt necessary to de elop a transparent guidance document that would see to expand and explain the d comple or multi component substances: such substances, for e ample, hydrocarbon mi tures, criteria in such a way that a common understanding of their rationale and a common approach to data interpretation may fre uently cannot be dissolved into a homogeneous solution, and the multiple components ma e be achie ed. This is of particular importance since any harmonized system applied to the “universe of chemicals” will monitoring impossible. onsideration therefore needs to be given to using the data derived from rely hea ily on self classification by manufacturers and suppliers, classifications that must be accepted across national the testing of water accommodated fractions s for a uatic to icity, and the utilization of boundaries without always recei ing regulatory scrutiny. his guidance document, therefore, see s to inform the reader, such data in the classification scheme. iodegradation, bioaccumulation, partitioning behaviour in a number of ey areas, and as a result lead to classification in a consistent manner, thus ensuring a truly harmonized and water solubility all present problems of interpretation, where each component of the mi ture and self operating system. may behave differently
. . irstly, it will pro ide a detailed description of the criteria, a rationale for the criteria selected, and an e polymers: such substances fre uently have a wide range of molecular masses, with only a fraction o er iew of how the scheme will wor in practice ection . . his section will address the common sources of being water soluble. pecial methods are available to determine the water soluble fraction and data, the need to apply quality criteria, how to classify when the data set is incomplete or when a large data set leads to these data will need to be used in interpreting the test data against the classification criteria an ambiguous classification, and other commonly encountered classification problems. f inorganic compounds and metals: such substances, which can interact with the media, can produce . . econdly, the guidance will pro ide detailed expert ad ice on the interpretation of data deri ed from a range of a uatic to icities dependant on such factors as p , water hardness etc. ifficult the a ailable databases, including how to use non standard data, and specific quality criteria that may apply for interpretational problems also arise from the testing of essential elements that are beneficial at individual properties. The problems of data interpretation for “difficult substances”, those substances for which standard certain levels. or metals and inorganic metal compounds, the concept of degradability as applied testing methods either do not apply or gi e difficult interpretational problems, will be described and ad ice pro ided on to organic compounds has limited or no meaning. ually the use of bioaccumulation data should suitable solutions. he emphasis will be on data interpretation rather than testing since the system will, as far as be treated with care possible, rely on the best a ailable existing data and data required for regulatory purposes. he four core properties, acute and chronic aquatic toxicity ection . , degradability ection . and bioaccumulation ection . are g surface active substances: such substances can form emulsions in which the bioavailablity is treated separately. difficult to ascertain, even with careful solution preparation. icelle formation can result in an overestimation of the bioavailable fraction even when “solutions” are apparently formed. This . . he range of interpretational problems can be extensi e and as a result such interpretation will always presents significant problems of interpretation in each of the water solubility, partition coefficient, rely on the ability and expertise of the indi iduals responsible for classification. owe er, it is possible to identify some bioaccumulation and a uatic to icity studies commonly occurring difficulties and pro ide guidance that distils accepted expert udgement that can act as an aid to achie ing a reliable and consistent result. uch difficulties can fall into a number of o erlapping issues h ionizable substances: such substances can change the e tent of ionization according to the level of counter ions in the media. cids and bases, for e ample, will show radically different partitioning a he difficulty in applying the current test procedures to a number of types of substance behaviour depending on the p
b The difficulty in interpreting the data derived both from these “difficult to test” substances and i coloured substances: such substance can cause problems in the algal a uatic plant testing because from other substances of the bloc ing of incident light
c he difficulty in interpretation of di erse data sets deri ed from a wide ariety of sources. impurities: some substances can contain impurities that can change in and in chemical nature between production batches. Interpretational problems can arise where either or both the to icity . . or many organic substances, the testing and interpretation of data present no problems when and water solubility of the impurities are greater than the parent substance, thus potentially applying both the rele ant uideline and the classification criteria. here are a number of typical interpretational influencing the to icity data in a significant way. problems, howe er, that can be characterized by the type of substance being studied. hese are commonly called “difficult substances”: . . These represent some of the problems encountered in establishing the ade uacy of data, interpreting the data and applying that data to the classification scheme. etailed guidance on how to deal with these problems, as a poorly soluble substances these substances are difficult to test because they present problems in well as other issues related will be presented in the following sections. The interpretation of data on acute and on solution preparation, and in concentration maintenance and erification during aquatic toxicity chronic a uatic to icity will be covered in ection . . This section will deal with the specific interpretational testing. In addition, many available data for such substances have been produced using “solutions” problems encountered for the above “difficult substances”, including providing some advice on when and how such in excess of the water solubility resulting in ma or interpretational problems in defining the true data can be used within the classification scheme. lso covered will be a general description of the test data used and or for the purposes of classification. Interpretation of the partitioning beha iour the testing methodologies suitable for producing such data. can also be problematic where the poor solubility in water and octanol may be compounded by insufficient sensiti ity in the analytical method. ater solubility may be difficult to determine and . . wide range of degradation data are available that must be interpreted according to the criteria for is frequently recorded as simply being less than the detection limit, creating problems in rapid degradability. uidance is thus needed on how to use these data obtained by employing non standard test interpreting both aquatic toxicity and bioaccumulation studies. In biodegradation studies, poor methods, including the use of half lives where these are available, of primary degradation, of soil degradation rates and solubility may result in low bioa ailability and thus lower than expected biodegradation rates. he their suitability for e trapolation to a uatic degradation and of environmental degradation rates. short description of specific test method or the choice of procedures used can thus be of ey importance estimation techni ues for evaluating degradability in relation to the classification criteria is also included. This guidance will be provided in ection . . b unstable substances such substances that degrade or react rapidly in the test system present both testing and interpretational problems. It will be necessary to determine whether the correct . . ethods by which the potential to bioaccumulate can be determined will be described in ection methodology has been used, whether it is the substance or the degradation reaction product that . . This section will describe the relationship between the partition coefficient criteria and the bioconcentration factor has been tested, and whether the data produced is rele ant to the classification of the parent , provide guidance on the interpretation of e isting data, how to estimate the partition coefficient by the use of substance s when no e perimental data are available and in particular deal with the specific problems identified above for difficult substances. The problems encountered when dealing with substances of high molecular mass will also be c olatile substances such substances that can clearly present testing problems when used in open covered. systems should be e aluated to ensure adequate maintenance of exposure concentrations. oss of test material during biodegradation testing is ine itable in certain methods and will lead to . . section is also included which covers general issues concerning the use of s within the misinterpretation of the results system, when and how they may be used, for each of the three properties of concern. s a general approach, it is widely accepted that e perimental data should be used rather than data when such data are available. The use of s
- 451 - Copyright@United Nations, 2017. All rights reserved
will thus be limited to such times when no reliable data are available. ot all substances are suitable for the application readily available, or can be generated to highly standardised protocols. t is this acute to icity which has therefore been of estimations, however, and the guidance in ection . will address this issue. used as the core property in defining both the acute and the long term chronic ha ard if no ade uate chronic test data are available. evertheless, it has been recogni ed that chronic to icity data, if available should be preferred in defining . . inally, a section is devoted to the special problems associated with the classification of metals and the long term chronic ha ard category. their compounds. learly, for these compounds, a number of the specific criteria such as biodegradability and octanol water partition coefficient cannot be applied although the principle of lac of destruction via degradation, and . . . The combination of chronic to icity and intrinsic fate properties reflects the potential ha ard of a bioaccumulation remain important concepts. Thus it is necessary to adopt a different approach. etals and metal substance. ubstances that do not rapidly degrade have a higher potential for longer term e posures and therefore compounds can undergo interactions with the media which affect the solubility of the metal ion, partitioning from the should be classified in a more severe category than substances which are rapidly degradable see . . . . . water column, and the species of metal ion that e ists in the water column. n the water column, it is generally the dissolved metal ions which are of concern for to icity. The interaction of the substance with the media may either . . . hile recogni ing that acute to icity itself is not a sufficiently accurate predictor of chronic to icity increase or decrease the level of ions and hence to icity. t is thus necessary to consider whether metal ions are li ely to to be used solely and directly for establishing ha ard, it is considered that, in combination with either a potential to be formed from the substance and dissolve in the water, and if so whether they are formed rapidly enough to cause bioaccumulate i.e. a log ow unless or potential longer term e posure i.e. lac of rapid degradation it concern. scheme for interpreting the results from this type of study is presented in ection . . can be used as a suitable surrogate for classification purposes. ubstances rapidly biodegrading that show acute to icity with a significant degree of bioaccumulation will normally show chronic to icity at a significantly lower concentration. . . hile the uidance ocument provides useful advice on how to apply the criteria to a wide variety of ually substances that do not rapidly degrade have a higher potential for giving rise to longer term e posures which situations, it remains a guidance only. t cannot hope to cover all situations that arise in classification. t should again may result in long term to icity being reali ed. Thus, for e ample, in absence of ade uate chronic test data, therefore be seen as a living document that in part describes the fundamental principles of the system, e.g. ha ard based category hronic should be assigned if either of the following criteria are met rather than ris based, and the fi ed criteria. t must also, in part, be a repository for the accumulated e perience in using the scheme to include the interpretations which allow the apparently fi ed criteria to be applied in a wide variety a for any appropriate a uatic species mg l and a potential to bioaccumulate of non standard situations. log ow unless b for any appropriate a uatic species mg l and a lac of rapid degradation. e r oni e c i ic tion c e e . . . The precise definitions of the core elements of this system are described in detail in sections . , cope . and . . The criteria were developed ta ing into account e isting systems for ha ard classification, such as . . . or some poorly soluble substances, which are normally considered as those having a water solubility upply and se ystem, the anadian and esticide systems, ha ard evaluation procedure, mg l, no acute to icity is e pressed in to icity tests performed at the solubility limit. f for such a substance, cheme for arine ollutant, the uropean oad and ail Transport cheme , and the and Transport. however, the , or if absent, the log indicating a bioaccumulating potential and the substance is also These systems include supply and subse uent use of chemicals, the sea transport of chemicals as well as transport of ow not rapidly degradable, a safety net classification is applied, hronic . or these types of substance the e posure chemicals by road and rail. The harmoni ed criteria are therefore intended to identify ha ardous chemicals in a common duration in short term tests may well be too short for a steady state concentration of the substance to be reached in the way for use throughout all these systems. To address the needs for all different sectors transport, supply and use it was test organisms. Thus, even though no acute to icity has been measured in a short term acute test, it remains a real necessary to create two different sub classes, one sub class for short term acute a uatic ha ards, consisting of three possibility that such non rapidly degradable and bioaccumulative substances may e ert chronic effects, particularly categories and one sub class for lont term chronic a uatic ha ards, consisting of categories. The short term acute since such low degradability may lead to an e tended e posure period in the a uatic environment. classification sub class ma es provision for two short term acute ha ard categories cute and not normally used when considering pac aged goods. or chemicals transported in bul , there are a number of regulatory decisions that . . . n defining a uatic to icity, it is not possible to test all species present in an a uatic ecosystem. epresentative can uni uely arise because of the bul uantities being considered. or these situations, for e ample where decisions are species are therefore chosen which cover a range of trophic levels and ta onomic groupings. The ta a chosen, fish, re uired on the ship type to be used, consideration of all short term acute ha ard categories as well as the long term crustacea and aquatic plants that represent the “base set” in most ha ard profiles, represent a minimum data set for a chronic ha ard categories are considered important. The following paragraphs describe in detail the criteria to be used fully valid description of ha ard. The lowest of the available to icity values will normally be used to define the ha ard in defining each of these ha ard categories. category. iven the wide range of species in the environment, the three tested can only be a poor surrogate and the lowest value is therefore ta en for cautious reasons to define the ha ard category. n doing so, it is recogni ed that the Classification cate o ies an c ite ia distribution of species sensitivity can be several orders of magnitude wide and that there will thus be both more and less sensitive species in the environment. Thus, when data are limited, the use of the most sensitive species tested gives a The ha ard categories for acute and chronic a uatic to icity and their related criteria are set out in cautious but acceptable definition of the ha ard. There are some circumstances where it may not be appropriate to use hapter . , paragraph . . . and table . . . the lowest to icity value as the basis for classification. This will usually only arise where it is possible to define the sensitivity distribution with more accuracy than would normally be possible, such as when large data sets are available. ationale uch large data sets should be evaluated with due caution.
. . . The harmoni ed system for classification recogni es that the intrinsic ha ard to a uatic organisms is pplication represented by both the acute and chronic or long term to icity of a substance, the relative importance of which is determined by the specific regulatory regimes in operation. istinction can be made between the short term acute . . . enerally spea ing, in deciding whether a substance should be classified, a search of appropriate ha ard and the long term chronic ha ard and therefore ha ard classes are defined for both properties representing a databases and other sources of data should be made for the following data elements gradation in the level of ha ard identified. learly the ha ard identified by hronic is more severe than hronic . ince the acute short term ha ard and long term chronic ha ard represent distinct types of ha ard, they are not a water solubility comparable in terms of their relative severity. oth ha ard sub classes should be applied independently for the classification of substances to establish a basis for all regulatory systems. b acute a uatic to icity s c chronic a uatic to icity s and or e uivalent . . . The principal ha ard classes defined by the criteria relate largely to the potential for long term chronic ha ard. This reflects the overriding concern with respect to chemicals in the environment, namely that the d available degradation and specifically evidence of ready biodegradability effects caused are usually sub lethal, e.g. effects on reproduction, and caused by longer term e posure. hile e stability data, in water recogni ing that the long term chronic ha ard represents the principal concern, particularly for pac aged goods where environmental release would be limited in scope, it must also be recogni ed that chronic to icity data are e pensive to f fish bioconcentration factor generate and generally not readily available for most substances. n the other hand, acute to icity data are fre uently g octanol water partition coefficient log ow - 452 - Copyright@United Nations, 2017. All rights reserved
will thus be limited to such times when no reliable data are available. ot all substances are suitable for the application readily available, or can be generated to highly standardised protocols. t is this acute to icity which has therefore been of estimations, however, and the guidance in ection . will address this issue. used as the core property in defining both the acute and the long term chronic ha ard if no ade uate chronic test data are available. evertheless, it has been recogni ed that chronic to icity data, if available should be preferred in defining . . inally, a section is devoted to the special problems associated with the classification of metals and the long term chronic ha ard category. their compounds. learly, for these compounds, a number of the specific criteria such as biodegradability and octanol water partition coefficient cannot be applied although the principle of lac of destruction via degradation, and . . . The combination of chronic to icity and intrinsic fate properties reflects the potential ha ard of a bioaccumulation remain important concepts. Thus it is necessary to adopt a different approach. etals and metal substance. ubstances that do not rapidly degrade have a higher potential for longer term e posures and therefore compounds can undergo interactions with the media which affect the solubility of the metal ion, partitioning from the should be classified in a more severe category than substances which are rapidly degradable see . . . . . water column, and the species of metal ion that e ists in the water column. n the water column, it is generally the dissolved metal ions which are of concern for to icity. The interaction of the substance with the media may either . . . hile recogni ing that acute to icity itself is not a sufficiently accurate predictor of chronic to icity increase or decrease the level of ions and hence to icity. t is thus necessary to consider whether metal ions are li ely to to be used solely and directly for establishing ha ard, it is considered that, in combination with either a potential to be formed from the substance and dissolve in the water, and if so whether they are formed rapidly enough to cause bioaccumulate i.e. a log ow unless or potential longer term e posure i.e. lac of rapid degradation it concern. scheme for interpreting the results from this type of study is presented in ection . . can be used as a suitable surrogate for classification purposes. ubstances rapidly biodegrading that show acute to icity with a significant degree of bioaccumulation will normally show chronic to icity at a significantly lower concentration. . . hile the uidance ocument provides useful advice on how to apply the criteria to a wide variety of ually substances that do not rapidly degrade have a higher potential for giving rise to longer term e posures which situations, it remains a guidance only. t cannot hope to cover all situations that arise in classification. t should again may result in long term to icity being reali ed. Thus, for e ample, in absence of ade uate chronic test data, therefore be seen as a living document that in part describes the fundamental principles of the system, e.g. ha ard based category hronic should be assigned if either of the following criteria are met rather than ris based, and the fi ed criteria. t must also, in part, be a repository for the accumulated e perience in using the scheme to include the interpretations which allow the apparently fi ed criteria to be applied in a wide variety a for any appropriate a uatic species mg l and a potential to bioaccumulate of non standard situations. log ow unless b for any appropriate a uatic species mg l and a lac of rapid degradation. e r oni e c i ic tion c e e . . . The precise definitions of the core elements of this system are described in detail in sections . , cope . and . . The criteria were developed ta ing into account e isting systems for ha ard classification, such as . . . or some poorly soluble substances, which are normally considered as those having a water solubility upply and se ystem, the anadian and esticide systems, ha ard evaluation procedure, mg l, no acute to icity is e pressed in to icity tests performed at the solubility limit. f for such a substance, cheme for arine ollutant, the uropean oad and ail Transport cheme , and the and Transport. however, the , or if absent, the log indicating a bioaccumulating potential and the substance is also These systems include supply and subse uent use of chemicals, the sea transport of chemicals as well as transport of ow not rapidly degradable, a safety net classification is applied, hronic . or these types of substance the e posure chemicals by road and rail. The harmoni ed criteria are therefore intended to identify ha ardous chemicals in a common duration in short term tests may well be too short for a steady state concentration of the substance to be reached in the way for use throughout all these systems. To address the needs for all different sectors transport, supply and use it was test organisms. Thus, even though no acute to icity has been measured in a short term acute test, it remains a real necessary to create two different sub classes, one sub class for short term acute a uatic ha ards, consisting of three possibility that such non rapidly degradable and bioaccumulative substances may e ert chronic effects, particularly categories and one sub class for lont term chronic a uatic ha ards, consisting of categories. The short term acute since such low degradability may lead to an e tended e posure period in the a uatic environment. classification sub class ma es provision for two short term acute ha ard categories cute and not normally used when considering pac aged goods. or chemicals transported in bul , there are a number of regulatory decisions that . . . n defining a uatic to icity, it is not possible to test all species present in an a uatic ecosystem. epresentative can uni uely arise because of the bul uantities being considered. or these situations, for e ample where decisions are species are therefore chosen which cover a range of trophic levels and ta onomic groupings. The ta a chosen, fish, re uired on the ship type to be used, consideration of all short term acute ha ard categories as well as the long term crustacea and aquatic plants that represent the “base set” in most ha ard profiles, represent a minimum data set for a chronic ha ard categories are considered important. The following paragraphs describe in detail the criteria to be used fully valid description of ha ard. The lowest of the available to icity values will normally be used to define the ha ard in defining each of these ha ard categories. category. iven the wide range of species in the environment, the three tested can only be a poor surrogate and the lowest value is therefore ta en for cautious reasons to define the ha ard category. n doing so, it is recogni ed that the Classification cate o ies an c ite ia distribution of species sensitivity can be several orders of magnitude wide and that there will thus be both more and less sensitive species in the environment. Thus, when data are limited, the use of the most sensitive species tested gives a The ha ard categories for acute and chronic a uatic to icity and their related criteria are set out in cautious but acceptable definition of the ha ard. There are some circumstances where it may not be appropriate to use hapter . , paragraph . . . and table . . . the lowest to icity value as the basis for classification. This will usually only arise where it is possible to define the sensitivity distribution with more accuracy than would normally be possible, such as when large data sets are available. ationale uch large data sets should be evaluated with due caution.
. . . The harmoni ed system for classification recogni es that the intrinsic ha ard to a uatic organisms is pplication represented by both the acute and chronic or long term to icity of a substance, the relative importance of which is determined by the specific regulatory regimes in operation. istinction can be made between the short term acute . . . enerally spea ing, in deciding whether a substance should be classified, a search of appropriate ha ard and the long term chronic ha ard and therefore ha ard classes are defined for both properties representing a databases and other sources of data should be made for the following data elements gradation in the level of ha ard identified. learly the ha ard identified by hronic is more severe than hronic . ince the acute short term ha ard and long term chronic ha ard represent distinct types of ha ard, they are not a water solubility comparable in terms of their relative severity. oth ha ard sub classes should be applied independently for the classification of substances to establish a basis for all regulatory systems. b acute a uatic to icity s c chronic a uatic to icity s and or e uivalent . . . The principal ha ard classes defined by the criteria relate largely to the potential for long term chronic ha ard. This reflects the overriding concern with respect to chemicals in the environment, namely that the d available degradation and specifically evidence of ready biodegradability effects caused are usually sub lethal, e.g. effects on reproduction, and caused by longer term e posure. hile e stability data, in water recogni ing that the long term chronic ha ard represents the principal concern, particularly for pac aged goods where environmental release would be limited in scope, it must also be recogni ed that chronic to icity data are e pensive to f fish bioconcentration factor generate and generally not readily available for most substances. n the other hand, acute to icity data are fre uently g octanol water partition coefficient log ow - 453 - Copyright@United Nations, 2017. All rights reserved
he ater s lubilit and stabilit data alth u h n t used directl in the criteria are ne ertheless c ata deri ed r testin hich hile n t strictl acc rdin t a uideline detailed ab e i p rtant since the are a aluable help in the data interpretati n the ther pr perties see ll s accepted scienti ic principles and pr cedures and r has been peer re ie ed pri r t publicati n r such data here all the e peri ental detail is n t rec rded s e ud e ent classi a re ie sh uld irst be ade the a ailable aquatic t icit data t ill be necessar t a be required t deter ine alidit r all such data a be used ithin the c nsider all the a ailable data and select th se hich eet the necessar qualit criteria r classi icati n there are n classi icati n sche e data a ailable that eet the qualit criteria required b the internati nall standardi ed eth ds it ill be necessar t d ata deri ed r testin pr cedures hich de iate si ni icantl r standard uidelines and e a ine an a ailable data t deter ine hether a classi icati n can be ade the data indicate that the acute aquatic are c nsidered as unreliable sh uld n t be used in classi icati n t icit is reater than l r s luble substances and the chr nic aquatic t icit is reater than l then the substance is n t classi ied as ha ard us here are a nu ber cases here n e ects are bser ed in the test e data he circu stances use and alidit data are discussed in the rele ant and the aquatic t icit is thus rec rded as a ater s lubilit alue i e there is n acute t icit ithin the ran e secti ns the ater s lubilit in the test edia here this is the case and the ater s lubilit in the test edia is l a ain ata deri ed r sec ndar s urces such as handb s re ie s citati n etc here the data n classi icati n need be applied qualit cann t be directl e aluated uch data sh uld be e a ined here data r qualit chr nic aquatic t icit data are a ailable cut alues ill depend n hether the substance is rapidl and are n t a ailable t deter ine hether it can be used uch data sh uld ha e su icient de radable r n t here re r n n rapidl de radable substances and th se r hich n in r ati n n de radati n detail t all qualit t be assessed n deter inin the acceptabilit these data r the is a ailable the cut le els are hi her than r th se substances here rapid de radabilit can be c n ir ed see purp ses classi icati n due re ard sh uld be i en t the di iculties in testin that a hapter ables and ha e a ected data qualit and the si ni icance the rep rted result in ter s the le el ha ard identi ied see here the l est acute aquatic t icit data are bel l and n adequate chr nic t icit data lassi icati n a als be ade n inc plete t icit data sets e here data are n t a ailable n are a ailable it is necessar t irst decide hich ha ard cate r the t icit alls in and then t deter ine hether the all three tr phic levels. In these cases, the classification may be considered as “provisional” and subject to further chr nic and r the acute sub class sh uld be applied his can si pl be achie ed b e a inin the a ailable data n in r ati n bec in a ailable n eneral all the data a ailable ill need t be c nsidered pri r t assi nin a the partiti n c e icient l and the a ailable data n de radati n either the l r the substance cann t classi icati n here d qualit data are n t a ailable l er qualit data ill need t be c nsidered n these be c nsidered as rapidl de radable then the appr priate l n ter chr nic ha ard cate r and the c rresp ndin circu stances a ud e ent ill need t be ade re ardin the true le el ha ard r e a ple here d qualit acute sh rt ter ha ard cate r are applied independentl t sh uld be n ted that alth u h the l is the st data are a ailable r a particular species r ta a this sh uld be used in pre erence t an l er qualit data hich readil a ailable indicati n a p tential t bi accu ulate an e peri entall deri ed is pre erred here this is i ht als be a ailable r that species r ta a e er d qualit data a n t al a s be a ailable r all the basic a ailable this sh uld be used rather than the partiti n c e icient n these circu stances a uld indicate data set tr phic le els t ill be necessar t c nsider data l er qualit r th se tr phic le els r hich d bi accu ulati n su icient t classi in the appr priate l n ter chr nic ha ard cate r the substance is b th qualit data are n t a ailable nsiderati n such data h e er ill als need t c nsider the di iculties that a rapidl de radable and has a l p tential t bi accu ulate r i absent l then it sh uld n t be ha e a ected the li elih d achie in a alid result r e a ple the test details and e peri ental desi n a be assi ned t a l n ter chr nic ha ard cate r unless the chr nic t icit data indicate ther ise see critical t the assess ent the usabilit s e data such as that r h dr l ticall unstable che icals hile less s r ther che icals uch di iculties are described urther in ecti n r p rl s luble substances enerall spea in th se ith a ater s lubilit in the test edia l r hich n aquatic t icit has been und sh uld be urther e a ined t deter ine hether hr nic r all the identi icati n ha ard and hence the classi icati n ill be based n in r ati n ate r needs t be applied hus i the substance is b th n t rapidl de radable and has a p tential t directl btained r testin the substance bein c nsidered here are ccasi ns h e er here this can create bi accu ulate r i absent l the hr nic sh uld be applied di iculties in the testin r the utc es d n t c n r t c n sense r e a ple s e che icals alth u h stable in the b ttle ill react rapidl r sl l in ater i in rise t de radati n pr ducts that a ha e di erent ata a aila ilit pr perties here such de radati n is rapid the a ailable test data ill requentl de ine the ha ard the de radati n pr ducts since it ill be these that ha e been tested hese data a be used t classi the parent substance in the he data used t classi a substance can be dra n r data required r re ulat r purp ses as ell n r al a e er here de radati n is sl er it a be p ssible t test the parent substance and thus enerate as the rele ant literature alth u h a nu ber internati nall rec ni ed data bases e ist hich can act as a d ha ard data in the n r al anner he subsequent de radati n a then be c nsidered in deter inin hether a sh rt startin p int uch databases ar idel in qualit and c prehensi eness and it is unli el that an ne database ter acute r l n ter chr nic ha ard cate r sh uld appl here a be ccasi ns h e er hen a substance ill h ld all he in r ati n necessar r classi icati n t be ade e databases speciali e in aquatic t icit and s tested a de rade t i e rise t a re ha ard us pr duct n these circu stances the classi icati n the parent thers in en ir n ental ate here is an bli ati n n the che ical supplier t a e the necessar searches and chec s sh uld ta e due acc unt the ha ard the de radati n pr duct and the rate at hich it can be r ed under n r al t deter ine the e tent and qualit the data a ailable and t use it in assi nin the appr priate ha ard cate r en ir n ental c nditi ns ata alit tic to icit he precise use the a ailable data ill be described in the rele ant secti n but as a eneral rule nt o ction data enerated t standard internati nal uidelines and t is t be pre erred er ther t pes data quall h e er it is i p rtant t appreciate that classi icati n can be ade based n the best a ailable data hus i n data is he basis r the identi icati n ha ard t the aquatic en ir n ent r a substance is the aquatic t icit a ailable hich c n r s t the qualit standard detailed ab e classi icati n can still be ade pr ided the data used that substance lassi icati n is predicated n ha in t icit data r ish crustacea and al ae aquatic plant a ailable is n t c nsidered in alid assist this pr cess a qualit sc rin uide has been de el ped and used e tensi el in a hese ta a are enerall accepted as representati e aquatic auna and l ra r ha ard identi icati n ata n these nu ber ra and enerall c n r s t the ll in cate ries particular ta a are re li el t be und because this eneral acceptance b re ulat r auth rities and the che ical a ata deri ed r icial data s urces that ha e been alidated b re ulat r auth rities such industr ther in r ati n n the de radati n and bi accu ulati n beha i ur is used t better delineate the aquatic ha ard as ater ualit n raphs ater ualit riteria hese data can be his secti n describes the appr priate tests r ec t icit pr ides s e basic c ncepts in e aluatin the data and usin c nsidered as alid r classi icati n purp ses assu pti n sh uld be ade that these are c binati ns testin results r classi icati n su ari es appr aches r dealin ith di icult substances and includes the nl data a ailable h e er and due re ard sh uld be i en t the date the rele ant a brie discussi n n interpretati n data qualit rep rt e l a ailable data a n t ha e been c nsidered esc iption of tests b ata deri ed r rec ni ed internati nal uidelines e uidelines r nati nal uidelines equi alent qualit ub ect t the data interpretati n issues raised in the ll in r classi in substances in the har ni ed s ste resh ater and arine species t icit data can be secti ns these data can be used r classi icati n c nsidered as equi alent data t sh uld be n ted that s e t pes substances e i ni able r anic che icals r
- 454 - Copyright@United Nations, 2017. All rights reserved
he ater s lubilit and stabilit data alth u h n t used directl in the criteria are ne ertheless c ata deri ed r testin hich hile n t strictl acc rdin t a uideline detailed ab e i p rtant since the are a aluable help in the data interpretati n the ther pr perties see ll s accepted scienti ic principles and pr cedures and r has been peer re ie ed pri r t publicati n r such data here all the e peri ental detail is n t rec rded s e ud e ent classi a re ie sh uld irst be ade the a ailable aquatic t icit data t ill be necessar t a be required t deter ine alidit r all such data a be used ithin the c nsider all the a ailable data and select th se hich eet the necessar qualit criteria r classi icati n there are n classi icati n sche e data a ailable that eet the qualit criteria required b the internati nall standardi ed eth ds it ill be necessar t d ata deri ed r testin pr cedures hich de iate si ni icantl r standard uidelines and e a ine an a ailable data t deter ine hether a classi icati n can be ade the data indicate that the acute aquatic are c nsidered as unreliable sh uld n t be used in classi icati n t icit is reater than l r s luble substances and the chr nic aquatic t icit is reater than l then the substance is n t classi ied as ha ard us here are a nu ber cases here n e ects are bser ed in the test e data he circu stances use and alidit data are discussed in the rele ant and the aquatic t icit is thus rec rded as a ater s lubilit alue i e there is n acute t icit ithin the ran e secti ns the ater s lubilit in the test edia here this is the case and the ater s lubilit in the test edia is l a ain ata deri ed r sec ndar s urces such as handb s re ie s citati n etc here the data n classi icati n need be applied qualit cann t be directl e aluated uch data sh uld be e a ined here data r qualit chr nic aquatic t icit data are a ailable cut alues ill depend n hether the substance is rapidl and are n t a ailable t deter ine hether it can be used uch data sh uld ha e su icient de radable r n t here re r n n rapidl de radable substances and th se r hich n in r ati n n de radati n detail t all qualit t be assessed n deter inin the acceptabilit these data r the is a ailable the cut le els are hi her than r th se substances here rapid de radabilit can be c n ir ed see purp ses classi icati n due re ard sh uld be i en t the di iculties in testin that a hapter ables and ha e a ected data qualit and the si ni icance the rep rted result in ter s the le el ha ard identi ied see here the l est acute aquatic t icit data are bel l and n adequate chr nic t icit data lassi icati n a als be ade n inc plete t icit data sets e here data are n t a ailable n are a ailable it is necessar t irst decide hich ha ard cate r the t icit alls in and then t deter ine hether the all three tr phic levels. In these cases, the classification may be considered as “provisional” and subject to further chr nic and r the acute sub class sh uld be applied his can si pl be achie ed b e a inin the a ailable data n in r ati n bec in a ailable n eneral all the data a ailable ill need t be c nsidered pri r t assi nin a the partiti n c e icient l and the a ailable data n de radati n either the l r the substance cann t classi icati n here d qualit data are n t a ailable l er qualit data ill need t be c nsidered n these be c nsidered as rapidl de radable then the appr priate l n ter chr nic ha ard cate r and the c rresp ndin circu stances a ud e ent ill need t be ade re ardin the true le el ha ard r e a ple here d qualit acute sh rt ter ha ard cate r are applied independentl t sh uld be n ted that alth u h the l is the st data are a ailable r a particular species r ta a this sh uld be used in pre erence t an l er qualit data hich readil a ailable indicati n a p tential t bi accu ulate an e peri entall deri ed is pre erred here this is i ht als be a ailable r that species r ta a e er d qualit data a n t al a s be a ailable r all the basic a ailable this sh uld be used rather than the partiti n c e icient n these circu stances a uld indicate data set tr phic le els t ill be necessar t c nsider data l er qualit r th se tr phic le els r hich d bi accu ulati n su icient t classi in the appr priate l n ter chr nic ha ard cate r the substance is b th qualit data are n t a ailable nsiderati n such data h e er ill als need t c nsider the di iculties that a rapidl de radable and has a l p tential t bi accu ulate r i absent l then it sh uld n t be ha e a ected the li elih d achie in a alid result r e a ple the test details and e peri ental desi n a be assi ned t a l n ter chr nic ha ard cate r unless the chr nic t icit data indicate ther ise see critical t the assess ent the usabilit s e data such as that r h dr l ticall unstable che icals hile less s r ther che icals uch di iculties are described urther in ecti n r p rl s luble substances enerall spea in th se ith a ater s lubilit in the test edia l r hich n aquatic t icit has been und sh uld be urther e a ined t deter ine hether hr nic r all the identi icati n ha ard and hence the classi icati n ill be based n in r ati n ate r needs t be applied hus i the substance is b th n t rapidl de radable and has a p tential t directl btained r testin the substance bein c nsidered here are ccasi ns h e er here this can create bi accu ulate r i absent l the hr nic sh uld be applied di iculties in the testin r the utc es d n t c n r t c n sense r e a ple s e che icals alth u h stable in the b ttle ill react rapidl r sl l in ater i in rise t de radati n pr ducts that a ha e di erent ata a aila ilit pr perties here such de radati n is rapid the a ailable test data ill requentl de ine the ha ard the de radati n pr ducts since it ill be these that ha e been tested hese data a be used t classi the parent substance in the he data used t classi a substance can be dra n r data required r re ulat r purp ses as ell n r al a e er here de radati n is sl er it a be p ssible t test the parent substance and thus enerate as the rele ant literature alth u h a nu ber internati nall rec ni ed data bases e ist hich can act as a d ha ard data in the n r al anner he subsequent de radati n a then be c nsidered in deter inin hether a sh rt startin p int uch databases ar idel in qualit and c prehensi eness and it is unli el that an ne database ter acute r l n ter chr nic ha ard cate r sh uld appl here a be ccasi ns h e er hen a substance ill h ld all he in r ati n necessar r classi icati n t be ade e databases speciali e in aquatic t icit and s tested a de rade t i e rise t a re ha ard us pr duct n these circu stances the classi icati n the parent thers in en ir n ental ate here is an bli ati n n the che ical supplier t a e the necessar searches and chec s sh uld ta e due acc unt the ha ard the de radati n pr duct and the rate at hich it can be r ed under n r al t deter ine the e tent and qualit the data a ailable and t use it in assi nin the appr priate ha ard cate r en ir n ental c nditi ns ata alit tic to icit he precise use the a ailable data ill be described in the rele ant secti n but as a eneral rule nt o ction data enerated t standard internati nal uidelines and t is t be pre erred er ther t pes data quall h e er it is i p rtant t appreciate that classi icati n can be ade based n the best a ailable data hus i n data is he basis r the identi icati n ha ard t the aquatic en ir n ent r a substance is the aquatic t icit a ailable hich c n r s t the qualit standard detailed ab e classi icati n can still be ade pr ided the data used that substance lassi icati n is predicated n ha in t icit data r ish crustacea and al ae aquatic plant a ailable is n t c nsidered in alid assist this pr cess a qualit sc rin uide has been de el ped and used e tensi el in a hese ta a are enerall accepted as representati e aquatic auna and l ra r ha ard identi icati n ata n these nu ber ra and enerall c n r s t the ll in cate ries particular ta a are re li el t be und because this eneral acceptance b re ulat r auth rities and the che ical a ata deri ed r icial data s urces that ha e been alidated b re ulat r auth rities such industr ther in r ati n n the de radati n and bi accu ulati n beha i ur is used t better delineate the aquatic ha ard as ater ualit n raphs ater ualit riteria hese data can be his secti n describes the appr priate tests r ec t icit pr ides s e basic c ncepts in e aluatin the data and usin c nsidered as alid r classi icati n purp ses assu pti n sh uld be ade that these are c binati ns testin results r classi icati n su ari es appr aches r dealin ith di icult substances and includes the nl data a ailable h e er and due re ard sh uld be i en t the date the rele ant a brie discussi n n interpretati n data qualit rep rt e l a ailable data a n t ha e been c nsidered esc iption of tests b ata deri ed r rec ni ed internati nal uidelines e uidelines r nati nal uidelines equi alent qualit ub ect t the data interpretati n issues raised in the ll in r classi in substances in the har ni ed s ste resh ater and arine species t icit data can be secti ns these data can be used r classi icati n c nsidered as equi alent data t sh uld be n ted that s e t pes substances e i ni able r anic che icals r
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or anometallic substances may e press different to icities in fresh ater and marine environments. ince the purpose of Crustacea tests classification is to characteri e ha ard in the a uatic environment, the result sho in the hi hest to icity should be chosen. cute testin . . . he criteria for determinin health and environmental ha ards should be test method neutral, allo in different approaches as lon as they are scientifically sound and validated accordin to international procedures cute tests ith crustacea enera be in ith irst instar u eni es or a hni s, a test uration o hours is use and criteria already referred to in e istin systems for the endpoints of concern and produce mutually acceptable data. or other crustacea, such as si s or others, a uration o hours is t ica he obser ationa en oint is orta it ccordin to the proposed system or i obi i ation as a surro ate to orta it obi i ation is e ine as unres onsi e to ent e ro in ests consistent ith est ui e ine art a hnia acute or si acute to icit “Acute toxicity would normally be determined using a fish 96 hour LC50 (OECD Test Guideline or or their e ui a ents shou be use or c assi ication equi alent a crustacea species hour Test Guideline or equi alent and or an algal species or hour Test Guideline or equi alent . These species are considered as hronic testin surrogate for all aquatic organisms and data on other species such as the duck eed emna may also be considered if the test methodology is suitable.” hronic tests ith crustacea a so enera be in ith irst instar u eni es an continue throu h aturation an re ro uction or a hni s, a s is su icient or aturation an the ro uction o broo s or si s, a s hronic testin enerally involves an e posure that is lin erin or continues for a lon er time the term is necessar bser ationa en oints inc u e ti e to irst broo , nu ber o o s rin ro uce er e a e, ro th, an can si nify periods from days to a year, or more dependin on the reproductive cycle of the a uatic or anism. hronic tests sur i a t is reco en e that tests consistent ith est ui e ine art a hnia re ro uction or can be done to assess certain endpoints relatin to ro th, survival, reproduction and development. si chronic or their e ui a ents be use in the c assi ication sche e
“Chronic toxicity data are less available than acute data and the range of testing procedures less Algae lant tests standardised. ata generated according to the Test Guidelines ish arly ife Stage art or aphnia Reproduction and lgal Gro th nhibition can be accepted. ther ests in a ae alidated and internationally accepted tests could also be used. The N s or other equi alent should be used.” ae are cu ture an e ose to the test substance in a nutrient enriche e iu ests consistent ith est ui e ine a ro th inhibition shou be use tan ar test etho s e o a ce ensit in the n document describes the main statistical methods for the analysis of data of standardi ed inocu u in or er to ensure e onentia ro th throu h the test, usua to a s uration ecoto icity tests . he a a test is a short ter test that ro i es both acute an chronic en oints he re erre . . . It should be noted that several of the uidelines cited as e amples for classification are bein obser ationa en oint in this stu is a a ro th rate inhibition because it is not e en ent on the test esi n, hereas revised or are bein planned for updatin . uch revisions may lead to minor modifications of test conditions. herefore, the bio ass e en s both on ro th rate o the test s ecies as e as test uration an other e e ents o test esi n the e pert roup that developed the harmoni ed criteria for classification intended some fle ibility in test duration or even en oint is re orte on as re uction in bio ass or is not s eci ie , then this a ue a be inter rete as an e ui a ent species used. en oint
. . . uidelines for conductin acceptable tests ith fish, crustacea, and al ae can be found in many sources ests in a uatic acro h tes , , , I . he mono raph o. , etailed evie aper on uatic o icity estin for Industrial hemicals and esticides, is a ood compilation of pela ic test methods and sources of he ost co on use ascu ar ants or a uatic to icit tests are uc ee s e na ibba an testin uidance. his document is also a source of appropriate test methodolo ies. e na inor he e na test is a short ter test an , a thou h it ro i es both acute an sub chronic en oints, on the acute is use or c assi ication in the har oni e s ste he tests ast or u to a s an are er or e in nutrient . . . ish tests enriche e ia si i ar to that use or a ae, but a be increase in stren th he obser ationa en oint is base on chan e in the nu ber o ron s ro uce ests consistent ith est ui e ine on e na in re aration an . . . . cute testin a uatic ant to icit , e na shou be use
cute tests are enerally performed ith youn juveniles . in si e for a period of hours. he atic to icit concepts observational endpoint in these tests is mortality. ish lar er than this ran e and or durations shorter than hours are enerally less sensitive. o ever, for classification, they could be used if no acceptable data ith the smaller fish for his section a resses the use o acute an chronic to icit ata in c assi ication, an s ecia hours are available or the results of these tests ith different si e fish or test durations ould influence classification in a consi erations or e osure re i es, a a to icit testin , an use o s or a ore etai e iscussion o a uatic more ha ardous cate ory. ests consistent ith est uideline ish hour or e uivalent should be to icit conce ts, one can re er to an used for classification. Acute toxicity . . . . hronic testin cute to icit or ur oses o c assi ication re ers to the intrinsic ro ert o a substance to be in urious to hronic or lon term tests ith fish can be initiated ith fertili ed e s, embryos, juveniles, or an or anis in a short ter e osure to that substance cute to icit is enera e resse in ter s o a concentration reproductively active adults. ests consistent ith est uideline ish arly ife ta e , the fish life cycle test hich is etha to o the test or anis s , causes a easurab e a erse e ect to o the test or anis s e . , or e uivalent can be used in the classification scheme. urations can vary idely dependin on the test i obi i ation o a hni s , or ea s to a re uction in test treate or anis res onses ro contro untreate purpose any here from days to over days . bservational endpoints can include hatchin success, ro th len th or anis res onses e ro th rate in a ae and ei ht chan es , spa nin success, and survival. echnically, the uideline ish arly ife ta e is not a “chronic” test, but a sub chronic test on sensitive life sta es. It is idely accepted as a predictor of chronic to icity and is ubstances ith an acute to icit eter ine to be ess than one art er i ion are enera used as such for purposes of classification in the harmoni ed system. ish early life sta e to icity data are much more reco ni e as bein er to ic he han in , use, or ischar e into the en iron ent o these substances oses a hi h e ree available than fish life cycle or reproduction studies. o ha ar an the are c assi ie in hronic an or cute eci a ban s are acce te or cate ori in acute to icit abo e this cate or ubstances ith an acute to icit easure ro one to ten arts er i ion are c assi ie in cute , ro ten to one hun re arts er i ion are c assi ie in cute , an those o er one hun re arts er i ion are re ar e as ractica non to ic
- 456 - Copyright@United Nations, 2017. All rights reserved
or anometallic substances may e press different to icities in fresh ater and marine environments. ince the purpose of Crustacea tests classification is to characteri e ha ard in the a uatic environment, the result sho in the hi hest to icity should be chosen. cute testin . . . he criteria for determinin health and environmental ha ards should be test method neutral, allo in different approaches as lon as they are scientifically sound and validated accordin to international procedures cute tests ith crustacea enera be in ith irst instar u eni es or a hni s, a test uration o hours is use and criteria already referred to in e istin systems for the endpoints of concern and produce mutually acceptable data. or other crustacea, such as si s or others, a uration o hours is t ica he obser ationa en oint is orta it ccordin to the proposed system or i obi i ation as a surro ate to orta it obi i ation is e ine as unres onsi e to ent e ro in ests consistent ith est ui e ine art a hnia acute or si acute to icit “Acute toxicity would normally be determined using a fish 96 hour LC50 (OECD Test Guideline or or their e ui a ents shou be use or c assi ication equi alent a crustacea species hour Test Guideline or equi alent and or an algal species or hour Test Guideline or equi alent . These species are considered as hronic testin surrogate for all aquatic organisms and data on other species such as the duck eed emna may also be considered if the test methodology is suitable.” hronic tests ith crustacea a so enera be in ith irst instar u eni es an continue throu h aturation an re ro uction or a hni s, a s is su icient or aturation an the ro uction o broo s or si s, a s hronic testin enerally involves an e posure that is lin erin or continues for a lon er time the term is necessar bser ationa en oints inc u e ti e to irst broo , nu ber o o s rin ro uce er e a e, ro th, an can si nify periods from days to a year, or more dependin on the reproductive cycle of the a uatic or anism. hronic tests sur i a t is reco en e that tests consistent ith est ui e ine art a hnia re ro uction or can be done to assess certain endpoints relatin to ro th, survival, reproduction and development. si chronic or their e ui a ents be use in the c assi ication sche e
“Chronic toxicity data are less available than acute data and the range of testing procedures less Algae lant tests standardised. ata generated according to the Test Guidelines ish arly ife Stage art or aphnia Reproduction and lgal Gro th nhibition can be accepted. ther ests in a ae alidated and internationally accepted tests could also be used. The N s or other equi alent should be used.” ae are cu ture an e ose to the test substance in a nutrient enriche e iu ests consistent ith est ui e ine a ro th inhibition shou be use tan ar test etho s e o a ce ensit in the n document describes the main statistical methods for the analysis of data of standardi ed inocu u in or er to ensure e onentia ro th throu h the test, usua to a s uration ecoto icity tests . he a a test is a short ter test that ro i es both acute an chronic en oints he re erre . . . It should be noted that several of the uidelines cited as e amples for classification are bein obser ationa en oint in this stu is a a ro th rate inhibition because it is not e en ent on the test esi n, hereas revised or are bein planned for updatin . uch revisions may lead to minor modifications of test conditions. herefore, the bio ass e en s both on ro th rate o the test s ecies as e as test uration an other e e ents o test esi n the e pert roup that developed the harmoni ed criteria for classification intended some fle ibility in test duration or even en oint is re orte on as re uction in bio ass or is not s eci ie , then this a ue a be inter rete as an e ui a ent species used. en oint
. . . uidelines for conductin acceptable tests ith fish, crustacea, and al ae can be found in many sources ests in a uatic acro h tes , , , I . he mono raph o. , etailed evie aper on uatic o icity estin for Industrial hemicals and esticides, is a ood compilation of pela ic test methods and sources of he ost co on use ascu ar ants or a uatic to icit tests are uc ee s e na ibba an testin uidance. his document is also a source of appropriate test methodolo ies. e na inor he e na test is a short ter test an , a thou h it ro i es both acute an sub chronic en oints, on the acute is use or c assi ication in the har oni e s ste he tests ast or u to a s an are er or e in nutrient . . . ish tests enriche e ia si i ar to that use or a ae, but a be increase in stren th he obser ationa en oint is base on chan e in the nu ber o ron s ro uce ests consistent ith est ui e ine on e na in re aration an . . . . cute testin a uatic ant to icit , e na shou be use
cute tests are enerally performed ith youn juveniles . in si e for a period of hours. he atic to icit concepts observational endpoint in these tests is mortality. ish lar er than this ran e and or durations shorter than hours are enerally less sensitive. o ever, for classification, they could be used if no acceptable data ith the smaller fish for his section a resses the use o acute an chronic to icit ata in c assi ication, an s ecia hours are available or the results of these tests ith different si e fish or test durations ould influence classification in a consi erations or e osure re i es, a a to icit testin , an use o s or a ore etai e iscussion o a uatic more ha ardous cate ory. ests consistent ith est uideline ish hour or e uivalent should be to icit conce ts, one can re er to an used for classification. Acute toxicity . . . . hronic testin cute to icit or ur oses o c assi ication re ers to the intrinsic ro ert o a substance to be in urious to hronic or lon term tests ith fish can be initiated ith fertili ed e s, embryos, juveniles, or an or anis in a short ter e osure to that substance cute to icit is enera e resse in ter s o a concentration reproductively active adults. ests consistent ith est uideline ish arly ife ta e , the fish life cycle test hich is etha to o the test or anis s , causes a easurab e a erse e ect to o the test or anis s e . , or e uivalent can be used in the classification scheme. urations can vary idely dependin on the test i obi i ation o a hni s , or ea s to a re uction in test treate or anis res onses ro contro untreate purpose any here from days to over days . bservational endpoints can include hatchin success, ro th len th or anis res onses e ro th rate in a ae and ei ht chan es , spa nin success, and survival. echnically, the uideline ish arly ife ta e is not a “chronic” test, but a sub chronic test on sensitive life sta es. It is idely accepted as a predictor of chronic to icity and is ubstances ith an acute to icit eter ine to be ess than one art er i ion are enera used as such for purposes of classification in the harmoni ed system. ish early life sta e to icity data are much more reco ni e as bein er to ic he han in , use, or ischar e into the en iron ent o these substances oses a hi h e ree available than fish life cycle or reproduction studies. o ha ar an the are c assi ie in hronic an or cute eci a ban s are acce te or cate ori in acute to icit abo e this cate or ubstances ith an acute to icit easure ro one to ten arts er i ion are c assi ie in cute , ro ten to one hun re arts er i ion are c assi ie in cute , an those o er one hun re arts er i ion are re ar e as ractica non to ic
- 457 - Copyright@United Nations, 2017. All rights reserved
Chronic toxicity Exposure regimes
hronic to icit , or ur oses o c assi ication, re ers to the intrinsic ro ert o a substance to cause our t es o e osure con itions are e o e in both acute an chronic tests an in both resh ater an a erse e ects to a uatic or anis s urin e osures hich are eter ine in re ation to the i e c c e o the or anis sa t ater e ia static, static rene a se i static , recircu ation, an o throu h he choice or hich test t e to use uch chronic e ects usua inc u e a ran e o sub etha en oints an are enera e resse in ter s o a o bser ab e usua e en s on test substance characteristics, test uration, test s ecies, an re u ator re uire ents ect oncentration , or an e ui a ent bser ab e en oints t ica inc u e sur i a , ro th an or re ro uction hronic to icit e osure urations can ar i e e en in on test en oint easure an test s ecies Test media for algae use a tests are er or e in nutrient enriche e ia an the use o one co on constituent, , or or the c assi ication base on chronic to icit a i erentiation is a e bet een ra i e ra ab e other che ators, shou be consi ere care u hen testin the to icit o or anic che ica s, trace a ounts o a che ator an non ra i e ra ab e substances ubstances that o ra i e ra e are c assi ie in cate or hronic hen a i e are nee e to co e icronutrients in the cu ture e iu i o itte , a a ro th can be si ni icant chronic toxicity determined to be ≤ 0.01 mg/l. Decimal bands are accepted for categorizing chronic toxicity above this re uce an co ro ise test uti it o e er, che ators can re uce the obser e to icit o eta test substances cate or ubstances ith a chronic to icit easure ro to are c assi ie in cate or hronic or here ore, or eta co oun s, it is esirab e that ata ro tests ith hi h concentration o che ators an or tests chronic to icit , ro to are c assi ie in cate or hronic or chronic to icit , an those o er ith stoichio etrica e cess o che ator re ati e to iron shou be critica e a uate ree che ator a as hea are re ar e as ractica non to ic or substances that o not ra i e ra e or here no in or ation on ra i eta to icit consi erab , in articu ar ith stron che ators i e o e er, in the absence o a ai ab e iron in e radation is available two chronic categories are used: Chronic 1 when a chronic toxicity determined to be ≤ 0.1 mg/l the e iu the ro th o a ae can beco e iron i ite , an conse uent ata ro tests ith no or ith re uce iron an hronic hen chronic to icit is easure ro to an shou be treate ith caution
ince chronic to icit ata are ess co on in certain sectors than acute ata, or c assi ication se of A s sche es, the otentia or chronic to icit is, in absence o a e uate chronic to icit ata, i enti ie b a ro riate co binations o acute to icit , ac o e ra abi it an or the otentia or actua bioaccu u ation o e er, here or ur ose o c assi ication, an in the absence o e eri enta ata, s can be re ie u on to a e uate chronic to icit ata e ist, this sha be use in re erence o er the c assi ication base on the co bination o ro i e re ictions o acute to icit or ish, a hnia, an a ae or non e ectro te, non e ectro hi ic, an other ise non acute to icit ith e ra abi it an or bioaccu u ation n this conte t, the o o in enera a roach shou be use reacti e substances ee ection on se of A rob e s re ain or substances such as or ano hos hates hich o erate b eans o s ecia echanis s such as unctiona rou s hich interact ith bio o ica rece tors, or hich can a a e uate chronic to icit ata are a ai ab e or a three tro hic e e s this can be use or su h r bon s ith ce u ar roteins e iab e s ha e been eri e or che ica s actin b a basic narcosis irect to eter ine an a ro riate on ter chronic ha ar cate or echanis hese che ica s are none ectro tes o o reacti it such as h rocarbons, a coho s, etones an certain a i hatic ch orinate h rocarbons hich ro uce their bio o ica e ects as a unction o their artition coe icients er b a e uate chronic to icit ata are a ai ab e or one or t o tro hic e e s, it shou be or anic che ica can ro uce narcosis o e er, i the che ica is an e ectro te or contains s eci ic unctiona rou s e a ine i acute to icit ata are a ai ab e or the other tro hic e e s otentia ea in to non narcotic echanis s as e , an ca cu ations o to icit base on artition coe icient a one ou se ere c assi ication is a e or the tro hic e e s ith chronic ata an co are ith that a e un eresti ate the to icit s or acute a uatic to icit o arent co oun s cannot be use to re ict the e ects o usin the acute to icit ata or the other tro hic e e s he ina c assi ication sha be a e to ic etabo ites or e ra ates, hen these arise a ter a on er ti e erio than the uration o acute tests accor in to the ost strin ent outco e ei t of e i ence c n or er to re o e or o er a chronic c assi ication, usin chronic to icit ata, it ust be e onstrate that the s or e ui a ent use ou be suitab e to re o e or o er he best ua it ata shou be use as the un a enta basis or c assi ication assi ication shou the concern or a ta a hich resu te in c assi ication base on acute ata in co bination ith re erab be base on ri ar ata sources t is essentia that test con itions are c ear an co ete articu ate e ra abi it , an or bioaccu u ation his can o ten be achie e b usin a on ter or the ost sensiti e s ecies i enti ie b the acute to icit hus, i a c assi ication has been here u ti e stu ies or a ta ono ic rou are a ai ab e, a ecision on hat is the ost sensiti e an base on a ish acute , it ou enera not be ossib e to re o e or o er this hi hest ua it ust be a e u e ent has to be a e on a case b case basis hether a non stu ith a ore c assi ication usin a on ter ro an in ertebrate to icit test n this case, the sensiti e obser ation is use in ieu o a stu t ou a ear that resu ts that in icate hi h to icit ro tests ou nor a nee to be eri e ro a on ter ish test o the sa e s ecies or one er or e accor in to non stan ar or non ui e ines shou be ab e to be use or c assi ication, hereas stu ies, o e ui a ent or reater sensiti it ua , i c assi ication has resu te ro the acute hich e onstrate ne i ib e to icit , ou re uire ore care u consi eration ubstances, hich are i icu t to test, a to icit to ore than one ta a, it is i e that s ro each ta a i be nee e n case ie a arent resu ts that are ore or ess se ere than the true to icit ert u e ent ou a so be nee e or o c assi ication o a substance as hronic , su icient e i ence shou be ro i e that the c assi ication in these cases or e ui a ent or each ta a is reater than or reater than the ater so ubi it o the substances un er consi eration here ore than one acce tab e test is a ai ab e or the sa e ta ono ic rou , the ost sensiti e the one ith the o est or is enera use or c assi ication o e er, this ust be ea t ith on a case b estin ith a ae e na cannot be use or re o in or o erin a c assi ication because case basis hen ar er ata sets or ore a ues are a ai ab e or the sa e s ecies, the eo etric ean o to icit a ues a be use as the re resentati e to icit a ue or that s ecies n esti atin a ean a ue, it is not a isab e to co bine a the a ae an e na tests are not on ter stu ies tests o i erent s ecies ithin a ta a rou or in i erent i e sta es or teste un er i erent con itions or uration b the acute to chronic ratio is enera narro an iffic lt to test s stances c the en oints are ore consistent ith the acute en oints or other or anis s a i a uatic to icit tests re uire the isso ution o the test substance in the ater e ia un er the test o e er here c assi ication is a ie so e ue to the acute to icit obser e in sin e con itions reco en e b the ui e ine n a ition, a bioa ai ab e e osure concentration shou be aintaine or the a ae a uatic ant tests, but there is e i ence ro a ran e o other a ae tests that the chronic to icit s or uration o the test o e substances are i icu t to test in a uatic s ste s an ui ance has been e e o e to assist in this ta ono ic rou is in the to icit ban corres on in to a ess strin ent c assi ication cate or or abo e , this testin these ateria s o an ui ance ocu ent on a uatic to icit e i ence cou be use to consi er re o in or o erin a c assi ication t resent this a roach cannot be a ie to testin o i icu t substances an i tures , is a oo source o in or ation on the t es o substances that are a uatic ants since no stan ar i e chronic to icit tests ha e been e e o e i icu t to test an the ste s nee e to ensure a i conc usions ro tests ith these ateria s
e erthe ess, uch test ata e ist that a ha e use testin etho o o ies hich, hi e not in con or it ith hat i ht be consi ere best ractice to a , can sti ie in or ation suitab e or a ication o the
- 458 - Copyright@United Nations, 2017. All rights reserved
Chronic toxicity Exposure regimes
hronic to icit , or ur oses o c assi ication, re ers to the intrinsic ro ert o a substance to cause our t es o e osure con itions are e o e in both acute an chronic tests an in both resh ater an a erse e ects to a uatic or anis s urin e osures hich are eter ine in re ation to the i e c c e o the or anis sa t ater e ia static, static rene a se i static , recircu ation, an o throu h he choice or hich test t e to use uch chronic e ects usua inc u e a ran e o sub etha en oints an are enera e resse in ter s o a o bser ab e usua e en s on test substance characteristics, test uration, test s ecies, an re u ator re uire ents ect oncentration , or an e ui a ent bser ab e en oints t ica inc u e sur i a , ro th an or re ro uction hronic to icit e osure urations can ar i e e en in on test en oint easure an test s ecies Test media for algae use a tests are er or e in nutrient enriche e ia an the use o one co on constituent, , or or the c assi ication base on chronic to icit a i erentiation is a e bet een ra i e ra ab e other che ators, shou be consi ere care u hen testin the to icit o or anic che ica s, trace a ounts o a che ator an non ra i e ra ab e substances ubstances that o ra i e ra e are c assi ie in cate or hronic hen a i e are nee e to co e icronutrients in the cu ture e iu i o itte , a a ro th can be si ni icant chronic toxicity determined to be ≤ 0.01 mg/l. Decimal bands are accepted for categorizing chronic toxicity above this re uce an co ro ise test uti it o e er, che ators can re uce the obser e to icit o eta test substances cate or ubstances ith a chronic to icit easure ro to are c assi ie in cate or hronic or here ore, or eta co oun s, it is esirab e that ata ro tests ith hi h concentration o che ators an or tests chronic to icit , ro to are c assi ie in cate or hronic or chronic to icit , an those o er ith stoichio etrica e cess o che ator re ati e to iron shou be critica e a uate ree che ator a as hea are re ar e as ractica non to ic or substances that o not ra i e ra e or here no in or ation on ra i eta to icit consi erab , in articu ar ith stron che ators i e o e er, in the absence o a ai ab e iron in e radation is available two chronic categories are used: Chronic 1 when a chronic toxicity determined to be ≤ 0.1 mg/l the e iu the ro th o a ae can beco e iron i ite , an conse uent ata ro tests ith no or ith re uce iron an hronic hen chronic to icit is easure ro to an shou be treate ith caution
ince chronic to icit ata are ess co on in certain sectors than acute ata, or c assi ication se of A s sche es, the otentia or chronic to icit is, in absence o a e uate chronic to icit ata, i enti ie b a ro riate co binations o acute to icit , ac o e ra abi it an or the otentia or actua bioaccu u ation o e er, here or ur ose o c assi ication, an in the absence o e eri enta ata, s can be re ie u on to a e uate chronic to icit ata e ist, this sha be use in re erence o er the c assi ication base on the co bination o ro i e re ictions o acute to icit or ish, a hnia, an a ae or non e ectro te, non e ectro hi ic, an other ise non acute to icit ith e ra abi it an or bioaccu u ation n this conte t, the o o in enera a roach shou be use reacti e substances ee ection on se of A rob e s re ain or substances such as or ano hos hates hich o erate b eans o s ecia echanis s such as unctiona rou s hich interact ith bio o ica rece tors, or hich can a a e uate chronic to icit ata are a ai ab e or a three tro hic e e s this can be use or su h r bon s ith ce u ar roteins e iab e s ha e been eri e or che ica s actin b a basic narcosis irect to eter ine an a ro riate on ter chronic ha ar cate or echanis hese che ica s are none ectro tes o o reacti it such as h rocarbons, a coho s, etones an certain a i hatic ch orinate h rocarbons hich ro uce their bio o ica e ects as a unction o their artition coe icients er b a e uate chronic to icit ata are a ai ab e or one or t o tro hic e e s, it shou be or anic che ica can ro uce narcosis o e er, i the che ica is an e ectro te or contains s eci ic unctiona rou s e a ine i acute to icit ata are a ai ab e or the other tro hic e e s otentia ea in to non narcotic echanis s as e , an ca cu ations o to icit base on artition coe icient a one ou se ere c assi ication is a e or the tro hic e e s ith chronic ata an co are ith that a e un eresti ate the to icit s or acute a uatic to icit o arent co oun s cannot be use to re ict the e ects o usin the acute to icit ata or the other tro hic e e s he ina c assi ication sha be a e to ic etabo ites or e ra ates, hen these arise a ter a on er ti e erio than the uration o acute tests accor in to the ost strin ent outco e ei t of e i ence c n or er to re o e or o er a chronic c assi ication, usin chronic to icit ata, it ust be e onstrate that the s or e ui a ent use ou be suitab e to re o e or o er he best ua it ata shou be use as the un a enta basis or c assi ication assi ication shou the concern or a ta a hich resu te in c assi ication base on acute ata in co bination ith re erab be base on ri ar ata sources t is essentia that test con itions are c ear an co ete articu ate e ra abi it , an or bioaccu u ation his can o ten be achie e b usin a on ter or the ost sensiti e s ecies i enti ie b the acute to icit hus, i a c assi ication has been here u ti e stu ies or a ta ono ic rou are a ai ab e, a ecision on hat is the ost sensiti e an base on a ish acute , it ou enera not be ossib e to re o e or o er this hi hest ua it ust be a e u e ent has to be a e on a case b case basis hether a non stu ith a ore c assi ication usin a on ter ro an in ertebrate to icit test n this case, the sensiti e obser ation is use in ieu o a stu t ou a ear that resu ts that in icate hi h to icit ro tests ou nor a nee to be eri e ro a on ter ish test o the sa e s ecies or one er or e accor in to non stan ar or non ui e ines shou be ab e to be use or c assi ication, hereas stu ies, o e ui a ent or reater sensiti it ua , i c assi ication has resu te ro the acute hich e onstrate ne i ib e to icit , ou re uire ore care u consi eration ubstances, hich are i icu t to test, a to icit to ore than one ta a, it is i e that s ro each ta a i be nee e n case ie a arent resu ts that are ore or ess se ere than the true to icit ert u e ent ou a so be nee e or o c assi ication o a substance as hronic , su icient e i ence shou be ro i e that the c assi ication in these cases or e ui a ent or each ta a is reater than or reater than the ater so ubi it o the substances un er consi eration here ore than one acce tab e test is a ai ab e or the sa e ta ono ic rou , the ost sensiti e the one ith the o est or is enera use or c assi ication o e er, this ust be ea t ith on a case b estin ith a ae e na cannot be use or re o in or o erin a c assi ication because case basis hen ar er ata sets or ore a ues are a ai ab e or the sa e s ecies, the eo etric ean o to icit a ues a be use as the re resentati e to icit a ue or that s ecies n esti atin a ean a ue, it is not a isab e to co bine a the a ae an e na tests are not on ter stu ies tests o i erent s ecies ithin a ta a rou or in i erent i e sta es or teste un er i erent con itions or uration b the acute to chronic ratio is enera narro an iffic lt to test s stances c the en oints are ore consistent ith the acute en oints or other or anis s a i a uatic to icit tests re uire the isso ution o the test substance in the ater e ia un er the test o e er here c assi ication is a ie so e ue to the acute to icit obser e in sin e con itions reco en e b the ui e ine n a ition, a bioa ai ab e e osure concentration shou be aintaine or the a ae a uatic ant tests, but there is e i ence ro a ran e o other a ae tests that the chronic to icit s or uration o the test o e substances are i icu t to test in a uatic s ste s an ui ance has been e e o e to assist in this ta ono ic rou is in the to icit ban corres on in to a ess strin ent c assi ication cate or or abo e , this testin these ateria s o an ui ance ocu ent on a uatic to icit e i ence cou be use to consi er re o in or o erin a c assi ication t resent this a roach cannot be a ie to testin o i icu t substances an i tures , is a oo source o in or ation on the t es o substances that are a uatic ants since no stan ar i e chronic to icit tests ha e been e e o e i icu t to test an the ste s nee e to ensure a i conc usions ro tests ith these ateria s
e erthe ess, uch test ata e ist that a ha e use testin etho o o ies hich, hi e not in con or it ith hat i ht be consi ere best ractice to a , can sti ie in or ation suitab e or a ication o the
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c assi ication criteria uch ata re uire s ecia ui ance on inter retation, a thou h u ti ate , e ert u e ent ust be n ost i icu t to test con itions, the actua test concentration is i e to be ess than the no ina or use in eter inin ata a i it uch i icu t to test substances a be oor so ub e, o ati e, or sub ect to ra i e ecte test concentration here acute to icities s are esti ate to be or a i icu t to test substance, e ra ation ue to such rocesses as hototrans or ation, h ro sis, o i ation, or biotic e ra ation hen testin a ae, one can be air con i ent the c assi ication in the cute an hronic i a ro riate is arrante o e er, i the co oure ateria s a inter ere ith the test en oint b attenuatin the i ht nee e or ce ro th n a si i ar anner, esti ate acute to icit is reater than , the esti ate to icit is i e to un er re resent the to icit n these substances teste as c ou is ersions abo e so ubi it a i e rise to a se to icit easure ents oa in o the ater circu stances, e ert u e ent is nee e to eter ine the acce tabi it o a test ith a i icu t to test substance or use in co u n ith test ateria can be an issue or articu ates or so i s such as eta s etro eu isti ate ractions can a so ose c assi ication here the nature o the testin i icu t is be ie e to ha e a si ni icant in uence on the actua test oa in rob e s, as e as i icu t inter retationa rob e s hen eci in on the a ro riate concentrations or concentration hen acute to icit is esti ate to be reater than an the test concentration is not easure , then the eter inin a ues he ra t ui ance ocu ent on uatic o icit estin o i icu t ubstances an test shou be use ith ue caution in c assi ication i tures escribes the ore co on ro erties o an t es o substances hich are i e to ose testin i icu ties he o o in ara ra hs ro i e so e etai e ui ance on so e o these inter retationa rob e s n a tabi it test che ica concentrations are e ecte to a be o o no ina , testin , in oin so it shou be re e bere that this is ui ance an har an ast ru es cannot be a ie he nature o an o the or er to be a i , a re uire e osure re i es hich ro i e or rene a o the test ateria i icu ties ean that e ert u e ent ust a a s be a ie both in eter inin hether there is su icient in or ation e i static or o throu h con itions are re erre ecia rob e s arise, there ore, ith in a test or a u e ent to be a e on its a i it , an a so hether a to icit e e can be eter ine suitab e or use in res ect to testin on a ae, here the stan ar ui e ines enera inc u e static tests to be a in the c assi ication criteria con ucte hi e a ternati e e osure re i es are ossib e or crustacea an ish, these tests are re uent con ucte on static con itions as inc u e in the internationa a ree ui e ines n nstable substances these tests, a certain e e o e ra ation as e as other re e ant actors ha e to be to erate an a ro riate account ust be ta en in ca cu ations o to ic concentrations o e a roaches on hi e testin roce ures shou i ea ha e been a o te hich ini i e the i acts o instabi it in ho this can be ea t ith are co ere in here e ra ation occurs, it is a so i ortant the test e ia, in ractice, in certain tests, it can be a ost i ossib e to aintain a concentration throu hout the test to consi er the in uence o the to icit o the e ra ation ro ucts on the recor e to icit in the o on causes o such instabi it are o i ation, h ro sis, hoto e ra ation an bio e ra ation hi e the atter or s o test ert u e ent i nee to be e ercise hen eci in i the ata can be use or e ra ation can ore rea i be contro e , such contro s are re uent absent in uch e istin testin e erthe ess, or c assi ication so e testin , articu ar acute an chronic ish to icit testin , a choice o e osure re i es is a ai ab e to he ini i e osses ue to instabi it , an this shou be ta en into account in eci in on the test ata a i it b e ra ation hen a co oun brea s o n or e ra es un er test con ition, e ert u e ent shou be use in ca cu atin to icit or c assi ication, inc u in consi eration o here instabi it is a actor in eter inin the e e o e osure urin the test, an essentia rere uisite no n or i e brea o n ro ucts oncentrations o the arent ateria an a si ni icant or ata inter retation is the e istence o easure e osure concentrations at suitab e ti e oints throu hout the test n the to ic e ra ates are esirab e e ra ates are e ecte to be re ati e non to ic, rene ab e absence o ana tica easure concentrations at east at the start an en o test, no a i inter retation can be a e an e osure re i es are esirab e in or er to ensure that e e s o the arent co oun s are the test shou be consi ere as in a i or c assi ication ur oses here easure ata are a ai ab e, a nu ber o ractica aintaine ru es can be consi ere b a o ui ance in inter retation
c aturation or sin e co onent substances, c assi ication shou be base on on to ic a here easure ata are a ai ab e or the start an en o test as is nor a or the acute a hnia res onses obser e in the so ub e ran e, an not on tota che ica oa in abo e so ubi it an a a tests , the , or c assi ication ur oses, a be ca cu ate base on the eo etric re uent , ata are a ai ab e hich in icate to icit at e e s in e cess o ater so ubi it an , ean o the start an en o test concentrations here the en o test concentrations are be o the hi e these ata i o ten be re ar e as not a i , so e inter retation a be ossib e hese ana tica etection i it, such concentrations sha be consi ere to be ha that etection i it rob e s enera a hen testin oor so ub e substances, an ui ance on ho to inter ret such ata is inc u e in see a so the ui ance ocu ent on a uatic to icit testin o b here easure ata are a ai ab e at the start an en o e ia rene a erio s as a be i icu t substances an i tures a ai ab e or the se i static tests , the eo etric ean or each rene a erio shou be ca cu ate , an the ean e osure o er the ho e e osure erio ca cu ate ro these ata erturbation o test e ia ecia ro isions a be nee e to ensure isso ution o i icu t to c here the to icit can be attribute to a e ra ation brea o n ro uct, an the concentrations o test substances uch easures shou not ea to si ni icant chan es in the test e ia hen such this are no n, the or c assi ication ur oses, a be ca cu ate base on the eo etric chan es are i e to ea to an increase or ecrease in the a arent to icit an hence the ean o the e ra ation ro uct concentration, bac ca cu ate to the arent substance c assi ication e e o the test substance si i ar rinci es a be a ie to easure ata in chronic to icit testin e o e substances an substances co ere b the c assi ication sche e are in act i tures, or hich easure ent o e osure concentrations is i icu t, an in so e cases oorly soluble substances i ossib e ubstances such as etro eu isti ate ractions, o ers, substances ith si ni icant e e s o i urities, etc can ose s ecia rob e s since the to ic concentration is hese substances, usua ta en to be those ith a so ubi it in ater , are re uent i icu t to i icu t to e ine an i ossib e to eri ica testin roce ures o ten re on the isso e in the test e ia, an the isso e concentrations i o ten ro e i icu t to easure at the o concentrations or ation o a ater o ub e raction or ater cco o ate raction an antici ate or an substances, the true so ubi it in the test e ia i be un no n, an i o ten be recor e as ata are re orte in ter s o oa in rates hese ata a be use in a in the etection i it in uri ie ater e erthe ess such substances can sho to icit , an here no to icit is oun , u e ent c assi ication criteria ust be a ie to hether the resu t can be consi ere a i or c assi ication u e ent shou err on the si e o caution an shou not un eresti ate the ha ar or c assi ication o or anic co oun s, it is esirab e to ha e stabi i e an ana tica easure test concentrations thou h easure concentrations are re erre , c assi ication a be base on no ina concentration ea , tests usin a ro riate isso ution techni ues an ith accurate easure concentrations ithin stu ies hen these are the on a i ata a ai ab e un er certain circu stances the ateria is i e to substantia the ran e o ater so ubi it shou be use here such test ata are a ai ab e, the shou be use in re erence to other e ra e or other ise be ost ro the ater co u n, care ust be ta en in ata inter retation an c assi ication shou be ata t is nor a , ho e er, articu ar hen consi erin o er ata, to in such substances ith to icit e e s recor e in one ta in the oss o the to icant urin the test into account, i re e ant an ossib e itiona , eta s resent their e cess o the ater so ubi it , or here the isso e e e s are be o the etection i it o the ana tica etho hus, in o n set o i icu ties an are iscusse se arate ab e ists se era ro erties o i icu t to test substances an both circu stances, it is not ossib e to eri the actua e osure concentrations usin easure ata here these are the their re e ance or c assi ication on ata a ai ab e on hich to c assi , so e ractica ru es can be consi ere b a o enera ui ance
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c assi ication criteria uch ata re uire s ecia ui ance on inter retation, a thou h u ti ate , e ert u e ent ust be n ost i icu t to test con itions, the actua test concentration is i e to be ess than the no ina or use in eter inin ata a i it uch i icu t to test substances a be oor so ub e, o ati e, or sub ect to ra i e ecte test concentration here acute to icities s are esti ate to be or a i icu t to test substance, e ra ation ue to such rocesses as hototrans or ation, h ro sis, o i ation, or biotic e ra ation hen testin a ae, one can be air con i ent the c assi ication in the cute an hronic i a ro riate is arrante o e er, i the co oure ateria s a inter ere ith the test en oint b attenuatin the i ht nee e or ce ro th n a si i ar anner, esti ate acute to icit is reater than , the esti ate to icit is i e to un er re resent the to icit n these substances teste as c ou is ersions abo e so ubi it a i e rise to a se to icit easure ents oa in o the ater circu stances, e ert u e ent is nee e to eter ine the acce tabi it o a test ith a i icu t to test substance or use in co u n ith test ateria can be an issue or articu ates or so i s such as eta s etro eu isti ate ractions can a so ose c assi ication here the nature o the testin i icu t is be ie e to ha e a si ni icant in uence on the actua test oa in rob e s, as e as i icu t inter retationa rob e s hen eci in on the a ro riate concentrations or concentration hen acute to icit is esti ate to be reater than an the test concentration is not easure , then the eter inin a ues he ra t ui ance ocu ent on uatic o icit estin o i icu t ubstances an test shou be use ith ue caution in c assi ication i tures escribes the ore co on ro erties o an t es o substances hich are i e to ose testin i icu ties he o o in ara ra hs ro i e so e etai e ui ance on so e o these inter retationa rob e s n a tabi it test che ica concentrations are e ecte to a be o o no ina , testin , in oin so it shou be re e bere that this is ui ance an har an ast ru es cannot be a ie he nature o an o the or er to be a i , a re uire e osure re i es hich ro i e or rene a o the test ateria i icu ties ean that e ert u e ent ust a a s be a ie both in eter inin hether there is su icient in or ation e i static or o throu h con itions are re erre ecia rob e s arise, there ore, ith in a test or a u e ent to be a e on its a i it , an a so hether a to icit e e can be eter ine suitab e or use in res ect to testin on a ae, here the stan ar ui e ines enera inc u e static tests to be a in the c assi ication criteria con ucte hi e a ternati e e osure re i es are ossib e or crustacea an ish, these tests are re uent con ucte on static con itions as inc u e in the internationa a ree ui e ines n nstable substances these tests, a certain e e o e ra ation as e as other re e ant actors ha e to be to erate an a ro riate account ust be ta en in ca cu ations o to ic concentrations o e a roaches on hi e testin roce ures shou i ea ha e been a o te hich ini i e the i acts o instabi it in ho this can be ea t ith are co ere in here e ra ation occurs, it is a so i ortant the test e ia, in ractice, in certain tests, it can be a ost i ossib e to aintain a concentration throu hout the test to consi er the in uence o the to icit o the e ra ation ro ucts on the recor e to icit in the o on causes o such instabi it are o i ation, h ro sis, hoto e ra ation an bio e ra ation hi e the atter or s o test ert u e ent i nee to be e ercise hen eci in i the ata can be use or e ra ation can ore rea i be contro e , such contro s are re uent absent in uch e istin testin e erthe ess, or c assi ication so e testin , articu ar acute an chronic ish to icit testin , a choice o e osure re i es is a ai ab e to he ini i e osses ue to instabi it , an this shou be ta en into account in eci in on the test ata a i it b e ra ation hen a co oun brea s o n or e ra es un er test con ition, e ert u e ent shou be use in ca cu atin to icit or c assi ication, inc u in consi eration o here instabi it is a actor in eter inin the e e o e osure urin the test, an essentia rere uisite no n or i e brea o n ro ucts oncentrations o the arent ateria an a si ni icant or ata inter retation is the e istence o easure e osure concentrations at suitab e ti e oints throu hout the test n the to ic e ra ates are esirab e e ra ates are e ecte to be re ati e non to ic, rene ab e absence o ana tica easure concentrations at east at the start an en o test, no a i inter retation can be a e an e osure re i es are esirab e in or er to ensure that e e s o the arent co oun s are the test shou be consi ere as in a i or c assi ication ur oses here easure ata are a ai ab e, a nu ber o ractica aintaine ru es can be consi ere b a o ui ance in inter retation
c aturation or sin e co onent substances, c assi ication shou be base on on to ic a here easure ata are a ai ab e or the start an en o test as is nor a or the acute a hnia res onses obser e in the so ub e ran e, an not on tota che ica oa in abo e so ubi it an a a tests , the , or c assi ication ur oses, a be ca cu ate base on the eo etric re uent , ata are a ai ab e hich in icate to icit at e e s in e cess o ater so ubi it an , ean o the start an en o test concentrations here the en o test concentrations are be o the hi e these ata i o ten be re ar e as not a i , so e inter retation a be ossib e hese ana tica etection i it, such concentrations sha be consi ere to be ha that etection i it rob e s enera a hen testin oor so ub e substances, an ui ance on ho to inter ret such ata is inc u e in see a so the ui ance ocu ent on a uatic to icit testin o b here easure ata are a ai ab e at the start an en o e ia rene a erio s as a be i icu t substances an i tures a ai ab e or the se i static tests , the eo etric ean or each rene a erio shou be ca cu ate , an the ean e osure o er the ho e e osure erio ca cu ate ro these ata erturbation o test e ia ecia ro isions a be nee e to ensure isso ution o i icu t to c here the to icit can be attribute to a e ra ation brea o n ro uct, an the concentrations o test substances uch easures shou not ea to si ni icant chan es in the test e ia hen such this are no n, the or c assi ication ur oses, a be ca cu ate base on the eo etric chan es are i e to ea to an increase or ecrease in the a arent to icit an hence the ean o the e ra ation ro uct concentration, bac ca cu ate to the arent substance c assi ication e e o the test substance si i ar rinci es a be a ie to easure ata in chronic to icit testin e o e substances an substances co ere b the c assi ication sche e are in act i tures, or hich easure ent o e osure concentrations is i icu t, an in so e cases oorly soluble substances i ossib e ubstances such as etro eu isti ate ractions, o ers, substances ith si ni icant e e s o i urities, etc can ose s ecia rob e s since the to ic concentration is hese substances, usua ta en to be those ith a so ubi it in ater , are re uent i icu t to i icu t to e ine an i ossib e to eri ica testin roce ures o ten re on the isso e in the test e ia, an the isso e concentrations i o ten ro e i icu t to easure at the o concentrations or ation o a ater o ub e raction or ater cco o ate raction an antici ate or an substances, the true so ubi it in the test e ia i be un no n, an i o ten be recor e as ata are re orte in ter s o oa in rates hese ata a be use in a in the etection i it in uri ie ater e erthe ess such substances can sho to icit , an here no to icit is oun , u e ent c assi ication criteria ust be a ie to hether the resu t can be consi ere a i or c assi ication u e ent shou err on the si e o caution an shou not un eresti ate the ha ar or c assi ication o or anic co oun s, it is esirab e to ha e stabi i e an ana tica easure test concentrations thou h easure concentrations are re erre , c assi ication a be base on no ina concentration ea , tests usin a ro riate isso ution techni ues an ith accurate easure concentrations ithin stu ies hen these are the on a i ata a ai ab e un er certain circu stances the ateria is i e to substantia the ran e o ater so ubi it shou be use here such test ata are a ai ab e, the shou be use in re erence to other e ra e or other ise be ost ro the ater co u n, care ust be ta en in ata inter retation an c assi ication shou be ata t is nor a , ho e er, articu ar hen consi erin o er ata, to in such substances ith to icit e e s recor e in one ta in the oss o the to icant urin the test into account, i re e ant an ossib e itiona , eta s resent their e cess o the ater so ubi it , or here the isso e e e s are be o the etection i it o the ana tica etho hus, in o n set o i icu ties an are iscusse se arate ab e ists se era ro erties o i icu t to test substances an both circu stances, it is not ossib e to eri the actua e osure concentrations usin easure ata here these are the their re e ance or c assi ication on ata a ai ab e on hich to c assi , so e ractica ru es can be consi ere b a o enera ui ance
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a here the acute to icit is recor e at e e s in e cess o the ater so ubi it , the or Complex substances c assi ication ur oses, a be consi ere to be e ua to or be o the easure ater so ubi it n such circu stances it is i e that hronic an or cute shou be a ie n a in this o e substances are characteri e b a ran e o che ica structures, re uent in a ho o o ous ecision, ue attention shou be ai to the ossibi it that the e cess un isso e substance a series, but co erin a i e ran e o ater so ubi ities an other h sico che ica characteristics n a ition to ater, ha e i en rise to h sica e ects on the test or anis s here this is consi ere the i e cause an e ui ibriu i be reache bet een the isso e an un isso e ractions hich i be characteristic o the o the e ects obser e , the test shou be consi ere as in a i or c assi ication ur oses oa in o the substance or this reason, such co e substances are usua teste as a or , an the recor e base on the oa in or no ina concentrations na tica su ort ata are not nor a a ai ab e b here no acute to icit is recor e at e e s in e cess o the ater so ubi it , the or since the isso e raction i itse be a co e i tures o co onents he to icit ara eter is so eti es c assi ication ur oses a be consi ere to be reater than the easure ater so ubi it n such re erre to as , re ate to the etha oa in e e his oa in e e ro the or a be use irect in circu stances, consi eration shou be i en to hether the hronic shou a n a in a the c assi ication criteria ecision that the substance sho s no acute to icit , ue account shou be ta en o the techni ues use to achie e the a i u isso e concentrations here these are not consi ere as a e uate, the test shou be consi ere as in a i or c assi ication ur oses o ers re resent a s ecia in o co e substance, re uirin consi eration o the o er t e an their isso ution is ersa beha iour o ers a isso e as such ithout chan e, true so ubi it re ate to c here the ater so ubi it is be o the etection i it o the ana tica etho or a substance, an artic e si e , be is ersib e, or ortions consistin o o o ecu ar ei ht ractions a o into so ution n the atter acute to icit is recor e , the or c assi ication ur oses, a be consi ere to be ess case, in e ect, the testin o a o er is a test o the abi it o o o ecu ar ass ateria to each ro the bu than the ana tica etection i it here no to icit is obser e , the or c assi ication o er, an hether this eachate is to ic t can thus be consi ere in the sa e a as a co e i ture in that a ur oses, a be consi ere to be reater than the ater so ubi it ue consi eration shou a so oa in o o er can best characteri e the resu tant eachate, an hence the to icit can be re ate to this oa in be i en to the ua it criteria entione abo e here chronic to icit ata are a ai ab e, the sa e enera ru es shou a ain, here these ata cannot be a i ate b consi eration o easure concentrations, the techni ues use to achie e the a i u isso e concentrations ust be consi ere as a ro riate
Other factors contributing to concentration loss
nu ber o other actors can a so contribute to osses o concentration an , hi e so e can be a oi e b correct stu esi n, inter retation o ata here these actors ha e contribute a , ro ti e to ti e, be necessar
a se i entation this can occur urin a test or a nu ber o reasons co on e anation is that the substance has not tru isso e es ite the a arent absence o articu ates, an a o eration occurs urin the test ea in to reci itation n these circu stances, the or or c assi ication ur oses, a be consi ere to be base on the en o test concentrations ua , reci itation can occur throu h reaction ith the e ia his is consi ere un er instabi it abo e b a sor tion this can occur or substances o hi h a sor tion characteristics such as hi h o o substances here this occurs, the oss o concentration is usua ra i an e osure a best be characteri e b the en o test concentrations c bioaccu u ation osses a occur throu h the bioaccu u ation o a substance into the test or anis s his a be articu ar i ortant here the ater so ubi it is o an o o corres on in hi h he or or c assi ication ur oses, a be ca cu ate base on the eo etric ean o the start an en o test concentrations
erturbation of the test media
tron aci s an bases a a ear to ic because the a a ter enera ho e er chan es o the in a uatic s ste s are nor a re ente b bu er s ste s in the test e iu no ata are a ai ab e on a sa t, the sa t shou enera be c assi ie in the sa e a as the anion or cation, i e as the ion that recei es the ost strin ent c assi ication the e ect concentration is re ate to on one o the ions, the c assi ication o the sa t shou ta e the o ecu ar ei ht i erence into consi eration b correctin the e ect concentration b u ti in ith the ratio sa t ion
o ers are t ica not a ai ab e in a uatic s ste s is ersib e o ers an other hi h o ecu ar ass ateria s can erturb the test s ste an inter ere ith u ta e o o en, an i e rise to echanica or secon ar e ects hese actors nee to be ta en into account hen consi erin ata ro these substances an o ers beha e i e co e substances, ho e er, ha in a si ni icant o o ecu ar ass raction hich can each ro the bu o er his is consi ere urther be o
- 462 - Copyright@United Nations, 2017. All rights reserved
a here the acute to icit is recor e at e e s in e cess o the ater so ubi it , the or Complex substances c assi ication ur oses, a be consi ere to be e ua to or be o the easure ater so ubi it n such circu stances it is i e that hronic an or cute shou be a ie n a in this o e substances are characteri e b a ran e o che ica structures, re uent in a ho o o ous ecision, ue attention shou be ai to the ossibi it that the e cess un isso e substance a series, but co erin a i e ran e o ater so ubi ities an other h sico che ica characteristics n a ition to ater, ha e i en rise to h sica e ects on the test or anis s here this is consi ere the i e cause an e ui ibriu i be reache bet een the isso e an un isso e ractions hich i be characteristic o the o the e ects obser e , the test shou be consi ere as in a i or c assi ication ur oses oa in o the substance or this reason, such co e substances are usua teste as a or , an the recor e base on the oa in or no ina concentrations na tica su ort ata are not nor a a ai ab e b here no acute to icit is recor e at e e s in e cess o the ater so ubi it , the or since the isso e raction i itse be a co e i tures o co onents he to icit ara eter is so eti es c assi ication ur oses a be consi ere to be reater than the easure ater so ubi it n such re erre to as , re ate to the etha oa in e e his oa in e e ro the or a be use irect in circu stances, consi eration shou be i en to hether the hronic shou a n a in a the c assi ication criteria ecision that the substance sho s no acute to icit , ue account shou be ta en o the techni ues use to achie e the a i u isso e concentrations here these are not consi ere as a e uate, the test shou be consi ere as in a i or c assi ication ur oses o ers re resent a s ecia in o co e substance, re uirin consi eration o the o er t e an their isso ution is ersa beha iour o ers a isso e as such ithout chan e, true so ubi it re ate to c here the ater so ubi it is be o the etection i it o the ana tica etho or a substance, an artic e si e , be is ersib e, or ortions consistin o o o ecu ar ei ht ractions a o into so ution n the atter acute to icit is recor e , the or c assi ication ur oses, a be consi ere to be ess case, in e ect, the testin o a o er is a test o the abi it o o o ecu ar ass ateria to each ro the bu than the ana tica etection i it here no to icit is obser e , the or c assi ication o er, an hether this eachate is to ic t can thus be consi ere in the sa e a as a co e i ture in that a ur oses, a be consi ere to be reater than the ater so ubi it ue consi eration shou a so oa in o o er can best characteri e the resu tant eachate, an hence the to icit can be re ate to this oa in be i en to the ua it criteria entione abo e here chronic to icit ata are a ai ab e, the sa e enera ru es shou a ain, here these ata cannot be a i ate b consi eration o easure concentrations, the techni ues use to achie e the a i u isso e concentrations ust be consi ere as a ro riate
Other factors contributing to concentration loss
nu ber o other actors can a so contribute to osses o concentration an , hi e so e can be a oi e b correct stu esi n, inter retation o ata here these actors ha e contribute a , ro ti e to ti e, be necessar
a se i entation this can occur urin a test or a nu ber o reasons co on e anation is that the substance has not tru isso e es ite the a arent absence o articu ates, an a o eration occurs urin the test ea in to reci itation n these circu stances, the or or c assi ication ur oses, a be consi ere to be base on the en o test concentrations ua , reci itation can occur throu h reaction ith the e ia his is consi ere un er instabi it abo e b a sor tion this can occur or substances o hi h a sor tion characteristics such as hi h o o substances here this occurs, the oss o concentration is usua ra i an e osure a best be characteri e b the en o test concentrations c bioaccu u ation osses a occur throu h the bioaccu u ation o a substance into the test or anis s his a be articu ar i ortant here the ater so ubi it is o an o o corres on in hi h he or or c assi ication ur oses, a be ca cu ate base on the eo etric ean o the start an en o test concentrations
erturbation of the test media
tron aci s an bases a a ear to ic because the a a ter enera ho e er chan es o the in a uatic s ste s are nor a re ente b bu er s ste s in the test e iu no ata are a ai ab e on a sa t, the sa t shou enera be c assi ie in the sa e a as the anion or cation, i e as the ion that recei es the ost strin ent c assi ication the e ect concentration is re ate to on one o the ions, the c assi ication o the sa t shou ta e the o ecu ar ei ht i erence into consi eration b correctin the e ect concentration b u ti in ith the ratio sa t ion
o ers are t ica not a ai ab e in a uatic s ste s is ersib e o ers an other hi h o ecu ar ass ateria s can erturb the test s ste an inter ere ith u ta e o o en, an i e rise to echanica or secon ar e ects hese actors nee to be ta en into account hen consi erin ata ro these substances an o ers beha e i e co e substances, ho e er, ha in a si ni icant o o ecu ar ass raction hich can each ro the bu o er his is consi ere urther be o
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e i ic tion o i ic t te t t nce nte p etin ata alit
ropert t re o i ic t e e nce or c i ic tion tandardi ation
oor ater so ub e chie in aintainin re uire e osure hen to ic res onses are obser e abo e an actors can in uence the resu ts o to icit tests ith a uatic or anis s hese actors inc u e concentration na sin e osure a arent so ubi it , e ert u e ent is characteristics o the test ater, e eri enta esi n, che ica characteristics o the test ateria , an bio o ica re uire to con ir hether e ects are ue to characteristics o the test or anis s here ore, it is i ortant in con uctin a uatic to icit tests to use stan ar i e test che ica to icit or a h sica e ect i no roce ures to re uce the in uence o these sources o e traneous ariabi it he oa o test stan ar i ation an e ects are obser e , it shou be internationa har oni ation o these stan ar s is to re uce test ariabi it an i ro e recision, re ro ucibi it , an e onstrate that u , saturate isso ution consistenc o test resu ts has been achie e o ic at o chie in aintainin re uire e osure assi ie base on to icit Data hierarchies concentrations concentration na sin e osure assi ication shou be base on ri ar ata o oo ua it re erence is i en to ata con or in to est ui e ines or e ui a ent an oo aborator ractices hi e ata ro internationa har oni e o ati e aintainin an easurin e osure assi ication shou be base on re iab e test etho s er or e on stan ar test s ecies are re erre , resu ts o tests er or e usin i e reco ni e concentration easure ent o concentrations internationa or nationa etho s or their e ui a ent a a so be use , e or etho s ata ro tests that hoto e ra ab e aintainin e osure concentrations assi ication re uires e ert u e ent an a ear to con or to acce te ui e ines but hich ac s ro isions or can be use in the absence o ertinent o icit o brea o n ro ucts shou be base on easure concentrations ata o icit o si ni icant brea o n ro ucts shou be characteri e e ersen et a ro i es a ata ua it scorin s ste , hich is co atib e ith an others in ro tica aintainin e osure concentrations assi ication re uires e ert u e ent, current use, inc u in that, use b the or its atabase ee a so ensin et a or iscussions o ata ua it he ata ua it scorin s ste escribe in e ersen et al inc u es a re iabi it ran in sche e, hich can be unstab e o icit o brea o n ro ucts o arison shou be base on easure concentrations, a o e or use ith in c assi in un er the har oni e sche e he irst three e e s o ata escribe b e ersen are or o e ra ation ha i es to the e osure an nee s to a ress the to icit o si ni icant re erre ata re i en use in testin brea o n ro ucts i i ab e chie in , aintainin an easurin assi ication re uires e ert u e ent, ata or c assi ication un er the har oni e sche e shou co e ro ri ar sources o e er, since e osure concentration o icit o o i ie shou be base on easure concentrations, an nations an re u ator authorities i er or c assi ication usin the oba har oni e sche e, c assi ication che ica structures or brea o n ro ucts an nee s to a ress the to icit o si ni icant shou a o or use o re ie s ro nationa authorities an e ert ane s as on as the re ie s are base on ri ar o arison o e ra ation ha i es to the brea o n ro ucts sources uch re ie s shou inc u e su aries o test con itions, hich are su icient etai e or ei ht o e i ence e osure re i en use in testin an c assi ication ecisions to be a e t a be ossib e to use the re ie s, hich ere a e b a e reco ni e rou ub ect to corrosion chie in , aintainin an easurin assi ication re uires e ert u e ent, such as or hich the ri ar ata are accessib e trans or ation e osure concentration o arison o shou be base on easure concentrations, this re ers to eta s artitionin ro the ater co u n ha i es an nee s to a ress the to icit o si ni icant n the absence o e irica test ata, a i ate uantitati e tructure cti it e ationshi s s or eta co oun s to the e osure re i en use in testin brea o n ro ucts a uatic to icit a be use est ata a a s ta e rece ence o er re ictions, ro i in the test ata are a i
io e ra ab e aintainin e osure concentrations o icit assi ication re uires e ert u e ent, e r tion o brea o n ro ucts o arison o shou be base on easure concentrations, e ra ation ha i es to the e osure re i en an nee s to a ress the to icit o si ni icant nt o ction use in testin brea o n ro ucts sorbin aintainin e osure concentrations assi ication shou use easure e ra abi it is one o the i ortant intrinsic ro erties o substances that eter ine their otentia na sin e osure o icit iti ation ue concentration o a ai ab e ateria en iron enta ha ar on e ra ab e substances i ersist in the en iron ent an a conse uent ha e a to re uce a ai abi it o test substance otentia or causin on ter a erse e ects on biota n contrast, e ra ab e substances a be re o e in the se ers, in se a e treat ent ants or in the en iron ent he atin istin uishin che ate an non che ate assi ication shou use easure ent o ractions in e ia concentration o bioa ai ab e ateria assi ication o substances is ri ari base on their intrinsic ro erties o e er, the e ree o o oure i ht attenuation an a a rob e assi ication ust istin uish to ic e ects e ra ation e en s not on on the intrinsic reca citrance o the o ecu e, but a so on the actua con itions in the ro re uce ro th ue to i ht attenuation recei in en iron enta co art ent as e re o otentia , , resence o suitab e icro or anis s, concentration ro hobic aintainin constant e osure concentrations assi ication shou use easure o the substances an occurrence an concentration o other substrates he inter retation o the e ra ation ro erties concentration in an a uatic ha ar c assi ication conte t there ore re uires etai e criteria that ba ance the intrinsic ro erties o the substance an the re ai in en iron enta con itions into a conc u in state ent on the otentia or on ter oni e aintainin e osure concentrations o icit assi ication re uires e ert u e ent, a erse e ects he ur ose o the resent section is to resent ui ance or inter retation o ata on e ra abi it o o brea o n ro ucts o arison o shou be base on easure concentrations, or anic substances he ui ance is base on an ana sis o the abo e entione as ects re ar in e ra ation in the e ra ation ha i es to the e osure re i e an nee s to a ress the to icit o si ni icant a uatic en iron ent ase on the ui ance a etai e ecision sche e or use o e istin e ra ation ata or use in testin brea o n ro ucts c assi ication ur oses is ro ose he t es o e ra ation ata inc u e in this ui ance ocu ent are rea u ti co onent re arin re resentati e test batches onsi ere sa e as co e i ture bio e ra abi it ata, si u ation ata or trans or ation in ater, a uatic se i ent an soi , ata an techni ues or esti ation o ra i e ra abi it in the a uatic en iron ent so consi ere are anaerobic e ra abi it , inherent bio e ra abi it , se a e treat ent ant si u ation test ata, abiotic trans or ation ata such as h ro sis an hoto sis, re o a rocess such as o ati i ation an ina , ata obtaine ro ie in esti ations an onitorin stu ies
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e i ic tion o i ic t te t t nce nte p etin ata alit
ropert t re o i ic t e e nce or c i ic tion tandardi ation
oor ater so ub e chie in aintainin re uire e osure hen to ic res onses are obser e abo e an actors can in uence the resu ts o to icit tests ith a uatic or anis s hese actors inc u e concentration na sin e osure a arent so ubi it , e ert u e ent is characteristics o the test ater, e eri enta esi n, che ica characteristics o the test ateria , an bio o ica re uire to con ir hether e ects are ue to characteristics o the test or anis s here ore, it is i ortant in con uctin a uatic to icit tests to use stan ar i e test che ica to icit or a h sica e ect i no roce ures to re uce the in uence o these sources o e traneous ariabi it he oa o test stan ar i ation an e ects are obser e , it shou be internationa har oni ation o these stan ar s is to re uce test ariabi it an i ro e recision, re ro ucibi it , an e onstrate that u , saturate isso ution consistenc o test resu ts has been achie e o ic at o chie in aintainin re uire e osure assi ie base on to icit Data hierarchies concentrations concentration na sin e osure assi ication shou be base on ri ar ata o oo ua it re erence is i en to ata con or in to est ui e ines or e ui a ent an oo aborator ractices hi e ata ro internationa har oni e o ati e aintainin an easurin e osure assi ication shou be base on re iab e test etho s er or e on stan ar test s ecies are re erre , resu ts o tests er or e usin i e reco ni e concentration easure ent o concentrations internationa or nationa etho s or their e ui a ent a a so be use , e or etho s ata ro tests that hoto e ra ab e aintainin e osure concentrations assi ication re uires e ert u e ent an a ear to con or to acce te ui e ines but hich ac s ro isions or can be use in the absence o ertinent o icit o brea o n ro ucts shou be base on easure concentrations ata o icit o si ni icant brea o n ro ucts shou be characteri e e ersen et a ro i es a ata ua it scorin s ste , hich is co atib e ith an others in ro tica aintainin e osure concentrations assi ication re uires e ert u e ent, current use, inc u in that, use b the or its atabase ee a so ensin et a or iscussions o ata ua it he ata ua it scorin s ste escribe in e ersen et al inc u es a re iabi it ran in sche e, hich can be unstab e o icit o brea o n ro ucts o arison shou be base on easure concentrations, a o e or use ith in c assi in un er the har oni e sche e he irst three e e s o ata escribe b e ersen are or o e ra ation ha i es to the e osure an nee s to a ress the to icit o si ni icant re erre ata re i en use in testin brea o n ro ucts i i ab e chie in , aintainin an easurin assi ication re uires e ert u e ent, ata or c assi ication un er the har oni e sche e shou co e ro ri ar sources o e er, since e osure concentration o icit o o i ie shou be base on easure concentrations, an nations an re u ator authorities i er or c assi ication usin the oba har oni e sche e, c assi ication che ica structures or brea o n ro ucts an nee s to a ress the to icit o si ni icant shou a o or use o re ie s ro nationa authorities an e ert ane s as on as the re ie s are base on ri ar o arison o e ra ation ha i es to the brea o n ro ucts sources uch re ie s shou inc u e su aries o test con itions, hich are su icient etai e or ei ht o e i ence e osure re i en use in testin an c assi ication ecisions to be a e t a be ossib e to use the re ie s, hich ere a e b a e reco ni e rou ub ect to corrosion chie in , aintainin an easurin assi ication re uires e ert u e ent, such as or hich the ri ar ata are accessib e trans or ation e osure concentration o arison o shou be base on easure concentrations, this re ers to eta s artitionin ro the ater co u n ha i es an nee s to a ress the to icit o si ni icant n the absence o e irica test ata, a i ate uantitati e tructure cti it e ationshi s s or eta co oun s to the e osure re i en use in testin brea o n ro ucts a uatic to icit a be use est ata a a s ta e rece ence o er re ictions, ro i in the test ata are a i
io e ra ab e aintainin e osure concentrations o icit assi ication re uires e ert u e ent, e r tion o brea o n ro ucts o arison o shou be base on easure concentrations, e ra ation ha i es to the e osure re i en an nee s to a ress the to icit o si ni icant nt o ction use in testin brea o n ro ucts sorbin aintainin e osure concentrations assi ication shou use easure e ra abi it is one o the i ortant intrinsic ro erties o substances that eter ine their otentia na sin e osure o icit iti ation ue concentration o a ai ab e ateria en iron enta ha ar on e ra ab e substances i ersist in the en iron ent an a conse uent ha e a to re uce a ai abi it o test substance otentia or causin on ter a erse e ects on biota n contrast, e ra ab e substances a be re o e in the se ers, in se a e treat ent ants or in the en iron ent he atin istin uishin che ate an non che ate assi ication shou use easure ent o ractions in e ia concentration o bioa ai ab e ateria assi ication o substances is ri ari base on their intrinsic ro erties o e er, the e ree o o oure i ht attenuation an a a rob e assi ication ust istin uish to ic e ects e ra ation e en s not on on the intrinsic reca citrance o the o ecu e, but a so on the actua con itions in the ro re uce ro th ue to i ht attenuation recei in en iron enta co art ent as e re o otentia , , resence o suitab e icro or anis s, concentration ro hobic aintainin constant e osure concentrations assi ication shou use easure o the substances an occurrence an concentration o other substrates he inter retation o the e ra ation ro erties concentration in an a uatic ha ar c assi ication conte t there ore re uires etai e criteria that ba ance the intrinsic ro erties o the substance an the re ai in en iron enta con itions into a conc u in state ent on the otentia or on ter oni e aintainin e osure concentrations o icit assi ication re uires e ert u e ent, a erse e ects he ur ose o the resent section is to resent ui ance or inter retation o ata on e ra abi it o o brea o n ro ucts o arison o shou be base on easure concentrations, or anic substances he ui ance is base on an ana sis o the abo e entione as ects re ar in e ra ation in the e ra ation ha i es to the e osure re i e an nee s to a ress the to icit o si ni icant a uatic en iron ent ase on the ui ance a etai e ecision sche e or use o e istin e ra ation ata or use in testin brea o n ro ucts c assi ication ur oses is ro ose he t es o e ra ation ata inc u e in this ui ance ocu ent are rea u ti co onent re arin re resentati e test batches onsi ere sa e as co e i ture bio e ra abi it ata, si u ation ata or trans or ation in ater, a uatic se i ent an soi , ata an techni ues or esti ation o ra i e ra abi it in the a uatic en iron ent so consi ere are anaerobic e ra abi it , inherent bio e ra abi it , se a e treat ent ant si u ation test ata, abiotic trans or ation ata such as h ro sis an hoto sis, re o a rocess such as o ati i ation an ina , ata obtaine ro ie in esti ations an onitorin stu ies
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he ter e ra ation is e ine in ha ter as the eco osition o or anic o ecu es to s a er different chain lengths, degree and or site of branching or stereo isomers, even in their most purified commercial forms. o ecu es an e entua to carbon io i e, ater an sa ts or inor anic co oun s an eta s, the conce t o Testing of each individual component may be costly and impractical. If a test on the complex, multi component e ra abi it as a ie to or anic co oun s has i ite or no eanin ather the substance a be trans or e b substance is performed and it is anticipated that a sequential biodegradation of the individual structures is ta ing place, nor a en iron enta rocesses to either increase or ecrease the bioa ai abi it o the to ic s ecies here ore, the then the 10 day indo should not be applied to interpret the results of the test. A case by case evaluation should resent section ea s on ith or anic substances an or ano eta s n iron enta artitionin ro the ater ho ever ta e place on hether a biodegradability test on such a substance ould give valuable information regarding co u n is iscusse in ection its biodegradability as such (i.e. regarding the degradability of all the constituents) or hether instead an investigation of the degradability of carefully selected individual components of the complex, multi component substance is required. ata on e ra ation ro erties o a substance a be a ai ab e ro stan ar i e tests or ro other t es o in esti ations, or the a be esti ate ro the structure o the o ecu es he inter retation o such A9.4.2.3 OD5 COD e ra ation ata or c assi ication ur oses o ten re uires etai e e a uation o the test ata ui ance is i en in the resent section an ore etai s can be oun in t o ara ra hs escribin a ai ab e etho s en i an Information on the 5 day biochemical oxygen demand ( OD5) ill be used for classification purposes actors in uencin e ra ation in a uatic en iron ents en i only hen no other measured degradability data are available. Thus, priority is given to data from ready biodegradability tests and from simulation studies regarding degradability in the aquatic environment. The OD5 test is nte p etation of e a a ilit ata a traditional biodegradation test that is no replaced by the ready biodegradability tests. Therefore, this test should not be performed today for assessment of the ready biodegradability of substances. Older test data may, ho ever, be used apid degradability hen no other degradability data are available. or substances here the chemical structure is no n, the theoretical oxygen demand (ThOD) can be calculated and this value should be used instead of the chemical oxygen demand uatic ha ar c assi ication o substances is nor a base on e istin ata on their en iron enta (COD). ro erties n se o i test ata be ro uce ith the ain ur ose o aci itatin a c assi ication ten a i erse ran e o test ata is a ai ab e that oes not necessari its irect ith the c assi ication criteria onse uent , A9.4.2.4 Other convincing scientific evidence ui ance is nee e on inter retation o e istin test ata in the conte t o the a uatic ha ar c assi ication ase on the har oni e criteria, ui ance or inter retation o e ra ation ata is re are be o or the three t es o ata A9.4.2.4.1 Rapid degradation in the aquatic environment may be demonstrated by other data than referred to in comprised by the expression “rapid degradation” in the aquatic environment (see A9.1.8, A9.1.9, A9.1.2. to Chapter 4.1, paragraph 4.1.2.11.3 (a) and (b). These may be data on biotic and or abiotic degradation. Data on primary an the e inition in ha ter , ara degradation can only be used here it is demonstrated that the degradation products shall not be classified as ha ardous to the aquatic environment, i.e. that they do not fulfil the classification criteria. eady biodegradability A9.4.2.4.2 The fulfilment of paragraph 4.1.2.11.3 (c) requires that the substance is degraded in the aquatic ea bio e ra abi it is e ine in the est ui e ines o or anic environment to a level of 0 ithin a 28 day period. If first order inetics are assumed, hich is reasonable at the substances that e ra e to a e e hi her than the ass e e in a stan ar rea bio e ra abi it test or in a lo substance concentrations prevailing in most aquatic environments, the degradation rate ill be relatively constant si i ar test shou be consi ere rea i bio e ra ab e an conse uent a so ra i e ra ab e an iterature test for the 28 day period. Thus, the degradation requirement ill be fulfilled ith an average degradation rate constant, 1 ata, ho e er, o not s eci a o the con itions that shou be e a uate to e onstrate hether or not the test u i s (ln 0.3 ln 1) 28 0.043 day . This corresponds to a degradation half life, t ln 2 0.043 1 days. the re uire ents o a rea bio e ra abi it test ert u e ent is there ore nee e as re ar s the a i it o the ata be ore use or c assi ication ur oses e ore conc u in on the rea bio e ra abi it o a test substance, A9.4.2.4.3 oreover, as degradation processes are temperature dependent, this parameter should also be ta en ho e er, at east the o o in ara eters shou be consi ere into account hen assessing degradation in the environment. Data from studies employing environmentally realistic temperatures should be used for the evaluation. hen data from studies performed at different temperatures need to be oncentration o test substance compared, the traditional 10 approach could be used, i.e. that the degradation rate is halved hen the temperature decreases by 10 C. e ati e hi h concentrations o test substance are use in the rea bio e ra abi it tests an substances a , ho e er, be to ic to the inocu a at such hi h concentrations causin a o A9.4.2.4.4 The evaluation of data on fulfilment of this criterion should be conducted on a case by case basis by e ra ation in the tests a thou h the substances i ht be ra i e ra ab e at o er non to ic concentrations expert udgement. o ever, guidance on the interpretation of various types of data that may be used for demonstrating to icit test ith icro or anis s as e the OECD Test Guideline 209 “Activated Sludge, Respiration Inhibition a rapid degradation in the aquatic environment is given belo . In general, only data from aquatic biodegradation Test”, the ISO 9509 nitrification inhibition test, or the ISO 11348 luminescent bacteria inhibition test) may demonstrate simulation tests are considered directly applicable. o ever simulation test data from other environmental the to icit o the test substance hen it is i e that inhibition is the reason or a substance bein not rea i compartments could be considered as ell, but such data require in general more scientific udgement before use. e ra ab e, resu ts ro a test e o in o er non to ic concentrations o the test substance shou be use hen a ai ab e uch test resu ts cou on a case b case basis be consi ere in re ation to the c assi ication criteria or ra i A9.4.2.4.5 Aquatic simulation tests e ra ation, e en thou h sur ace ater e ra ation test ata ith en iron enta rea istic icrobia bio ass an non to ic rea istic o concentration o the test substance in enera are re erre , i a ai ab e Aquatic simulation tests are tests conducted in laboratory, but simulating environmental conditions and employing natural samples as inoculum. Results of aquatic simulation tests may be used directly for classification i e in o purposes, hen realistic environmental conditions in surface aters are simulated, i.e.
he har oni e criteria see inc u e a enera re uire ent or a o the rea (a) substance concentration that is realistic for the general aquatic environment (often in the lo bio e ra abi it tests on achie e ent o the ass e e ithin a s his is not in ine ith the est ui e ine g l range) in hich the a s ti e in o a ies to the rea bio e ra abi it tests e ce t to the test (b) inoculum from a relevant aquatic environment est ui e ine n the ose ott e test est ui e ine , a a s in o a be use 3 instea hen easure ents ha e not been a e a ter a s oreo er, o ten on i ite in or ation is a ai ab e in (c) realistic concentration of inoculum (10 10 cells ml) re erences o bio e ra ation tests hus, as a ra atic a roach the ercenta e o e ra ation reache a ter a s a be use irect or assess ent o rea bio e ra abi it hen no in or ation on the a s ti e in o is (d) realistic temperature (e.g. 5 C to 25 C) and a ai ab e his shou , ho e er, on be acce te or e istin test ata an ata ro tests here the a s in o (e) ultimate degradation is determined (i.e. determination of the minerali ation rate or the oes not a individual degradation rates of the total biodegradation path ay).
here there is su icient usti ication, the a in o con ition a be ai e or co e , Substances that under these conditions are degraded at least 0 ithin 28 days, i.e. ith a half life u ti co onent substances an the ass e e a ie at a s he constituents o such substances a ha e 1 days, are considered rapidly degradable.
4 - 466 - Copyright@United Nations, 2017. All rights reserved
he ter e ra ation is e ine in ha ter as the eco osition o or anic o ecu es to s a er different chain lengths, degree and or site of branching or stereo isomers, even in their most purified commercial forms. o ecu es an e entua to carbon io i e, ater an sa ts or inor anic co oun s an eta s, the conce t o Testing of each individual component may be costly and impractical. If a test on the complex, multi component e ra abi it as a ie to or anic co oun s has i ite or no eanin ather the substance a be trans or e b substance is performed and it is anticipated that a sequential biodegradation of the individual structures is ta ing place, nor a en iron enta rocesses to either increase or ecrease the bioa ai abi it o the to ic s ecies here ore, the then the 10 day indo should not be applied to interpret the results of the test. A case by case evaluation should resent section ea s on ith or anic substances an or ano eta s n iron enta artitionin ro the ater ho ever ta e place on hether a biodegradability test on such a substance ould give valuable information regarding co u n is iscusse in ection its biodegradability as such (i.e. regarding the degradability of all the constituents) or hether instead an investigation of the degradability of carefully selected individual components of the complex, multi component substance is required. ata on e ra ation ro erties o a substance a be a ai ab e ro stan ar i e tests or ro other t es o in esti ations, or the a be esti ate ro the structure o the o ecu es he inter retation o such A9.4.2.3 OD5 COD e ra ation ata or c assi ication ur oses o ten re uires etai e e a uation o the test ata ui ance is i en in the resent section an ore etai s can be oun in t o ara ra hs escribin a ai ab e etho s en i an Information on the 5 day biochemical oxygen demand ( OD5) ill be used for classification purposes actors in uencin e ra ation in a uatic en iron ents en i only hen no other measured degradability data are available. Thus, priority is given to data from ready biodegradability tests and from simulation studies regarding degradability in the aquatic environment. The OD5 test is nte p etation of e a a ilit ata a traditional biodegradation test that is no replaced by the ready biodegradability tests. Therefore, this test should not be performed today for assessment of the ready biodegradability of substances. Older test data may, ho ever, be used apid degradability hen no other degradability data are available. or substances here the chemical structure is no n, the theoretical oxygen demand (ThOD) can be calculated and this value should be used instead of the chemical oxygen demand uatic ha ar c assi ication o substances is nor a base on e istin ata on their en iron enta (COD). ro erties n se o i test ata be ro uce ith the ain ur ose o aci itatin a c assi ication ten a i erse ran e o test ata is a ai ab e that oes not necessari its irect ith the c assi ication criteria onse uent , A9.4.2.4 Other convincing scientific evidence ui ance is nee e on inter retation o e istin test ata in the conte t o the a uatic ha ar c assi ication ase on the har oni e criteria, ui ance or inter retation o e ra ation ata is re are be o or the three t es o ata A9.4.2.4.1 Rapid degradation in the aquatic environment may be demonstrated by other data than referred to in comprised by the expression “rapid degradation” in the aquatic environment (see A9.1.8, A9.1.9, A9.1.2. to Chapter 4.1, paragraph 4.1.2.11.3 (a) and (b). These may be data on biotic and or abiotic degradation. Data on primary an the e inition in ha ter , ara degradation can only be used here it is demonstrated that the degradation products shall not be classified as ha ardous to the aquatic environment, i.e. that they do not fulfil the classification criteria. eady biodegradability A9.4.2.4.2 The fulfilment of paragraph 4.1.2.11.3 (c) requires that the substance is degraded in the aquatic ea bio e ra abi it is e ine in the est ui e ines o or anic environment to a level of 0 ithin a 28 day period. If first order inetics are assumed, hich is reasonable at the substances that e ra e to a e e hi her than the ass e e in a stan ar rea bio e ra abi it test or in a lo substance concentrations prevailing in most aquatic environments, the degradation rate ill be relatively constant si i ar test shou be consi ere rea i bio e ra ab e an conse uent a so ra i e ra ab e an iterature test for the 28 day period. Thus, the degradation requirement ill be fulfilled ith an average degradation rate constant, 1 ata, ho e er, o not s eci a o the con itions that shou be e a uate to e onstrate hether or not the test u i s (ln 0.3 ln 1) 28 0.043 day . This corresponds to a degradation half life, t ln 2 0.043 1 days. the re uire ents o a rea bio e ra abi it test ert u e ent is there ore nee e as re ar s the a i it o the ata be ore use or c assi ication ur oses e ore conc u in on the rea bio e ra abi it o a test substance, A9.4.2.4.3 oreover, as degradation processes are temperature dependent, this parameter should also be ta en ho e er, at east the o o in ara eters shou be consi ere into account hen assessing degradation in the environment. Data from studies employing environmentally realistic temperatures should be used for the evaluation. hen data from studies performed at different temperatures need to be oncentration o test substance compared, the traditional 10 approach could be used, i.e. that the degradation rate is halved hen the temperature decreases by 10 C. e ati e hi h concentrations o test substance are use in the rea bio e ra abi it tests an substances a , ho e er, be to ic to the inocu a at such hi h concentrations causin a o A9.4.2.4.4 The evaluation of data on fulfilment of this criterion should be conducted on a case by case basis by e ra ation in the tests a thou h the substances i ht be ra i e ra ab e at o er non to ic concentrations expert udgement. o ever, guidance on the interpretation of various types of data that may be used for demonstrating to icit test ith icro or anis s as e the OECD Test Guideline 209 “Activated Sludge, Respiration Inhibition a rapid degradation in the aquatic environment is given belo . In general, only data from aquatic biodegradation Test”, the ISO 9509 nitrification inhibition test, or the ISO 11348 luminescent bacteria inhibition test) may demonstrate simulation tests are considered directly applicable. o ever simulation test data from other environmental the to icit o the test substance hen it is i e that inhibition is the reason or a substance bein not rea i compartments could be considered as ell, but such data require in general more scientific udgement before use. e ra ab e, resu ts ro a test e o in o er non to ic concentrations o the test substance shou be use hen a ai ab e uch test resu ts cou on a case b case basis be consi ere in re ation to the c assi ication criteria or ra i A9.4.2.4.5 Aquatic simulation tests e ra ation, e en thou h sur ace ater e ra ation test ata ith en iron enta rea istic icrobia bio ass an non to ic rea istic o concentration o the test substance in enera are re erre , i a ai ab e Aquatic simulation tests are tests conducted in laboratory, but simulating environmental conditions and employing natural samples as inoculum. Results of aquatic simulation tests may be used directly for classification i e in o purposes, hen realistic environmental conditions in surface aters are simulated, i.e.
he har oni e criteria see inc u e a enera re uire ent or a o the rea (a) substance concentration that is realistic for the general aquatic environment (often in the lo bio e ra abi it tests on achie e ent o the ass e e ithin a s his is not in ine ith the est ui e ine g l range) in hich the a s ti e in o a ies to the rea bio e ra abi it tests e ce t to the test (b) inoculum from a relevant aquatic environment est ui e ine n the ose ott e test est ui e ine , a a s in o a be use 3 instea hen easure ents ha e not been a e a ter a s oreo er, o ten on i ite in or ation is a ai ab e in (c) realistic concentration of inoculum (10 10 cells ml) re erences o bio e ra ation tests hus, as a ra atic a roach the ercenta e o e ra ation reache a ter a s a be use irect or assess ent o rea bio e ra abi it hen no in or ation on the a s ti e in o is (d) realistic temperature (e.g. 5 C to 25 C) and a ai ab e his shou , ho e er, on be acce te or e istin test ata an ata ro tests here the a s in o (e) ultimate degradation is determined (i.e. determination of the minerali ation rate or the oes not a individual degradation rates of the total biodegradation path ay).
here there is su icient usti ication, the a in o con ition a be ai e or co e , Substances that under these conditions are degraded at least 0 ithin 28 days, i.e. ith a half life u ti co onent substances an the ass e e a ie at a s he constituents o such substances a ha e 1 days, are considered rapidly degradable.
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A9.4.2.4. ield investigations environment. It is therefore proposed that an experimentally determined rapid degradation in soil is sufficient documentation for a rapid degradation in surface aters hen arallels to laboratory simulation tests are field investigations or mesocosm experiments. In such studies, fate and or effects of chemicals in environments or environmental enclosures may be investigated. ate data (a) no pre exposure (pre adaptation) of the soil micro organisms has ta en place and from such experiments might be used for assessing the potential for a rapid degradation. This may, ho ever, often be (b) an environmentally realistic concentration of substance is tested and difficult, as it requires that an ultimate degradation can be demonstrated. This may be documented by preparing mass balances sho ing that no non degradable intermediates are formed, and hich ta e the fractions into account that are (c) the substance is ultimately degraded ithin 28 days ith a half life 1 days corresponding to removed from the aqueous system due to other processes such as sorption to sediment or volatili ation from the aquatic a degradation rate 0.043 day 1 . environment. The same argumentation is considered valid for data on degradation in sediment under aerobic conditions. A9.4.2.4. onitoring data A9.4.2.4.11 Anaerobic degradation data onitoring data may demonstrate the removal of contaminants from the aquatic environment. Such data are, ho ever, very difficult to use for classification purposes. The follo ing aspects should be considered before use Data regarding anaerobic degradation cannot be used in relation to deciding hether a substance should be regarded as rapidly degradable, because the aquatic environment is generally regarded as the aerobic compartment here the aquatic organisms, such as those employed for aquatic ha ard classification, live. (a) Is the removal a result of degradation, or is it a result of other processes such as dilution or distribution bet een compartments (sorption, volatili ation) A9.4.2.4.12 ydrolysis (b) Is formation of non degradable intermediates excluded Data on hydrolysis (e.g. OECD Test Guideline 111) might be considered for classification purposes only hen the longest half life t determined ithin the p range 4 9 is shorter than 1 days. o ever, hydrolysis is Only hen it can be demonstrated that removal as a result of ultimate degradation fulfils the criteria not an ultimate degradation and various intermediate degradation products may be formed, some of hich may be only for rapid degradability, such data be considered for use for classification purposes. In general, monitoring data should slo ly degradable. Only hen it can be satisfactorily demonstrated that the hydrolysis products formed do not fulfil the only be used as supporting evidence for demonstration of either persistence in the aquatic environment or a rapid criteria for classification as ha ardous for the aquatic environment, data from hydrolysis studies could be considered. degradation. hen a substance is quic ly hydrolysed (e.g. ith t a fe days), this process is a part of the A9.4.2.4.8 Inherent biodegradability tests degradation determined in biodegradation tests. ydrolysis may be the initial transformation process in biodegradation. Substances that are degraded more than 0 in tests for inherent biodegradability (OECD Test A9.4.2.4.13 hotochemical degradation Guidelines 302) have the potential for ultimate biodegradation. o ever, because of the optimum conditions in these tests, the rapid biodegradability of inherently biodegradable substances in the environment cannot be assumed. The optimum conditions in inherent biodegradability tests stimulate adaptation of the micro organisms thus increasing the Information on photochemical degradation (e.g. OECD, 199 ) is difficult to use for classification biodegradation potential, compared to natural environments. Therefore, positive results in general should not be purposes. The actual degree of photochemical degradation in the aquatic environment depends on local conditions (e.g. 2 ater depth, suspended solids, turbidity) and the ha ard of the degradation products is usually not no n. robably interpreted as evidence for rapid degradation in the environment . only seldom ill enough information be available for a thorough evaluation based on photochemical degradation. A9.4.2.4.9 Se age treatment plant simulation tests A9.4.2.4.14 Estimation of degradation Results from tests simulating the conditions in a se age treatment plant (ST ) (e.g. the OECD Test A9.4.2.4.14.1 Certain SARs have been developed for prediction of an approximate hydrolysis half life, hich Guideline 303) cannot be used for assessing the degradation in the aquatic environment. The main reasons for this are that the microbial biomass in a ST is significantly different from the biomass in the environment, that there is a should only be considered hen no experimental data are available. o ever, a hydrolysis half life can only be used in considerably different composition of substrates, and that the presence of rapidly minerali ed organic matter in aste relation to classification with great care, because hydrolysis does not concern ultimate degradability (see “Hydrolysis” of this Section). urthermore the SARs developed until no have a rather limited applicability and are only able to ater facilitates degradation of the test substance by co metabolism. predict the potential for hydrolysis on a limited number of chemical classes. The SAR program DRO I (version 1. , Syracuse Research Corporation) is for example only able to predict the potential for hydrolysis on less than 1 5th A9.4.2.4.10 Soil and sediment degradation data of the existing E substances hich have a defined (precise) molecular structure ( iemel , 2000). It has been argued that for many non sorptive (non lipophilic) substances more or less the same A9.4.2.4.14.2 In general, no quantitative estimation method ( SAR) for estimating the degree of biodegradability of degradation rates are found in soil and in surface ater. or lipophilic substances, a lo er degradation rate may organic substances is yet sufficiently accurate to predict rapid degradation. o ever, results from such methods may be generally be expected in soil than in ater due to partial immobili ation caused by sorption. Thus, hen a substance has used to predict that a substance is not rapidly degradable. or example, hen in the iodegradation robability rogram been sho n to be degraded rapidly in a soil simulation study, it is most li ely also rapidly degradable in the aquatic (e.g. IO I version 3. , Syracuse Research Corporation) the probability is 0.5 estimated by the linear or non linear methods, the substances should be regarded as not rapidly degradable (OECD, 1994 edersen et al., 1995 2 n relation to interpretation of degradation data e uivalent with the harmonised OECD criteria for Chronic the angenberg et al., 199 ). Also other ( )SAR methods may be used as ell as expert udgement, for example, hen standing E wor ing group for environmental ha ard classification of substances is discussing whether certain types of degradation data for structurally analogue compounds are available, but such udgement should be conducted ith great data from inherent biodegradability tests may be used in a case by case evaluation as a basis for not classifying care. In general, a SAR prediction that a substance is not rapidly degradable is considered a better documentation for a substances otherwise fulfilling this classification criterion. classification than application of a default classification, hen no useful degradation data are available. The inherent biodegradability tests concerned are the ahn ellens test (OECD TG 0 and the T test (OECD TG 0 C . The conditions for use in this regard are A9.4.2.4.15 olatili ation (a The methods must not employ pre exposed (pre adapted micro organisms Chemicals may be removed from some aquatic environments by volatili ation. The intrinsic potential (b The time for adaptation within each test should be limited the test endpoint should refer to the minerali ation for volatili ation is determined by the enry s a constant ( ) of the substance. olatili ation from the aquatic only and the pass level and time for reaching these should be respectively environment is highly dependent on the environmental conditions of the specific ater body in question, such as the (i T pass level 60 within days ater depth, the gas exchange coefficients (depending on ind speed and ater flo ) and stratification of the ater (ii ahn ellens Test 0 within days. body. ecause volatili ation only represents removal of a chemical from ater phase, the enry s a constant cannot 4 8 4 9 - 468 - Copyright@United Nations, 2017. All rights reserved
A9.4.2.4. ield investigations environment. It is therefore proposed that an experimentally determined rapid degradation in soil is sufficient documentation for a rapid degradation in surface aters hen arallels to laboratory simulation tests are field investigations or mesocosm experiments. In such studies, fate and or effects of chemicals in environments or environmental enclosures may be investigated. ate data (a) no pre exposure (pre adaptation) of the soil micro organisms has ta en place and from such experiments might be used for assessing the potential for a rapid degradation. This may, ho ever, often be (b) an environmentally realistic concentration of substance is tested and difficult, as it requires that an ultimate degradation can be demonstrated. This may be documented by preparing mass balances sho ing that no non degradable intermediates are formed, and hich ta e the fractions into account that are (c) the substance is ultimately degraded ithin 28 days ith a half life 1 days corresponding to removed from the aqueous system due to other processes such as sorption to sediment or volatili ation from the aquatic a degradation rate 0.043 day 1 . environment. The same argumentation is considered valid for data on degradation in sediment under aerobic conditions. A9.4.2.4. onitoring data A9.4.2.4.11 Anaerobic degradation data onitoring data may demonstrate the removal of contaminants from the aquatic environment. Such data are, ho ever, very difficult to use for classification purposes. The follo ing aspects should be considered before use Data regarding anaerobic degradation cannot be used in relation to deciding hether a substance should be regarded as rapidly degradable, because the aquatic environment is generally regarded as the aerobic compartment here the aquatic organisms, such as those employed for aquatic ha ard classification, live. (a) Is the removal a result of degradation, or is it a result of other processes such as dilution or distribution bet een compartments (sorption, volatili ation) A9.4.2.4.12 ydrolysis (b) Is formation of non degradable intermediates excluded Data on hydrolysis (e.g. OECD Test Guideline 111) might be considered for classification purposes only hen the longest half life t determined ithin the p range 4 9 is shorter than 1 days. o ever, hydrolysis is Only hen it can be demonstrated that removal as a result of ultimate degradation fulfils the criteria not an ultimate degradation and various intermediate degradation products may be formed, some of hich may be only for rapid degradability, such data be considered for use for classification purposes. In general, monitoring data should slo ly degradable. Only hen it can be satisfactorily demonstrated that the hydrolysis products formed do not fulfil the only be used as supporting evidence for demonstration of either persistence in the aquatic environment or a rapid criteria for classification as ha ardous for the aquatic environment, data from hydrolysis studies could be considered. degradation. hen a substance is quic ly hydrolysed (e.g. ith t a fe days), this process is a part of the A9.4.2.4.8 Inherent biodegradability tests degradation determined in biodegradation tests. ydrolysis may be the initial transformation process in biodegradation. Substances that are degraded more than 0 in tests for inherent biodegradability (OECD Test A9.4.2.4.13 hotochemical degradation Guidelines 302) have the potential for ultimate biodegradation. o ever, because of the optimum conditions in these tests, the rapid biodegradability of inherently biodegradable substances in the environment cannot be assumed. The optimum conditions in inherent biodegradability tests stimulate adaptation of the micro organisms thus increasing the Information on photochemical degradation (e.g. OECD, 199 ) is difficult to use for classification biodegradation potential, compared to natural environments. Therefore, positive results in general should not be purposes. The actual degree of photochemical degradation in the aquatic environment depends on local conditions (e.g. 2 ater depth, suspended solids, turbidity) and the ha ard of the degradation products is usually not no n. robably interpreted as evidence for rapid degradation in the environment . only seldom ill enough information be available for a thorough evaluation based on photochemical degradation. A9.4.2.4.9 Se age treatment plant simulation tests A9.4.2.4.14 Estimation of degradation Results from tests simulating the conditions in a se age treatment plant (ST ) (e.g. the OECD Test A9.4.2.4.14.1 Certain SARs have been developed for prediction of an approximate hydrolysis half life, hich Guideline 303) cannot be used for assessing the degradation in the aquatic environment. The main reasons for this are that the microbial biomass in a ST is significantly different from the biomass in the environment, that there is a should only be considered hen no experimental data are available. o ever, a hydrolysis half life can only be used in considerably different composition of substrates, and that the presence of rapidly minerali ed organic matter in aste relation to classification with great care, because hydrolysis does not concern ultimate degradability (see “Hydrolysis” of this Section). urthermore the SARs developed until no have a rather limited applicability and are only able to ater facilitates degradation of the test substance by co metabolism. predict the potential for hydrolysis on a limited number of chemical classes. The SAR program DRO I (version 1. , Syracuse Research Corporation) is for example only able to predict the potential for hydrolysis on less than 1 5th A9.4.2.4.10 Soil and sediment degradation data of the existing E substances hich have a defined (precise) molecular structure ( iemel , 2000). It has been argued that for many non sorptive (non lipophilic) substances more or less the same A9.4.2.4.14.2 In general, no quantitative estimation method ( SAR) for estimating the degree of biodegradability of degradation rates are found in soil and in surface ater. or lipophilic substances, a lo er degradation rate may organic substances is yet sufficiently accurate to predict rapid degradation. o ever, results from such methods may be generally be expected in soil than in ater due to partial immobili ation caused by sorption. Thus, hen a substance has used to predict that a substance is not rapidly degradable. or example, hen in the iodegradation robability rogram been sho n to be degraded rapidly in a soil simulation study, it is most li ely also rapidly degradable in the aquatic (e.g. IO I version 3. , Syracuse Research Corporation) the probability is 0.5 estimated by the linear or non linear methods, the substances should be regarded as not rapidly degradable (OECD, 1994 edersen et al., 1995 2 n relation to interpretation of degradation data e uivalent with the harmonised OECD criteria for Chronic the angenberg et al., 199 ). Also other ( )SAR methods may be used as ell as expert udgement, for example, hen standing E wor ing group for environmental ha ard classification of substances is discussing whether certain types of degradation data for structurally analogue compounds are available, but such udgement should be conducted ith great data from inherent biodegradability tests may be used in a case by case evaluation as a basis for not classifying care. In general, a SAR prediction that a substance is not rapidly degradable is considered a better documentation for a substances otherwise fulfilling this classification criterion. classification than application of a default classification, hen no useful degradation data are available. The inherent biodegradability tests concerned are the ahn ellens test (OECD TG 0 and the T test (OECD TG 0 C . The conditions for use in this regard are A9.4.2.4.15 olatili ation (a The methods must not employ pre exposed (pre adapted micro organisms Chemicals may be removed from some aquatic environments by volatili ation. The intrinsic potential (b The time for adaptation within each test should be limited the test endpoint should refer to the minerali ation for volatili ation is determined by the enry s a constant ( ) of the substance. olatili ation from the aquatic only and the pass level and time for reaching these should be respectively environment is highly dependent on the environmental conditions of the specific ater body in question, such as the (i T pass level 60 within days ater depth, the gas exchange coefficients (depending on ind speed and ater flo ) and stratification of the ater (ii ahn ellens Test 0 within days. body. ecause volatili ation only represents removal of a chemical from ater phase, the enry s a constant cannot 4 8 4 9 - 469 - Copyright@United Nations, 2017. All rights reserved
be used for assessment of degradation in relation to a uatic ha ard classification of substances ubstances that are rimary biodegradation gases at ambient tem erature may howe er for e am le be considered further in this regard (see also edersen et al., n some tests, only the disa earance of the arent com ound (i e rimary degradation is determined for e am le by following the degradation by s ecific or grou s ecific chemical analyses of the test substance ata on o degradation data available rimary biodegradability may be used for demonstrating ra id degradability only when it can be satisfactorily demonstrated that the degradation roducts formed do not fulfil the criteria for classification as ha ardous to the a uatic hen no useful data on degradability are a ailable either e erimentally determined or estimated en ironment data the substance should be regarded as not ra idly degradable Conflicting results from screening tests ene al inte p etation p o le s he situation where more degradation data are a ailable for the same substance introduces the Complex substances ossibility of conflicting results n general, conflicting results for a substance which has been tested se eral times with an appropriate biodegradability test could be interpreted by a “weight of evidence approach”. This implies that if both he harmoni ed criteria for classification of chemicals as ha ardous for the a uatic en ironment focus ositi e (i e higher degradation than the ass le el and negati e results ha e been obtained for a substance in ready on single substances certain ty e of intrinsically com le substance are multi com onent substances hey are biodegradability tests, then the data of the highest uality and the best documentation should be used for determining ty ically of natural origin and need occasionally to be considered his may be the case for chemicals that are roduced the ready biodegradability of the substance Howe er, ositi e results in ready biodegradability tests could be or e tracted from mineral oil or lant material uch com le chemicals are normally considered as single substances in considered alid, irres ecti e of negati e results, when the scientific uality is good and the test conditions are well a regulatory conte t n most cases they are defined as a homologous series of substances within a certain range of documented, i e guideline criteria are fulfilled, including the use of non re e osed (non ada ted inoculum one of carbon chain length and or degree of substitution hen this is the case, no ma or difference in degradability is foreseen the arious screening tests are suitable for the testing of all ty es of substances, and results obtained by the use of a test and the degree of degradability can be established from tests of the com le chemical ne e ce tion would be when a rocedure which is not suitable for the s ecific substance should be e aluated carefully before a decision on the use is borderline degradation is found because in this case some of the indi idual substances may be ra idly degradable and ta en other may be not ra idly degradable his re uires a more detailed assessment of the degradability of the indi idual hus, there are a number of factors that may e lain conflicting biodegradability data from screening com onents in the com le substance hen not ra idly degradable com onents constitute a significant art of the tests com le substance (e g more than , or for a ha ardous com onent, an e en lower content , the substance should be regarded as not ra idly degradable (a inoculum Availability of the substance (b to icity of test substance (c test conditions egradation of organic substances in the en ironment ta es lace mostly in the a uatic com artments or in a uatic hases in soil or sediment Hydrolysis, of course, re uires the resence of water he acti ity of micro (d solubility of the test substance and organisms de ends on the resence of water oreo er, biodegradation re uires that the micro organisms are directly in (e olatili ation of the test substance contact with the substance issolution of the substance in the water hase that surrounds the micro organisms is therefore the most direct way for contact between the bacteria and fungi and the substrate he suitability of the inoculum for degrading the test substance de ends on the resence and amount of com etent degraders hen the inoculum is obtained from an en ironment that has re iously been e osed to the he resent standard methods for in estigating degradability of substances are de elo ed for readily test substance, the inoculum may be ada ted as e idenced by a degradation ca acity, which is greater than that of an soluble test com ounds Howe er, many organic substances are only slightly soluble in water s the standard tests inoculum from a non e osed en ironment s far as ossible the inoculum must be sam led from an une osed re uire mg l of the test substance, sufficient a ailability may not be reached for substances with a low water en ironment, but for substances that are used ubi uitously in high olumes and released wides read or more or less solubility ests with continuous mi ing and or an increased e osure time, or tests with a s ecial design where continuously, this may be difficult or im ossible hen conflicting results are obtained, the origin of the inoculum concentrations of the test substance lower than the water solubility ha e been em loyed, may be a ailable on slightly should be chec ed in order to clarify whether or not differences in the ada tation of the microbial community may be soluble com ounds the reason Test duration less than days s mentioned abo e, many substances may be to ic or inhibitory to the inoculum at the relati ely high concentrations tested in ready biodegradability tests s ecially in the odified ( test ( est ometimes degradation is re orted for tests terminated before the day eriod s ecified in the uideline and the anometric es irometry test ( est uideline high concentrations ( mg l standards (e g the , hese data are of course directly a licable when a degradation greater than or e ual to are rescribed he lowest test substance concentrations are rescribed in the losed ottle test ( est uideline the ass le el is obtained hen a lower degradation le el is reached, the results need to be inter reted with caution where mg l is used he ossibility of to ic effects may be e aluated by including a to icity control in the ne ossibility is that the duration of the test was too short and that the chemical structure would robably ha e been ready biodegradability test or by com aring the test concentration with to icity test data on micro organisms, e g the degraded in a day biodegradability test f substantial degradation occurs within a short time eriod, the situation res iration inhibition tests ( est uideline , the nitrification inhibition test ( or, if other microbial /COD ≥ 0.5 or with the requirements on degradation within the 10 days time may be com ared with the criterion to icity tests are not a ailable, the bioluminescence inhibition test ( hen conflicting results are found, this window n these cases, a substance may be considered readily degradable (and hence ra idly degradable , if may becaused by to icity of the test substance f the substance is not inhibitory at en ironmentally realistic concentrations, the greatest degradation measured in screening tests may be used as a basis for classification f (a the ultimate biodegradability within days or simulation test data are a ailable in such cases, consideration of these data may be es ecially im ortant, because a low non inhibitory concentration of the substance may ha e been em loyed, thus gi ing a more reliable indication of the (b the ultimate degradation rate constant in this eriod is day corres onding to a half life biodegradation half life of the substance under en ironmentally realistic conditions of days hen the solubility of the test substance is lower than the concentrations em loyed in a test, this hese criteria are ro osed in order to ensure that ra id minerali ation did occur, although the test arameter may be the limiting factor for the actual degradation measured n these cases, results from tests em loying was ended before days and before the ass le el was attained nter retation of test data that do not com ly with the the lowest concentrations of test substance should re ail, i e often the losed ottle test ( est uideline rescribed ass le els must be made with great caution t is mandatory to consider whether a biodegradability below n general, the ie way test ( est uideline and the odified creening test the ass le el was due to a artial degradation of the substance and not a com lete minerali ation f artial degradation ( est uideline are not suitable for testing the biodegradability of oorly soluble substances (e g is the robable e lanation for the obser ed biodegradability, the substance should be considered not readily est uideline biodegradable
- 470 - Copyright@United Nations, 2017. All rights reserved
be used for assessment of degradation in relation to a uatic ha ard classification of substances ubstances that are rimary biodegradation gases at ambient tem erature may howe er for e am le be considered further in this regard (see also edersen et al., n some tests, only the disa earance of the arent com ound (i e rimary degradation is determined for e am le by following the degradation by s ecific or grou s ecific chemical analyses of the test substance ata on o degradation data available rimary biodegradability may be used for demonstrating ra id degradability only when it can be satisfactorily demonstrated that the degradation roducts formed do not fulfil the criteria for classification as ha ardous to the a uatic hen no useful data on degradability are a ailable either e erimentally determined or estimated en ironment data the substance should be regarded as not ra idly degradable Conflicting results from screening tests ene al inte p etation p o le s he situation where more degradation data are a ailable for the same substance introduces the Complex substances ossibility of conflicting results n general, conflicting results for a substance which has been tested se eral times with an appropriate biodegradability test could be interpreted by a “weight of evidence approach”. This implies that if both he harmoni ed criteria for classification of chemicals as ha ardous for the a uatic en ironment focus ositi e (i e higher degradation than the ass le el and negati e results ha e been obtained for a substance in ready on single substances certain ty e of intrinsically com le substance are multi com onent substances hey are biodegradability tests, then the data of the highest uality and the best documentation should be used for determining ty ically of natural origin and need occasionally to be considered his may be the case for chemicals that are roduced the ready biodegradability of the substance Howe er, ositi e results in ready biodegradability tests could be or e tracted from mineral oil or lant material uch com le chemicals are normally considered as single substances in considered alid, irres ecti e of negati e results, when the scientific uality is good and the test conditions are well a regulatory conte t n most cases they are defined as a homologous series of substances within a certain range of documented, i e guideline criteria are fulfilled, including the use of non re e osed (non ada ted inoculum one of carbon chain length and or degree of substitution hen this is the case, no ma or difference in degradability is foreseen the arious screening tests are suitable for the testing of all ty es of substances, and results obtained by the use of a test and the degree of degradability can be established from tests of the com le chemical ne e ce tion would be when a rocedure which is not suitable for the s ecific substance should be e aluated carefully before a decision on the use is borderline degradation is found because in this case some of the indi idual substances may be ra idly degradable and ta en other may be not ra idly degradable his re uires a more detailed assessment of the degradability of the indi idual hus, there are a number of factors that may e lain conflicting biodegradability data from screening com onents in the com le substance hen not ra idly degradable com onents constitute a significant art of the tests com le substance (e g more than , or for a ha ardous com onent, an e en lower content , the substance should be regarded as not ra idly degradable (a inoculum Availability of the substance (b to icity of test substance (c test conditions egradation of organic substances in the en ironment ta es lace mostly in the a uatic com artments or in a uatic hases in soil or sediment Hydrolysis, of course, re uires the resence of water he acti ity of micro (d solubility of the test substance and organisms de ends on the resence of water oreo er, biodegradation re uires that the micro organisms are directly in (e olatili ation of the test substance contact with the substance issolution of the substance in the water hase that surrounds the micro organisms is therefore the most direct way for contact between the bacteria and fungi and the substrate he suitability of the inoculum for degrading the test substance de ends on the resence and amount of com etent degraders hen the inoculum is obtained from an en ironment that has re iously been e osed to the he resent standard methods for in estigating degradability of substances are de elo ed for readily test substance, the inoculum may be ada ted as e idenced by a degradation ca acity, which is greater than that of an soluble test com ounds Howe er, many organic substances are only slightly soluble in water s the standard tests inoculum from a non e osed en ironment s far as ossible the inoculum must be sam led from an une osed re uire mg l of the test substance, sufficient a ailability may not be reached for substances with a low water en ironment, but for substances that are used ubi uitously in high olumes and released wides read or more or less solubility ests with continuous mi ing and or an increased e osure time, or tests with a s ecial design where continuously, this may be difficult or im ossible hen conflicting results are obtained, the origin of the inoculum concentrations of the test substance lower than the water solubility ha e been em loyed, may be a ailable on slightly should be chec ed in order to clarify whether or not differences in the ada tation of the microbial community may be soluble com ounds the reason Test duration less than days s mentioned abo e, many substances may be to ic or inhibitory to the inoculum at the relati ely high concentrations tested in ready biodegradability tests s ecially in the odified ( test ( est ometimes degradation is re orted for tests terminated before the day eriod s ecified in the uideline and the anometric es irometry test ( est uideline high concentrations ( mg l standards (e g the , hese data are of course directly a licable when a degradation greater than or e ual to are rescribed he lowest test substance concentrations are rescribed in the losed ottle test ( est uideline the ass le el is obtained hen a lower degradation le el is reached, the results need to be inter reted with caution where mg l is used he ossibility of to ic effects may be e aluated by including a to icity control in the ne ossibility is that the duration of the test was too short and that the chemical structure would robably ha e been ready biodegradability test or by com aring the test concentration with to icity test data on micro organisms, e g the degraded in a day biodegradability test f substantial degradation occurs within a short time eriod, the situation res iration inhibition tests ( est uideline , the nitrification inhibition test ( or, if other microbial /COD ≥ 0.5 or with the requirements on degradation within the 10 days time may be com ared with the criterion to icity tests are not a ailable, the bioluminescence inhibition test ( hen conflicting results are found, this window n these cases, a substance may be considered readily degradable (and hence ra idly degradable , if may becaused by to icity of the test substance f the substance is not inhibitory at en ironmentally realistic concentrations, the greatest degradation measured in screening tests may be used as a basis for classification f (a the ultimate biodegradability within days or simulation test data are a ailable in such cases, consideration of these data may be es ecially im ortant, because a low non inhibitory concentration of the substance may ha e been em loyed, thus gi ing a more reliable indication of the (b the ultimate degradation rate constant in this eriod is day corres onding to a half life biodegradation half life of the substance under en ironmentally realistic conditions of days hen the solubility of the test substance is lower than the concentrations em loyed in a test, this hese criteria are ro osed in order to ensure that ra id minerali ation did occur, although the test arameter may be the limiting factor for the actual degradation measured n these cases, results from tests em loying was ended before days and before the ass le el was attained nter retation of test data that do not com ly with the the lowest concentrations of test substance should re ail, i e often the losed ottle test ( est uideline rescribed ass le els must be made with great caution t is mandatory to consider whether a biodegradability below n general, the ie way test ( est uideline and the odified creening test the ass le el was due to a artial degradation of the substance and not a com lete minerali ation f artial degradation ( est uideline are not suitable for testing the biodegradability of oorly soluble substances (e g is the robable e lanation for the obser ed biodegradability, the substance should be considered not readily est uideline biodegradable
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. . . . olatile substances should only be tested in closed systems as the losed ottle test Test iii the substance is considered to be not rapidly degradable based on indirect evidence as uideline the T test Test uideline and the anometric espirometry test Test e.g. nowledge from structurally similar substances or uideline . esults from other tests should be evaluated carefully and only considered if it can be demonstrated e.g. by mass balance estimates that the removal of the test substance is not a result of volatili ation. iv no other data regarding degradability are available.
. . . ariation in simulation test data io cc tion
number of simulation test data may be available for certain high priority chemicals. ften such data nt o ction provide a range of half lives in environmental media such as soil sediment and or surface water. The observed differences in half lives from simulation tests performed on the same substance may reflect differences in test . . . ioaccumulation is one of the important intrinsic properties of substances that determine the potential conditions all of which may be environmentally relevant. suitable half life in the higher end of the observed range of environmental ha ard. ioaccumulation of a substance into an organism is not a ha ard in itself but bioconcentration half lives from such investigations should be selected for classification by employing a weight of evidence approach and bioaccumulation will result in a body burden which may or may not lead to to ic effects. n the harmoni ed and ta ing the realism and relevance of the employed tests into account in relation to environmental conditions. n integrated ha ard classification system for human health and environmental effects of chemical substances general simulation test data of surface water are preferred relative to a uatic sediment or soil simulation test data in 1998), the wording “potential for bioaccumulation” is given. A distinction should however be drawn between relation to the evaluation of rapid degradability in the a uatic environment. bioconcentration and bioaccumulation. ere bioconcentration is defined as the net result of upta e transformation and elimination of a substance in an organism due to waterborne e posure whereas bioaccumulation includes all routes of ecision sc e e e posure i.e. via air water sediment soil and food . inally biomagnification is defined as accumulation and transfer of substances via the food chain resulting in an increase of internal concentrations in organisms on higher levels of the The following decision scheme may be used as a general guidance to facilitate decisions in relation to trophic chain uropean ommission . or most organic chemicals upta e from water bioconcentration is rapid degradability in the a uatic environment and classification of chemicals ha ardous to the a uatic environment. believed to be the predominant route of upta e. nly for very hydrophobic substances does upta e from food becomes important. lso the harmoni ed classification criteria use the bioconcentration factor or the octanol water partition substance is considered to be not rapidly degradable unless at least one of the following is fulfilled coefficient as the measure of the potential for bioaccumulation. or these reasons the present guidance document only a the substance is demonstrated to be readily biodegradable in a day test for ready considers bioconcentration and does not discuss upta e via food or other routes. biodegradability. The pass level of the test removal or theoretical o ygen demand must be achieved within days from the onset of biodegradation if it is possible to . . . lassification of a substance is primarily based on its intrinsic properties. owever the degree of evaluate this according to the available test data. f this is not possible then the pass level bioconcentration also depends on factors such as the degree of bioavailability the physiology of test organism should be evaluated within a days time window if possible or after the end of the test or maintenance of constant e posure concentration e posure duration metabolism inside the body of the target organism and e cretion from the body. The interpretation of the bioconcentration potential in a chemical classification conte t b the substance is demonstrated to be ultimately degraded in a surface water simulation test with therefore re uires an evaluation of the intrinsic properties of the substance as well as of the e perimental conditions a half life of days corresponding to a degradation of within days or under which bioconcentration factor has been determined. ased on the guide a decision scheme for application of bioconcentration data or log ow data for classification purposes has been developed. The emphasis of the present c the substance is demonstrated to be primarily degraded biotically or abiotically in the a uatic section is organic substances and organo metals. ioaccumulation of metals is also discussed in ection . . environment with a half life days corresponding to a degradation of within days and it can be demonstrated that the degradation products do not fulfil the criteria for . . . ata on bioconcentration properties of a substance may be available from standardi ed tests or may be classification as ha ardous to the a uatic environment. estimated from the structure of the molecule. The interpretation of such bioconcentration data for classification purposes often re uires detailed evaluation of test data. n order to facilitate this evaluation two additional appendi es hen these data are not available rapid degradation may be demonstrated if either of the following are enclosed. These appendi es describe available methods ppendi of nne and factors influencing the criteria are ustified bioconcentration potential ppendi of nne . inally a list of standardi ed e perimental methods for determination of bioconcentration and ow are attached ppendi of nne together with a list of references d the substance is demonstrated to be ultimately degraded in an a uatic sediment or soil ppendi of nne . simulation test with a half life of days corresponding to a degradation of within days or nte p etation of ioconcent ation ata
e in those cases where only and data are available the ratio of /COD is ≥ 0.5. . . . nvironmental ha ard classification of a substance is normally based on e isting data on its The same criterion applies to ready biodegradability tests of a shorter duration than days if environmental properties. Test data will only seldom be produced with the main purpose of facilitating a classification. the half life furthermore is days. ften a diverse range of test data is available which does not necessarily match the classification criteria. onse uently guidance is needed on interpretation of e isting test data in the conte t of ha ard classification. f none of the above types of data are available then the substance is considered as not rapidly degradable. This decision may be supported by fulfilment of at least one of the following . . . ioconcentration of an organic substance can be e perimentally determined in bioconcentration criteria e periments during which is measured as the concentration in the organism relative to the concentration in water under steady state conditions and or estimated from the upta e rate constant and the elimination rate constant i the substance is not inherently degradable in an inherent biodegradability test or . n general the potential of an organic substance to bioconcentrate is primarily related to the ii the substances is predicted to be slowly biodegradable by scientifically valid s lipophilicity of the substance. measure of lipophilicity is the n octanol water partition coefficient ow which for e.g. for the iodegradation robability rogram the score for rapid degradation linear lipophilic non ionic organic substances undergoing minimal metabolism or biotransformation within the organism is or non linear model . or correlated with the bioconcentration factor. Therefore ow is often used for estimating the bioconcentration of organic substances based on the empirical relationship between log and log ow. or most organic substances estimation methods are available for calculating the ow. ata on the bioconcentration properties of a substance may thus be i e perimentally determined ii estimated from e perimentally determined ow or iii estimated from ow values derived by use of uantitative tructure ctivity elationships s . uidance for interpretation of such data is given below together with guidance on assessment of chemical classes which need special attention. imulations tests should reflect realistic environmental conditions such as low concentration of the chemical realistic temperature and employment of ambient microbial biomass not pre exposed to the chemical. - 472 - Copyright@United Nations, 2017. All rights reserved
. . . . olatile substances should only be tested in closed systems as the losed ottle test Test iii the substance is considered to be not rapidly degradable based on indirect evidence as uideline the T test Test uideline and the anometric espirometry test Test e.g. nowledge from structurally similar substances or uideline . esults from other tests should be evaluated carefully and only considered if it can be demonstrated e.g. by mass balance estimates that the removal of the test substance is not a result of volatili ation. iv no other data regarding degradability are available.
. . . ariation in simulation test data io cc tion
number of simulation test data may be available for certain high priority chemicals. ften such data nt o ction provide a range of half lives in environmental media such as soil sediment and or surface water. The observed differences in half lives from simulation tests performed on the same substance may reflect differences in test . . . ioaccumulation is one of the important intrinsic properties of substances that determine the potential conditions all of which may be environmentally relevant. suitable half life in the higher end of the observed range of environmental ha ard. ioaccumulation of a substance into an organism is not a ha ard in itself but bioconcentration half lives from such investigations should be selected for classification by employing a weight of evidence approach and bioaccumulation will result in a body burden which may or may not lead to to ic effects. n the harmoni ed and ta ing the realism and relevance of the employed tests into account in relation to environmental conditions. n integrated ha ard classification system for human health and environmental effects of chemical substances general simulation test data of surface water are preferred relative to a uatic sediment or soil simulation test data in 1998), the wording “potential for bioaccumulation” is given. A distinction should however be drawn between relation to the evaluation of rapid degradability in the a uatic environment. bioconcentration and bioaccumulation. ere bioconcentration is defined as the net result of upta e transformation and elimination of a substance in an organism due to waterborne e posure whereas bioaccumulation includes all routes of ecision sc e e e posure i.e. via air water sediment soil and food . inally biomagnification is defined as accumulation and transfer of substances via the food chain resulting in an increase of internal concentrations in organisms on higher levels of the The following decision scheme may be used as a general guidance to facilitate decisions in relation to trophic chain uropean ommission . or most organic chemicals upta e from water bioconcentration is rapid degradability in the a uatic environment and classification of chemicals ha ardous to the a uatic environment. believed to be the predominant route of upta e. nly for very hydrophobic substances does upta e from food becomes important. lso the harmoni ed classification criteria use the bioconcentration factor or the octanol water partition substance is considered to be not rapidly degradable unless at least one of the following is fulfilled coefficient as the measure of the potential for bioaccumulation. or these reasons the present guidance document only a the substance is demonstrated to be readily biodegradable in a day test for ready considers bioconcentration and does not discuss upta e via food or other routes. biodegradability. The pass level of the test removal or theoretical o ygen demand must be achieved within days from the onset of biodegradation if it is possible to . . . lassification of a substance is primarily based on its intrinsic properties. owever the degree of evaluate this according to the available test data. f this is not possible then the pass level bioconcentration also depends on factors such as the degree of bioavailability the physiology of test organism should be evaluated within a days time window if possible or after the end of the test or maintenance of constant e posure concentration e posure duration metabolism inside the body of the target organism and e cretion from the body. The interpretation of the bioconcentration potential in a chemical classification conte t b the substance is demonstrated to be ultimately degraded in a surface water simulation test with therefore re uires an evaluation of the intrinsic properties of the substance as well as of the e perimental conditions a half life of days corresponding to a degradation of within days or under which bioconcentration factor has been determined. ased on the guide a decision scheme for application of bioconcentration data or log ow data for classification purposes has been developed. The emphasis of the present c the substance is demonstrated to be primarily degraded biotically or abiotically in the a uatic section is organic substances and organo metals. ioaccumulation of metals is also discussed in ection . . environment with a half life days corresponding to a degradation of within days and it can be demonstrated that the degradation products do not fulfil the criteria for . . . ata on bioconcentration properties of a substance may be available from standardi ed tests or may be classification as ha ardous to the a uatic environment. estimated from the structure of the molecule. The interpretation of such bioconcentration data for classification purposes often re uires detailed evaluation of test data. n order to facilitate this evaluation two additional appendi es hen these data are not available rapid degradation may be demonstrated if either of the following are enclosed. These appendi es describe available methods ppendi of nne and factors influencing the criteria are ustified bioconcentration potential ppendi of nne . inally a list of standardi ed e perimental methods for determination of bioconcentration and ow are attached ppendi of nne together with a list of references d the substance is demonstrated to be ultimately degraded in an a uatic sediment or soil ppendi of nne . simulation test with a half life of days corresponding to a degradation of within days or nte p etation of ioconcent ation ata
e in those cases where only and data are available the ratio of /COD is ≥ 0.5. . . . nvironmental ha ard classification of a substance is normally based on e isting data on its The same criterion applies to ready biodegradability tests of a shorter duration than days if environmental properties. Test data will only seldom be produced with the main purpose of facilitating a classification. the half life furthermore is days. ften a diverse range of test data is available which does not necessarily match the classification criteria. onse uently guidance is needed on interpretation of e isting test data in the conte t of ha ard classification. f none of the above types of data are available then the substance is considered as not rapidly degradable. This decision may be supported by fulfilment of at least one of the following . . . ioconcentration of an organic substance can be e perimentally determined in bioconcentration criteria e periments during which is measured as the concentration in the organism relative to the concentration in water under steady state conditions and or estimated from the upta e rate constant and the elimination rate constant i the substance is not inherently degradable in an inherent biodegradability test or . n general the potential of an organic substance to bioconcentrate is primarily related to the ii the substances is predicted to be slowly biodegradable by scientifically valid s lipophilicity of the substance. measure of lipophilicity is the n octanol water partition coefficient ow which for e.g. for the iodegradation robability rogram the score for rapid degradation linear lipophilic non ionic organic substances undergoing minimal metabolism or biotransformation within the organism is or non linear model . or correlated with the bioconcentration factor. Therefore ow is often used for estimating the bioconcentration of organic substances based on the empirical relationship between log and log ow. or most organic substances estimation methods are available for calculating the ow. ata on the bioconcentration properties of a substance may thus be i e perimentally determined ii estimated from e perimentally determined ow or iii estimated from ow values derived by use of uantitative tructure ctivity elationships s . uidance for interpretation of such data is given below together with guidance on assessment of chemical classes which need special attention. imulations tests should reflect realistic environmental conditions such as low concentration of the chemical realistic temperature and employment of ambient microbial biomass not pre exposed to the chemical. - 473 - Copyright@United Nations, 2017. All rights reserved
A9. . . ioconcentration factor ( C A9. . . .8. enerall , the highest valid value e pressed on this common lipid basis is used to determine the wet weight based value in relation to the cut off value for of of the harmoni ed classification criteria see A9. . . .1 he bioconcentration factor is defined as the ratio on a weight basis between the concentration of the hapter .1, able .1.1). chemical in biota and the concentration in the surrounding medium, here water, at stead state. can thus be e perimentall derived under stead state conditions, on the basis of measured concentrations. owever, can also A9. . . .9 se of radiolabelled substances be calculated as the ratio between the first order upta e and elimination rate constants a method which does not re uire e uilibrium conditions. A9. . . .9.1 he use of radiolabelled test substances can facilitate the anal sis of water and fish samples. owever, unless combined with a specific anal tical method, the total radioactivit measurements potentiall reflect the presence A9. . . . ifferent test guidelines for the e perimental determination of bioconcentration in fish have been of the parent substance as well as possible metabolite s) and possible metaboli ed carbon, which have been documented and adopted, the most generall applied being the test guideline , 199 ). incorporated in the fish tissue in organic molecules. values determined b use of radiolabelled test substances are therefore normall overestimated. A9. . . . perimentall derived values of high ualit are ultimatel preferred for classification purposes as such data override surrogate data, e.g. ow. A9. . . .9. hen using radiolabelled substances, the labelling is most often placed in the stable part of the molecule, for which reason the measured value includes the of the metabolites. or some substances it is the A9. . . . igh ualit data are defined as data where the validit criteria for the test method applied are metabolite which is the most to ic and which has the highest bioconcentration potential. easurements of the parent fulfilled and described, e.g. maintenance of constant e posure concentration o gen and temperature variations, and substance as well as the metabolites ma thus be important for the interpretation of the a uatic ha ard including the documentation that stead state conditions have been reached, etc. he e periment will be regarded as a high ualit bioconcentration potential) of such substances. stud , if a proper description is provided e.g. b ood aborator ractice )) allowing verification that validit criteria are fulfilled. n addition, an appropriate anal tical method must be used to uantif the chemical and its to ic A9. . . .9. n e periments where radiolabelled substances have been used, high radiolabel concentrations are metabolites in the water and fish tissue see section 1, Appendi for further details). often found in the gall bladder of fish. his is interpreted to be caused b biotransformation in the liver and subse uentl b e cretion of metabolites in the gall bladder omotto et al., 19 9 a aba ashi et al., 198 oodrich A9. . . . values of low or uncertain ualit ma give a false and too low value e.g. application of et al., 1991 oshima et al., 199 ). hen fish do not eat, the content of the gall bladder is not emptied into the gut, and measured concentrations of the test substance in fish and water, but measured after a too short e posure period in which high concentrations of metabolites ma build up in the gall bladder. he feeding regime ma thus have a pronounced stead state conditions have not been reached cf. , 199 , regarding estimation of time to e uilibrium). effect on the measured . n the literature man studies are found where radiolabelled compounds are used, and herefore, such data should be carefull evaluated before use and consideration should be given to using ow instead. where the fish are not fed. As a result high concentrations of radioactive material are found in the gall bladder. n these studies the bioconcentration ma in most cases have been overestimated. hus when evaluating e periments, in which A9. . . . f there is no value for fish species, high ualit data on the value for other species ma be radiolabelled compounds are used, it is essential to evaluate the feeding regime as well. used e.g. determined on blue mussel, o ster, scallop A 1 9 )). eported s for microalgae should be used with caution. A9. . . .9. f the in terms of radiolabelled residues is documented to be 1 , identification and uantification of degradation products, representing 1 of total residues in fish tissues at stead state, are for e.g. A9. . . . or highl lipophilic substances, e.g. with log ow above , e perimentall derived values tend pesticides strongl recommended in the guideline o. 199 ). f no identification and uantification of to decrease with increasing log ow. onceptual e planations of this non linearit mainl refer to either reduced metabolites are available, the assessment of bioconcentration should be based on the measured radiolabelled value. membrane permeation inetics or reduced biotic lipid solubilit for large molecules. A low bioavailabilit and upta e of f, for highl bioaccumulative substances ), onl s based on the parent compound and on radiolabelled these substances in the organism will thus occur. ther factors comprise e perimental artefacts, such as e uilibrium not measurements are available, the latter should thus be used in relation to classification. being reached, reduced bioavailabilit due to sorption to organic matter in the a ueous phase, and anal tical errors. pecial care should thus be ta en when evaluating e perimental data on for highl lipophilic substances as these A9. . . Octanol water partitioning coefficient ( data will have a much higher level of uncertaint than values determined for less lipophilic substances. ow A9. . . .1 or organic substances e perimentall derived high ualit values, or values which are evaluated A9. . . .8 in different test species ow in reviews and assigned as the “recommended values”, are preferred over other determinations of ow. hen no e perimental data of high ualit are available, validated uantitative tructure Activit elationships A s) for log A9. . . .8.1 values used for classification are based on whole bod measurements. As stated previousl , the ow ma be used in the classification process. uch validated A s ma be used without modification to the agreed optimal data for classification are values derived using the test method or internationall e uivalent criteria if the are restricted to chemicals for which their applicabilit is well characteri ed. or substances li e strong methods, which uses small fish. ue to the higher gill surface to weight ratio for smaller organisms than larger acids and bases, substances which react with the eluent, or surface active substances, a A estimated value of or organisms, stead state conditions will be reached sooner in smaller organisms than in larger ones. he si e of the ow an estimate based on individual n octanol and water solubilities should be provided instead of an anal tical organisms fish) used in bioconcentration studies is thus of considerable importance in relation to the time used in the determination of ow A.8., 199 11 , 1989). easurements should be ta en on ioni able substances in upta e phase, when the reported value is based solel on measured concentrations in fish and water at stead state. their non ioni ed form free acid or free base) onl b using an appropriate buffer with p below p for free acid or hus, if large fish, e.g. adult salmon, have been used in bioconcentration studies, it should be evaluated whether the above the p for free base. upta e period was sufficientl long for stead state to be reached or to allow for a inetic upta e rate constant to be determined precisel . A9. . . . perimental determination of ow
A9. . . .8. urthermore, when using e isting data for classification, it is possible that the values could be or e perimental determination of ow values, several different methods, ha e flas , and , are derived from several different fish or other a uatic species e.g. clams) and for different organs in the fish. hus, to described in standard guidelines, e.g. est uideline 1 199 ) est uideline 11 1989) A.8. compare these data to each other and to the criteria, some common basis or normali ation will be re uired. t has been 199 ) A 198 ) A A 198 ) A 199 ). he sha e flas method is recommended when the log noted that there is a close relationship between the lipid content of a fish or an a uatic organism and the observed ow value falls within the range from – to . he sha e flas method applies onl to essential pure substances soluble value. herefore, when comparing values across different fish species or when converting values for specific in water and n octanol. or highl lipophilic substances, which slowl dissolve in water, data obtained b emplo ing a organs to whole bod s, the common approach is to e press the values on a common lipid content. f e.g. slow stirring method are generall more reliable. urthermore, the e perimental difficulties, associated with the whole bod values or values for specific organs are found in the literature, the first step is to calculate the formation of microdroplets during the sha e flas e periment, can to some degree be overcome b a slow stirring on a lipid basis using the relative content of fat in the fish cf. literature test guideline for t pical fat content of method where water, octanol, and test compound are e uilibrated in a gentl stirred reactor. ith the slow stirring the test species) or the organ. n the second step the for the whole bod for a t pical a uatic organism i.e. small method est uideline 1 ) a precise and accurate determination of ow of compounds with log ow of up to fish) is calculated assuming a common default lipid content. A default value of is most commonl used edersen et 8. is allowed. As for the sha e flas method, the slow stirring method applies onl to essentiall pure substances al., 199 ) as this represents the average lipid content of the small fish used in 199 ). soluble in water and n octanol. he method, which is performed on anal tical columns, is recommended when the log ow value falls within the range to . he method is less sensitive to the presence of impurities in the
- 474 - Copyright@United Nations, 2017. All rights reserved
A9. . . ioconcentration factor ( C A9. . . .8. enerall , the highest valid value e pressed on this common lipid basis is used to determine the wet weight based value in relation to the cut off value for of of the harmoni ed classification criteria see A9. . . .1 he bioconcentration factor is defined as the ratio on a weight basis between the concentration of the hapter .1, able .1.1). chemical in biota and the concentration in the surrounding medium, here water, at stead state. can thus be e perimentall derived under stead state conditions, on the basis of measured concentrations. owever, can also A9. . . .9 se of radiolabelled substances be calculated as the ratio between the first order upta e and elimination rate constants a method which does not re uire e uilibrium conditions. A9. . . .9.1 he use of radiolabelled test substances can facilitate the anal sis of water and fish samples. owever, unless combined with a specific anal tical method, the total radioactivit measurements potentiall reflect the presence A9. . . . ifferent test guidelines for the e perimental determination of bioconcentration in fish have been of the parent substance as well as possible metabolite s) and possible metaboli ed carbon, which have been documented and adopted, the most generall applied being the test guideline , 199 ). incorporated in the fish tissue in organic molecules. values determined b use of radiolabelled test substances are therefore normall overestimated. A9. . . . perimentall derived values of high ualit are ultimatel preferred for classification purposes as such data override surrogate data, e.g. ow. A9. . . .9. hen using radiolabelled substances, the labelling is most often placed in the stable part of the molecule, for which reason the measured value includes the of the metabolites. or some substances it is the A9. . . . igh ualit data are defined as data where the validit criteria for the test method applied are metabolite which is the most to ic and which has the highest bioconcentration potential. easurements of the parent fulfilled and described, e.g. maintenance of constant e posure concentration o gen and temperature variations, and substance as well as the metabolites ma thus be important for the interpretation of the a uatic ha ard including the documentation that stead state conditions have been reached, etc. he e periment will be regarded as a high ualit bioconcentration potential) of such substances. stud , if a proper description is provided e.g. b ood aborator ractice )) allowing verification that validit criteria are fulfilled. n addition, an appropriate anal tical method must be used to uantif the chemical and its to ic A9. . . .9. n e periments where radiolabelled substances have been used, high radiolabel concentrations are metabolites in the water and fish tissue see section 1, Appendi for further details). often found in the gall bladder of fish. his is interpreted to be caused b biotransformation in the liver and subse uentl b e cretion of metabolites in the gall bladder omotto et al., 19 9 a aba ashi et al., 198 oodrich A9. . . . values of low or uncertain ualit ma give a false and too low value e.g. application of et al., 1991 oshima et al., 199 ). hen fish do not eat, the content of the gall bladder is not emptied into the gut, and measured concentrations of the test substance in fish and water, but measured after a too short e posure period in which high concentrations of metabolites ma build up in the gall bladder. he feeding regime ma thus have a pronounced stead state conditions have not been reached cf. , 199 , regarding estimation of time to e uilibrium). effect on the measured . n the literature man studies are found where radiolabelled compounds are used, and herefore, such data should be carefull evaluated before use and consideration should be given to using ow instead. where the fish are not fed. As a result high concentrations of radioactive material are found in the gall bladder. n these studies the bioconcentration ma in most cases have been overestimated. hus when evaluating e periments, in which A9. . . . f there is no value for fish species, high ualit data on the value for other species ma be radiolabelled compounds are used, it is essential to evaluate the feeding regime as well. used e.g. determined on blue mussel, o ster, scallop A 1 9 )). eported s for microalgae should be used with caution. A9. . . .9. f the in terms of radiolabelled residues is documented to be 1 , identification and uantification of degradation products, representing 1 of total residues in fish tissues at stead state, are for e.g. A9. . . . or highl lipophilic substances, e.g. with log ow above , e perimentall derived values tend pesticides strongl recommended in the guideline o. 199 ). f no identification and uantification of to decrease with increasing log ow. onceptual e planations of this non linearit mainl refer to either reduced metabolites are available, the assessment of bioconcentration should be based on the measured radiolabelled value. membrane permeation inetics or reduced biotic lipid solubilit for large molecules. A low bioavailabilit and upta e of f, for highl bioaccumulative substances ), onl s based on the parent compound and on radiolabelled these substances in the organism will thus occur. ther factors comprise e perimental artefacts, such as e uilibrium not measurements are available, the latter should thus be used in relation to classification. being reached, reduced bioavailabilit due to sorption to organic matter in the a ueous phase, and anal tical errors. pecial care should thus be ta en when evaluating e perimental data on for highl lipophilic substances as these A9. . . Octanol water partitioning coefficient ( data will have a much higher level of uncertaint than values determined for less lipophilic substances. ow A9. . . .1 or organic substances e perimentall derived high ualit values, or values which are evaluated A9. . . .8 in different test species ow in reviews and assigned as the “recommended values”, are preferred over other determinations of ow. hen no e perimental data of high ualit are available, validated uantitative tructure Activit elationships A s) for log A9. . . .8.1 values used for classification are based on whole bod measurements. As stated previousl , the ow ma be used in the classification process. uch validated A s ma be used without modification to the agreed optimal data for classification are values derived using the test method or internationall e uivalent criteria if the are restricted to chemicals for which their applicabilit is well characteri ed. or substances li e strong methods, which uses small fish. ue to the higher gill surface to weight ratio for smaller organisms than larger acids and bases, substances which react with the eluent, or surface active substances, a A estimated value of or organisms, stead state conditions will be reached sooner in smaller organisms than in larger ones. he si e of the ow an estimate based on individual n octanol and water solubilities should be provided instead of an anal tical organisms fish) used in bioconcentration studies is thus of considerable importance in relation to the time used in the determination of ow A.8., 199 11 , 1989). easurements should be ta en on ioni able substances in upta e phase, when the reported value is based solel on measured concentrations in fish and water at stead state. their non ioni ed form free acid or free base) onl b using an appropriate buffer with p below p for free acid or hus, if large fish, e.g. adult salmon, have been used in bioconcentration studies, it should be evaluated whether the above the p for free base. upta e period was sufficientl long for stead state to be reached or to allow for a inetic upta e rate constant to be determined precisel . A9. . . . perimental determination of ow
A9. . . .8. urthermore, when using e isting data for classification, it is possible that the values could be or e perimental determination of ow values, several different methods, ha e flas , and , are derived from several different fish or other a uatic species e.g. clams) and for different organs in the fish. hus, to described in standard guidelines, e.g. est uideline 1 199 ) est uideline 11 1989) A.8. compare these data to each other and to the criteria, some common basis or normali ation will be re uired. t has been 199 ) A 198 ) A A 198 ) A 199 ). he sha e flas method is recommended when the log noted that there is a close relationship between the lipid content of a fish or an a uatic organism and the observed ow value falls within the range from – to . he sha e flas method applies onl to essential pure substances soluble value. herefore, when comparing values across different fish species or when converting values for specific in water and n octanol. or highl lipophilic substances, which slowl dissolve in water, data obtained b emplo ing a organs to whole bod s, the common approach is to e press the values on a common lipid content. f e.g. slow stirring method are generall more reliable. urthermore, the e perimental difficulties, associated with the whole bod values or values for specific organs are found in the literature, the first step is to calculate the formation of microdroplets during the sha e flas e periment, can to some degree be overcome b a slow stirring on a lipid basis using the relative content of fat in the fish cf. literature test guideline for t pical fat content of method where water, octanol, and test compound are e uilibrated in a gentl stirred reactor. ith the slow stirring the test species) or the organ. n the second step the for the whole bod for a t pical a uatic organism i.e. small method est uideline 1 ) a precise and accurate determination of ow of compounds with log ow of up to fish) is calculated assuming a common default lipid content. A default value of is most commonl used edersen et 8. is allowed. As for the sha e flas method, the slow stirring method applies onl to essentiall pure substances al., 199 ) as this represents the average lipid content of the small fish used in 199 ). soluble in water and n octanol. he method, which is performed on anal tical columns, is recommended when the log ow value falls within the range to . he method is less sensitive to the presence of impurities in the
- 475 - Copyright@United Nations, 2017. All rights reserved
test compound compared to the sha e flas method nother techni ue for measuring log ow is the generator column o ioconcentrate organic compounds, a su stance needs to e solu le in lipids, present in the water, method and availa le for transfer across the fish gills roperties which alter this availa ilit will thus change the actual ioconcentration of a su stance, when compared with the prediction or e ample, readil iodegrada le su stances s an e perimental determination of the ow is not alwa s possi le, e g for ver water solu le ma onl e present in the a uatic compartment for short periods of time imilarl , volatilit , and h drol sis will su stances, ver lipophilic su stances, and surfactants, a derived ow ma e used reduce the concentration and the time during which a su stance is availa le for ioconcentration further important parameter, which ma reduce the actual e posure concentration of a su stance, is adsorption, either to particulate matter se of s for determination of log ow or to surfaces in general here are a num er of su stances, which have shown to e rapidl transformed in the organism, thus leading to a lower value than e pected u stances that form micelles or aggregates ma hen an estimated ow value is found, the estimation method has to e ta en into account umerous ioconcentrate to a lower e tent than would e predicted from simple ph sico chemical properties his is also the case s have een and continue to e developed for the estimation of ow our commerciall availa le programmes for h dropho ic su stances that are contained in micelles formed as a conse uence of the use of dispersants herefore, , , , are fre uentl used for ris assessment if no e perimentall the use of dispersants in ioaccumulation tests is discouraged derived data are availa le , and are ased upon the addition of group contri utions, while is ased upon a more fundamental chemical structure algorithm nl can e emplo ed in a n general, for difficult to test su stances, measured and ow values – ased on the parent general wa for inorganic or organometallic compounds pecial methods are needed for estimating log ow for surface su stance – are a prere uisite for the determination of the ioconcentration potential urthermore, proper active compounds, chelating compounds and mi tures is recommended in the oint pro ect on documentation of the test concentration is a prere uisite for the validation of the given value validation of estimation methods edersen et al recommended the and the programmes for classification purposes ecause of their relia ilit , commercial availa ilit , and oorly soluble and complex substances convenience of use he following estimation methods are recommended for classification purposes a le e eco en e or e ti tion o pecial attention should e paid to poorl solu le su stances re uentl the solu ilit of these o su stances is recorded as less than the detection limit, which creates pro lems in interpreting the ioconcentration
o e o o r n e t nce ti it potential or such su stances the ioconcentration potential should e ased on e perimental determination of log ow or estimations of log ow log ow he program calculates log ow for organic compounds containing , , , , al, , and or hen a multi component su stance is not full solu le in water, it is important to attempt to identif log ow he program calculates log ow for organic compounds containing , , the components of the mi ture as far as practicall possi le and to e amine the possi ilit of determining its , , al, i, , e, i, a, , and or g ome surfactants e g ioaccumulation potential using availa le information on its components hen ioaccumulating components alcohol etho lates, d estuffs, and dissociated su stances ma e constitute a significant part of the comple su stance e g more than or for ha ardous components an even lower predicted the program as well content , the comple su stance should e regarded as eing ioaccumulating
log ow he programme calculates log ow for organic compounds containing , , , , al, and mprovements are in progress in order to e tend igh molecular weight substances the applica ilit of ove certain molecular dimensions, the potential of a su stance to ioconcentrate decreases his is rovides improved results is a mechanistic model ased on chemical thermod namic possi l due to steric hindrance of the passage of the su stance through gill mem ranes t has een proposed that a cut over and principles rather than a deterministic model rooted in nowledge off limit of for the molecular weight could e applied e g uropean ommission, owever, this cut off has for compounds o tained from o servational data herefore, differs from een su ect to criticism and an alternative cut off of has een proposed in relation to e clusion of consideration with models that use s i e , , in that of su stances with possi le indirect a uatic effects , n general, ioconcentration of possi le meta olites no measured log data are needed for a training set of chemicals log ow ow or environmental degradation products of large molecules should e considered ata on ioconcentration of molecules nl can e emplo ed in a general wa for inorganic or with a high molecular weight should therefore e carefull evaluated and onl e used if such data are considered to e organometallic compounds full valid in respect to oth the parent compound and its possi le meta olites and environmental degradation products a A validation study performed by iemel who compared experimental determined log ow values with estimated values showed that the program precisely predicts the log ow for a great number of organic chemicals in the log ow urface active agents range from below 0 to above 9 (n 50 r 0.96 (Tema ord 995 5 . urfactants consist of a lipophilic most often an al l chain and a h drophilic part the polar headgroup ased on a scatter plot of estimated vs. experimental log ( yracuse esearch Corporation 999 where 05 ow ccording to the charge of the headgroup, surfactants are su divided into classes of anionic, cationic, non ionic, or compound have been tested the LOG O is evaluated being valid for compounds with a log in the interval . ow amphoteric surfactants ue to the variet of different headgroups, surfactants are a structurall diverse class of compounds, which is defined surface activit rather than chemical structure he ioaccumulation potential of C e ical classes t at nee special attention it espect to C an o al es surfactants should thus e considered in relation to the different su classes anionic, cationic, non ionic, or amphoteric instead of to the group as a whole urface active su stances ma form emulsions, in which the ioavaila ilit is here are certain ph sico chemical properties, which can ma e the determination of or its difficult to ascertain icelle formation can result in a change of the ioavaila le fraction even when the solutions are measurement difficult hese ma e su stances, which do not ioconcentrate in a manner consistent with their other apparentl formed, thus giving pro lems in interpretation of the ioaccumulation potential ph sico chemical properties, e g steric hindrance or su stances which ma e the use of descriptors inappropriate, e g surface activit , which ma es oth the measurement and use of log inappropriate ow perimentall derived ioconcentration factors Difficult substances easured values on surfactants show that ma increase with increasing al l chain length ome su stances are difficult to test in a uatic s stems and guidance has een developed to assist in and e dependant of the site of attachment of the head group, and other structural features testing these materials o , he uidance document on a uatic to icit testing of difficult su stances and mi tures , is also a good source of information for ctanol water partition coefficient ow ioconcentration studies, on the t pes of su stances that are difficult to test and the steps needed to ensure valid conclusions from tests with these su stances ifficult to test su stances ma e poorl solu le, volatile, or su ect to he octanol water partition coefficient for surfactants can not e determined using the sha e flas or rapid degradation due to such processes as phototransformation, h drol sis, o idation, or iotic degradation slow stirring method ecause of the formation of emulsions n addition, the surfactant molecules will e ist in the water phase almost e clusivel as ions, whereas the will have to pair with a counter ion in order to e dissolved in octanol herefore, e perimental determination of ow does not characteri e the partition of ionic surfactants olls, n
- 476 - Copyright@United Nations, 2017. All rights reserved
test compound compared to the sha e flas method nother techni ue for measuring log ow is the generator column o ioconcentrate organic compounds, a su stance needs to e solu le in lipids, present in the water, method and availa le for transfer across the fish gills roperties which alter this availa ilit will thus change the actual ioconcentration of a su stance, when compared with the prediction or e ample, readil iodegrada le su stances s an e perimental determination of the ow is not alwa s possi le, e g for ver water solu le ma onl e present in the a uatic compartment for short periods of time imilarl , volatilit , and h drol sis will su stances, ver lipophilic su stances, and surfactants, a derived ow ma e used reduce the concentration and the time during which a su stance is availa le for ioconcentration further important parameter, which ma reduce the actual e posure concentration of a su stance, is adsorption, either to particulate matter se of s for determination of log ow or to surfaces in general here are a num er of su stances, which have shown to e rapidl transformed in the organism, thus leading to a lower value than e pected u stances that form micelles or aggregates ma hen an estimated ow value is found, the estimation method has to e ta en into account umerous ioconcentrate to a lower e tent than would e predicted from simple ph sico chemical properties his is also the case s have een and continue to e developed for the estimation of ow our commerciall availa le programmes for h dropho ic su stances that are contained in micelles formed as a conse uence of the use of dispersants herefore, , , , are fre uentl used for ris assessment if no e perimentall the use of dispersants in ioaccumulation tests is discouraged derived data are availa le , and are ased upon the addition of group contri utions, while is ased upon a more fundamental chemical structure algorithm nl can e emplo ed in a n general, for difficult to test su stances, measured and ow values – ased on the parent general wa for inorganic or organometallic compounds pecial methods are needed for estimating log ow for surface su stance – are a prere uisite for the determination of the ioconcentration potential urthermore, proper active compounds, chelating compounds and mi tures is recommended in the oint pro ect on documentation of the test concentration is a prere uisite for the validation of the given value validation of estimation methods edersen et al recommended the and the programmes for classification purposes ecause of their relia ilit , commercial availa ilit , and oorly soluble and complex substances convenience of use he following estimation methods are recommended for classification purposes a le e eco en e or e ti tion o pecial attention should e paid to poorl solu le su stances re uentl the solu ilit of these o su stances is recorded as less than the detection limit, which creates pro lems in interpreting the ioconcentration
o e o o r n e t nce ti it potential or such su stances the ioconcentration potential should e ased on e perimental determination of log ow or estimations of log ow log ow he program calculates log ow for organic compounds containing , , , , al, , and or hen a multi component su stance is not full solu le in water, it is important to attempt to identif log ow he program calculates log ow for organic compounds containing , , the components of the mi ture as far as practicall possi le and to e amine the possi ilit of determining its , , al, i, , e, i, a, , and or g ome surfactants e g ioaccumulation potential using availa le information on its components hen ioaccumulating components alcohol etho lates, d estuffs, and dissociated su stances ma e constitute a significant part of the comple su stance e g more than or for ha ardous components an even lower predicted the program as well content , the comple su stance should e regarded as eing ioaccumulating
log ow he programme calculates log ow for organic compounds containing , , , , al, and mprovements are in progress in order to e tend igh molecular weight substances the applica ilit of ove certain molecular dimensions, the potential of a su stance to ioconcentrate decreases his is rovides improved results is a mechanistic model ased on chemical thermod namic possi l due to steric hindrance of the passage of the su stance through gill mem ranes t has een proposed that a cut over and principles rather than a deterministic model rooted in nowledge off limit of for the molecular weight could e applied e g uropean ommission, owever, this cut off has for compounds o tained from o servational data herefore, differs from een su ect to criticism and an alternative cut off of has een proposed in relation to e clusion of consideration with models that use s i e , , in that of su stances with possi le indirect a uatic effects , n general, ioconcentration of possi le meta olites no measured log data are needed for a training set of chemicals log ow ow or environmental degradation products of large molecules should e considered ata on ioconcentration of molecules nl can e emplo ed in a general wa for inorganic or with a high molecular weight should therefore e carefull evaluated and onl e used if such data are considered to e organometallic compounds full valid in respect to oth the parent compound and its possi le meta olites and environmental degradation products a A validation study performed by iemel who compared experimental determined log ow values with estimated values showed that the program precisely predicts the log ow for a great number of organic chemicals in the log ow urface active agents range from below 0 to above 9 (n 50 r 0.96 (Tema ord 995 5 . urfactants consist of a lipophilic most often an al l chain and a h drophilic part the polar headgroup ased on a scatter plot of estimated vs. experimental log ( yracuse esearch Corporation 999 where 05 ow ccording to the charge of the headgroup, surfactants are su divided into classes of anionic, cationic, non ionic, or compound have been tested the LOG O is evaluated being valid for compounds with a log in the interval . ow amphoteric surfactants ue to the variet of different headgroups, surfactants are a structurall diverse class of compounds, which is defined surface activit rather than chemical structure he ioaccumulation potential of C e ical classes t at nee special attention it espect to C an o al es surfactants should thus e considered in relation to the different su classes anionic, cationic, non ionic, or amphoteric instead of to the group as a whole urface active su stances ma form emulsions, in which the ioavaila ilit is here are certain ph sico chemical properties, which can ma e the determination of or its difficult to ascertain icelle formation can result in a change of the ioavaila le fraction even when the solutions are measurement difficult hese ma e su stances, which do not ioconcentrate in a manner consistent with their other apparentl formed, thus giving pro lems in interpretation of the ioaccumulation potential ph sico chemical properties, e g steric hindrance or su stances which ma e the use of descriptors inappropriate, e g surface activit , which ma es oth the measurement and use of log inappropriate ow perimentall derived ioconcentration factors Difficult substances easured values on surfactants show that ma increase with increasing al l chain length ome su stances are difficult to test in a uatic s stems and guidance has een developed to assist in and e dependant of the site of attachment of the head group, and other structural features testing these materials o , he uidance document on a uatic to icit testing of difficult su stances and mi tures , is also a good source of information for ctanol water partition coefficient ow ioconcentration studies, on the t pes of su stances that are difficult to test and the steps needed to ensure valid conclusions from tests with these su stances ifficult to test su stances ma e poorl solu le, volatile, or su ect to he octanol water partition coefficient for surfactants can not e determined using the sha e flas or rapid degradation due to such processes as phototransformation, h drol sis, o idation, or iotic degradation slow stirring method ecause of the formation of emulsions n addition, the surfactant molecules will e ist in the water phase almost e clusivel as ions, whereas the will have to pair with a counter ion in order to e dissolved in octanol herefore, e perimental determination of ow does not characteri e the partition of ionic surfactants olls, n
- 477 - Copyright@United Nations, 2017. All rights reserved
a alid high quality experimentally determined value LOGKOW could represent the bioaccumulation potential; however, for other surfactants some ‘correction’ to the i BCF ≥ 500: The substance has a potential for bioconcentration ii The substance does not have a potential for bioconcentration. b alid high quality experimentally determined value O alid high quality experimentally determined log Kow value
Conflictin ata an lac of ata i log Kow ≥ 4: The substance has a potential for bioconcentration ii log Kow The substance does not have a potential for bioconcentration. Conflicting C data c alid high quality experimentally determined value O
alid high quality experimentally determined log K value O ow an appropriate bioconcentration test, should be interpreted by a “weight of evidence approach”. This implies that if se of validated for estimating a log Kow value i log Kow ≥ 4: The substance has a potential for bioconcentration ii log Kow The substance does not have a potential for bioconcentration. e o
isto . . . uantitative tructure ctivity elationships in aquatic toxicology can be traced to the wor Conflicting log ow data of Overton in rich Lipnic , and eyer in arburg Lipnic , a . They demonstrated that the potency of substances producing narcosis in tadpoles and small fish is in direct proportion to their partition coefficients measured between olive oil and water. Overton postulated in his 1901 monograph “Studien über die Narkose,” that this correlation reflects toxicity ta ing place at a standard molar concentration or molar volume within some molecular site within the organism Lipnic , a . In addition, he concluded that this corresponds to the same concentration or volume for a various organisms, regardless of whether upta e is from water or via gaseous inhalation. This correlation became nown in anaesthesia as the eyer Overton theory.
Expert udgement . . . orwin ansch and co wor ers at omona ollege proposed the use of n octanol water as a standard partitioning system, and found that these partition coefficients were an additive, constitutive property that can be directly estimated from chemical structure. In addition, they found that regression analysis could be used to derive models, providing a statistical analysis of the findings. sing this approach, in these wor ers reported models in the form log log Kow , where Kow is the n octanol water partition coefficient, and is the molar concentration of a chemical yielding a standard biological response for the effect of simple non electrolyte non reactive organic compounds on whole animals, organs, cells, or even pure en ymes. ive of these equations, which ecision sc e e relate to the toxicity of five simple monohydric alcohols to five species of fish, have almost identical slopes and intercepts that are in fact virtually the same as those found by K nemann in , who appears to have been unaware of ansch s earlier wor . K nemann and others have demonstrated that such simple non reactive non electrolytes all act by a narcosis mechanism in an acute fish toxicity test, giving rise to minimum or baseline toxicity Lipnic , b .
pe i ental a tifacts ca sin n e esti ation of a a . . . Other non electrolytes can be more toxic than predicted by such a , but not less toxic, except as a result of a testing artefact. uch testing artefacts include data obtained for compounds such as hydrocarbons which tend to volatili e during the experiment, as well as very hydrophobic compounds for which the acute testing duration may be inadequate to achieve steady state equilibrium partitioning between the concentration in the aquatic phase aquarium test solution , and the internal hydrophobic site of narcosis action. plot of log Kow vs log for such in reviews and assigned as the “recommended values”, are preferred. If no experimentally data of high quality are simple non reactive non electrolytes exhibits a linear relationship so long as such equilibrium is established within the test duration. eyond this point, a bilinear relationship is observed, with the most toxic chemical being the one with the highest log Kow value for which such equilibrium is established Lipnic , . n . . . nother testing problem is posed by water solubility cut off. If the toxic concentration required to produce the effect is above the compound s water solubility, no effect will be observed even at water saturation. ompounds for which the predicted toxic concentration is close to water solubility will also show no effect if the test duration is insufficient to achieve equilibrium partitioning. similar cut off is observed for surfactants if toxicity is predicted at a concentration beyond the critical micelle concentration. lthough such compounds may show no toxicity under these conditions when tested alone, their toxic contributions to mixtures are still present. or compounds with the same log Kow value, differences in water solubility reflect differences in enthalpy of fusion related to melting point. elting point is a reflection of the degree of stability of the crystal lattice and is controlled by intermolecular hydrogen - 478 - Copyright@United Nations, 2017. All rights reserved
a alid high quality experimentally determined value LOGKOW could represent the bioaccumulation potential; however, for other surfactants some ‘correction’ to the i BCF ≥ 500: The substance has a potential for bioconcentration ii The substance does not have a potential for bioconcentration. b alid high quality experimentally determined value O alid high quality experimentally determined log Kow value
Conflictin ata an lac of ata i log Kow ≥ 4: The substance has a potential for bioconcentration ii log Kow The substance does not have a potential for bioconcentration. Conflicting C data c alid high quality experimentally determined value O
alid high quality experimentally determined log K value O ow an appropriate bioconcentration test, should be interpreted by a “weight of evidence approach”. This implies that if se of validated for estimating a log Kow value i log Kow ≥ 4: The substance has a potential for bioconcentration ii log Kow The substance does not have a potential for bioconcentration. e o
isto . . . uantitative tructure ctivity elationships in aquatic toxicology can be traced to the wor Conflicting log ow data of Overton in rich Lipnic , and eyer in arburg Lipnic , a . They demonstrated that the potency of substances producing narcosis in tadpoles and small fish is in direct proportion to their partition coefficients measured between olive oil and water. Overton postulated in his 1901 monograph “Studien über die Narkose,” that this correlation reflects toxicity ta ing place at a standard molar concentration or molar volume within some molecular site within the organism Lipnic , a . In addition, he concluded that this corresponds to the same concentration or volume for a various organisms, regardless of whether upta e is from water or via gaseous inhalation. This correlation became nown in anaesthesia as the eyer Overton theory.
Expert udgement . . . orwin ansch and co wor ers at omona ollege proposed the use of n octanol water as a standard partitioning system, and found that these partition coefficients were an additive, constitutive property that can be directly estimated from chemical structure. In addition, they found that regression analysis could be used to derive models, providing a statistical analysis of the findings. sing this approach, in these wor ers reported models in the form log log Kow , where Kow is the n octanol water partition coefficient, and is the molar concentration of a chemical yielding a standard biological response for the effect of simple non electrolyte non reactive organic compounds on whole animals, organs, cells, or even pure en ymes. ive of these equations, which ecision sc e e relate to the toxicity of five simple monohydric alcohols to five species of fish, have almost identical slopes and intercepts that are in fact virtually the same as those found by K nemann in , who appears to have been unaware of ansch s earlier wor . K nemann and others have demonstrated that such simple non reactive non electrolytes all act by a narcosis mechanism in an acute fish toxicity test, giving rise to minimum or baseline toxicity Lipnic , b .
pe i ental a tifacts ca sin n e esti ation of a a . . . Other non electrolytes can be more toxic than predicted by such a , but not less toxic, except as a result of a testing artefact. uch testing artefacts include data obtained for compounds such as hydrocarbons which tend to volatili e during the experiment, as well as very hydrophobic compounds for which the acute testing duration may be inadequate to achieve steady state equilibrium partitioning between the concentration in the aquatic phase aquarium test solution , and the internal hydrophobic site of narcosis action. plot of log Kow vs log for such in reviews and assigned as the “recommended values”, are preferred. If no experimentally data of high quality are simple non reactive non electrolytes exhibits a linear relationship so long as such equilibrium is established within the test duration. eyond this point, a bilinear relationship is observed, with the most toxic chemical being the one with the highest log Kow value for which such equilibrium is established Lipnic , . n . . . nother testing problem is posed by water solubility cut off. If the toxic concentration required to produce the effect is above the compound s water solubility, no effect will be observed even at water saturation. ompounds for which the predicted toxic concentration is close to water solubility will also show no effect if the test duration is insufficient to achieve equilibrium partitioning. similar cut off is observed for surfactants if toxicity is predicted at a concentration beyond the critical micelle concentration. lthough such compounds may show no toxicity under these conditions when tested alone, their toxic contributions to mixtures are still present. or compounds with the same log Kow value, differences in water solubility reflect differences in enthalpy of fusion related to melting point. elting point is a reflection of the degree of stability of the crystal lattice and is controlled by intermolecular hydrogen - 479 - Copyright@United Nations, 2017. All rights reserved
bonding, lac of conformational flexibility, and symmetry. The more highly symmetric a compound, the higher the elease and e osu e may in itsel be ade uate to t igge testing o the de elo ment o a new o a class o melting point Lipnic , . chemicals o which such decisions a e needed model can be de i ed by statistical analysis e g eg ession analysis om such a data set he most commonly em loyed molecula desc i to log Kow may be t ied as a i st o ellin iss es attem t
. . . hoosing an appropriate implies that the model will yield a reliable prediction for the toxicity y cont ast de i ation o a mechanism based model e ui es an unde standing o wo ing or biological activity of an untested chemical. Generally spea ing, reliability decreases with increasing complexity of hy othesis o molecula mechanism and what a amete o a amete s would a o iately model these actions t is chemical structure, unless a has been derived for a narrowly defined set of chemicals similar in structure to the im o tant to ee in mind that this is di e ent om a hy othesis ega ding mode o action which elates to candidate substance. models derived from narrowly defined classes of chemicals are commonly employed in the biological hysiological es onse but not molecula mechanism development of pharmaceuticals once a new lead compound is identified and there is a need to ma e minor structural modifications to optimi e activity and decrease toxicity . Overall, the ob ective is ma e estimates by interpolation se of s in a atic classification rather than extrapolation. he ollowing inhe ent o e ties o substances a e ele ant o classi ication u oses conce ning the . . . or example, if h L test data for fathead minnow are available for ethanol, n butanol, n a uatic en i onment hexanol, and n nonanol, there is some confidence in ma ing a prediction for this endpoint for n propanol and n pentanol. In contrast, there is would have less confidence in ma ing such a prediction for methanol, which is an a a tition coe icient n octanol wate log Kow extrapolation, with fewer carbon atoms than any of the tested chemicals. In fact, the behaviour of the first member of such a homologous is typically the most anomalous, and should not be predicted using data from remaining members of b bioconcent ation acto the series. ven the toxicity of branched chain alcohols may be an unreasonable extrapolation, depending upon the endpoint in question. uch extrapolation becomes more unreliable to the extent that toxicity is related to production of c deg adability abiotic and biodeg adation metabolites for a particular endpoint, as opposed to the properties of the parent compound. lso, if toxicity is mediated by a specific receptor binding mechanism, dramatic effects may be observed with small changes in chemical structure. d acute a uatic to icity o ish da hnia and algae
. . . What ultimately governs the validity of such predictions is the degree to which the compounds used to e olonged to icity o ish and da hnia derive the for a specific biological endpoint, are acting by a common molecular mechanism. In many and perhaps most cases, a does not represent such a mechanistic model, but merely a correlative one. truly valid est data always ta e ecedence o e edications o iding the test data a e alid with mechanistic model must be derived from a series of chemicals all acting by a common molecular mechanism, and fit to s used o illing data ga s o u oses o classi ication ince the a ailable s a e o a ying eliability and an equation using one or more parameters that relate directly to one or more steps of the mechanism in question. uch a lication ange di e ent est ictions a ly o the ediction o each o these end oints e e theless i a tested parameters or properties are more generally nown as molecular descriptors. It is also important to eep in mind that com ound belongs to a chemical class o st uctu e ty e see abo e o which the e is some con idence in the edicti e many such molecular descriptors in common use may not have a direct physical interpretation. or a correlative model, utility o the model it is wo thwhile to com a e this ediction with the e e imental data as it is not unusual to the statistical fit of the data are li ely to be poorer than a mechanistic one given these limitations. echanisms are not use this a oach to detect some o the e e imental a te acts olatili ation insu icient test du ation to achie e necessarily completely understood, but enough information may be nown to provide confidence in this approach. or e uilib ium and wate solubility cut o in the measu ed data which would mostly esult in classi ying substances as correlative models, the predictive reliability increases with the narrowness with which each is defined, e.g. categories of lowe than actual to icity electrophiles, such as acrylates, in which the degree of reactivity may be similar and toxicity can be estimated for a “new” chemical using a model based solely on the log Kow parameter. hen two o mo e s a e a licable o a ea to be a licable it is use ul to com a e the edictions o these a ious models in the same way that edicted data should be com a ed with measu ed as . . . s an example, primary and secondary alcohols containing a double or triple bond that is con ugated discussed abo e the e is no disc e ancy between these models the esult o ides encou agement o the alidity o with the hydroxyl function i.e. allylic or propargylic are more toxic than would be predicted for a for the the edictions cou se it may also mean that the models we e all de elo ed using data on simila com ounds and corresponding saturated compounds. This behaviour has been ascribed to a proelectrophile mechanism involving statistical methods n the othe hand i the edictions a e uite di e ent this esult needs to be e amined u the metabolic activation by the ubiquitous enzyme alcohol dehydrogenase to the corresponding α,β unsaturated aldehydes he e is always the ossibility that none o the models used o ides a alid ediction s a i st ste the st uctu es and etones which can act as electrophiles via a ichael type acceptor mechanism eith et al., . In the presence and o e ties o the chemicals used to de i e each o the edicti e models should be e amined to dete mine i any of an alcohol dehydrogenase inhibitor, these compounds behave li e other alcohols and do not show excess toxicity, models a e based u on chemicals simila in both o these es ects to the one o which a ediction is needed one consistent with the mechanistic hypothesis. data set contains such an a o iate analogue used to de i e the model the measu ed alue in the database o that com ound s model ediction should be tested the esults it well with the o e all model it is li ely the most . . . The situation quic ly becomes more complex once one goes beyond such a homologous series of eliable one to use i ewise i none o the models contain test data o such an analogue testing o the chemical in compounds. onsider, for example, simple ben ene derivatives. series of chloroben enes may be viewed as similar to uestion is ecommended a homologous series. ot much difference is li ely in the toxicities of the three isomeric dichloroben enes, so that a for chloroben enes based upon test data for one of these isomers is li ely to be adequate. What about the The U.S. EPA has recently posted a draft document on its website “Development of Chemical substitution of other functional groups on ben ene ring nli e an aliphatic alcohol, addition of a hydroxyl Categories in the HPV Challenge Program,” that proposes the use of chemical categories to “... voluntarily compile a functionality to a ben ene ring produces a phenol which is no longer neutral, but an ioni able acidic compound, due to c eening n o mation ata et on all chemicals on the list to o ide basic sc eening data needed the resonance stabili ation of the resulting negative charge. or this reason, phenol does not act as a true narcotic agent. o an initial assessment o the hysicochemical o e ties en i onmental ate and human and en i onmental e ects o With the addition of electron withdrawing substituents to phenol e.g. chlorine atoms , there is a shift to these chemicals” (US EPA, 1999). This list consists of “...about 2,800 HPV chemicals which were reported for the Toxic compounds acting as uncouplers of oxidative phosphorylation e.g. the herbicide dinoseb . ubstitution of an aldehyde Substances Control Act’s 1990 Inventory Update Rule (IUR)”. group leads to increased toxicity via an electrophile mechanism for such compounds react with amino groups, such as the lysine ε amino group to produce a chiff ase adduct. imilarly, a ben ylic chloride acts as an electrophile to form One approach being proposed “...where this is scientifically usti iable is to conside closely elated covalent abducts with sulfhydryl groups. In tac ling a prediction for an untested compound, the chemical reactivity of chemicals as a g ou o catego y athe than test them as indi idual chemicals n the catego y a oach not e e y these and many other functional groups and their interaction with one another should be carefully studied, and attempts chemical needs to be tested for every SIDS endpoint”. Such limited testing could be justified providing that the “...final made to document these from the chemical literature Lipnic , b . data set must allow one to assess the untested end oints ideally by inte olation between and among the catego y members.” The process for defining such categories and in the development of such data are described in the o osal . . . Given these limitations in using s for ma ing predictions, it is best employed as a means of establishing testing priorities, rather than as a means of substituting for testing, unless some mechanistic information is A second potentially less data intensive approach being considered (US EPA, 2000a) is “... applying available on the untested compound itself. In fact, the inability to ma e a prediction along with nown environmental SAR principles to a single chemical that is closely related to one or more better characterized chemicals (“analogs”).” A
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bonding, lac of conformational flexibility, and symmetry. The more highly symmetric a compound, the higher the elease and e osu e may in itsel be ade uate to t igge testing o the de elo ment o a new o a class o melting point Lipnic , . chemicals o which such decisions a e needed model can be de i ed by statistical analysis e g eg ession analysis om such a data set he most commonly em loyed molecula desc i to log Kow may be t ied as a i st o ellin iss es attem t
. . . hoosing an appropriate implies that the model will yield a reliable prediction for the toxicity y cont ast de i ation o a mechanism based model e ui es an unde standing o wo ing or biological activity of an untested chemical. Generally spea ing, reliability decreases with increasing complexity of hy othesis o molecula mechanism and what a amete o a amete s would a o iately model these actions t is chemical structure, unless a has been derived for a narrowly defined set of chemicals similar in structure to the im o tant to ee in mind that this is di e ent om a hy othesis ega ding mode o action which elates to candidate substance. models derived from narrowly defined classes of chemicals are commonly employed in the biological hysiological es onse but not molecula mechanism development of pharmaceuticals once a new lead compound is identified and there is a need to ma e minor structural modifications to optimi e activity and decrease toxicity . Overall, the ob ective is ma e estimates by interpolation se of s in a atic classification rather than extrapolation. he ollowing inhe ent o e ties o substances a e ele ant o classi ication u oses conce ning the . . . or example, if h L test data for fathead minnow are available for ethanol, n butanol, n a uatic en i onment hexanol, and n nonanol, there is some confidence in ma ing a prediction for this endpoint for n propanol and n pentanol. In contrast, there is would have less confidence in ma ing such a prediction for methanol, which is an a a tition coe icient n octanol wate log Kow extrapolation, with fewer carbon atoms than any of the tested chemicals. In fact, the behaviour of the first member of such a homologous is typically the most anomalous, and should not be predicted using data from remaining members of b bioconcent ation acto the series. ven the toxicity of branched chain alcohols may be an unreasonable extrapolation, depending upon the endpoint in question. uch extrapolation becomes more unreliable to the extent that toxicity is related to production of c deg adability abiotic and biodeg adation metabolites for a particular endpoint, as opposed to the properties of the parent compound. lso, if toxicity is mediated by a specific receptor binding mechanism, dramatic effects may be observed with small changes in chemical structure. d acute a uatic to icity o ish da hnia and algae
. . . What ultimately governs the validity of such predictions is the degree to which the compounds used to e olonged to icity o ish and da hnia derive the for a specific biological endpoint, are acting by a common molecular mechanism. In many and perhaps most cases, a does not represent such a mechanistic model, but merely a correlative one. truly valid est data always ta e ecedence o e edications o iding the test data a e alid with mechanistic model must be derived from a series of chemicals all acting by a common molecular mechanism, and fit to s used o illing data ga s o u oses o classi ication ince the a ailable s a e o a ying eliability and an equation using one or more parameters that relate directly to one or more steps of the mechanism in question. uch a lication ange di e ent est ictions a ly o the ediction o each o these end oints e e theless i a tested parameters or properties are more generally nown as molecular descriptors. It is also important to eep in mind that com ound belongs to a chemical class o st uctu e ty e see abo e o which the e is some con idence in the edicti e many such molecular descriptors in common use may not have a direct physical interpretation. or a correlative model, utility o the model it is wo thwhile to com a e this ediction with the e e imental data as it is not unusual to the statistical fit of the data are li ely to be poorer than a mechanistic one given these limitations. echanisms are not use this a oach to detect some o the e e imental a te acts olatili ation insu icient test du ation to achie e necessarily completely understood, but enough information may be nown to provide confidence in this approach. or e uilib ium and wate solubility cut o in the measu ed data which would mostly esult in classi ying substances as correlative models, the predictive reliability increases with the narrowness with which each is defined, e.g. categories of lowe than actual to icity electrophiles, such as acrylates, in which the degree of reactivity may be similar and toxicity can be estimated for a “new” chemical using a model based solely on the log Kow parameter. hen two o mo e s a e a licable o a ea to be a licable it is use ul to com a e the edictions o these a ious models in the same way that edicted data should be com a ed with measu ed as . . . s an example, primary and secondary alcohols containing a double or triple bond that is con ugated discussed abo e the e is no disc e ancy between these models the esult o ides encou agement o the alidity o with the hydroxyl function i.e. allylic or propargylic are more toxic than would be predicted for a for the the edictions cou se it may also mean that the models we e all de elo ed using data on simila com ounds and corresponding saturated compounds. This behaviour has been ascribed to a proelectrophile mechanism involving statistical methods n the othe hand i the edictions a e uite di e ent this esult needs to be e amined u the metabolic activation by the ubiquitous enzyme alcohol dehydrogenase to the corresponding α,β unsaturated aldehydes he e is always the ossibility that none o the models used o ides a alid ediction s a i st ste the st uctu es and etones which can act as electrophiles via a ichael type acceptor mechanism eith et al., . In the presence and o e ties o the chemicals used to de i e each o the edicti e models should be e amined to dete mine i any of an alcohol dehydrogenase inhibitor, these compounds behave li e other alcohols and do not show excess toxicity, models a e based u on chemicals simila in both o these es ects to the one o which a ediction is needed one consistent with the mechanistic hypothesis. data set contains such an a o iate analogue used to de i e the model the measu ed alue in the database o that com ound s model ediction should be tested the esults it well with the o e all model it is li ely the most . . . The situation quic ly becomes more complex once one goes beyond such a homologous series of eliable one to use i ewise i none o the models contain test data o such an analogue testing o the chemical in compounds. onsider, for example, simple ben ene derivatives. series of chloroben enes may be viewed as similar to uestion is ecommended a homologous series. ot much difference is li ely in the toxicities of the three isomeric dichloroben enes, so that a for chloroben enes based upon test data for one of these isomers is li ely to be adequate. What about the The U.S. EPA has recently posted a draft document on its website “Development of Chemical substitution of other functional groups on ben ene ring nli e an aliphatic alcohol, addition of a hydroxyl Categories in the HPV Challenge Program,” that proposes the use of chemical categories to “... voluntarily compile a functionality to a ben ene ring produces a phenol which is no longer neutral, but an ioni able acidic compound, due to c eening n o mation ata et on all chemicals on the list to o ide basic sc eening data needed the resonance stabili ation of the resulting negative charge. or this reason, phenol does not act as a true narcotic agent. o an initial assessment o the hysicochemical o e ties en i onmental ate and human and en i onmental e ects o With the addition of electron withdrawing substituents to phenol e.g. chlorine atoms , there is a shift to these chemicals” (US EPA, 1999). This list consists of “...about 2,800 HPV chemicals which were reported for the Toxic compounds acting as uncouplers of oxidative phosphorylation e.g. the herbicide dinoseb . ubstitution of an aldehyde Substances Control Act’s 1990 Inventory Update Rule (IUR)”. group leads to increased toxicity via an electrophile mechanism for such compounds react with amino groups, such as the lysine ε amino group to produce a chiff ase adduct. imilarly, a ben ylic chloride acts as an electrophile to form One approach being proposed “...where this is scientifically usti iable is to conside closely elated covalent abducts with sulfhydryl groups. In tac ling a prediction for an untested compound, the chemical reactivity of chemicals as a g ou o catego y athe than test them as indi idual chemicals n the catego y a oach not e e y these and many other functional groups and their interaction with one another should be carefully studied, and attempts chemical needs to be tested for every SIDS endpoint”. Such limited testing could be justified providing that the “...final made to document these from the chemical literature Lipnic , b . data set must allow one to assess the untested end oints ideally by inte olation between and among the catego y members.” The process for defining such categories and in the development of such data are described in the o osal . . . Given these limitations in using s for ma ing predictions, it is best employed as a means of establishing testing priorities, rather than as a means of substituting for testing, unless some mechanistic information is A second potentially less data intensive approach being considered (US EPA, 2000a) is “... applying available on the untested compound itself. In fact, the inability to ma e a prediction along with nown environmental SAR principles to a single chemical that is closely related to one or more better characterized chemicals (“analogs”).” A
- 481 - Copyright@United Nations, 2017. All rights reserved
third approach proposed consists of using “... a combination of the analogue and category approaches ... [for] individual Also, caution should be applied in comparing predicted CF values with those using radiolabeled compounds, where chemicals ... similar to that used in ECOSAR (US EPA, 2000b), a SAR based computer program that generates the tissue concentration thus detected may represent a mix of parent compound and metabolites or even covalently ecotoxicity values. “. The document also details the history of the use of SARs within the U.S. EPA new chemicals bound parent or metabolite. program, and how to go about collecting and analysing data for the sa e of such SAR approaches. A . . . .2 Experimental log ow values are to be used preferentially. However, older shake flask values above A9. . . The ordic Council of inisters issued a report (Pederson et al., 199 ) entitled “Environmental 5.5 are not reliable and in many cases it is better to use some average of calculated values or to have these remeasured Hazard Classification,” that includes information on data collection and interpretation, as well as a section (5.2.8) using the slow stirring method ( rui n et al., 8 ). f there is reasonable doubt about the accuracy of the measured entitled “QSAR estimates of water solubility and acute aquatic toxicity”. This section also discusses the estimation of data, calculated log ow values shall be used. physicochemical properties, including log ow. or the sa e of classification purposes, estimation methods are recommended for prediction of “minimum acute aquatic toxicity,” for “...neutral, organic, non reactive and non A . . . Degradability abiotic and biodegradation ionizable compounds such as alcohols, etones, ethers, al yl, and aryl halides, and can also be used for aromatic hydrocarbons, halogenated aromatic and aliphatic hydrocarbons as well as sulphides and disulphides,” as cited in an QSARs for abiotic degradation in water phases are narrowly defined linear free energy relationships earlier OECD uidance Document (OECD, 199 ). The ordic document also includes dis ettes for a computerized ( FERs) for specific classes of chemicals and mechanisms. For example, such FERs are available for hydrolysis of application of some of these methods. benzylic chlorides with various substituents on the aromatic ring. Such narrowly defined FER models tend to be very reliable if the needed parameters are available for the Substituent(s) in question. hoto degradation, i.e. reaction with A9. . .8 The European Centre for Ecotoxicology and Toxicology of Chemicals (ECETOC) has published a produced reactive species, may be extrapolated from estimates for the air compartment. hile these abiotic report entitled “QSARs in the Assessment of the Environmental Fate and Effects of Chemicals,” which describes the processes do not usually result in complete degradation of organic compounds, they are frequently significant starting use of QSARs to “...check the validity of data or to fill data gaps for priority setting, risk assessment and classification” points, and may be rate limiting. QSARs for calculating biodegradability are either compound specific ( EC , 5) or (ECETOC, 1998). SARs are described for predicting environmental fate and a uatic toxicity. The report notes that “a group contribution models like the E program (Hansch and eo, 5 eylan and Howard 5 Hilal et al., consistent dataset for [an endpoint] covered ... for a well defined scope of chemical structures (“domain”) [is needed] ... Howard et al., 2 oethling et al., Howard and eylan 2 oonen et al., ). hile validated from which a training set is developed. The document also discusses the advantage of mechanism based models, the use compound class specific models are very limited in their application range, the application range of group contribution of statistical analysis in the development of QSARs, and how to assess “outliers”. models is potentially much broader, but limited to compounds containing the model substructures. alidation studies have suggested that the biodegradability predictions by currently available group contribution models may be used for A9. . .9 Octanol water partition coefficient ( ow prediction of “not ready biodegradability” (Pedersen et al., 5 angenberg et al., SE A, ) – and thus in relation to aquatic hazard classification “not rapid degradability.” A9. . .9.1 Computerized methods such as C O P (US EPA, 1999), O O (US EPA, 2000a) and SPARC (US EPA, 2000b) are available to calculate log ow directly from chemical structure. C O P and O O are A . . . 2 Acute a uatic toxicity for fish daphnia and algae based upon the addition of group contributions, while SPARC is based upon a more fundamental chemical structure algorithm. Caution should be used in using calculated values for compounds that can undergo hydrolysis in water or The acute aquatic toxicity of non reactive, non electrolyte organic chemicals (baseline toxicity) can be some other reaction, since these transformations need to be considered in the interpretation of a uatic toxicity test data predicted from their log ow value with a quite high level of confidence, provided the presence of electrophile, for such reactive chemicals. Only SPARC can be employed in a general way for inorganic or organometallic compounds. proelectrophile, or special mechanism functional groups (see above) were not detected. roblems remain for such Special methods are needed in ma ing estimates of log ow or a uatic toxicity for surface active compounds, chelating specific toxicants, for which the appropriate QSAR has to be selected in a prospective manner. Since straightforward compounds, and mixtures. criteria for the identification of the relevant modes of action are still lacking, empirical expert udgement needs to be applied for selecting a suitable model. Thus, if an inappropriate QSAR is employed, the predictions may be in error by A9. . .9.2 Values of log ow can be calculated for pentachlorophenol and similar compounds, both for the several orders of magnitude, and in the case of baseline toxicity, will be predicted less toxic, rather than more. ionized and unionized (neutral) forms. These values can potentially be calculated for certain reactive molecules (e.g. benzotrichloride), but the reactivity and subse uent hydrolysis also need to be considered. Also, for such ionizable A . . . rolonged toxicity for fish and Daphnia phenols, p a is a second parameter. Specific models can be used to calculate log ow values for organometallic compounds, but they need to be applied with caution since some of these compounds really exist in the form of ion Calculated values for chronic toxicity to fish and aphnia should not be used to overrule classification pairs in water. based on experimental acute toxicity data. nly a few validated models are available for calculating prolonged toxicity for fish and aphnia. These models are based solely on log ow correlations and are limited in their application to non A9. . .9. or compounds of extremely high lipophilicity, measurements up to about to . can be made by reactive, non electrolyte organic compounds, and are not suitable for chemicals with specific modes of action under sha e flas , and can be extended up to about log ow of 8 using the slow stirring approach ( ruijn et al., 1989). prolonged exposure conditions. The reliable estimation of chronic toxicity values depends on the correct discrimination Calculations are considered useful even in extrapolating beyond what can be measured by either of these methods. Of between non specific and specific chronic toxicity mechanisms otherwise, the predicted toxicity can be wrong by course, it should be ept in mind that if the SAR models for toxicity, etc. are based on chemicals with lower log ow orders of magnitude. t should be noted that although for many compounds, excess toxicity in a chronic test correlates values, the prediction itself will also be an extrapolation in fact, it is nown that in the case of bioconcentration, the with excess toxicity in an acute test, this is not always the case. relationship with log ow becomes non linear at higher values. or compounds with low log ow values, the group contribution can also be applied, but this is not very useful for hazard purposes since for such substances, particularly i ic tion o et n et co po n with negative log ow values, little if any partitioning can ta e place into lipophilic sites and as Overton reported, these substances produce toxicity through osmotic effects ( ipnic , 198 ). nt o ction
A9. . .10 ioconcentration factor C A . . . The harmonized system for classifying substances is a hazard based system, and the basis of the identification of hazard is the aquatic toxicity of the substances, and information on the degradation and A9. . .10.1 If experimentally determined C values are available, these values should be used for classification. bioaccumulation behaviour ( EC 8). Since this document deals only with the hazards associated with a given ioconcentration measurements must be performed using pure samples at test concentrations within water solubility, substance when the substance is dissolved in the water column, exposure from this source is limited by the solubility of and for an ade uate test duration to achieve steady state e uilibrium between the a ueous concentration and that in the the substance in water and bioavailability of the substance in species in the aquatic environment. Thus, the hazard fish tissue. oreover, with bioconcentration tests of extended duration, the correlation with log ow levels off and classification schemes for metals and metal compounds are limited to the hazards posed by metals and metal ultimately decreases. Under environmental conditions, bioconcentration of highly lipophilic chemicals ta es place by a compounds when they are available (i.e. exist as dissolved metal ions, for example, as when present as ), and combination of upta e from food and water, with the switch to food ta ing place at log ow ≈ 6. Otherwise log Kow do not take into account exposures to metals and metal compounds that are not dissolved in the water column but may values can be used with a SAR model as a predictor of the bioaccumulation potential of organic compounds. still be bioavailable, such as metals in foods. This section does not take into account the non metallic ion (e.g. C ) of Deviations from these SARs tend to reflect differences in the extent to which the chemicals undergo metabolism in the fish. Thus, some chemicals, such as phthalate, can bioconcentrate significantly less than predicted for this reason. Excess toxicity Te ( redicted baseline toxicity Observed toxicity. 82 8 - 482 - Copyright@United Nations, 2017. All rights reserved
third approach proposed consists of using “... a combination of the analogue and category approaches ... [for] individual Also, caution should be applied in comparing predicted CF values with those using radiolabeled compounds, where chemicals ... similar to that used in ECOSAR (US EPA, 2000b), a SAR based computer program that generates the tissue concentration thus detected may represent a mix of parent compound and metabolites or even covalently ecotoxicity values. “. The document also details the history of the use of SARs within the U.S. EPA new chemicals bound parent or metabolite. program, and how to go about collecting and analysing data for the sa e of such SAR approaches. A . . . .2 Experimental log ow values are to be used preferentially. However, older shake flask values above A9. . . The ordic Council of inisters issued a report (Pederson et al., 199 ) entitled “Environmental 5.5 are not reliable and in many cases it is better to use some average of calculated values or to have these remeasured Hazard Classification,” that includes information on data collection and interpretation, as well as a section (5.2.8) using the slow stirring method ( rui n et al., 8 ). f there is reasonable doubt about the accuracy of the measured entitled “QSAR estimates of water solubility and acute aquatic toxicity”. This section also discusses the estimation of data, calculated log ow values shall be used. physicochemical properties, including log ow. or the sa e of classification purposes, estimation methods are recommended for prediction of “minimum acute aquatic toxicity,” for “...neutral, organic, non reactive and non A . . . Degradability abiotic and biodegradation ionizable compounds such as alcohols, etones, ethers, al yl, and aryl halides, and can also be used for aromatic hydrocarbons, halogenated aromatic and aliphatic hydrocarbons as well as sulphides and disulphides,” as cited in an QSARs for abiotic degradation in water phases are narrowly defined linear free energy relationships earlier OECD uidance Document (OECD, 199 ). The ordic document also includes dis ettes for a computerized ( FERs) for specific classes of chemicals and mechanisms. For example, such FERs are available for hydrolysis of application of some of these methods. benzylic chlorides with various substituents on the aromatic ring. Such narrowly defined FER models tend to be very reliable if the needed parameters are available for the Substituent(s) in question. hoto degradation, i.e. reaction with A9. . .8 The European Centre for Ecotoxicology and Toxicology of Chemicals (ECETOC) has published a produced reactive species, may be extrapolated from estimates for the air compartment. hile these abiotic report entitled “QSARs in the Assessment of the Environmental Fate and Effects of Chemicals,” which describes the processes do not usually result in complete degradation of organic compounds, they are frequently significant starting use of QSARs to “...check the validity of data or to fill data gaps for priority setting, risk assessment and classification” points, and may be rate limiting. QSARs for calculating biodegradability are either compound specific ( EC , 5) or (ECETOC, 1998). SARs are described for predicting environmental fate and a uatic toxicity. The report notes that “a group contribution models like the E program (Hansch and eo, 5 eylan and Howard 5 Hilal et al., consistent dataset for [an endpoint] covered ... for a well defined scope of chemical structures (“domain”) [is needed] ... Howard et al., 2 oethling et al., Howard and eylan 2 oonen et al., ). hile validated from which a training set is developed. The document also discusses the advantage of mechanism based models, the use compound class specific models are very limited in their application range, the application range of group contribution of statistical analysis in the development of QSARs, and how to assess “outliers”. models is potentially much broader, but limited to compounds containing the model substructures. alidation studies have suggested that the biodegradability predictions by currently available group contribution models may be used for A9. . .9 Octanol water partition coefficient ( ow prediction of “not ready biodegradability” (Pedersen et al., 5 angenberg et al., SE A, ) – and thus in relation to aquatic hazard classification “not rapid degradability.” A9. . .9.1 Computerized methods such as C O P (US EPA, 1999), O O (US EPA, 2000a) and SPARC (US EPA, 2000b) are available to calculate log ow directly from chemical structure. C O P and O O are A . . . 2 Acute a uatic toxicity for fish daphnia and algae based upon the addition of group contributions, while SPARC is based upon a more fundamental chemical structure algorithm. Caution should be used in using calculated values for compounds that can undergo hydrolysis in water or The acute aquatic toxicity of non reactive, non electrolyte organic chemicals (baseline toxicity) can be some other reaction, since these transformations need to be considered in the interpretation of a uatic toxicity test data predicted from their log ow value with a quite high level of confidence, provided the presence of electrophile, for such reactive chemicals. Only SPARC can be employed in a general way for inorganic or organometallic compounds. proelectrophile, or special mechanism functional groups (see above) were not detected. roblems remain for such Special methods are needed in ma ing estimates of log ow or a uatic toxicity for surface active compounds, chelating specific toxicants, for which the appropriate QSAR has to be selected in a prospective manner. Since straightforward compounds, and mixtures. criteria for the identification of the relevant modes of action are still lacking, empirical expert udgement needs to be applied for selecting a suitable model. Thus, if an inappropriate QSAR is employed, the predictions may be in error by A9. . .9.2 Values of log ow can be calculated for pentachlorophenol and similar compounds, both for the several orders of magnitude, and in the case of baseline toxicity, will be predicted less toxic, rather than more. ionized and unionized (neutral) forms. These values can potentially be calculated for certain reactive molecules (e.g. benzotrichloride), but the reactivity and subse uent hydrolysis also need to be considered. Also, for such ionizable A . . . rolonged toxicity for fish and Daphnia phenols, p a is a second parameter. Specific models can be used to calculate log ow values for organometallic compounds, but they need to be applied with caution since some of these compounds really exist in the form of ion Calculated values for chronic toxicity to fish and aphnia should not be used to overrule classification pairs in water. based on experimental acute toxicity data. nly a few validated models are available for calculating prolonged toxicity for fish and aphnia. These models are based solely on log ow correlations and are limited in their application to non A9. . .9. or compounds of extremely high lipophilicity, measurements up to about to . can be made by reactive, non electrolyte organic compounds, and are not suitable for chemicals with specific modes of action under sha e flas , and can be extended up to about log ow of 8 using the slow stirring approach ( ruijn et al., 1989). prolonged exposure conditions. The reliable estimation of chronic toxicity values depends on the correct discrimination Calculations are considered useful even in extrapolating beyond what can be measured by either of these methods. Of between non specific and specific chronic toxicity mechanisms otherwise, the predicted toxicity can be wrong by course, it should be ept in mind that if the SAR models for toxicity, etc. are based on chemicals with lower log ow orders of magnitude. t should be noted that although for many compounds, excess toxicity in a chronic test correlates values, the prediction itself will also be an extrapolation in fact, it is nown that in the case of bioconcentration, the with excess toxicity in an acute test, this is not always the case. relationship with log ow becomes non linear at higher values. or compounds with low log ow values, the group contribution can also be applied, but this is not very useful for hazard purposes since for such substances, particularly i ic tion o et n et co po n with negative log ow values, little if any partitioning can ta e place into lipophilic sites and as Overton reported, these substances produce toxicity through osmotic effects ( ipnic , 198 ). nt o ction
A9. . .10 ioconcentration factor C A . . . The harmonized system for classifying substances is a hazard based system, and the basis of the identification of hazard is the aquatic toxicity of the substances, and information on the degradation and A9. . .10.1 If experimentally determined C values are available, these values should be used for classification. bioaccumulation behaviour ( EC 8). Since this document deals only with the hazards associated with a given ioconcentration measurements must be performed using pure samples at test concentrations within water solubility, substance when the substance is dissolved in the water column, exposure from this source is limited by the solubility of and for an ade uate test duration to achieve steady state e uilibrium between the a ueous concentration and that in the the substance in water and bioavailability of the substance in species in the aquatic environment. Thus, the hazard fish tissue. oreover, with bioconcentration tests of extended duration, the correlation with log ow levels off and classification schemes for metals and metal compounds are limited to the hazards posed by metals and metal ultimately decreases. Under environmental conditions, bioconcentration of highly lipophilic chemicals ta es place by a compounds when they are available (i.e. exist as dissolved metal ions, for example, as when present as ), and combination of upta e from food and water, with the switch to food ta ing place at log ow ≈ 6. Otherwise log Kow do not take into account exposures to metals and metal compounds that are not dissolved in the water column but may values can be used with a SAR model as a predictor of the bioaccumulation potential of organic compounds. still be bioavailable, such as metals in foods. This section does not take into account the non metallic ion (e.g. C ) of Deviations from these SARs tend to reflect differences in the extent to which the chemicals undergo metabolism in the fish. Thus, some chemicals, such as phthalate, can bioconcentrate significantly less than predicted for this reason. Excess toxicity Te ( redicted baseline toxicity Observed toxicity. 82 8 - 483 - Copyright@United Nations, 2017. All rights reserved
etal co pounds hich ay be to ic or hich ay be organic and ay pose bioaccu ulation or persistence hazards. A . .1. his section deals with metals and metal compounds. ithin the conte t of this uidance ocument, or such etal co pounds the hazards of the non etallic ions ust also be considered. metals and metal compounds are characteri ed as follows, and therefore, organo metals are outside the scope of this section . . . he le el of the etal ion hich ay be present in solution follo ing the addition of the etal and or its co pounds ill largely be deter ined by t o processes the e tent to hich it can be dissol ed i.e. its ater a metals, , in their elemental state are not soluble in water but ma transform to ield the solubility and the e tent to hich it can react ith the edia to transfor to ater soluble for s. he rate and e tent at a ailable form. his means that a metal in the elemental state ma react with water or a dilute which this latter process, known as “transformation” for the purposes of this guidance ta es place can ary e tensi ely a ueous electrol te to form soluble cationic or anionic products, and in the process the metal bet een different co pounds and the etal itself and is an i portant factor in deter ining the appropriate hazard will o idi e, or transform, from the neutral or ero o idation state to a higher one class. here data on transfor ation are a ailable they should be ta en into account in deter ining the classification. he Protocol for deter ining this rate is a ailable in nne . b in a simple metal compound, such as an o ide or sulphide, the metal alread e ists in the o idi ed state, so that further metal o idation is unlikel to occur when the compound is . . . enerally spea ing the rate at hich a substance dissol es is not considered rele ant to the introduced into an a ueous medium. deter ination of its intrinsic to icity. o e er for etals and any poorly soluble inorganic etal co pounds the difficulties in achie ing dissolution through nor al solubilization techniques is so se ere that the t o processes of owe er, while o idi ation ma not change, interaction with the media ma ield more soluble solubilization and transfor ation beco e indistinguishable. hus here the co pound is sufficiently poorly soluble forms. A sparingl soluble metal compound can be considered as one for which a solubilit product can be calculated, that the le els dissol ed follo ing nor al atte pts at solubilization do not e ceed the a ailable ( it is the rate and which will ield a small amount of the a ailable form b dissolution. owe er, it should be recogni ed that the and e tent of transfor ation hich ust be considered. he transfor ation ill be affected by a nu ber of factors not final solution concentration ma be influenced b a number of factors, including the solubilit product of some metal least of hich ill be the properties of the edia ith respect to p ater hardness te perature etc. n addition to compounds precipitated during the transformation dissolution test, e.g. aluminium h dro ide. these properties other factors such as the size and specific surface area of the particles hich ha e been tested the length of ti e o er hich e posure to the edia ta es place and of course the ass or surface area loading of the pplication of a atic to icit ata an sol ilit ata fo classification substance in the edia ill all play a part in deter ining the le el of dissol ed etal ions in the ater. ransfor ation data can generally therefore only be considered as reliable for the purposes of classification if conducted according to A . . .1 nterpretation of a uatic toxicity data the standard Protocol in nne . A . . .1.1 A uatic to icit studies carried out according to a recogni ed protocol should normall be acceptable . . . his Protocol ai s at standardizing the principal ariables such that the le el of dissol ed ion can be as alid for the purposes of classification. ection A . should also be consulted for generic issues that are common to directly related to the loading of the substance added. t is this loading le el hich yields the le el of etal ion assessing an a uatic to icit data point for the purposes of classification. equi alent to the a ailable ( that can then be used to deter ine the hazard category appropriate for classification. he testing ethodology is detailed in nne . he strategy to be adopted in using the data fro the testing protocol A . . .1. etal comple ation and speciation and the data require ents needed to a e that strategy or ill be described. A . . .1. .1 he to icit of a particular metal in solution, appears to depend primaril on but is not strictl limited . . . n considering the classification of etals and etal co pounds both readily and poorly soluble to the le el of dissol ed free metal ions. Abiotic factors including alkalinit , ionic strength and p can influence the recognition has to be paid to a number of factors. As defined in Chapter 4.1, the term “degradation” refers to the to icit of metals in two wa s i b influencing the chemical speciation of the metal in water and hence affecting the deco position of organic olecules. or inorganic co pounds and etals clearly the concept of degradability as it a ailabilit and ii b influencing the uptake and binding of a ailable metal b biological tissues. has been considered and used for organic substances has li ited or no eaning. ather the substance ay be transfor ed by nor al en iron ental processes to either increase or decrease the bioa ailability of the to ic species. A . . .1. . here speciation is important, it ma be possible to model the concentrations of the different forms of qually the log o cannot be considered as a easure of the potential to accu ulate. e ertheless the concepts that a the metal, including those that are likel to cause to icit . Anal sis methods for uantif ing e posure concentrations, substance or a to ic etabolite reaction product ay not be rapidly lost fro the en iron ent and or ay which are capable of distinguishing between the comple ed and uncomple ed fractions of a test substance, ma not bioaccu ulate are as applicable to etals and etal co pounds as they are to organic substances. alwa s be a ailable or economic.
. . . peciation of the soluble for can be affected by p ater hardness and other ariables and ay A . . .1. . Comple ation of metals to organic and inorganic ligands in test media and natural en ironments can yield particular for s of the etal ion hich are ore or less to ic. n addition etal ions could be ade non a ailable be estimated from metal speciation models. peciation models for metals, including p , hardness, C, and inorganic fro the ater colu n by a nu ber of processes (e.g. ineralization and partitioning . o eti es these processes can substances such as rown and Allison, 1 , A ipping, 1 4 and C antore and riscoll, be sufficiently rapid to be analogous to degradation in assessing chronic classification. o e er partitioning of the 1 can be used to calculate the uncomple ed and comple ed fractions of the metal ions. Alternati el , the iotic etal ion fro the ater colu n to other en iron ental edia does not necessarily ean that it is no longer igand odel , allows for the calculation of the concentration of metal ion responsible for the to ic effect at the bioa ailable nor does it ean that the etal has been ade per anently una ailable. le el of the organism. he model has at present onl been alidated for a limited number of metals, organisms, and end points antore and i oro, 1 . he models and formula used for the characteri ation of metal . . . nfor ation pertaining to the e tent of the partitioning of a etal ion fro the ater colu n or the comple ation in the media should alwa s be clearl reported, allowing for their translation back to natural en ironments e tent to hich a etal has been or can be con erted to a for that is less to ic or non to ic is frequently not a ailable C , . o er a sufficiently ide range of en iron entally rele ant conditions and thus a nu ber of assu ptions ill need to be ade as an aid in classification. hese assu ptions ay be odified if a ailable data sho other ise. n the first A . . . nterpretation of solubility data instance it should be assu ed that the etal ions once in the ater are not rapidly partitioned fro the ater colu n and thus these co pounds do not eet the criteria. nderlying this is the assu ption that although speciation can A . . . .1 hen considering the a ailable data on solubilit , their alidit and applicabilit to the identification occur the species ill re ain a ailable under en iron entally rele ant conditions. his ay not al ays be the case as of the ha ard of metal compounds should be assessed. n particular, a knowledge of the p at which the data were described abo e and any e idence a ailable that ould suggest changes to the bioa ailability o er the course of generated should be known. days should be carefully e a ined. he bioaccu ulation of etals and inorganic etal co pounds is a co ple process and bioaccu ulation data should be used ith care. he application of bioaccu ulation criteria ill need to be A . . . . Assessment of e isting data considered on a case by case basis ta ing due account of all the a ailable data. isting data will be in one of three forms. or some well studied metals, there will be solubilit . . . further assu ption that can be ade hich represents a cautious approach is that in the absence of products and or solubilit data for the arious inorganic metal compounds. t is also possible that the p relationship of any solubility data for a particular etal co pound either easured or calculated the substance ill be sufficiently the solubilit will be known. owe er, for man metals or metal compounds, it is probable that the a ailable soluble to cause to icity at the le el of the ( and thus ay be classified in the sa e ay as other soluble salts. information will be descripti e onl , e.g. poorl soluble. nfortunatel there appears to be er little consistent gain this is clearly not al ays the case and it ay be ise to generate appropriate solubility data.
4 - 484 - Copyright@United Nations, 2017. All rights reserved
etal co pounds hich ay be to ic or hich ay be organic and ay pose bioaccu ulation or persistence hazards. A . .1. his section deals with metals and metal compounds. ithin the conte t of this uidance ocument, or such etal co pounds the hazards of the non etallic ions ust also be considered. metals and metal compounds are characteri ed as follows, and therefore, organo metals are outside the scope of this section . . . he le el of the etal ion hich ay be present in solution follo ing the addition of the etal and or its co pounds ill largely be deter ined by t o processes the e tent to hich it can be dissol ed i.e. its ater a metals, , in their elemental state are not soluble in water but ma transform to ield the solubility and the e tent to hich it can react ith the edia to transfor to ater soluble for s. he rate and e tent at a ailable form. his means that a metal in the elemental state ma react with water or a dilute which this latter process, known as “transformation” for the purposes of this guidance ta es place can ary e tensi ely a ueous electrol te to form soluble cationic or anionic products, and in the process the metal bet een different co pounds and the etal itself and is an i portant factor in deter ining the appropriate hazard will o idi e, or transform, from the neutral or ero o idation state to a higher one class. here data on transfor ation are a ailable they should be ta en into account in deter ining the classification. he Protocol for deter ining this rate is a ailable in nne . b in a simple metal compound, such as an o ide or sulphide, the metal alread e ists in the o idi ed state, so that further metal o idation is unlikel to occur when the compound is . . . enerally spea ing the rate at hich a substance dissol es is not considered rele ant to the introduced into an a ueous medium. deter ination of its intrinsic to icity. o e er for etals and any poorly soluble inorganic etal co pounds the difficulties in achie ing dissolution through nor al solubilization techniques is so se ere that the t o processes of owe er, while o idi ation ma not change, interaction with the media ma ield more soluble solubilization and transfor ation beco e indistinguishable. hus here the co pound is sufficiently poorly soluble forms. A sparingl soluble metal compound can be considered as one for which a solubilit product can be calculated, that the le els dissol ed follo ing nor al atte pts at solubilization do not e ceed the a ailable ( it is the rate and which will ield a small amount of the a ailable form b dissolution. owe er, it should be recogni ed that the and e tent of transfor ation hich ust be considered. he transfor ation ill be affected by a nu ber of factors not final solution concentration ma be influenced b a number of factors, including the solubilit product of some metal least of hich ill be the properties of the edia ith respect to p ater hardness te perature etc. n addition to compounds precipitated during the transformation dissolution test, e.g. aluminium h dro ide. these properties other factors such as the size and specific surface area of the particles hich ha e been tested the length of ti e o er hich e posure to the edia ta es place and of course the ass or surface area loading of the pplication of a atic to icit ata an sol ilit ata fo classification substance in the edia ill all play a part in deter ining the le el of dissol ed etal ions in the ater. ransfor ation data can generally therefore only be considered as reliable for the purposes of classification if conducted according to A . . .1 nterpretation of a uatic toxicity data the standard Protocol in nne . A . . .1.1 A uatic to icit studies carried out according to a recogni ed protocol should normall be acceptable . . . his Protocol ai s at standardizing the principal ariables such that the le el of dissol ed ion can be as alid for the purposes of classification. ection A . should also be consulted for generic issues that are common to directly related to the loading of the substance added. t is this loading le el hich yields the le el of etal ion assessing an a uatic to icit data point for the purposes of classification. equi alent to the a ailable ( that can then be used to deter ine the hazard category appropriate for classification. he testing ethodology is detailed in nne . he strategy to be adopted in using the data fro the testing protocol A . . .1. etal comple ation and speciation and the data require ents needed to a e that strategy or ill be described. A . . .1. .1 he to icit of a particular metal in solution, appears to depend primaril on but is not strictl limited . . . n considering the classification of etals and etal co pounds both readily and poorly soluble to the le el of dissol ed free metal ions. Abiotic factors including alkalinit , ionic strength and p can influence the recognition has to be paid to a number of factors. As defined in Chapter 4.1, the term “degradation” refers to the to icit of metals in two wa s i b influencing the chemical speciation of the metal in water and hence affecting the deco position of organic olecules. or inorganic co pounds and etals clearly the concept of degradability as it a ailabilit and ii b influencing the uptake and binding of a ailable metal b biological tissues. has been considered and used for organic substances has li ited or no eaning. ather the substance ay be transfor ed by nor al en iron ental processes to either increase or decrease the bioa ailability of the to ic species. A . . .1. . here speciation is important, it ma be possible to model the concentrations of the different forms of qually the log o cannot be considered as a easure of the potential to accu ulate. e ertheless the concepts that a the metal, including those that are likel to cause to icit . Anal sis methods for uantif ing e posure concentrations, substance or a to ic etabolite reaction product ay not be rapidly lost fro the en iron ent and or ay which are capable of distinguishing between the comple ed and uncomple ed fractions of a test substance, ma not bioaccu ulate are as applicable to etals and etal co pounds as they are to organic substances. alwa s be a ailable or economic.
. . . peciation of the soluble for can be affected by p ater hardness and other ariables and ay A . . .1. . Comple ation of metals to organic and inorganic ligands in test media and natural en ironments can yield particular for s of the etal ion hich are ore or less to ic. n addition etal ions could be ade non a ailable be estimated from metal speciation models. peciation models for metals, including p , hardness, C, and inorganic fro the ater colu n by a nu ber of processes (e.g. ineralization and partitioning . o eti es these processes can substances such as rown and Allison, 1 , A ipping, 1 4 and C antore and riscoll, be sufficiently rapid to be analogous to degradation in assessing chronic classification. o e er partitioning of the 1 can be used to calculate the uncomple ed and comple ed fractions of the metal ions. Alternati el , the iotic etal ion fro the ater colu n to other en iron ental edia does not necessarily ean that it is no longer igand odel , allows for the calculation of the concentration of metal ion responsible for the to ic effect at the bioa ailable nor does it ean that the etal has been ade per anently una ailable. le el of the organism. he model has at present onl been alidated for a limited number of metals, organisms, and end points antore and i oro, 1 . he models and formula used for the characteri ation of metal . . . nfor ation pertaining to the e tent of the partitioning of a etal ion fro the ater colu n or the comple ation in the media should alwa s be clearl reported, allowing for their translation back to natural en ironments e tent to hich a etal has been or can be con erted to a for that is less to ic or non to ic is frequently not a ailable C , . o er a sufficiently ide range of en iron entally rele ant conditions and thus a nu ber of assu ptions ill need to be ade as an aid in classification. hese assu ptions ay be odified if a ailable data sho other ise. n the first A . . . nterpretation of solubility data instance it should be assu ed that the etal ions once in the ater are not rapidly partitioned fro the ater colu n and thus these co pounds do not eet the criteria. nderlying this is the assu ption that although speciation can A . . . .1 hen considering the a ailable data on solubilit , their alidit and applicabilit to the identification occur the species ill re ain a ailable under en iron entally rele ant conditions. his ay not al ays be the case as of the ha ard of metal compounds should be assessed. n particular, a knowledge of the p at which the data were described abo e and any e idence a ailable that ould suggest changes to the bioa ailability o er the course of generated should be known. days should be carefully e a ined. he bioaccu ulation of etals and inorganic etal co pounds is a co ple process and bioaccu ulation data should be used ith care. he application of bioaccu ulation criteria ill need to be A . . . . Assessment of e isting data considered on a case by case basis ta ing due account of all the a ailable data. isting data will be in one of three forms. or some well studied metals, there will be solubilit . . . further assu ption that can be ade hich represents a cautious approach is that in the absence of products and or solubilit data for the arious inorganic metal compounds. t is also possible that the p relationship of any solubility data for a particular etal co pound either easured or calculated the substance ill be sufficiently the solubilit will be known. owe er, for man metals or metal compounds, it is probable that the a ailable soluble to cause to icity at the le el of the ( and thus ay be classified in the sa e ay as other soluble salts. information will be descripti e onl , e.g. poorl soluble. nfortunatel there appears to be er little consistent gain this is clearly not al ays the case and it ay be ise to generate appropriate solubility data.
4 - 485 - Copyright@United Nations, 2017. All rights reserved
guidance about the solubilit ranges for such descripti e terms. here these are the onl information a ailable it is ioacc lation probable that solubilit data will need to be generated using the ransformation issolution rotocol Anne 1 . . . .1 hile log ow is a good predictor of for certain types of organic compounds e.g. non polar A . . . . creening test for assessing solubilit of metal compounds organic substances, it is of course irrele ant for inorganic substances such as inorganic metal compounds.
n the absence of solubility data, a simple “Screening Test” for assessing solubility, based on the high . . . The mechanisms for upta e and depuration rates of metals are ery comple and ariable and there is rate of loading for 4 h, can be used for metal compounds as described in the ransformation issolution rotocol at present no general model to describe this. nstead the bioaccumulation of metals according to the classification Anne 1 . he function of the screening test is to identif those metal compounds which undergo either dissolution or criteria should be e aluated on a case by case basis using e pert udgement. rapid transformation such that the are indistinguishable from soluble forms and hence ma be classified based on the dissol ed ion concentration. here data are a ailable from the screening test detailed in the ransformation issolution . . . hile s are indicati e of the potential for bioaccumulation there may be a number of rotocol, the ma imum solubilit obtained o er the tested p range should be used. here data are not a ailable o er complications in interpreting measured alues for metals and inorganic metal compounds. or some metals and the full p range, a check should be made that this ma imum solubilit has been achie ed b reference to suitable inorganic metal compounds the relationship between water concentration and in some a uatic organisms is thermod namic speciation models or other suitable methods see A . . .1. . . t should be noted that this test is onl in erse, and bioconcentration data should be used with care. This is particularly rele ant for metals that are biologically intended to be used for metal compounds. essential. etals that are biologically essential are acti ely regulated in organisms in which the metal is essential. Since nutritional re uirement of the organisms can be higher than the en ironmental concentration, this acti e regulation can A . . . .4 ull test for assessing solubilit of metals and metal compounds result in high s and an in erse relationship between s and the concentration of the metal in water. hen en ironmental concentrations are low, high s may be e pected as a natural conse uence of metal upta e to meet he first step in this part of the stud is, as with the screening test, an assessment of the p s at nutritional re uirements and in these instances can be iewed as a normal phenomenon. dditionally, if internal which the stud should be conducted. ormall , the ull est should ha e been carried out at the p that ma imi es the concentration is regulated by the organism, then measured s may decline as e ternal concentration increases. hen concentration of dissol ed metal ions in solution. n such cases, the p ma be chosen following the same guidance as e ternal concentrations are so high that they e ceed a threshold le el or o erwhelm the regulatory mechanism, this can gi en for the screening test. cause harm to the organism. lso, while a metal may be essential in a particular organism, it may not be essential in other organisms. Therefore, where the metal is not essential or when the bioconcentration of an essential metal is abo e ased on the data from the ull est, it is possible to generate a concentration of the metal ions in nutritional le els special consideration should be gi en to the potential for bioconcentration and en ironmental concern. solution after da s for each of the three loadings (i.e. 1 mg/l as “low”, 10 mg/l as “medium” and 100 mg/l as “high”) used in the test. f the purpose of the test is to assess the long term chronic ha ard of the substance, then the test at the pplication of classification c ite ia to etals an etal co po n s low loading ma be e tended to da s, at an appropriate p . . . .1 ntroduction to the classification strategy for metals and metal compounds A . . . Comparison of a uatic toxicity data and solubility data . . .1.1 The schemes for the classification of metals and metal compounds are described below and A decision whether or not the substance be classified will be made b comparing a uatic to icit data summari ed diagrammatically in igure . .1. There are se eral stages in these schemes where data are used for and solubilit data. f the C is e ceeded, irrespecti e of whether the to icit and dissolution data are at the same decision purposes. t is not the intention of the classification schemes to generate new data. n the absence of alid data, p and if this is the onl data a ailable then the substance should be classified. f other solubilit data are a ailable to it will be necessary to use all a ailable data and e pert udgement. show that the dissolution concentration would not e ceed the C across the entire p range then the substance should not be classified on its soluble form. his ma in ol e the use of additional data either from ecoto icological n the following sections, the reference to the ( ) 0 refers to the data point(s) that will be used to testing or from applicable bioa ailabilit effect models. select the class for the metal or metal compound.
ssess ent of en i on ental t ansfo ation . . .1. hen considering ( ) 0 data for metal compounds, it is important to ensure that the data point to be used as the ustification for the classification is e pressed in the weight of the molecule of the metal compound to be A . . .1 n ironmental transformation of one species of a metal to another species of the same does not classified. This is nown as correcting for molecular weight. Thus while most metal data is e pressed in, for e ample, constitute degradation as applied to organic compounds and ma increase or decrease the a ailabilit and bioa ailabilit mg/l of the metal, this alue will need to be ad usted to the corresponding weight of the metal compound. Thus of the to ic species. owe er as a result of naturall occurring geochemical processes metal ions can partition from the water column. ata on water column residence time, the processes in ol ed at the water – sediment interface i.e. ) ( 0 met al compounds ( ) 0of met a (l olecular weight of met al compound/ t o mic weight of met al) deposition and re mobili ation are fairl e tensi e, but ha e not been integrated into a meaningful database. e ertheless, using the principles and assumptions discussed abo e in A . .1, it ma be possible to incorporate this data may also need to be ad usted to the corresponding weight of the metal compounds. approach into classification. . . . Classification strategy for metals A . . . uch assessments are er difficult to gi e guidance for and will normall be addressed on a case b case approach. owe er, the following ma be taken into account . . . .1 here the ( ) 0 for the metal ions of concern is greater than 100 mg/l, the metals need not be considered further in the classification scheme. a Changes in speciation if the are to non a ailable forms, howe er, the potential for the re erse change to occur must also be considered . . . . here the ( ) 0 for the metal ions of concern is ≤100 mg/l, consideration must be gi en to the data a ailable on the rate and e tent to which these ions can be generated from the metal. Such data, to be alid and useable b Changes to a metal compound which is considerabl less soluble than that of the metal should ha e been generated using the Transformation/ issolution rotocol ( nne 10). compound being considered. . . . . here such data are una ailable, i.e. there is no clear data of sufficient alidity to show that the ome caution is recommended, see A . .1. and A . .1. . transformation to metal ions will not occur, the safety net classification ( hronic ) should be applied since the nown classifiable to icity of these soluble forms is considered to produce sufficient concern.
. . . . here data from dissolution protocol are a ailable, then, the results should be used to aid classification according to the following rules
4 - 486 - Copyright@United Nations, 2017. All rights reserved
guidance about the solubilit ranges for such descripti e terms. here these are the onl information a ailable it is ioacc lation probable that solubilit data will need to be generated using the ransformation issolution rotocol Anne 1 . . . .1 hile log ow is a good predictor of for certain types of organic compounds e.g. non polar A . . . . creening test for assessing solubilit of metal compounds organic substances, it is of course irrele ant for inorganic substances such as inorganic metal compounds.
n the absence of solubility data, a simple “Screening Test” for assessing solubility, based on the high . . . The mechanisms for upta e and depuration rates of metals are ery comple and ariable and there is rate of loading for 4 h, can be used for metal compounds as described in the ransformation issolution rotocol at present no general model to describe this. nstead the bioaccumulation of metals according to the classification Anne 1 . he function of the screening test is to identif those metal compounds which undergo either dissolution or criteria should be e aluated on a case by case basis using e pert udgement. rapid transformation such that the are indistinguishable from soluble forms and hence ma be classified based on the dissol ed ion concentration. here data are a ailable from the screening test detailed in the ransformation issolution . . . hile s are indicati e of the potential for bioaccumulation there may be a number of rotocol, the ma imum solubilit obtained o er the tested p range should be used. here data are not a ailable o er complications in interpreting measured alues for metals and inorganic metal compounds. or some metals and the full p range, a check should be made that this ma imum solubilit has been achie ed b reference to suitable inorganic metal compounds the relationship between water concentration and in some a uatic organisms is thermod namic speciation models or other suitable methods see A . . .1. . . t should be noted that this test is onl in erse, and bioconcentration data should be used with care. This is particularly rele ant for metals that are biologically intended to be used for metal compounds. essential. etals that are biologically essential are acti ely regulated in organisms in which the metal is essential. Since nutritional re uirement of the organisms can be higher than the en ironmental concentration, this acti e regulation can A . . . .4 ull test for assessing solubilit of metals and metal compounds result in high s and an in erse relationship between s and the concentration of the metal in water. hen en ironmental concentrations are low, high s may be e pected as a natural conse uence of metal upta e to meet he first step in this part of the stud is, as with the screening test, an assessment of the p s at nutritional re uirements and in these instances can be iewed as a normal phenomenon. dditionally, if internal which the stud should be conducted. ormall , the ull est should ha e been carried out at the p that ma imi es the concentration is regulated by the organism, then measured s may decline as e ternal concentration increases. hen concentration of dissol ed metal ions in solution. n such cases, the p ma be chosen following the same guidance as e ternal concentrations are so high that they e ceed a threshold le el or o erwhelm the regulatory mechanism, this can gi en for the screening test. cause harm to the organism. lso, while a metal may be essential in a particular organism, it may not be essential in other organisms. Therefore, where the metal is not essential or when the bioconcentration of an essential metal is abo e ased on the data from the ull est, it is possible to generate a concentration of the metal ions in nutritional le els special consideration should be gi en to the potential for bioconcentration and en ironmental concern. solution after da s for each of the three loadings (i.e. 1 mg/l as “low”, 10 mg/l as “medium” and 100 mg/l as “high”) used in the test. f the purpose of the test is to assess the long term chronic ha ard of the substance, then the test at the pplication of classification c ite ia to etals an etal co po n s low loading ma be e tended to da s, at an appropriate p . . . .1 ntroduction to the classification strategy for metals and metal compounds A . . . Comparison of a uatic toxicity data and solubility data . . .1.1 The schemes for the classification of metals and metal compounds are described below and A decision whether or not the substance be classified will be made b comparing a uatic to icit data summari ed diagrammatically in igure . .1. There are se eral stages in these schemes where data are used for and solubilit data. f the C is e ceeded, irrespecti e of whether the to icit and dissolution data are at the same decision purposes. t is not the intention of the classification schemes to generate new data. n the absence of alid data, p and if this is the onl data a ailable then the substance should be classified. f other solubilit data are a ailable to it will be necessary to use all a ailable data and e pert udgement. show that the dissolution concentration would not e ceed the C across the entire p range then the substance should not be classified on its soluble form. his ma in ol e the use of additional data either from ecoto icological n the following sections, the reference to the ( ) 0 refers to the data point(s) that will be used to testing or from applicable bioa ailabilit effect models. select the class for the metal or metal compound.
ssess ent of en i on ental t ansfo ation . . .1. hen considering ( ) 0 data for metal compounds, it is important to ensure that the data point to be used as the ustification for the classification is e pressed in the weight of the molecule of the metal compound to be A . . .1 n ironmental transformation of one species of a metal to another species of the same does not classified. This is nown as correcting for molecular weight. Thus while most metal data is e pressed in, for e ample, constitute degradation as applied to organic compounds and ma increase or decrease the a ailabilit and bioa ailabilit mg/l of the metal, this alue will need to be ad usted to the corresponding weight of the metal compound. Thus of the to ic species. owe er as a result of naturall occurring geochemical processes metal ions can partition from the water column. ata on water column residence time, the processes in ol ed at the water – sediment interface i.e. ) ( 0 met al compounds ( ) 0of met a (l olecular weight of met al compound/ t o mic weight of met al) deposition and re mobili ation are fairl e tensi e, but ha e not been integrated into a meaningful database. e ertheless, using the principles and assumptions discussed abo e in A . .1, it ma be possible to incorporate this data may also need to be ad usted to the corresponding weight of the metal compounds. approach into classification. . . . Classification strategy for metals A . . . uch assessments are er difficult to gi e guidance for and will normall be addressed on a case b case approach. owe er, the following ma be taken into account . . . .1 here the ( ) 0 for the metal ions of concern is greater than 100 mg/l, the metals need not be considered further in the classification scheme. a Changes in speciation if the are to non a ailable forms, howe er, the potential for the re erse change to occur must also be considered . . . . here the ( ) 0 for the metal ions of concern is ≤100 mg/l, consideration must be gi en to the data a ailable on the rate and e tent to which these ions can be generated from the metal. Such data, to be alid and useable b Changes to a metal compound which is considerabl less soluble than that of the metal should ha e been generated using the Transformation/ issolution rotocol ( nne 10). compound being considered. . . . . here such data are una ailable, i.e. there is no clear data of sufficient alidity to show that the ome caution is recommended, see A . .1. and A . .1. . transformation to metal ions will not occur, the safety net classification ( hronic ) should be applied since the nown classifiable to icity of these soluble forms is considered to produce sufficient concern.
. . . . here data from dissolution protocol are a ailable, then, the results should be used to aid classification according to the following rules
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. . . . .1 day Transformation test . . . . . day transformation test
f the dissol ed metal ion concentration after a period of days (or earlier) e ceeds that of the or poorly soluble metal compounds classified with the default safety net classification further ( ) 0, then the default classification for the metals is replaced by the following classification information that may be a ailable from the day transformation/dissolution test can also be used. Such data should include transformation le els at low, medium and high loading le els.
(a) If the dissolved metal ion concentration at the low loading rate is ≥ L(E)C 0, then classify as cute 1. lassify also as hronic 1, unless there is e idence of both rapid partitioning from the f the dissol ed metal ion concentration after a period of days (or earlier) e ceeds that of the water column and no bioaccumulation ( ) 0, then the default classification for the metals is replaced by the following classification
(b) f the dissol ed metal ion concentration at the medium loading rate is ≥ L(E)C 0, then classify (a) If the dissolved metal ion concentration at the low loading rate is ≥L(E)C 0, then classify as as cute . lassify also as hronic unless there is e idence of both rapid partitioning from cute 1. lassify also as hronic 1, unless there is e idence of both rapid partitioning from the the water column and no bioaccumulation water column and no bioaccumulation
(c) f the dissol ed metal ion concentration at the high loading rate is ≥ L(E)C 0, then classify as cute . lassify also as hronic unless there is e idence of both rapid partitioning from the (b) f the dissol ed metal ion concentration at the medium loading rate is ≥ L(E)C 0, then classify water column and no bioaccumulation. as cute . lassify also as hronic unless there is e idence of both rapid partitioning from the water column and no bioaccumulation . . . . . day transformation test (c) f the dissol ed metal ion concentration at the high loading rate is ≥ L(E)C 0, then classify as cute . lassify also as hronic unless there is e idence of both rapid partitioning from the f the process described in . . . . .1 results in classification as hronic 1, no further assessment is water column and no bioaccumulation. re uired, as the metal will be classified irrespecti e of any further information. . . . . . day transformation test n all other cases, further data may ha e been generated through the dissolution/transformation test in order to show that the classification may be amended. f for substances classified as hronic , or , the dissol ed f the process described in . . . . . results in classification as hronic 1, no further assessment is metal ion concentration at the low loading rate after a total period of 28 days is ≤ long term s, then the re uired as the metal compound will be classified irrespecti e of any further information. classification is remo ed. n all other cases, further data may ha e been generated through the dissolution/transformation test for . . . Classification strategy for metal compounds days in order to show that the classification may be amended. f for poorly soluble metal compounds classified as hronic , or , the dissol ed metal ion concentration at the low loading rate after a total period of days is less than . . . .1 here the ( ) 0 for the metal ions of concern is 100 mg/l, the metal compounds need not be or e ual to the long term s, then classification is remo ed. considered further in the classification scheme. . . . article si e and surface area
. . . . f solubility ( ) 0, classify on the basis of soluble ion. . . . .1 article si e, or moreo er surface area, is a crucial parameter in that any ariation in the si e or . . . . .1 ll metal compounds with a water solubility (either measured e.g. through hour issolution surface area tested may cause a significant change in the le els of metals ions released in a gi en time window. Thus, this particle si e or surface area is fi ed for the purposes of the transformation test, allowing the comparati e Screening test or estimated e.g. from the solubility product) ≥ L(E)C 0 of the dissol ed metal ion concentration are considered as readily soluble metal compounds. are should be e ercised for compounds whose solubility is close to classifications to be based solely on the loading le el. ormally, the classification data generated would ha e used the the acute to icity alue as the conditions under which solubility is measured could differ significantly from those of the smallest particle si e mar eted to determine the e tent of transformation. There may be cases where data generated for a acute to icity test. n these cases the results of the issolution Screening Test are preferred. particular metal powder is not considered as suitable for classification of the massi e forms. or e ample, where it can be shown that the tested powder is structurally a different material (e.g. different crystallographic structure) and/or it has been produced by a special process and cannot be generated from the massi e metal, classification of the massi e can . . . . . eadily soluble metal compounds are classified on the basis of the ( ) 0 (corrected where necessary for molecular weight) be based on testing of a more representati e particle si e or surface area, if such data are a ailable. The powder may be classified separately based on the data generated on the powder. owe er, in normal circumstances it is not anticipated that more than two classification proposals would be made for the same metal. (a) f the ( ) 0 of the dissolved metal ion is ≤ 1 mg/l then classify as Acute 1. Classify also as hronic 1 unless there is e idence of both rapid partitioning from the water column and no . . . . etals with a particle si e smaller than the default diameter alue of 1 mm can be tested on a case by bioaccumulation case basis. ne e ample of this is where metal powders are produced by a different production techni ue or where the (b) f the ( ) 0 of the dissolved metal ion is >1 mg/l but ≤ 10 mg/l then classify as cute . powders gi e rise to a higher dissolution (or reaction) rate than the massi e form leading to a more stringent lassify also as hronic unless there is e idence of both rapid partitioning from the water classification. column and no bioaccumulation . . . . The particle si es tested depend on the substance being assessed and are shown in the table below (c) f the ( ) 0 of the dissolved metal ion is > 10 mg/l and ≤ 100 mg/l then classify as Acute 3. lassify also as hronic unless there is e idence of both rapid partitioning from the water column and no bioaccumulation. pe rtic e i e o ent etal compounds Smallest representati e si e sold e er larger than 1 mm . . . . f solubility L(E C classify default Chronic 50 etals – powders Smallest representati e si e sold ay need to consider different sources if yielding different crystallographic/morphologic properties . . . . .1 n the conte t of the classification criteria, poorly soluble compounds of metals are defined as those with a nown solubility (either measured e.g. through hour issolution Screening test or estimated e.g. from the etals – massi e 1 mm efault alue may be altered if sufficient ustification solubility product) less than the ( ) 0 of the soluble metal ion. n those cases when the soluble forms of the metal of poorly soluble metal compounds ha e a ( ) 0 ≤ 100 mg/l and the substance can be considered as poorly soluble the . . . . or some forms of metals, it may be possible, using the Transformation/ issolution rotocol ( default safety net classification ( hronic ) should be applied. 001), to obtain a correlation between the concentration of the metal ion after a specified time inter al as a function of the surface area loadings of the forms tested. n such cases, it could then be possible to estimate the le el of dissol ed metal ion concentration of the metal with different particles, using the critical surface area approach as proposed by
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. . . . .1 day Transformation test . . . . . day transformation test
f the dissol ed metal ion concentration after a period of days (or earlier) e ceeds that of the or poorly soluble metal compounds classified with the default safety net classification further ( ) 0, then the default classification for the metals is replaced by the following classification information that may be a ailable from the day transformation/dissolution test can also be used. Such data should include transformation le els at low, medium and high loading le els.
(a) If the dissolved metal ion concentration at the low loading rate is ≥ L(E)C 0, then classify as cute 1. lassify also as hronic 1, unless there is e idence of both rapid partitioning from the f the dissol ed metal ion concentration after a period of days (or earlier) e ceeds that of the water column and no bioaccumulation ( ) 0, then the default classification for the metals is replaced by the following classification
(b) f the dissol ed metal ion concentration at the medium loading rate is ≥ L(E)C 0, then classify (a) If the dissolved metal ion concentration at the low loading rate is ≥L(E)C 0, then classify as as cute . lassify also as hronic unless there is e idence of both rapid partitioning from cute 1. lassify also as hronic 1, unless there is e idence of both rapid partitioning from the the water column and no bioaccumulation water column and no bioaccumulation
(c) f the dissol ed metal ion concentration at the high loading rate is ≥ L(E)C 0, then classify as cute . lassify also as hronic unless there is e idence of both rapid partitioning from the (b) f the dissol ed metal ion concentration at the medium loading rate is ≥ L(E)C 0, then classify water column and no bioaccumulation. as cute . lassify also as hronic unless there is e idence of both rapid partitioning from the water column and no bioaccumulation . . . . . day transformation test (c) f the dissol ed metal ion concentration at the high loading rate is ≥ L(E)C 0, then classify as cute . lassify also as hronic unless there is e idence of both rapid partitioning from the f the process described in . . . . .1 results in classification as hronic 1, no further assessment is water column and no bioaccumulation. re uired, as the metal will be classified irrespecti e of any further information. . . . . . day transformation test n all other cases, further data may ha e been generated through the dissolution/transformation test in order to show that the classification may be amended. f for substances classified as hronic , or , the dissol ed f the process described in . . . . . results in classification as hronic 1, no further assessment is metal ion concentration at the low loading rate after a total period of 28 days is ≤ long term s, then the re uired as the metal compound will be classified irrespecti e of any further information. classification is remo ed. n all other cases, further data may ha e been generated through the dissolution/transformation test for . . . Classification strategy for metal compounds days in order to show that the classification may be amended. f for poorly soluble metal compounds classified as hronic , or , the dissol ed metal ion concentration at the low loading rate after a total period of days is less than . . . .1 here the ( ) 0 for the metal ions of concern is 100 mg/l, the metal compounds need not be or e ual to the long term s, then classification is remo ed. considered further in the classification scheme. . . . article si e and surface area
. . . . f solubility ( ) 0, classify on the basis of soluble ion. . . . .1 article si e, or moreo er surface area, is a crucial parameter in that any ariation in the si e or . . . . .1 ll metal compounds with a water solubility (either measured e.g. through hour issolution surface area tested may cause a significant change in the le els of metals ions released in a gi en time window. Thus, this particle si e or surface area is fi ed for the purposes of the transformation test, allowing the comparati e Screening test or estimated e.g. from the solubility product) ≥ L(E)C 0 of the dissol ed metal ion concentration are considered as readily soluble metal compounds. are should be e ercised for compounds whose solubility is close to classifications to be based solely on the loading le el. ormally, the classification data generated would ha e used the the acute to icity alue as the conditions under which solubility is measured could differ significantly from those of the smallest particle si e mar eted to determine the e tent of transformation. There may be cases where data generated for a acute to icity test. n these cases the results of the issolution Screening Test are preferred. particular metal powder is not considered as suitable for classification of the massi e forms. or e ample, where it can be shown that the tested powder is structurally a different material (e.g. different crystallographic structure) and/or it has been produced by a special process and cannot be generated from the massi e metal, classification of the massi e can . . . . . eadily soluble metal compounds are classified on the basis of the ( ) 0 (corrected where necessary for molecular weight) be based on testing of a more representati e particle si e or surface area, if such data are a ailable. The powder may be classified separately based on the data generated on the powder. owe er, in normal circumstances it is not anticipated that more than two classification proposals would be made for the same metal. (a) f the ( ) 0 of the dissolved metal ion is ≤ 1 mg/l then classify as Acute 1. Classify also as hronic 1 unless there is e idence of both rapid partitioning from the water column and no . . . . etals with a particle si e smaller than the default diameter alue of 1 mm can be tested on a case by bioaccumulation case basis. ne e ample of this is where metal powders are produced by a different production techni ue or where the (b) f the ( ) 0 of the dissolved metal ion is >1 mg/l but ≤ 10 mg/l then classify as cute . powders gi e rise to a higher dissolution (or reaction) rate than the massi e form leading to a more stringent lassify also as hronic unless there is e idence of both rapid partitioning from the water classification. column and no bioaccumulation . . . . The particle si es tested depend on the substance being assessed and are shown in the table below (c) f the ( ) 0 of the dissolved metal ion is > 10 mg/l and ≤ 100 mg/l then classify as Acute 3. lassify also as hronic unless there is e idence of both rapid partitioning from the water column and no bioaccumulation. pe rtic e i e o ent etal compounds Smallest representati e si e sold e er larger than 1 mm . . . . f solubility L(E C classify default Chronic 50 etals – powders Smallest representati e si e sold ay need to consider different sources if yielding different crystallographic/morphologic properties . . . . .1 n the conte t of the classification criteria, poorly soluble compounds of metals are defined as those with a nown solubility (either measured e.g. through hour issolution Screening test or estimated e.g. from the etals – massi e 1 mm efault alue may be altered if sufficient ustification solubility product) less than the ( ) 0 of the soluble metal ion. n those cases when the soluble forms of the metal of poorly soluble metal compounds ha e a ( ) 0 ≤ 100 mg/l and the substance can be considered as poorly soluble the . . . . or some forms of metals, it may be possible, using the Transformation/ issolution rotocol ( default safety net classification ( hronic ) should be applied. 001), to obtain a correlation between the concentration of the metal ion after a specified time inter al as a function of the surface area loadings of the forms tested. n such cases, it could then be possible to estimate the le el of dissol ed metal ion concentration of the metal with different particles, using the critical surface area approach as proposed by
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et. al. Annex 9 APPENDIX I Determination of degradability of organic substances
i re i ic tion tr te or et n et co po n 1. Organic substances may be degraded by abiotic or biotic processes or by a combination of these. A et or et co po n number of standard procedures or tests for determination of the degradability are available. The general principles of some of these are described below. It is by no way the intention to present a comprehensive review of degradability test methods, but only to place the methods in the context of aquatic hazard classification. o c i ic tion 2. Abiotic degradability 2.1 Abiotic degradation comprises chemical transformation and photochemical transformation. Usually abiotic transformations will yield other organic compounds but will not cause a full mineralization (Schwarzenbach et al., 1993). Chemical transformation is defined as transformation that happens without light and without the mediation of organisms whereas photochemical transformations require light. 2.2 Examples of relevant chemical transformation processes in aqueous environment are hydrolysis, nucleophilic substitution, elimination, oxidation and reduction reactions (Schwarzenbach et al., 1993). Of these, hydrolysis is often considered the most important and it is the only chemical transformation process for which international test guidelines are generally available. The tests for abiotic degradation of chemicals are generally in the form of determination of transformation rates under standardized conditions. This box applies only to metal Hydrolysis 2.3 compounds - 2.3.1 Hydrolysis is the reaction of the nucleophiles H2O or OH with a chemical where a (leaving) group of the chemical is exchanged with an OH group. Many compounds, especially acid derivatives, are susceptible to hydrolysis. Hydrolysis can both be abiotic and biotic, but in regard to testing only abiotic hydrolysis is considered. Hydrolysis can take place by different mechanisms at different pHs, neutral, acid- or base-catalysed hydrolysis, and hydrolysis rates may be very dependent on pH. ronic 2.3.2 Currently two guidelines for evaluating abiotic hydrolysis are generally available, the OECD Test c te Guideline 111 Hydrolysis as a function of pH (corresponding to OPPTS 835.2110) and OPPTS 835.2130 Hydrolysis as a function of pH and temperature. In OECD Test Guideline 111, the overall hydrolysis rate at different pHs in pure buffered water is determined. The test is divided in two, a preliminary test that is performed for chemicals with unknown hydrolysis rates and a more detailed test that is performed for chemicals that are known to be hydrolytically ronic unstable and for chemicals for which the preliminary test shows fast hydrolysis. In the preliminary test the concentration of the chemical in buffered solutions at pHs in the range normally found in the environment (pHs of 4, 7 o and 9) at 50 C is measured after 5 days. If the concentration of the chemical has decreased less than 10 % it is c te considered hydrolytically stable, otherwise the detailed test may be performed. In the detailed test, the overall hydrolysis rate is determined at three pHs (4, 7 and 9) by measuring the concentration of the chemical as a function of time. The hydrolysis rate is determined at different temperatures so that interpolations or extrapolations to environmentally relevant temperatures can be made. The OPPTS 835.2130 test is almost identical in design to the OECD Test Guideline 111, the difference mainly being in the treatment of data. ronic 2.3.3 It should be noted that apart from hydrolysis the hydrolysis rate constants determined by the tests include all other abiotic transformations that may occur without light under the given test conditions. Good agreement c te has been found between hydrolysis rates in natural and in pure waters (OPPTS 835.2110). 2.4 Photolysis 2.4.1 According to the definitions set out in the OECD Guidance Document concerning aquatic direct photolysis (OECD, 1997), phototransformation of compounds in water can be in the form of primary or secondary ronic phototransformation, where the primary phototransformation (photolysis) can be divided further into direct and indirect photolysis. Direct phototransformation (photolysis) is the case where the chemical absorbs light and as a direct result hereof undergoes transformation. Indirect phototransformation is the case where other excited species transfer energy, electrons or H-atoms to the chemical and thereby induces a transformation (sensitized photolysis). Secondary phototransformation is the case where chemical reactions occur between the chemical and reactive short lived species like hydroxy radicals, peroxy radicals or singlet oxygen that are formed in the presence of light by reactions of excited species like excited humic or fulvic acids or nitrate.
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et. al. Annex 9 APPENDIX I Determination of degradability of organic substances
i re i ic tion tr te or et n et co po n 1. Organic substances may be degraded by abiotic or biotic processes or by a combination of these. A et or et co po n number of standard procedures or tests for determination of the degradability are available. The general principles of some of these are described below. It is by no way the intention to present a comprehensive review of degradability test methods, but only to place the methods in the context of aquatic hazard classification. o c i ic tion 2. Abiotic degradability 2.1 Abiotic degradation comprises chemical transformation and photochemical transformation. Usually abiotic transformations will yield other organic compounds but will not cause a full mineralization (Schwarzenbach et al., 1993). Chemical transformation is defined as transformation that happens without light and without the mediation of organisms whereas photochemical transformations require light. 2.2 Examples of relevant chemical transformation processes in aqueous environment are hydrolysis, nucleophilic substitution, elimination, oxidation and reduction reactions (Schwarzenbach et al., 1993). Of these, hydrolysis is often considered the most important and it is the only chemical transformation process for which international test guidelines are generally available. The tests for abiotic degradation of chemicals are generally in the form of determination of transformation rates under standardized conditions. This box applies only to metal Hydrolysis 2.3 compounds - 2.3.1 Hydrolysis is the reaction of the nucleophiles H2O or OH with a chemical where a (leaving) group of the chemical is exchanged with an OH group. Many compounds, especially acid derivatives, are susceptible to hydrolysis. Hydrolysis can both be abiotic and biotic, but in regard to testing only abiotic hydrolysis is considered. Hydrolysis can take place by different mechanisms at different pHs, neutral, acid- or base-catalysed hydrolysis, and hydrolysis rates may be very dependent on pH. ronic 2.3.2 Currently two guidelines for evaluating abiotic hydrolysis are generally available, the OECD Test c te Guideline 111 Hydrolysis as a function of pH (corresponding to OPPTS 835.2110) and OPPTS 835.2130 Hydrolysis as a function of pH and temperature. In OECD Test Guideline 111, the overall hydrolysis rate at different pHs in pure buffered water is determined. The test is divided in two, a preliminary test that is performed for chemicals with unknown hydrolysis rates and a more detailed test that is performed for chemicals that are known to be hydrolytically ronic unstable and for chemicals for which the preliminary test shows fast hydrolysis. In the preliminary test the concentration of the chemical in buffered solutions at pHs in the range normally found in the environment (pHs of 4, 7 o and 9) at 50 C is measured after 5 days. If the concentration of the chemical has decreased less than 10 % it is c te considered hydrolytically stable, otherwise the detailed test may be performed. In the detailed test, the overall hydrolysis rate is determined at three pHs (4, 7 and 9) by measuring the concentration of the chemical as a function of time. The hydrolysis rate is determined at different temperatures so that interpolations or extrapolations to environmentally relevant temperatures can be made. The OPPTS 835.2130 test is almost identical in design to the OECD Test Guideline 111, the difference mainly being in the treatment of data. ronic 2.3.3 It should be noted that apart from hydrolysis the hydrolysis rate constants determined by the tests include all other abiotic transformations that may occur without light under the given test conditions. Good agreement c te has been found between hydrolysis rates in natural and in pure waters (OPPTS 835.2110). 2.4 Photolysis 2.4.1 According to the definitions set out in the OECD Guidance Document concerning aquatic direct photolysis (OECD, 1997), phototransformation of compounds in water can be in the form of primary or secondary ronic phototransformation, where the primary phototransformation (photolysis) can be divided further into direct and indirect photolysis. Direct phototransformation (photolysis) is the case where the chemical absorbs light and as a direct result hereof undergoes transformation. Indirect phototransformation is the case where other excited species transfer energy, electrons or H-atoms to the chemical and thereby induces a transformation (sensitized photolysis). Secondary phototransformation is the case where chemical reactions occur between the chemical and reactive short lived species like hydroxy radicals, peroxy radicals or singlet oxygen that are formed in the presence of light by reactions of excited species like excited humic or fulvic acids or nitrate.
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Direct photolysis rate in water by sunlight hototransformation of chemicals in water direct photolysis ndirect photolysis screening test n e ent io e a a ilit
iotic e r i it ea io e a a ilit ) is a relatively “weak” inherent test. However, although the degradation potential in these tests is atic si lation tests
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Direct photolysis rate in water by sunlight hototransformation of chemicals in water direct photolysis ndirect photolysis screening test n e ent io e a a ilit
iotic e r i it ea io e a a ilit ) is a relatively “weak” inherent test. However, although the degradation potential in these tests is atic si lation tests
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si lation tests oint ro e t on the valuation o ) s , ), and y edersen et al. ). he latter is rie ly re erred elow. ests are also availa le or si ulating the degrada ility in a sewage treat ent plant ), e.g. the est uideline oupled nit test, tivated sludge si ulation test, and the . test. . . validation set o e peri entally deter ined iodegradation data was sele ted a ong the data ro e ently, a new si ulation test e ploying low on entrations o organi pollutants has een proposed yhol et. al., ), ut e luding su stan es or whi h no pre ise degradation data were availa le and su stan es already ). used or develop ent o the progra e. he validation set then onsisted o su stan es. he iodegrada ility o these su stan es were esti ated y use o the progra e s non linear esti ation odule the ost relia le) and the nae o ic e a a ilit results compared with the measured data. 162 substances were predicted to degrade “fast”, but only 41 (25%) were actually readily degradable in the MITI I test. 142 substances were predicted to degrade “slowly”, which was confirmed . . est ethods or anaero i iodegrada ility deter ine the intrinsi potential o the test su stan e to y ) su stan es eing not readily degrada le in the test. hus, it was on luded that the progra e undergo iodegradation under anaero i onditions. a ples o su h tests are the ) test, the ay e used or lassi i ation purposes only when no e peri ental degradation data an e o tained, and when the test and the . test. programme predicts a substance to be degraded “slowly”. In this case, the substance can be regarded as not rapidly degrada le. . . he potential or anaero i degradation is deter ined during a period o up to eight weeks and with the test onditions indi ated elow . . he sa e on lusion was rea hed in the oint ro e t on the valuation o ) s y use o e peri ental and data on new su stan es noti ied in the . he evaluation was ased on an analysis o a) per or an e o the test in sealed vessels in the a sen e o initially in a pure predi tions on new su stan es also tested e peri entally in ready iodegrada ility tests. nly o the at osphere) su stan es in luded in this analysis were readily iodegrada le. he e ployed ethodology is not ully spe i ied in the inal report o the oint pro e t , ), ut it is likely that the a ority o ) use o digested sludge predi tions were ade y using ethods whi h later have een integrated in the iodegradation ro a ility rogra . ) a test te perature o and . . lso in the , ) it is re o ended that esti ated iodegrada ility y use o the iodegradation ro a ility rogra is used only in a onservative way, i.e. when the progra e predi ts ast d) deter ination o head spa e gas pressure and H or ation). iodegradation, this result should not e taken into onsideration, whereas predi tions o slow iodegradation ay e . . he ulti ate degradation is deter ined y deter ining the gas produ tion. However, also pri ary onsidered , ). degradation ay e deter ined y easuring the re aining parent su stan e. . . hus, the use o results o the iodegrada ility ro a ility rogra in a onservative way ay ul il e a ation in soil an se i ent the needs or evaluating iodegrada ility o so e o the large nu er o su stan es or whi h no e peri ental degradation data are availa le. . . any su stan es end up in the soil or sedi ent o part ents and an assess ent o their degrada ility in these environ ents ay there ore e o i portan e. ong standard ethods ay e entioned the est uideline test on inherent iodegrada ility in soil, whi h orresponds to the . test.
. . he spe ial test hara teristi s ensuring the deter ination o the inherent degrada ility in soil are
a) natural soil sa ples are used without additional ino ulation
) radiola elled test su stan e is used and
) evolution o radiola elled is deter ined.
. . standard ethod or deter ining the iodegradation in sedi ent is the . edi ent water i ro os iodegradation test. i ro os s ontaining sedi ent and water are olle ted ro test sites and test o pounds are introdu ed into the syste . isappearan e o the parent o pound i.e. pri ary iodegradation) and, i easi le, appearan e o eta olites or easure ents o ulti ate iodegradation ay e ade.
. . wo test guidelines address aero i and anaero i trans or ation in soil and in a uati sedi ents est uidelines and respe tively). he e peri ents are per or ed to deter ine the rate o trans or ation o the test su stan e and the nature and rates o or ation and de line o trans or ation produ ts under environ entally realisti onditions in luding a realisti on entration o the test su stan e. ither o plete inerali ation or pri ary degrada ility ay e deter ined depending on the analyti al ethod e ployed or deter ining the trans or ation o the test su stan e.
et o s fo esti atin io e a a ilit
. . n re ent years, possi ilities or esti ating environ ental properties o su stan es have een developed and, a ong these, also ethods or predi ting the iodegrada ility potential o organi su stan es e.g. the yra use esear h orporation s iodegrada ility ro a ility rogra , ). eviews o ethods have een per or ed y ) and y angen erg et al. ). hey show that group ontri ution ethods see to e the ost su ess ul ethods. these, the iodegradation ro a ility rogra ) see s to have the roadest appli ation. t gives a ualitative esti ate o the pro a ility o slow or ast iodegradation in the presen e o a i ed population o environ ental i ro organis s. he appli a ility o this progra has een evaluated y the
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si lation tests oint ro e t on the valuation o ) s , ), and y edersen et al. ). he latter is rie ly re erred elow. ests are also availa le or si ulating the degrada ility in a sewage treat ent plant ), e.g. the est uideline oupled nit test, tivated sludge si ulation test, and the . test. . . validation set o e peri entally deter ined iodegradation data was sele ted a ong the data ro e ently, a new si ulation test e ploying low on entrations o organi pollutants has een proposed yhol et. al., ), ut e luding su stan es or whi h no pre ise degradation data were availa le and su stan es already ). used or develop ent o the progra e. he validation set then onsisted o su stan es. he iodegrada ility o these su stan es were esti ated y use o the progra e s non linear esti ation odule the ost relia le) and the nae o ic e a a ilit results compared with the measured data. 162 substances were predicted to degrade “fast”, but only 41 (25%) were actually readily degradable in the MITI I test. 142 substances were predicted to degrade “slowly”, which was confirmed . . est ethods or anaero i iodegrada ility deter ine the intrinsi potential o the test su stan e to y ) su stan es eing not readily degrada le in the test. hus, it was on luded that the progra e undergo iodegradation under anaero i onditions. a ples o su h tests are the ) test, the ay e used or lassi i ation purposes only when no e peri ental degradation data an e o tained, and when the test and the . test. programme predicts a substance to be degraded “slowly”. In this case, the substance can be regarded as not rapidly degrada le. . . he potential or anaero i degradation is deter ined during a period o up to eight weeks and with the test onditions indi ated elow . . he sa e on lusion was rea hed in the oint ro e t on the valuation o ) s y use o e peri ental and data on new su stan es noti ied in the . he evaluation was ased on an analysis o a) per or an e o the test in sealed vessels in the a sen e o initially in a pure predi tions on new su stan es also tested e peri entally in ready iodegrada ility tests. nly o the at osphere) su stan es in luded in this analysis were readily iodegrada le. he e ployed ethodology is not ully spe i ied in the inal report o the oint pro e t , ), ut it is likely that the a ority o ) use o digested sludge predi tions were ade y using ethods whi h later have een integrated in the iodegradation ro a ility rogra . ) a test te perature o and . . lso in the , ) it is re o ended that esti ated iodegrada ility y use o the iodegradation ro a ility rogra is used only in a onservative way, i.e. when the progra e predi ts ast d) deter ination o head spa e gas pressure and H or ation). iodegradation, this result should not e taken into onsideration, whereas predi tions o slow iodegradation ay e . . he ulti ate degradation is deter ined y deter ining the gas produ tion. However, also pri ary onsidered , ). degradation ay e deter ined y easuring the re aining parent su stan e. . . hus, the use o results o the iodegrada ility ro a ility rogra in a onservative way ay ul il e a ation in soil an se i ent the needs or evaluating iodegrada ility o so e o the large nu er o su stan es or whi h no e peri ental degradation data are availa le. . . any su stan es end up in the soil or sedi ent o part ents and an assess ent o their degrada ility in these environ ents ay there ore e o i portan e. ong standard ethods ay e entioned the est uideline test on inherent iodegrada ility in soil, whi h orresponds to the . test.
. . he spe ial test hara teristi s ensuring the deter ination o the inherent degrada ility in soil are
a) natural soil sa ples are used without additional ino ulation
) radiola elled test su stan e is used and
) evolution o radiola elled is deter ined.
. . standard ethod or deter ining the iodegradation in sedi ent is the . edi ent water i ro os iodegradation test. i ro os s ontaining sedi ent and water are olle ted ro test sites and test o pounds are introdu ed into the syste . isappearan e o the parent o pound i.e. pri ary iodegradation) and, i easi le, appearan e o eta olites or easure ents o ulti ate iodegradation ay e ade.
. . wo test guidelines address aero i and anaero i trans or ation in soil and in a uati sedi ents est uidelines and respe tively). he e peri ents are per or ed to deter ine the rate o trans or ation o the test su stan e and the nature and rates o or ation and de line o trans or ation produ ts under environ entally realisti onditions in luding a realisti on entration o the test su stan e. ither o plete inerali ation or pri ary degrada ility ay e deter ined depending on the analyti al ethod e ployed or deter ining the trans or ation o the test su stan e.
et o s fo esti atin io e a a ilit
. . n re ent years, possi ilities or esti ating environ ental properties o su stan es have een developed and, a ong these, also ethods or predi ting the iodegrada ility potential o organi su stan es e.g. the yra use esear h orporation s iodegrada ility ro a ility rogra , ). eviews o ethods have een per or ed y ) and y angen erg et al. ). hey show that group ontri ution ethods see to e the ost su ess ul ethods. these, the iodegradation ro a ility rogra ) see s to have the roadest appli ation. t gives a ualitative esti ate o the pro a ility o slow or ast iodegradation in the presen e o a i ed population o environ ental i ro organis s. he appli a ility o this progra has een evaluated y the
- 495 -
Copyright@United Nations, 2017. All rights reserved
nne
ctor in encin e r i it in t e tic en iron ent
ntro ction
1.1 The classification criteria are considering the ha ards to the a uatic en ironment only. owe er, the ha ard classification is primarily based on data prepared by conduction of tests under laboratory conditions that only seldom are similar to the conditions in the en ironment. Thus, the interpretation of laboratory test data for prediction of the ha ards in the a uatic en ironment should be considered.
1.2 Interpretation of test results on biodegradability of organic substances has been considered in the etailed e iew aper on iodegradability Testing ( , 1 5).
1. The conditions in the en ironment are typically ery different from the conditions in the standardi ed test systems, which ma e the e trapolation of degradation data from laboratory tests to the en ironment difficult. mong the differences, the following ha e significant influence on the degradability
(a) rganism related factors (presence of competent micro organisms)
(b) ubstrate related factors (concentration of the substance and presence of other substrates) and
(c) n ironment related factors (physico chemical conditions, presence of nutrients, bioa ailability of the substance).
These aspects will be discussed further below.
re ence o co petent icro or ni
2.1 iodegradation in the a uatic en ironment is dependent on the presence of competent micro organisms in sufficient numbers. The natural microbial communities consist of a ery di erse biomass and when a new substance is introduced in a sufficiently high concentration, the biomass may be adapted to degrade this substance. re uently, the adaptation of the microbial population is caused by the growth of specific degraders that by nature are competent to degrade the substance. owe er, also other processes as en yme induction, e change of genetic material and de elopment of tolerance to to icity may be in ol ed.
2.2 Adaptation takes place during a “lag” phase, which is the time period from the onset of the e posure until a significant degradation begins. It seems ob ious that the length of the lag phase will depend on the initial presence of competent degraders. This will again depend on the history of the microbial community, i.e. whether the community formerly has been e posed to the substance. This means that when a enobiotic substance has been used and emitted ubi uitously in a number of years, the li elihood of finding competent degraders will increase. This will especially be the case in en ironments recei ing emissions as e.g. biological wastewater treatment plants. ften more consistent degradation results are found in tests where inocula from polluted waters are used compared to tests with inocula from unpolluted water ( , 1 5 yholm and Ingersle , 1 ).
2. number of factors determine whether the potential for adaptation in the a uatic en ironment is comparable with the potential in laboratory tests. mong other things adaptation depends on
(a) initial number of competent degraders in the biomass (fraction and number)
(b) presence of surfaces for attachment
(c) concentration and a ailability of substrate and
(d) presence of other substrates.
2.4 The length of the lag phase depends on the initial number of competent degraders and, for to ic substances, the sur i al and reco ery of these. In standard ready biodegradability tests, the inoculum is sampled in sewage treatment plants. s the load with pollutants is normally higher than in the en ironment, both the fraction and 4 - 497 - Copyright@United Nations, 2017. All rights reserved
the num er o competent degraders ma e higher than in the less polluted a uatic en ironment t is, howe er, di icult esence of ot e s st ates to estimate how much longer the lag phase will e in the a uatic en ironment than in a la orator test due to the likel lower initial num er o competent degraders n the standard tests, the test su stance is applied as the sole su strate or the micro organisms while in the en ironment, a large num er o other su strates are present n natural waters, concentrations o dissol ed organic er long periods o time, the initial concentration o competent degraders is not important as the car on are o ten ound in the range mg l, i e up to a actor higher than a pollutant owe er, much o this will grow up when a suita le su strate is present in su icient concentrations owe er, i the degrada ilit in a short organic car on is relati el persistent with an increasing raction o persistent matter the longer the distance rom the period o time is o concern, the initial concentration o competent degrading micro organisms should e considered shore cow, acteria in natural waters are primaril nourishing on e udates rom algae hese e udates are he presence o locs, aggregates and attached micro organisms ma also enhance adaptation e g minerali ed er uickl within minutes demonstrating that there is a high degradation potential in the natural micro de elopment o micro ial niches with consortia o micro organisms his is o importance when considering the organism communities hus, as micro organisms compete or the ariet o su strates in natural waters, there is a capa ilit o adaptation in the di erse en ironments in sewage treatment plants or in sediment or soil owe er, the selection pressure among micro organisms resulting in growth o opportunistic species capa le o nourishing on uickl total num er o micro organisms in read iodegrada ilit tests and in the a uatic en ironment are o the same orders minerali ed su strates, while growth o more speciali ed species is suppressed periences rom isolation o acteria o magnitude cells ml in read iodegrada ilit tests and cells ml or more in sur ace water cow, capa le o degrading arious eno iotics ha e demonstrated that these organisms are o ten growing relati el slowl hus, this actor is pro a l o minor importance and sur i e on comple car on sources in competition with more rapidl growing acteria hen competent micro organisms are present in the en ironment, their num ers ma increase i the speci ic eno iotic su strate is hen discussing the e trapolation to en ironmental conditions it ma e alua le to discriminate continuousl released and reach a concentration in the en ironment su icient to support growth owe er, most o the etween oligotrophic and eutrophic en ironments icro organisms thri ing under oligotrophic conditions are a le to organic pollutants in the a uatic en ironment are present in low concentrations and will onl e degraded as secondar minerali e organic su strates at low concentrations ractions o mg , and the normall ha e a greater a init or su strates not supporting growth the su strate ut lower growth rates and higher generation times than eutrophic organisms , oreo er, oligotrophs are una le to degrade chemicals in concentrations higher than mg l and ma e en e inhi ited at high n the other hand, the presence o uickl minerali ed su strates in higher concentrations ma concentrations pposite to that, eutrophs re uire higher su strate concentrations e ore minerali ation egins and the acilitate an initial trans ormation o the eno iotic molecule co meta olism he co meta oli ed su stance ma thri e at higher concentrations than oligotrophs hus, the lower threshold limit or degradation in the a uatic then e a aila le or urther degradation and minerali ation hus, the presence o other su strates ma increase the en ironment will depend on whether the micro ial population is an oligotroph or an eutroph population t is, howe er, possi ilities or a su stance to e degraded not clear whether oligotrophs and eutrophs are di erent species or whether there is onl an oligotrophic and an eutrophic wa o li e , ost pollutants reach the a uatic en ironment directl through discharge o t ma then e concluded that the presence o a ariet o su strates in natural waters and among them wastewater and conse uentl , these recipients are mostl eutrophic uickl minerali ed su strates, ma on the one hand cause a selection pressure suppressing growth o micro organisms competent o degrading micro pollutants n the other hand it ma acilitate an increased degradation an initial co rom the a o e discussion it ma thus e concluded that the chance o presence o competent meta olism ollowed a urther minerali ation he relati e importance o these processes under natural conditions degraders is greatest in highl e posed en ironments, i e in en ironments continuousl recei ing su stances which ma ar depending on oth the en ironmental conditions and the su stance and no generali ation can et e more re uentl occurs or high production olume chemicals than or low production olume chemicals hese esta lished en ironments are o ten eutrophic and there ore, the degradation ma re uire relati el high concentrations o su stances e ore onset n the other hand, in pristine waters competent species ma e lacking, especiall species n iron ent re te ctor capa le o degradation o chemicals onl occasionall released as low production olume chemicals he en ironmental aria les control the general micro ial acti it rather than speci ic degradation tr te re te ctor processes owe er, the signi icance o the in luence aries etween di erent ecos stems and micro ial species cow, Concent ation of test s stance e o potential n most la orator tests, the test su stance is applied in er high concentrations mg l compared to the concentrations in the lower g l range that ma e e pected in the a uatic en ironment n general, ne o the most important en ironment related actors in luencing the degrada ilit is pro a l the growth o micro organisms is not supported when a su strate is present in concentrations elow a threshold le el o presence o o gen he o gen content and the related redo potential determines the presence o di erent t pes o around g l and at lower concentrations, e en the energ re uirement or maintenance is not met , micro organisms in a uatic en ironments with aero ic organisms present in the water phase, in the upper la er o he reason or this lower threshold le el is possi l a lack o su icient stimulus to initiate an en matic response sediments and in parts o sewage treatment plants, and anaero ic organisms present in sediments and parts o sewage cow, his means in general that the concentrations o man su stances in the a uatic en ironment are at a treatment plants n most parts o the water phase, aero ic conditions are pre ailing and the prediction o the le el where the can onl hardl e the primar su strate or degrading micro organisms iodegrada ilit should e ased on results rom aero ic tests owe er, in some a uatic en ironments the o gen content ma e er low in periods o the ear due to eutrophication and the ollowing deca o produced organic oreo er, the degradation kinetics depends on su stance concentration compared with the matter n these periods, aero ic organisms will not e a le to degrade the chemical, ut anaero ic processes ma take saturation constant s as descri ed in the onod e uation he saturation constant is the concentration o the su strate o er i the chemical is degrada le under anaero ic conditions resulting in a speci ic growth rate o o the ma imum speci ic growth rate At su strate concentrations much lower than the saturation constant, which is the normal situation in most o the a uatic en ironment, the degradation can e e pe at e descri ed irst order or logistic kinetics , hen a low densit o micro organisms lower than cells ml pre ails e g in oligotrophic waters , the population grows at e er decreasing rates which is t pical o logistic Another important parameter is the temperature ost la orator tests are per ormed at kinetics At a higher densit o micro organisms e g in eutrophic waters , the su strate concentration is not high standard aero ic read iodegrada ilit tests , ut anaero ic tests ma e per ormed at as this etter mimics the enough to support growth o the cells and irst order kinetics appl , i e the degradation rate is proportional with the conditions in a sludge reactor icro ial acti it is ound in the en ironment at temperatures ranging rom elow su stance concentration n practice, it ma e impossi le to distinguish etween the two t pes o degradation kinetics to owe er, optimum temperatures are pro a l in the range rom to and roughl , the degradation due to uncertaint o the data , rate dou les or e er increase o temperature in this range de enau, utside this optimum range the acti it o the degraders is reduced drasticall although some speciali ed species termo and ps crophilic acteria n conclusion, su stances in low concentrations i e elow g l are pro a l not degraded as ma thri e hen e trapolating rom la orator conditions, it should e considered that some a uatic en ironments are primar su strates in the a uatic en ironment At higher concentrations, readil degrada le su stances will pro a l e co ered ice in su stantial periods o the ear and that onl minor or e en no degradation can e e pected during the degraded as primar su strates in the en ironment at a degradation rate more or less proportional with the concentration winter season o the su stance he degradation o su stances as secondar su strates is discussed elow
- 498 - Copyright@United Nations, 2017. All rights reserved
the num er o competent degraders ma e higher than in the less polluted a uatic en ironment t is, howe er, di icult esence of ot e s st ates to estimate how much longer the lag phase will e in the a uatic en ironment than in a la orator test due to the likel lower initial num er o competent degraders n the standard tests, the test su stance is applied as the sole su strate or the micro organisms while in the en ironment, a large num er o other su strates are present n natural waters, concentrations o dissol ed organic er long periods o time, the initial concentration o competent degraders is not important as the car on are o ten ound in the range mg l, i e up to a actor higher than a pollutant owe er, much o this will grow up when a suita le su strate is present in su icient concentrations owe er, i the degrada ilit in a short organic car on is relati el persistent with an increasing raction o persistent matter the longer the distance rom the period o time is o concern, the initial concentration o competent degrading micro organisms should e considered shore cow, acteria in natural waters are primaril nourishing on e udates rom algae hese e udates are he presence o locs, aggregates and attached micro organisms ma also enhance adaptation e g minerali ed er uickl within minutes demonstrating that there is a high degradation potential in the natural micro de elopment o micro ial niches with consortia o micro organisms his is o importance when considering the organism communities hus, as micro organisms compete or the ariet o su strates in natural waters, there is a capa ilit o adaptation in the di erse en ironments in sewage treatment plants or in sediment or soil owe er, the selection pressure among micro organisms resulting in growth o opportunistic species capa le o nourishing on uickl total num er o micro organisms in read iodegrada ilit tests and in the a uatic en ironment are o the same orders minerali ed su strates, while growth o more speciali ed species is suppressed periences rom isolation o acteria o magnitude cells ml in read iodegrada ilit tests and cells ml or more in sur ace water cow, capa le o degrading arious eno iotics ha e demonstrated that these organisms are o ten growing relati el slowl hus, this actor is pro a l o minor importance and sur i e on comple car on sources in competition with more rapidl growing acteria hen competent micro organisms are present in the en ironment, their num ers ma increase i the speci ic eno iotic su strate is hen discussing the e trapolation to en ironmental conditions it ma e alua le to discriminate continuousl released and reach a concentration in the en ironment su icient to support growth owe er, most o the etween oligotrophic and eutrophic en ironments icro organisms thri ing under oligotrophic conditions are a le to organic pollutants in the a uatic en ironment are present in low concentrations and will onl e degraded as secondar minerali e organic su strates at low concentrations ractions o mg , and the normall ha e a greater a init or su strates not supporting growth the su strate ut lower growth rates and higher generation times than eutrophic organisms , oreo er, oligotrophs are una le to degrade chemicals in concentrations higher than mg l and ma e en e inhi ited at high n the other hand, the presence o uickl minerali ed su strates in higher concentrations ma concentrations pposite to that, eutrophs re uire higher su strate concentrations e ore minerali ation egins and the acilitate an initial trans ormation o the eno iotic molecule co meta olism he co meta oli ed su stance ma thri e at higher concentrations than oligotrophs hus, the lower threshold limit or degradation in the a uatic then e a aila le or urther degradation and minerali ation hus, the presence o other su strates ma increase the en ironment will depend on whether the micro ial population is an oligotroph or an eutroph population t is, howe er, possi ilities or a su stance to e degraded not clear whether oligotrophs and eutrophs are di erent species or whether there is onl an oligotrophic and an eutrophic wa o li e , ost pollutants reach the a uatic en ironment directl through discharge o t ma then e concluded that the presence o a ariet o su strates in natural waters and among them wastewater and conse uentl , these recipients are mostl eutrophic uickl minerali ed su strates, ma on the one hand cause a selection pressure suppressing growth o micro organisms competent o degrading micro pollutants n the other hand it ma acilitate an increased degradation an initial co rom the a o e discussion it ma thus e concluded that the chance o presence o competent meta olism ollowed a urther minerali ation he relati e importance o these processes under natural conditions degraders is greatest in highl e posed en ironments, i e in en ironments continuousl recei ing su stances which ma ar depending on oth the en ironmental conditions and the su stance and no generali ation can et e more re uentl occurs or high production olume chemicals than or low production olume chemicals hese esta lished en ironments are o ten eutrophic and there ore, the degradation ma re uire relati el high concentrations o su stances e ore onset n the other hand, in pristine waters competent species ma e lacking, especiall species n iron ent re te ctor capa le o degradation o chemicals onl occasionall released as low production olume chemicals he en ironmental aria les control the general micro ial acti it rather than speci ic degradation tr te re te ctor processes owe er, the signi icance o the in luence aries etween di erent ecos stems and micro ial species cow, Concent ation of test s stance e o potential n most la orator tests, the test su stance is applied in er high concentrations mg l compared to the concentrations in the lower g l range that ma e e pected in the a uatic en ironment n general, ne o the most important en ironment related actors in luencing the degrada ilit is pro a l the growth o micro organisms is not supported when a su strate is present in concentrations elow a threshold le el o presence o o gen he o gen content and the related redo potential determines the presence o di erent t pes o around g l and at lower concentrations, e en the energ re uirement or maintenance is not met , micro organisms in a uatic en ironments with aero ic organisms present in the water phase, in the upper la er o he reason or this lower threshold le el is possi l a lack o su icient stimulus to initiate an en matic response sediments and in parts o sewage treatment plants, and anaero ic organisms present in sediments and parts o sewage cow, his means in general that the concentrations o man su stances in the a uatic en ironment are at a treatment plants n most parts o the water phase, aero ic conditions are pre ailing and the prediction o the le el where the can onl hardl e the primar su strate or degrading micro organisms iodegrada ilit should e ased on results rom aero ic tests owe er, in some a uatic en ironments the o gen content ma e er low in periods o the ear due to eutrophication and the ollowing deca o produced organic oreo er, the degradation kinetics depends on su stance concentration compared with the matter n these periods, aero ic organisms will not e a le to degrade the chemical, ut anaero ic processes ma take saturation constant s as descri ed in the onod e uation he saturation constant is the concentration o the su strate o er i the chemical is degrada le under anaero ic conditions resulting in a speci ic growth rate o o the ma imum speci ic growth rate At su strate concentrations much lower than the saturation constant, which is the normal situation in most o the a uatic en ironment, the degradation can e e pe at e descri ed irst order or logistic kinetics , hen a low densit o micro organisms lower than cells ml pre ails e g in oligotrophic waters , the population grows at e er decreasing rates which is t pical o logistic Another important parameter is the temperature ost la orator tests are per ormed at kinetics At a higher densit o micro organisms e g in eutrophic waters , the su strate concentration is not high standard aero ic read iodegrada ilit tests , ut anaero ic tests ma e per ormed at as this etter mimics the enough to support growth o the cells and irst order kinetics appl , i e the degradation rate is proportional with the conditions in a sludge reactor icro ial acti it is ound in the en ironment at temperatures ranging rom elow su stance concentration n practice, it ma e impossi le to distinguish etween the two t pes o degradation kinetics to owe er, optimum temperatures are pro a l in the range rom to and roughl , the degradation due to uncertaint o the data , rate dou les or e er increase o temperature in this range de enau, utside this optimum range the acti it o the degraders is reduced drasticall although some speciali ed species termo and ps crophilic acteria n conclusion, su stances in low concentrations i e elow g l are pro a l not degraded as ma thri e hen e trapolating rom la orator conditions, it should e considered that some a uatic en ironments are primar su strates in the a uatic en ironment At higher concentrations, readil degrada le su stances will pro a l e co ered ice in su stantial periods o the ear and that onl minor or e en no degradation can e e pected during the degraded as primar su strates in the en ironment at a degradation rate more or less proportional with the concentration winter season o the su stance he degradation o su stances as secondar su strates is discussed elow
- 499 - Copyright@United Nations, 2017. All rights reserved
p nne
Acti e micro organisms are ound in the entire p range ound in the en ironment owe er, or acteria as a group, slightl alkaline conditions a our the acti it and the optimum p range is At a p lower than , the meta olic acti it in acteria is signi icantl decreased or ungi as a group, slightl acidic conditions a our the ic princip e o t e e peri ent n e ti tion et o or acti it with an optimum p range o cow, hus, an optimum or the degrading acti it o micro eter in tion o n o o or nic t nce organisms will pro a l e within the p range o , which is the range most o ten pre ailing in the a uatic en ironment ioconcentr tion ctor esence of n t ients efinition he presence o inorganic nutrients nitrogen and phosphorus is o ten re uired or micro ial growth owe er, these are onl seldom the acti it limiting actors in the a uatic en ironment where growth o micro he ioconcentration actor is de ined as the ratio etween the concentration o the chemical in iota organisms is o ten su strate limited owe er, the presence o nutrient in luences the growth o primar producers and and the concentration in the surrounding medium, here water, at stead state can e measured e perimentall then again the a aila ilit o readil minerali ed e udates directl under stead state conditions or calculated the ratio o the irst order uptake and elimination rate constants, a method that does not re uire e uili rium conditions
pp op iate et o s fo e pe i ental ete ination of C
i erent test guidelines or the e perimental determination o ioconcentration in ish ha e een documented and adopted the most generall applied eing the test guideline , and the A standard guide A was re ised and replaced the pre ious ersion A , Although low through test regimes are pre erred , , semi static regimes are allowed A , pro ided that the alidit criteria on mortalit and maintenance o test conditions are ul illed or lipophilic su stances log ow , low through methods are pre erred
he principles o the and the A guidelines are similar, ut the e perimental conditions descri ed are di erent, especiall concerning
a method o test water suppl static, semi static or low through
the re uirement or carr ing out a depuration stud
c the mathematical method or calculating
d sampling re uenc um er o measurements in water and num er o samples o ish
e re uirement or measuring the lipid content o the ish
the minimum duration o the uptake phase
n general, the test consists o two phases he e posure uptake and post e posure depuration phases uring the uptake phase, separate groups o ish o one species are e posed to at least two concentrations o the test su stance A da e posure phase is o ligator unless a stead state has een reached within this period he time needed or reaching stead state conditions ma e set on the asis o ow – correlations e g log – log ow pacie and amelink, or log – log ow o as et al., he e pected time d or e g stead state ma thus e calculated ln , pro ided that the ioconcentration ollows irst order kinetics uring the depuration phase the ish are trans erred to a medium ree o the test su stance he concentration o the test su stance in the ish is ollowed through oth phases o the test he is e pressed as a unction o the total wet weight o the ish As or man organic su stances, there is a signi icant relationship etween the potential or ioconcentration and the lipophilicit , and urthermore, there is a corresponding relationship etween the lipid content o the test ish and the o ser ed ioconcentration o such su stances here ore, to reduce this source o aria ilit in the test results or the su stances with high lipophilicit , ioconcentration should e e pressed in relation to the lipid content in addition to whole od weight , he guidelines mentioned are ased on the assumption that ioconcentration ma e appro imated a irst order process one compartment model and thus that irst order uptake rate, irst order depuration rate, descri ed a log linear appro imation the depuration ollows iphasic kinetics, i e two distinct depuration rates can e identi ied, the appro imation ma signi icantl underestimate a second order kinetic has een indicated, ma e estimated rom the relation ish ater, provided that “steady state” for the fish water s stem has een reached
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p nne
Acti e micro organisms are ound in the entire p range ound in the en ironment owe er, or acteria as a group, slightl alkaline conditions a our the acti it and the optimum p range is At a p lower than , the meta olic acti it in acteria is signi icantl decreased or ungi as a group, slightl acidic conditions a our the ic princip e o t e e peri ent n e ti tion et o or acti it with an optimum p range o cow, hus, an optimum or the degrading acti it o micro eter in tion o n o o or nic t nce organisms will pro a l e within the p range o , which is the range most o ten pre ailing in the a uatic en ironment ioconcentr tion ctor esence of n t ients efinition he presence o inorganic nutrients nitrogen and phosphorus is o ten re uired or micro ial growth owe er, these are onl seldom the acti it limiting actors in the a uatic en ironment where growth o micro he ioconcentration actor is de ined as the ratio etween the concentration o the chemical in iota organisms is o ten su strate limited owe er, the presence o nutrient in luences the growth o primar producers and and the concentration in the surrounding medium, here water, at stead state can e measured e perimentall then again the a aila ilit o readil minerali ed e udates directl under stead state conditions or calculated the ratio o the irst order uptake and elimination rate constants, a method that does not re uire e uili rium conditions
pp op iate et o s fo e pe i ental ete ination of C
i erent test guidelines or the e perimental determination o ioconcentration in ish ha e een documented and adopted the most generall applied eing the test guideline , and the A standard guide A was re ised and replaced the pre ious ersion A , Although low through test regimes are pre erred , , semi static regimes are allowed A , pro ided that the alidit criteria on mortalit and maintenance o test conditions are ul illed or lipophilic su stances log ow , low through methods are pre erred
he principles o the and the A guidelines are similar, ut the e perimental conditions descri ed are di erent, especiall concerning
a method o test water suppl static, semi static or low through
the re uirement or carr ing out a depuration stud
c the mathematical method or calculating
d sampling re uenc um er o measurements in water and num er o samples o ish
e re uirement or measuring the lipid content o the ish
the minimum duration o the uptake phase
n general, the test consists o two phases he e posure uptake and post e posure depuration phases uring the uptake phase, separate groups o ish o one species are e posed to at least two concentrations o the test su stance A da e posure phase is o ligator unless a stead state has een reached within this period he time needed or reaching stead state conditions ma e set on the asis o ow – correlations e g log – log ow pacie and amelink, or log – log ow o as et al., he e pected time d or e g stead state ma thus e calculated ln , pro ided that the ioconcentration ollows irst order kinetics uring the depuration phase the ish are trans erred to a medium ree o the test su stance he concentration o the test su stance in the ish is ollowed through oth phases o the test he is e pressed as a unction o the total wet weight o the ish As or man organic su stances, there is a signi icant relationship etween the potential or ioconcentration and the lipophilicit , and urthermore, there is a corresponding relationship etween the lipid content o the test ish and the o ser ed ioconcentration o such su stances here ore, to reduce this source o aria ilit in the test results or the su stances with high lipophilicit , ioconcentration should e e pressed in relation to the lipid content in addition to whole od weight , he guidelines mentioned are ased on the assumption that ioconcentration ma e appro imated a irst order process one compartment model and thus that irst order uptake rate, irst order depuration rate, descri ed a log linear appro imation the depuration ollows iphasic kinetics, i e two distinct depuration rates can e identi ied, the appro imation ma signi icantl underestimate a second order kinetic has een indicated, ma e estimated rom the relation ish ater, provided that “steady state” for the fish water s stem has een reached
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o ether ith detai s of sa p e preparatio a d stora e, a appropriate a a yti a ethod of o s rfa e a tive ateria s, or s sta es that rea t ith the e e t he ethod is app i a e he the o o a ra y, pre isio , a d se sitivity st e avai a e for the a tifi atio of the s sta e i the test so tio a d i va e fa s ithi the ra e to , he ethod is ess se sitive to the prese e of i p rities the io o i a ateria f these are a i it is i possi e to deter i e a tr e he se of radio a e ed test i the test o po d o pared to the sha e f as ethod s sta e a fa i itate the a a ysis of ater a d fish sa p es o ever, ess o i ed ith a spe ifi a a yti a ethod, the tota radioa tivity eas re e ts pote tia y ref e t the prese e of pare t s sta e, possi e eta o ite s , low stirring method a d possi e eta o i ed ar o , hi h have ee i orporated i the fish tiss e i or a i o e es or the deter i atio of a tr e it is esse tia to ear y dis ri i ate the pare t s sta e fro possi e eta o ites f ith the s o stirri ethod a pre ise a d a rate deter i atio of o of o po ds ith o o radio a e ed ateria s are sed i the test, it is possi e to a a yse for tota radio a e i e pare t a d eta o ites or p ti is a o ed e r i et al , or hi h y ipophi i o po ds the sha e f as ethod is pro e to the sa p es ay e p rified so that the pare t o po d a e a a ysed separate y prod e artefa ts for atio of i rodrop ets , a d ith the ethod o eeds to e e trapo ated eyo d the a i ratio ra e to o tai esti ates of o the o o ra e a ove , the eas red data te d to de rease ith i reasi o o o ept a e p a atio s of o i earity ai y refer to either iotra sfor atio , red ed e ra e per eatio order to deter i e a partitio oeffi ie t, ater, o ta o , a d test o po d are e i i rated ith i eti s or red ed ioti ipid so i ity for ar e o e es ther fa tors o sider e peri e ta artefa ts, s h as ea h other after hi h the o e tratio of the test o po d i the t o phases is deter i ed he e peri e ta e i i ri ot ei rea hed, red ed ioavai a i ity d e to sorptio to or a i atter i the a eo s phase, a d diffi ties asso iated ith the for atio of i rodrop ets d ri the sha e f as e peri e t a to so e de ree e a a yti a errors oreover, are sho d e ta e he eva ati e peri e ta data o for s sta es ith o over o e i the s o stirri e peri e t as ater, o ta o , a d the test o po d are e i i rated i a e t y stirred o a ove , as these data i have a h hi her eve of ertai ty tha va es deter i ed for s sta es ith rea tor he stirri reates a ore or ess a i ar f o et ee the o ta o a d the ater, a d e ha e et ee the o o e o phases is e ha ed itho t i rodrop ets ei for ed
o o Generator column method
efinition an ene al consi e ations other very versati e ethod for eas ri o o is the e erator o ethod this ethod, a e erator o ethod is sed to partitio the test s sta e et ee the o ta o a d ater phases he o is he o n o ta o ater partitio oeffi ie t o o is a eas re of the ipophi i ity of a s sta e pa ed ith a so id s pport a d is sat rated ith a fi ed o e tratio of the test s sta e i n o ta o he test s s h, o o is a ey para eter i the assess e t of e viro e ta fate a y distri tio pro esses are drive y s sta e is e ted fro the o ta o sat rated e erator o ith ater he a eo s so tio e iti the o o o , e sorptio to soi a d sedi e t a d io o e tratio i or a is s represe ts the e i i ri o e tratio of the test s sta e that has partitio ed fro the o ta o phase i to the ater phase he pri ary adva ta e of the e erator o ethod over the sha e f as ethod is that the for er he asis for the re atio ship et ee io o e tratio a d o o is the a a o y for the partitio o p ete y avoids the for atio of i ro e sio s herefore, this ethod is parti ar y sef for eas ri o for pro ess et ee the ipid phase of fish a d ater a d the partitio pro ess et ee o ta o a d ater he reaso for s sta es va es over o ette a d dre , a d hi et al., as e as for s sta es havi si o arises fro the a i ity of o ta o to a t as a satisfa tory s rro ate for ipids i fish tiss e i h y si ifi a t o o va es ess tha disadva ta e of the e erator o ethod is that it re ires sophisti ated e ip e t re atio ships et ee o o a d the so i ity of s sta es i od iver oi a d trio i e ist ii i, rio i is detai ed des riptio of the e erator olumn method is presented in the “Toxic Substances Control Act Test o e of the ost a da t tria y y ero s fo d i fresh ater fish ipids e derso a d o her, Guidelines” (USEPA 1985).
he deter i atio of the n o ta o ater partitio oeffi ie t o is a re ire e t of the ase data se of s fo ete ination of lo o (see also in A9.6 se of A s set to e s itted for otified e a d priority e isti s sta es ithi the s the e peri e ta deter i atio of the o is ot a ays possi e, e for very ater so e a d for very ipophi i s sta es, a derived o ero s s have ee a d o ti e to e deve oped for the esti atio of o o o y sed ay e sed o ever, e tre e a tio sho d e e er i ed he si s for s sta es here the e peri e ta ethods are ased o fra e t o sta ts he fra e ta approa hes are ased o a si p e additio of the ipophi i ity deter i atio is ot possi e as for e s rfa ta ts of the i divid a o e ar fra e ts of a ive o e e hree o er ia y avai a e pro ra s are re o e ded in the European Commission’s Technical Guidance Document (European Commission, 1996) for risk
pp op iate et o s fo e pe i ental ete ination of o al es assess e t, part , if o e peri e ta y derived data are avai a e
or e peri e ta deter i atio of o va es, t o differe t ethods, ha e f as a d , have ay i ht he i a for atio yste s, as i itia y deve oped for se i dr ee des ri ed i sta dard ide i es e desi he ode is ased o the a s h a d eo a atio pro ed re a s h a d eo, he pro ra ot o y data o tai ed y the e p oy e t of the sha e f as or the ethod a ates o o for or a i o po ds o tai i , , , , a , , a d or o o for sa ts a d for o po ds a ordi to sta dard ide i es are re o e ded or hi h y ipophi i s sta es, hi h are s o y so e i ater, ith for a har es a ot e a ated e ept for itro o po ds a d itro e o ides he a atio res ts of data o tai ed y e p oyi a s o stirri ethod are e era y ore re ia e e r i et al , o s a d i , o o for io i a e s sta es, i e phe o s, a i es, a d ar o y i a ids, represe t the e tra or io i ed for a d ide i e i e p depe de t e era , the pro ra res ts i ear esti ates i the ra e of o o et ee a d ropea o issio , , part o ever a va idatio st dy perfor ed y ie e , ho o pared ha e flas method e peri e ta deter i ed o o va es ith esti ated va es, sho ed that the pro ra pre ise y predi ts the o o for a reat er of or a i he i a s i the o o ra e fro e o to a ove , r a si i ar va idatio st dy o ore tha s sta es the res ts ith the pro ra versio , versio he asi pri ip e of the ethod is to eas re the disso tio of the s sta e i t o differe t phases, ater a d n o ta o order to deter i e the partitio oeffi ie t, e i i ri et ee a i tera ti ere r , s d , hese va idatio s sho that the pro ra ay e sed for esti ati o po e ts of the syste st e a hieved after hi h the o e tratio of the s sta es disso ved i the t o phases re ia e o o va es he o e peri e ta data are avai a e or he ati o po ds a d s rfa ta ts the pro ra is stated to e of i ited re ia i ity , o ever, as re ards a io i s rfa ta ts a orre tio is deter i ed he sha e f as ethod is app i a e he the o o va e fa s ithi the ra e fro to , he sha e f as ethod app ies o y to esse tia p re s sta es so e i ater a d n o ta o a d sho d ethod for esti ati ad sted va es has ee proposed o erts, e perfor ed at a o sta t te perat re i the ra e or yra se esear h orporatio ses str t ra fra e ts a d orre tio LC method fa tors he pro ra a ates o o for or a i o po ds o tai i the fo o i ato s , , , , a , i, , e, i, a, , a d or o o for o po ds ith for a har es i e itro e o ides a d itro o po ds a a so e a ated he a atio of o for io i a e s sta es, i e phe o s, a i es a d ar o y i a ids, is perfor ed o a a yti a o s pa ed ith a o er ia y avai a e so id phase o represe t the e tra or io i ed for , a d the va es i th s e p depe de t o e s rfa ta ts e a oho o tai i o hydro ar o hai s e , he i a y o d o to si i a he i a s i e ted o to s h a o etho y ates o s, , dyest ffs, a d disso iated s sta es ay e predi ted y the pro ra ederse ove a o at differe t rates e a se of the differe t de rees of partitio i et ee the o i e a eo s phase a d the et al, e era , the pro ra ives ear esti ates i the ra e of o et ee a d e a ord statio ary hydro ar o phase he ethod is ot app i a e to stro a ids a d ases, eta s o p e es, o
- 502 - Copyright@United Nations, 2017. All rights reserved
o ether ith detai s of sa p e preparatio a d stora e, a appropriate a a yti a ethod of o s rfa e a tive ateria s, or s sta es that rea t ith the e e t he ethod is app i a e he the o o a ra y, pre isio , a d se sitivity st e avai a e for the a tifi atio of the s sta e i the test so tio a d i va e fa s ithi the ra e to , he ethod is ess se sitive to the prese e of i p rities the io o i a ateria f these are a i it is i possi e to deter i e a tr e he se of radio a e ed test i the test o po d o pared to the sha e f as ethod s sta e a fa i itate the a a ysis of ater a d fish sa p es o ever, ess o i ed ith a spe ifi a a yti a ethod, the tota radioa tivity eas re e ts pote tia y ref e t the prese e of pare t s sta e, possi e eta o ite s , low stirring method a d possi e eta o i ed ar o , hi h have ee i orporated i the fish tiss e i or a i o e es or the deter i atio of a tr e it is esse tia to ear y dis ri i ate the pare t s sta e fro possi e eta o ites f ith the s o stirri ethod a pre ise a d a rate deter i atio of o of o po ds ith o o radio a e ed ateria s are sed i the test, it is possi e to a a yse for tota radio a e i e pare t a d eta o ites or p ti is a o ed e r i et al , or hi h y ipophi i o po ds the sha e f as ethod is pro e to the sa p es ay e p rified so that the pare t o po d a e a a ysed separate y prod e artefa ts for atio of i rodrop ets , a d ith the ethod o eeds to e e trapo ated eyo d the a i ratio ra e to o tai esti ates of o the o o ra e a ove , the eas red data te d to de rease ith i reasi o o o ept a e p a atio s of o i earity ai y refer to either iotra sfor atio , red ed e ra e per eatio order to deter i e a partitio oeffi ie t, ater, o ta o , a d test o po d are e i i rated ith i eti s or red ed ioti ipid so i ity for ar e o e es ther fa tors o sider e peri e ta artefa ts, s h as ea h other after hi h the o e tratio of the test o po d i the t o phases is deter i ed he e peri e ta e i i ri ot ei rea hed, red ed ioavai a i ity d e to sorptio to or a i atter i the a eo s phase, a d diffi ties asso iated ith the for atio of i rodrop ets d ri the sha e f as e peri e t a to so e de ree e a a yti a errors oreover, are sho d e ta e he eva ati e peri e ta data o for s sta es ith o over o e i the s o stirri e peri e t as ater, o ta o , a d the test o po d are e i i rated i a e t y stirred o a ove , as these data i have a h hi her eve of ertai ty tha va es deter i ed for s sta es ith rea tor he stirri reates a ore or ess a i ar f o et ee the o ta o a d the ater, a d e ha e et ee the o o e o phases is e ha ed itho t i rodrop ets ei for ed
o o Generator column method
efinition an ene al consi e ations other very versati e ethod for eas ri o o is the e erator o ethod this ethod, a e erator o ethod is sed to partitio the test s sta e et ee the o ta o a d ater phases he o is he o n o ta o ater partitio oeffi ie t o o is a eas re of the ipophi i ity of a s sta e pa ed ith a so id s pport a d is sat rated ith a fi ed o e tratio of the test s sta e i n o ta o he test s s h, o o is a ey para eter i the assess e t of e viro e ta fate a y distri tio pro esses are drive y s sta e is e ted fro the o ta o sat rated e erator o ith ater he a eo s so tio e iti the o o o , e sorptio to soi a d sedi e t a d io o e tratio i or a is s represe ts the e i i ri o e tratio of the test s sta e that has partitio ed fro the o ta o phase i to the ater phase he pri ary adva ta e of the e erator o ethod over the sha e f as ethod is that the for er he asis for the re atio ship et ee io o e tratio a d o o is the a a o y for the partitio o p ete y avoids the for atio of i ro e sio s herefore, this ethod is parti ar y sef for eas ri o for pro ess et ee the ipid phase of fish a d ater a d the partitio pro ess et ee o ta o a d ater he reaso for s sta es va es over o ette a d dre , a d hi et al., as e as for s sta es havi si o arises fro the a i ity of o ta o to a t as a satisfa tory s rro ate for ipids i fish tiss e i h y si ifi a t o o va es ess tha disadva ta e of the e erator o ethod is that it re ires sophisti ated e ip e t re atio ships et ee o o a d the so i ity of s sta es i od iver oi a d trio i e ist ii i, rio i is detai ed des riptio of the e erator olumn method is presented in the “Toxic Substances Control Act Test o e of the ost a da t tria y y ero s fo d i fresh ater fish ipids e derso a d o her, Guidelines” (USEPA 1985).
he deter i atio of the n o ta o ater partitio oeffi ie t o is a re ire e t of the ase data se of s fo ete ination of lo o (see also in A9.6 se of A s set to e s itted for otified e a d priority e isti s sta es ithi the s the e peri e ta deter i atio of the o is ot a ays possi e, e for very ater so e a d for very ipophi i s sta es, a derived o ero s s have ee a d o ti e to e deve oped for the esti atio of o o o y sed ay e sed o ever, e tre e a tio sho d e e er i ed he si s for s sta es here the e peri e ta ethods are ased o fra e t o sta ts he fra e ta approa hes are ased o a si p e additio of the ipophi i ity deter i atio is ot possi e as for e s rfa ta ts of the i divid a o e ar fra e ts of a ive o e e hree o er ia y avai a e pro ra s are re o e ded in the European Commission’s Technical Guidance Document (European Commission, 1996) for risk
pp op iate et o s fo e pe i ental ete ination of o al es assess e t, part , if o e peri e ta y derived data are avai a e
or e peri e ta deter i atio of o va es, t o differe t ethods, ha e f as a d , have ay i ht he i a for atio yste s, as i itia y deve oped for se i dr ee des ri ed i sta dard ide i es e desi he ode is ased o the a s h a d eo a atio pro ed re a s h a d eo, he pro ra ot o y data o tai ed y the e p oy e t of the sha e f as or the ethod a ates o o for or a i o po ds o tai i , , , , a , , a d or o o for sa ts a d for o po ds a ordi to sta dard ide i es are re o e ded or hi h y ipophi i s sta es, hi h are s o y so e i ater, ith for a har es a ot e a ated e ept for itro o po ds a d itro e o ides he a atio res ts of data o tai ed y e p oyi a s o stirri ethod are e era y ore re ia e e r i et al , o s a d i , o o for io i a e s sta es, i e phe o s, a i es, a d ar o y i a ids, represe t the e tra or io i ed for a d ide i e i e p depe de t e era , the pro ra res ts i ear esti ates i the ra e of o o et ee a d ropea o issio , , part o ever a va idatio st dy perfor ed y ie e , ho o pared ha e flas method e peri e ta deter i ed o o va es ith esti ated va es, sho ed that the pro ra pre ise y predi ts the o o for a reat er of or a i he i a s i the o o ra e fro e o to a ove , r a si i ar va idatio st dy o ore tha s sta es the res ts ith the pro ra versio , versio he asi pri ip e of the ethod is to eas re the disso tio of the s sta e i t o differe t phases, ater a d n o ta o order to deter i e the partitio oeffi ie t, e i i ri et ee a i tera ti ere r , s d , hese va idatio s sho that the pro ra ay e sed for esti ati o po e ts of the syste st e a hieved after hi h the o e tratio of the s sta es disso ved i the t o phases re ia e o o va es he o e peri e ta data are avai a e or he ati o po ds a d s rfa ta ts the pro ra is stated to e of i ited re ia i ity , o ever, as re ards a io i s rfa ta ts a orre tio is deter i ed he sha e f as ethod is app i a e he the o o va e fa s ithi the ra e fro to , he sha e f as ethod app ies o y to esse tia p re s sta es so e i ater a d n o ta o a d sho d ethod for esti ati ad sted va es has ee proposed o erts, e perfor ed at a o sta t te perat re i the ra e or yra se esear h orporatio ses str t ra fra e ts a d orre tio LC method fa tors he pro ra a ates o o for or a i o po ds o tai i the fo o i ato s , , , , a , i, , e, i, a, , a d or o o for o po ds ith for a har es i e itro e o ides a d itro o po ds a a so e a ated he a atio of o for io i a e s sta es, i e phe o s, a i es a d ar o y i a ids, is perfor ed o a a yti a o s pa ed ith a o er ia y avai a e so id phase o represe t the e tra or io i ed for , a d the va es i th s e p depe de t o e s rfa ta ts e a oho o tai i o hydro ar o hai s e , he i a y o d o to si i a he i a s i e ted o to s h a o etho y ates o s, , dyest ffs, a d disso iated s sta es ay e predi ted y the pro ra ederse ove a o at differe t rates e a se of the differe t de rees of partitio i et ee the o i e a eo s phase a d the et al, e era , the pro ra ives ear esti ates i the ra e of o et ee a d e a ord statio ary hydro ar o phase he ethod is ot app i a e to stro a ids a d ases, eta s o p e es, o
- 503 - Copyright@United Nations, 2017. All rights reserved
1995 581). ike the C GP pro ram, G has been alidated (Table ) and is recommended for classification nne purposes because of its reliabilit , commercial a ailabilit , and con enience of use. . . AUT GP (De illers et al., 1995) has been deri ed from a hetero eneous data set, comprisin 8 or anic chemicals collected from literature. The pro ram calculates lo o alues for or anic chemicals containin C, n ence o e tern n intern ctor on t e ioconcentr tion potenti o or nic t nce , , , al, P, and S. The lo o alues of salts cannot be calculated. Also the lo o of some compounds ith formal char es cannot be calculated, ith the exception of nitro compounds. The lo o alues of ioni able chemicals like phenols, amines, and corbox lic acids can be calculated althou h p dependencies should be noted. mpro ements ctor in encin t e pt e are in pro ress in order to extend the applicabilit of AUT GP. Accordin to the presentl a ailable information, AUT GP i es accurate alues especiall for hi hl lipophilic substances (lo o 5) (European Commission, The upt e r te or lipophili ompounds is m inly un tion o the si e o the or nism Si m nd 1996). inde . E tern l tors su h s the mole ul r si e tors in luen in the bio v il bility nd di erent environment l tors re o re t import n e to the upt e r te s ell. . .5 SPARC. The SPARC model is still under development by EPA’s Environmental esearch aborator in Athens, Geor ia, and is not et public a ailable. SPA C is a mechanistic model based on chemical thermod namic i e of o anis principles rather than a deterministic model rooted in kno led e obtained from obser ational data. Therefore, SPA C differs from models that use SA s (i.e. , GP) in that no measured lo o data are needed for a trainin Sin e l r er ish h ve rel tively lo er ill sur e to ei ht r tio lo er upt e r te onst nt is set of chemicals. EPA does occasionall run the model for a list of CAS numbers, if re uested. SPA C pro ides to be e pe ted or l r e ish omp red to sm ll ish Si m nd inde pperhui en nd Si m . The upt e impro ed results o er and C GP onl for compounds ith lo o alues reater than 5. nl SPA C o subst n es in ish is urther ontrolled by the ter lo throu h the ills the di usion throu h ueous di usion can be emplo ed in a eneral a for inor anic or or anometallic compounds. l yers t the ill epithelium the perme tion throu h the ill epithelium the r te o blood lo throu h the ills nd the bindin p ity o blood onstituents ECET C .
n Table 1, this Appendix, an o er ie of lo o estimation methods based on fra mentation olec la si e methodolo ies is presented. Also other methods for the estimation of lo o alues exist, but the should onl be used on a case b case basis and onl ith appropriate scientific ustification. oni ed subst n es do not re dily penetr te membr nes s ueous p n in luen e the subst n e upt e. oss o membr ne perme bility is e pe ted or subst n es ith onsider ble ross se tion l re e er ie o et o or e ti tion o o e on r ent tion et o o o ie o r o pperhui en et al. Anli er et al. or lon h in len th . nm pperhui en . oss o n e n membr ne perme bility due to the si e o the mole ules ill thus result in tot l loss o upt e. The e e t o mole ul r et o et o o o t ti tic ei ht on bio on entr tion is due to n in luen e on the di usion oe i ient o the subst n e hi h redu es the upt e r te onst nts ob s et al. . C GP ra ments correction Total n 89 , r ,91 , sd , 8 ansch and eo (19 9), factors alidation n 5 1, r ,96 aila ilit C GP Da li ht (1995) alidation n 1, r ,89, sd ,58 e ore subst n e is ble to bio on entr te in n or nism it needs to be present in ter nd G ( ) 1 fra ments Calibration n , r ,981, sd , 19, me ,161 v il ble or tr ns er ross ish ills. tors hi h e t this v il bility under both n tur l nd test onditions ill e lan and o ard (1995), 6 correction factors alidation n 8855, r ,95, sd , , me , lter the tu l bio on entr tion in omp rison to the estim ted v lue or C . As ish re ed durin bio on entr tion S C studies rel tively hi h on entr tions o dissolved nd p rti ul te or ni m tter m y be e pe ted thus redu in the r tion o hemi l th t is tu lly v il ble or dire t upt e vi the ills. C rthy nd imene h ve sho n AUT GP 66 atomic and roup Calibration n 8 , r ,96, sd , 8 th t dsorption o lipophili subst n es to dissolved humi m teri ls redu es the v il bility o the subst n e the more De illers et al. (1995) contributions from ekker lipophili the subst n e the l r er redu tion in v il bility S hr p nd pperhui en . urthermore dsorption to and anhold (199 ) dissolved or p rti ul te or ni m tter or sur es in ener l m y inter ere durin the me surement o C nd other SPA C ased upon fundamental o measured lo o data are needed for a trainin set physi l hemi l properties nd thus m e the determin tion o C or ppropri te des riptors di i ult. As Under de elopment b EPA, chemical structure of chemicals. bio on entr tion in ish is dire tly orrel ted ith the v il ble r tion o the hemi l in ter it is ne ess ry or Athens, Geor ia. al orithm. hi hly lipophili subst n es to eep the v il ble on entr tion o the test hemi l ithin rel tively n rro limits durin the upt e period. ekker and De ort (19 9) ra ments correction Calibration n 1 5 , r ,99 factors alidation n , r ,91 , sd ,5 , me , Subst n es hi h re re dily biode r d ble m y only be present in the test ter or short period nd bio on entr tion o these subst n es m y thus be insi ni i nt. Simil rly vol tility nd hydrolysis ill redu e the iemi et al. (199 ) C Calibration n 9, r , on entr tion nd time in hi h the subst n e is v il ble or bio on entr tion. alidation n 9, r , 9 lopman et al (199 ) 98 fra ments correction Calibration n 166 , r ,9 8, sd , 81 n i on ental facto s factors Environment l p r meters in luen in the physiolo y o the or nism m y lso e t the upt e o Su uki and udo (199 ) fra ments Total n 1686, me , 5 subst n es. or inst n e hen the o y en ontent o the ter is lo ered ish h ve to p ss more ter over their ills alidation n 1, me , 9 in order to meet respir tory dem nds im nd oeden . o ever there m y be spe ies dependen y s Ghose et al. (1988) 11 fra ments Calibration n 8 , r ,9 , sd , indi ted by pperhui en nd S hr p . t h s urthermore been sho n th t the temper ture m y h ve n in luen e on the upt e r te onst nt or lipophili subst n es Si m et al. here s other uthors h ve not ound AT GP alidation n 1 5, r ,8 , sd ,5 ny onsistent e e t o temper ture h n es l et al. . odor and uan (199 ) olecule orbital Calibration n , r ,96, sd , 1, me , alidation n 1 8, sd , 8 roto et al. (198 ) 11 fra ments Calibration n 1868, me ca. , Pro o P
5 - 504 - Copyright@United Nations, 2017. All rights reserved
1995 581). ike the C GP pro ram, G has been alidated (Table ) and is recommended for classification nne purposes because of its reliabilit , commercial a ailabilit , and con enience of use. . . AUT GP (De illers et al., 1995) has been deri ed from a hetero eneous data set, comprisin 8 or anic chemicals collected from literature. The pro ram calculates lo o alues for or anic chemicals containin C, n ence o e tern n intern ctor on t e ioconcentr tion potenti o or nic t nce , , , al, P, and S. The lo o alues of salts cannot be calculated. Also the lo o of some compounds ith formal char es cannot be calculated, ith the exception of nitro compounds. The lo o alues of ioni able chemicals like phenols, amines, and corbox lic acids can be calculated althou h p dependencies should be noted. mpro ements ctor in encin t e pt e are in pro ress in order to extend the applicabilit of AUT GP. Accordin to the presentl a ailable information, AUT GP i es accurate alues especiall for hi hl lipophilic substances (lo o 5) (European Commission, The upt e r te or lipophili ompounds is m inly un tion o the si e o the or nism Si m nd 1996). inde . E tern l tors su h s the mole ul r si e tors in luen in the bio v il bility nd di erent environment l tors re o re t import n e to the upt e r te s ell. . .5 SPARC. The SPARC model is still under development by EPA’s Environmental esearch aborator in Athens, Geor ia, and is not et public a ailable. SPA C is a mechanistic model based on chemical thermod namic i e of o anis principles rather than a deterministic model rooted in kno led e obtained from obser ational data. Therefore, SPA C differs from models that use SA s (i.e. , GP) in that no measured lo o data are needed for a trainin Sin e l r er ish h ve rel tively lo er ill sur e to ei ht r tio lo er upt e r te onst nt is set of chemicals. EPA does occasionall run the model for a list of CAS numbers, if re uested. SPA C pro ides to be e pe ted or l r e ish omp red to sm ll ish Si m nd inde pperhui en nd Si m . The upt e impro ed results o er and C GP onl for compounds ith lo o alues reater than 5. nl SPA C o subst n es in ish is urther ontrolled by the ter lo throu h the ills the di usion throu h ueous di usion can be emplo ed in a eneral a for inor anic or or anometallic compounds. l yers t the ill epithelium the perme tion throu h the ill epithelium the r te o blood lo throu h the ills nd the bindin p ity o blood onstituents ECET C .
n Table 1, this Appendix, an o er ie of lo o estimation methods based on fra mentation olec la si e methodolo ies is presented. Also other methods for the estimation of lo o alues exist, but the should onl be used on a case b case basis and onl ith appropriate scientific ustification. oni ed subst n es do not re dily penetr te membr nes s ueous p n in luen e the subst n e upt e. oss o membr ne perme bility is e pe ted or subst n es ith onsider ble ross se tion l re e er ie o et o or e ti tion o o e on r ent tion et o o o ie o r o pperhui en et al. Anli er et al. or lon h in len th . nm pperhui en . oss o n e n membr ne perme bility due to the si e o the mole ules ill thus result in tot l loss o upt e. The e e t o mole ul r et o et o o o t ti tic ei ht on bio on entr tion is due to n in luen e on the di usion oe i ient o the subst n e hi h redu es the upt e r te onst nts ob s et al. . C GP ra ments correction Total n 89 , r ,91 , sd , 8 ansch and eo (19 9), factors alidation n 5 1, r ,96 aila ilit C GP Da li ht (1995) alidation n 1, r ,89, sd ,58 e ore subst n e is ble to bio on entr te in n or nism it needs to be present in ter nd G ( ) 1 fra ments Calibration n , r ,981, sd , 19, me ,161 v il ble or tr ns er ross ish ills. tors hi h e t this v il bility under both n tur l nd test onditions ill e lan and o ard (1995), 6 correction factors alidation n 8855, r ,95, sd , , me , lter the tu l bio on entr tion in omp rison to the estim ted v lue or C . As ish re ed durin bio on entr tion S C studies rel tively hi h on entr tions o dissolved nd p rti ul te or ni m tter m y be e pe ted thus redu in the r tion o hemi l th t is tu lly v il ble or dire t upt e vi the ills. C rthy nd imene h ve sho n AUT GP 66 atomic and roup Calibration n 8 , r ,96, sd , 8 th t dsorption o lipophili subst n es to dissolved humi m teri ls redu es the v il bility o the subst n e the more De illers et al. (1995) contributions from ekker lipophili the subst n e the l r er redu tion in v il bility S hr p nd pperhui en . urthermore dsorption to and anhold (199 ) dissolved or p rti ul te or ni m tter or sur es in ener l m y inter ere durin the me surement o C nd other SPA C ased upon fundamental o measured lo o data are needed for a trainin set physi l hemi l properties nd thus m e the determin tion o C or ppropri te des riptors di i ult. As Under de elopment b EPA, chemical structure of chemicals. bio on entr tion in ish is dire tly orrel ted ith the v il ble r tion o the hemi l in ter it is ne ess ry or Athens, Geor ia. al orithm. hi hly lipophili subst n es to eep the v il ble on entr tion o the test hemi l ithin rel tively n rro limits durin the upt e period. ekker and De ort (19 9) ra ments correction Calibration n 1 5 , r ,99 factors alidation n , r ,91 , sd ,5 , me , Subst n es hi h re re dily biode r d ble m y only be present in the test ter or short period nd bio on entr tion o these subst n es m y thus be insi ni i nt. Simil rly vol tility nd hydrolysis ill redu e the iemi et al. (199 ) C Calibration n 9, r , on entr tion nd time in hi h the subst n e is v il ble or bio on entr tion. alidation n 9, r , 9 lopman et al (199 ) 98 fra ments correction Calibration n 166 , r ,9 8, sd , 81 n i on ental facto s factors Environment l p r meters in luen in the physiolo y o the or nism m y lso e t the upt e o Su uki and udo (199 ) fra ments Total n 1686, me , 5 subst n es. or inst n e hen the o y en ontent o the ter is lo ered ish h ve to p ss more ter over their ills alidation n 1, me , 9 in order to meet respir tory dem nds im nd oeden . o ever there m y be spe ies dependen y s Ghose et al. (1988) 11 fra ments Calibration n 8 , r ,9 , sd , indi ted by pperhui en nd S hr p . t h s urthermore been sho n th t the temper ture m y h ve n in luen e on the upt e r te onst nt or lipophili subst n es Si m et al. here s other uthors h ve not ound AT GP alidation n 1 5, r ,8 , sd ,5 ny onsistent e e t o temper ture h n es l et al. . odor and uan (199 ) olecule orbital Calibration n , r ,96, sd , 1, me , alidation n 1 8, sd , 8 roto et al. (198 ) 11 fra ments Calibration n 1868, me ca. , Pro o P
5 - 505 - Copyright@United Nations, 2017. All rights reserved
2. Factors influencing the elimination rate nne
The elimination rate is mainly a function of the size of the organism, the lipid content, the biotransformation process of the organism, and the lipophilicity of the test compound. e t i e ine 2.1 Size of organism
As for the uptake rate the elimination rate is dependent on the size of the organism. Due to the higher o t o t e i e ine entione re o n in co pi tion ro t e or ni tion i in t e e in gill surface to weight ratio for small organisms (e.g. fish larvae) than that of large organisms, steady-state and thus re erence to t e e re “toxic dose equilibrium” has shown to be reached sooner in early life stages than in juvenile/adult stages of fish (Petersen and Kristensen, 1998). As the time needed to reach steady-state conditions is dependent on k2, the size of fish EC uidelines Commission Re ul tion EC o o y l yin do n test methods used in bioconcentration studies has thus an important bearing on the time required for obtaining steady-state pursu nt to Re ul tion EC o o the Europe n P rli ment nd o the Coun il on the conditions. Re istr tion Ev lu tion Authoris tion nd Restri tion o Chemi ls REAC
2.2 Lipid content b S uidelines Av il ble rom the n tion l st nd rdis tion or nis tions or S omep e http .iso.or iso home.htm Due to partitioning relationships, organisms with a high fat content tend to accumulate higher concentrations of lipophilic substances than lean organisms under steady-state conditions. Body burdens are therefore EC uidelines or the testin o hemi ls. EC P ris ith re ul r upd tes often higher for “fatty” fish such as eel, compared to “lean” fish such as cod. In addition, lipid “pools” may act as http .oe d.or env test uidelines storage of highly lipophilic substances. Starvation or other physiological changes may change the lipid balance and release such substances and result in delayed impacts. d PPTS uidelines S EPA homep e http .ep . ov oppts rs home uidelin.htm
2.3 Metabolism e AST AST s homep e http . stm.or . Further search via “standards”.
2.3.1 In general, metabolism or biotransformation leads to the conversion of the parent compound into more e t i e ine or tic to icit water-soluble metabolites. As a result, the more hydrophilic metabolites may be more easily excreted from the body than the parent compound. When the chemical structure of a compound is altered, many properties of the compound are EC Test uideline Al ro th nhibition Test altered as well. Consequently the metabolites will behave differently within the organism with respect to tissue EC Test uideline phni sp. A ute mmobilis tion Test nd Reprodu tion Test distribution, bioaccumulation, persistence, and route and rate of excretion. Biotransformation may also alter the toxicity EC Test uideline ish A ute To i ity Test of a compound. This change in toxicity may either be beneficial or harmful to the organism. Biotransformation may prevent the concentration in the organism from becoming so high that a toxic response is expressed (detoxification). EC Test uideline ish E rly i e St e To i ity Test However, a metabolite may be formed which is more toxic than the parent compound (bioactivation) as known for e.g. EC Test uideline phni m n Reprodu tion Test benzo(a)pyrene. EC Test uideline ish Short term To i ity Test on Embryo nd S ry St es EC Test uideline ish uvenile ro th Test 2.3.2 Terrestrial organisms have a developed biotransformation system, which is generally better than that EC Test uideline emn sp. ro th inhibition test of organisms living in the aquatic environment. The reason for this difference may be the fact that biotransformation of xenobiotics may be of minor importance in gill breathing organisms as they can relatively easily excrete the compound EC C. A ute To i ity or ish into the water (Van Den Berg et al. 1995). Concerning the biotransformation capacity in aquatic organisms the capacity EC C. A ute To i ity or phni for biotransformation of xenobiotics increases in general as follows: Molluscs < crustaceans < fish (Wofford et al., 1981). EC C. Al l nhibition Test EC C. ish uvenile ro th Test 3. Lipophilicity of substance EC C. ish Short term To i ity Test on Embryo nd S ry St es EC C. phni n Reprodu tion Test A negative linear correlation between k2 (depuration constant) and log Kow (or BCF) has been shown in fish by several authors (e.g. Spacie and Hamelink, 1982; Gobas et al., 1989; Petersen and Kristensen, 1998), whereas PPTS Testin uidelines or Environment l E e ts Series Publi r ts k1 (uptake rate constant) is more or less independent of the lipophilicity of the substance (Connell, 1990). The resultant BCF will thus generally increase with increasing lipophilicity of the substances, i.e. log BCF and log Kow correlate for . Spe i l onsider tion or ondu tin u ti l bor tory studies substances which do not undergo extensive metabolism. . Spe i l onsider tion or ondu tin u ti l bor tory studies . A u ti invertebr te ute to i ity test resh ter d phnids . A u ti invertebr te ute to i ity test resh ter d phnids . mm rid ute to i ity test . mm rid ute to i ity test . ysid ute to i ity test . ysid ute to i ity test . Pen eid ute to i ity test . Pen eid ute to i ity test . ish ute to i ity test resh ter nd m rine . ish ute to i ity test resh ter nd m rine
The list below will need to be regularly updated as new guidelines are adopted or draft guidelines are elaborated.
- 506 - - 506 - Copyright@United Nations, 2017. All rights reserved
2. Factors influencing the elimination rate nne
The elimination rate is mainly a function of the size of the organism, the lipid content, the biotransformation process of the organism, and the lipophilicity of the test compound. e t i e ine 2.1 Size of organism
As for the uptake rate the elimination rate is dependent on the size of the organism. Due to the higher o t o t e i e ine entione re o n in co pi tion ro t e or ni tion i in t e e in gill surface to weight ratio for small organisms (e.g. fish larvae) than that of large organisms, steady-state and thus re erence to t e e re “toxic dose equilibrium” has shown to be reached sooner in early life stages than in juvenile/adult stages of fish (Petersen and Kristensen, 1998). As the time needed to reach steady-state conditions is dependent on k2, the size of fish EC uidelines Commission Re ul tion EC o o y l yin do n test methods used in bioconcentration studies has thus an important bearing on the time required for obtaining steady-state pursu nt to Re ul tion EC o o the Europe n P rli ment nd o the Coun il on the conditions. Re istr tion Ev lu tion Authoris tion nd Restri tion o Chemi ls REAC
2.2 Lipid content b S uidelines Av il ble rom the n tion l st nd rdis tion or nis tions or S omep e http .iso.or iso home.htm Due to partitioning relationships, organisms with a high fat content tend to accumulate higher concentrations of lipophilic substances than lean organisms under steady-state conditions. Body burdens are therefore EC uidelines or the testin o hemi ls. EC P ris ith re ul r upd tes often higher for “fatty” fish such as eel, compared to “lean” fish such as cod. In addition, lipid “pools” may act as http .oe d.or env test uidelines storage of highly lipophilic substances. Starvation or other physiological changes may change the lipid balance and release such substances and result in delayed impacts. d PPTS uidelines S EPA homep e http .ep . ov oppts rs home uidelin.htm
2.3 Metabolism e AST AST s homep e http . stm.or . Further search via “standards”.
2.3.1 In general, metabolism or biotransformation leads to the conversion of the parent compound into more e t i e ine or tic to icit water-soluble metabolites. As a result, the more hydrophilic metabolites may be more easily excreted from the body than the parent compound. When the chemical structure of a compound is altered, many properties of the compound are EC Test uideline Al ro th nhibition Test altered as well. Consequently the metabolites will behave differently within the organism with respect to tissue EC Test uideline phni sp. A ute mmobilis tion Test nd Reprodu tion Test distribution, bioaccumulation, persistence, and route and rate of excretion. Biotransformation may also alter the toxicity EC Test uideline ish A ute To i ity Test of a compound. This change in toxicity may either be beneficial or harmful to the organism. Biotransformation may prevent the concentration in the organism from becoming so high that a toxic response is expressed (detoxification). EC Test uideline ish E rly i e St e To i ity Test However, a metabolite may be formed which is more toxic than the parent compound (bioactivation) as known for e.g. EC Test uideline phni m n Reprodu tion Test benzo(a)pyrene. EC Test uideline ish Short term To i ity Test on Embryo nd S ry St es EC Test uideline ish uvenile ro th Test 2.3.2 Terrestrial organisms have a developed biotransformation system, which is generally better than that EC Test uideline emn sp. ro th inhibition test of organisms living in the aquatic environment. The reason for this difference may be the fact that biotransformation of xenobiotics may be of minor importance in gill breathing organisms as they can relatively easily excrete the compound EC C. A ute To i ity or ish into the water (Van Den Berg et al. 1995). Concerning the biotransformation capacity in aquatic organisms the capacity EC C. A ute To i ity or phni for biotransformation of xenobiotics increases in general as follows: Molluscs < crustaceans < fish (Wofford et al., 1981). EC C. Al l nhibition Test EC C. ish uvenile ro th Test 3. Lipophilicity of substance EC C. ish Short term To i ity Test on Embryo nd S ry St es EC C. phni n Reprodu tion Test A negative linear correlation between k2 (depuration constant) and log Kow (or BCF) has been shown in fish by several authors (e.g. Spacie and Hamelink, 1982; Gobas et al., 1989; Petersen and Kristensen, 1998), whereas PPTS Testin uidelines or Environment l E e ts Series Publi r ts k1 (uptake rate constant) is more or less independent of the lipophilicity of the substance (Connell, 1990). The resultant BCF will thus generally increase with increasing lipophilicity of the substances, i.e. log BCF and log Kow correlate for . Spe i l onsider tion or ondu tin u ti l bor tory studies substances which do not undergo extensive metabolism. . Spe i l onsider tion or ondu tin u ti l bor tory studies . A u ti invertebr te ute to i ity test resh ter d phnids . A u ti invertebr te ute to i ity test resh ter d phnids . mm rid ute to i ity test . mm rid ute to i ity test . ysid ute to i ity test . ysid ute to i ity test . Pen eid ute to i ity test . Pen eid ute to i ity test . ish ute to i ity test resh ter nd m rine . ish ute to i ity test resh ter nd m rine
The list below will need to be regularly updated as new guidelines are adopted or draft guidelines are elaborated. - 506 - - 507 - Copyright@United Nations, 2017. All rights reserved
. a hnid chr nic t icit test E est uideline ydrolysis as a function of p E guidelines for testing of chemicals . a hnid chr nic t icit test E est uideline ctivated sludge respiration inhibition test E guidelines for testing of . sid chr nic t icit test chemicals . sid chr nic t icit test E est uideline eady biodegradability E guidelines for testing of chemicals . Fish ear i e sta e t icit test E est uideline nherent biodegradability odified test E guidelines for testing of . Fish ear i e sta e t icit test chemicals . Fish i e c c e t icit E est uideline ahn ellens E test E guidelines for testing of chemicals . Fish i e c c e t icit E est uideline nherent biodegradability odified test E guidelines for testing of chemicals . Fish F E est uideline imulation test aerobic se age treatment oupled units test E guidelines . Fish F for testing of chemicals . uatic ant t icit test usin e na s . iers and E est uideline nherent biodegradability in soil E guidelines for testing of chemicals . uatic ant t icit test usin e na s . iers and E est uideline iodegradability in sea ater E guidelines for testing of chemicals . uatic ants ie d stud ier E est uideline erobic and anaerobic transformation in soil E guidelines for testing of chemicals . uatic ants ie d stud ier E est uideline erobic and anaerobic transformation in aquatic sediment systems E guidelines for . a t icit iers and testing of chemicals . a t icit iers and E est uideline erobic mineralisation in surface ater – imulation biodegradation test E guidelines for testing of chemicals e t i e ine or iotic n iotic e r tion ydrolysis as a function of p ydrolysis as a function of p and temperature tandard test eth d r i de radati n a sha e as die a a eth d irect photolysis rate in ater by sunlight tandard test eth d r deter inin i de rada i it r anic che ica s in se i c ntinu us eady biodegradability activated s ud e ha e flas die a ay test . . t F eter inati n read i de rada i it . irective nne . ediment ater microcosm biodegradability test . . e radati n i che ica en de and. irective nne . ahn ellens E test . . e radati n a i tic de radati n h dr sis as a uncti n . irective nne . odified test . . i de radati n ahn e ens test. irective nne . oil biodegradation . . i de radati n ctivated s ud e si u ati n tests. irective nne . naerobic biodegradability of organic chemicals . . i de radati n ctivated s ud e res irati n inhi iti n test. irective nne . ndirect photolysis screening test unlight photolysis in aters containing dissolved humic substances . . i de radati n di ied test. irective nne . . ater ua it Evaluation in an aqueous medium of the “ultimate” biodegradability of organic e t i e ine or io cc tion c unds eth d deter inin the en de and in a c sed res ir eter tandards on quatic o icology and a ard Evaluation ponsored by ommittee E . ater ua it va uati n in an a ue us ediu the “ultimate” biodegradability of organic on iological Effects and Environmental ate merican ociety for esting and aterials ace treet c unds eth d ana sis re eased car n di ide hiladelphia . ater ua it eth d r assessin the inhi iti n nitri icati n activated s ud e icr E tandard uide for onducting ioconcentration ests ith ishes and alt ater ivalve r anis s che ica s and aste aters olluscs merican ociety for esting and aterials . ater ua it va uati n the aer ic i de rada i it r anic c unds in an a ue us ediu e ic ntinu us activated s ud e eth d E E artition coefficient nne irective EE ethods for determination of physico . ater ua it va uati n the aer ic i de rada i it r anic c unds in an a ue us chemical properties to icity and ecoto icity ediu tatic test ahn e ens eth d E E ioconcentration lo through ish est . ater ua it Evaluation in an aqueous medium of the “ultimate” biodegradability of organic c unds eth d ana sis i che ica en de and c sed tt e test E uidelines and support documents for environmental effects testing hemical fate test guidelines and . ater ua it eter inati n the inhi it r e ect ater sa es n the i ht e issi n support documents nited tates Environmental rotection gency ffice of esticides and o ic ubstances i ri ischeri u inescent acteria test ashington E ugust and updates cf also ode of ederal egulations rotection of the Environment art to End evised as of uly E information regarding the latest . ater ua it va uati n the e i inati n and i de rada i it r anic c unds in an updates of these test guidelines ational echnical nformation ystem a ue us ediu ctivated s ud e si u ati n test . ater ua it Evaluation of the “ultimate” anaerobic biodegradability of organic compounds in E he ederal nsecticide ungicide and odenticide ct esticide ssessment uidelines di ested s ud e eth d easure ent the i as r ducti n subdivision chemistry Environmental fate and subdivision E a ard Evaluation ffice of esticide . ater ua it va uati n the aer ic i de rada i it r anic c unds at rograms Environmental rotection gency ashington and updates E information regarding c ncentrati ns in ater. art ha e as atch test ith sur ace ater r sur ace ater sedi ent sus ensi ns the latest updates of these test guidelines ational echnical nformation ystem . . E est uideline E uidelines for testing of chemicals artition oefficient n octanol ater ha e las ethod
The list below will need to be regularly updated as new guidelines are adopted or draft guidelines are elaborated. The list below will need to be regularly updated as new guidelines are adopted or draft guidelines are elaborated. - 508 - Copyright@United Nations, 2017. All rights reserved
. a hnid chr nic t icit test E est uideline ydrolysis as a function of p E guidelines for testing of chemicals . a hnid chr nic t icit test E est uideline ctivated sludge respiration inhibition test E guidelines for testing of . sid chr nic t icit test chemicals . sid chr nic t icit test E est uideline eady biodegradability E guidelines for testing of chemicals . Fish ear i e sta e t icit test E est uideline nherent biodegradability odified test E guidelines for testing of . Fish ear i e sta e t icit test chemicals . Fish i e c c e t icit E est uideline ahn ellens E test E guidelines for testing of chemicals . Fish i e c c e t icit E est uideline nherent biodegradability odified test E guidelines for testing of chemicals . Fish F E est uideline imulation test aerobic se age treatment oupled units test E guidelines . Fish F for testing of chemicals . uatic ant t icit test usin e na s . iers and E est uideline nherent biodegradability in soil E guidelines for testing of chemicals . uatic ant t icit test usin e na s . iers and E est uideline iodegradability in sea ater E guidelines for testing of chemicals . uatic ants ie d stud ier E est uideline erobic and anaerobic transformation in soil E guidelines for testing of chemicals . uatic ants ie d stud ier E est uideline erobic and anaerobic transformation in aquatic sediment systems E guidelines for . a t icit iers and testing of chemicals . a t icit iers and E est uideline erobic mineralisation in surface ater – imulation biodegradation test E guidelines for testing of chemicals e t i e ine or iotic n iotic e r tion ydrolysis as a function of p ydrolysis as a function of p and temperature tandard test eth d r i de radati n a sha e as die a a eth d irect photolysis rate in ater by sunlight tandard test eth d r deter inin i de rada i it r anic che ica s in se i c ntinu us eady biodegradability activated s ud e ha e flas die a ay test . . t F eter inati n read i de rada i it . irective nne . ediment ater microcosm biodegradability test . . e radati n i che ica en de and. irective nne . ahn ellens E test . . e radati n a i tic de radati n h dr sis as a uncti n . irective nne . odified test . . i de radati n ahn e ens test. irective nne . oil biodegradation . . i de radati n ctivated s ud e si u ati n tests. irective nne . naerobic biodegradability of organic chemicals . . i de radati n ctivated s ud e res irati n inhi iti n test. irective nne . ndirect photolysis screening test unlight photolysis in aters containing dissolved humic substances . . i de radati n di ied test. irective nne . . ater ua it Evaluation in an aqueous medium of the “ultimate” biodegradability of organic e t i e ine or io cc tion c unds eth d deter inin the en de and in a c sed res ir eter tandards on quatic o icology and a ard Evaluation ponsored by ommittee E . ater ua it va uati n in an a ue us ediu the “ultimate” biodegradability of organic on iological Effects and Environmental ate merican ociety for esting and aterials ace treet c unds eth d ana sis re eased car n di ide hiladelphia . ater ua it eth d r assessin the inhi iti n nitri icati n activated s ud e icr E tandard uide for onducting ioconcentration ests ith ishes and alt ater ivalve r anis s che ica s and aste aters olluscs merican ociety for esting and aterials . ater ua it va uati n the aer ic i de rada i it r anic c unds in an a ue us ediu e ic ntinu us activated s ud e eth d E E artition coefficient nne irective EE ethods for determination of physico . ater ua it va uati n the aer ic i de rada i it r anic c unds in an a ue us chemical properties to icity and ecoto icity ediu tatic test ahn e ens eth d E E ioconcentration lo through ish est . ater ua it Evaluation in an aqueous medium of the “ultimate” biodegradability of organic c unds eth d ana sis i che ica en de and c sed tt e test E uidelines and support documents for environmental effects testing hemical fate test guidelines and . ater ua it eter inati n the inhi it r e ect ater sa es n the i ht e issi n support documents nited tates Environmental rotection gency ffice of esticides and o ic ubstances i ri ischeri u inescent acteria test ashington E ugust and updates cf also ode of ederal egulations rotection of the Environment art to End evised as of uly E information regarding the latest . ater ua it va uati n the e i inati n and i de rada i it r anic c unds in an updates of these test guidelines ational echnical nformation ystem a ue us ediu ctivated s ud e si u ati n test . ater ua it Evaluation of the “ultimate” anaerobic biodegradability of organic compounds in E he ederal nsecticide ungicide and odenticide ct esticide ssessment uidelines di ested s ud e eth d easure ent the i as r ducti n subdivision chemistry Environmental fate and subdivision E a ard Evaluation ffice of esticide . ater ua it va uati n the aer ic i de rada i it r anic c unds at rograms Environmental rotection gency ashington and updates E information regarding c ncentrati ns in ater. art ha e as atch test ith sur ace ater r sur ace ater sedi ent sus ensi ns the latest updates of these test guidelines ational echnical nformation ystem . . E est uideline E uidelines for testing of chemicals artition oefficient n octanol ater ha e las ethod
The list below will need to be regularly updated as new guidelines are adopted or draft guidelines are elaborated. The list below will need to be regularly updated as new guidelines are adopted or draft guidelines are elaborated. - 509 - Copyright@United Nations, 2017. All rights reserved
nne e erence tic to icit
OECD 2006. “Current approaches in the statistical analysis of ecotoxicity data: A guidance to application”, OECD – –
iotic n iotic e r tion
. Environ. ual. Chemosphere
Environmental Toxicology and Chemistry
- 510 - Copyright@United Nations, 2017. All rights reserved
nne e erence tic to icit
OECD 2006. “Current approaches in the statistical analysis of ecotoxicity data: A guidance to application”, OECD – –
iotic n iotic e r tion
. Environ. ual. Chemosphere
Environmental Toxicology and Chemistry
- 511 - Copyright@United Nations, 2017. All rights reserved
angen erg . ., . . . . ei nen urg E. ori e 6 . On the usefulness and relia ility of existing s for C O P. 199 . Dayli ht Che ical nfor ation Syste s nf. Sys. nc. rvine Ca ris assess ent and priority setting. A and A in Environmental esearch , 6 CS EE (1999): D Scientific Co ittee for o icity an Ecoto icity an the Environ ent Opinion on revise oonen ., . indgren, . ansen . archer 6 . rediction of iodegrada ility fro che ical structure. n: proposal for a list of Priority su stances in the conte t of the ater fra e or irective (CO s Proce ure) prepare ei nen urg . . . . . Da ors y eds. . iodegrada ility rediction. lu er Acade ic u lishers y the rauenhofer nstitute er any . inal report opinion a opte at the 11th CS EE plenary eetin on th of Septe er 1999 2 . iodegradation and ioaccu ulation data on existing data ased on the C C apan. apan che ical industry, Ecology toxicology infor ation center. 0 0 Co otto R. . i erle R.A. S isher R.D. 19 9. ioconcentration an eta olis of linear al yl en enesulfonate y Daphni s an athea inno s. . . ar in R.A. i erle E s. A uatic o icolo y (AS 19 9) vol. AS ie el 2000 . ersonal co unication to OECD En iron ent Directorate, 20 arch 2000 S P yhol ., . . erg . ngersle 6 . Acti ated sludge iodegrada ility si ulation test. Danish E A, Connell D. . a er D. . 19 . Use of polyno ial e pressions to escri e the ioconcentration of hy ropho ic En iron ental eport o. che icals y fish. Ecoto icol. Environ. Saf. 1 : 4 yhol . . ngersle . inetic iodegradation tests ith lo test su stance concentrations: ha e flas test Connell D. . 199 . ioaccu ulation of eno iotic co poun s lori a: CRC Press nc. pp.1 1 ith surface ater and short ter rate easure ent in acti ated sludge. n: ales . . ed. . iodegradation inetics: eneration and use of data for regulatory decision a ing. ro the E AC Europe or shop. ort unlight. De ruijn J. usser . Seinen . er ens J. 19 9. Deter ination of octanol/ ater partition coefficients ith the epte er 6. pp. 0 . E AC Europe, russels “slow stirring” method. Environ. Toxicol. Chem. 8:499 1 yhol . . or ng . eport of ing test: hal e flas atch test ith surface ater or surface Devillers J. intein S. Do ine D. 199 . Co parison of C o els ase on lo P. Che osphere ( ):1 4 1 ater sedi ent suspensions. O CD 2 ater uality E aluation of the aero ic iodegrada ility of organic DoE 199 . ui ance on the a uatic to icity testin of ifficult su stance. Unites in o Depart ent of the co pounds at lo concentrations, O C C iodegrada ility Environ ent on on OECD . tructure Acti ity elationships for iodegradation. OECD En iron ent onographs o. 6 . aris Doucette .J. An ren A. . 19 . Correlation of octanol/ ater partition coefficients an total olecular surface area for hi hly hy ropho ic aro atic co poun s. Environ. Sci. echnol. 1 pa es 1 4 OECD (1994): “US EPA/EC Joint Project on the Evaluation of (Quantitative) Structure Activity Relationships.” OECD Doucette .J. An ren A. . 19 . Esti ation of octanol/ ater partition coefficients: evaluation of si etho s for En iron ent onograph o. . aris hi hly hy ropho ic aro atic co poun s. Che osphere 1 pa es 4 9 OECD . Detailed e ie aper on iodegrada ility esting. OECD En iron ental onograph o. . aris Driscoll S. . cElroy A.E. 199 . ioaccu ulation an eta olis of en o(a)pyrene in three species of polychaete OECD . uidance docu ent on direct phototransfor ation of che ical in ater. OECD D 2 . aris or s. Environ. o icol. Che . 1 ( ):14 1 141 OECD . ar oni ed integrated ha ard classification syste for hu an health and en iron ental effects of ECE OC 199 . he role of ioaccu ulation in environ ental ris assess ent: he a uatic environ ent an relate che ical su stances. aris. Docu ent E O O 200 6 foo e s russels el iu edersen ., . yle, . . ie el , . utt ann. . ander A. ede rand . En iron ental a ard Classification ECEOOC 199 . A uatic to icity testin of sparin ly solu le volatile an unsta le su stances. ECE OC ono raph – data collection and interpretation guide for su stances to e e aluated for classification as dangerous for the en iron ent. o. ECE OC russels ordic Council of inisters. 2nd edition. e a ord : , 66 pp European Co ission 199 . echnical ui ance Docu ent in support of Co ission Directive 9 /9 /EEC on Ris ch ar en ach . ., . . sch end D. . oden . En iron ental organic che istry st ed. ohn iley Assess ent for ne notifie su stances an Co ission Re ulation (EC) o 14 /94 on Ris Assess ent for E istin ons, nc. e or Su stances. russels co . . 2 . ate of iodegradation. n: y an . ., . . eehl D. . osen latt 2 : and oo of hose A. . Prottchet A. Crippen . . 19 . J. Co putational Che . 9: 9 Che ical roperty Esti ation ethods En iron ental eha iour of Organic Co pounds. A erican Che ical ociety. o as .A.P.C. Opperhui en A. ut in er O. 19 . ioconcentration of hy ropho ic che icals in fish: ashington DC 0 2 6 0 . Chapter Relationship ith e rane per eation. Environ. o icol. Che . : 4 trui s . . an den erg . tandardi ed iodegrada ility tests: Extrapolation to aero ic en iron ents. at. o as .A.P.C. Clar .E. Shiu . . ac ay D. 19 9. ioconcentration of poly ro inate en enes an es. 2 , 2 262 iphenyls an relate superhy ropho ic che icals in fish: Role of ioavaila ility an eli ination into feces. Environ. yracuse esearch Corporation. iodegradation ro a ility rogra O . yracuse. . . o icol. Che . : 1 4 http: esc.syrres.co esc iodeg.ht oo rich .S. elancon .J. Davis R.A. ech J.J. 1991. he to icity ioaccu ulation eta olis an ester ann ., . . Ahring .A. ah . e perature co pensation in ethanosarcina ar eri y odulation eli ination of ioctyl so iu sulfosuccinate DSS in rain o trout (Oncorhynchus my iss) ater Res. : 119 1 4 of hydrogen and acetate affinity. Applied and Environmental icrobiology , 262 266 ansch C. eo A. 19 9. Su stituent constants for correlation analysis in che istry an iolo y. iley e or 19 9 io cc tion en erson R.J. ocher D.R. 19 . he lipi co position an ioche istry of fresh ater fish. Pro . ipi . Res. Anli er, ., oser, ., oppinger, D. . ioaccu ulation of dyestuffs and organic pig ents in fish. elationships to : 1 4 hydropho icity and steric factors. Che . : 6 6 o ar P. . an eylan . . 199 . Pre iction of physical properties transport an e ra ation for environ ental intein, . De illers, . and archer, . . onlinear dependence of fish ioconcentration on n octanol ater fate an e posure assess ents QSAR in environ ental science . E s. Chen . an Sch r ann . pp. 1 partition coefficient. A and A in En iron ental esearch. ol. .pp.2 i erle R.A. S isher R.D. Schroe er Co otto R. . 19 . Surfactant structure an a uatic to icity Sy posiu on lac , .C., illsap, D. ., cCarthy, . . . Effects of acute te perature change on respiration and toxicant Structure Activity correlations in Stu ies on o icity an ioconcentration ith A uatic Or anis s urlin ton upta e y rain o trout, almo gairdneri ichardson . hysiol. ool. 6 : 6 Ontario Cana a pp. odor, ., uang, . . 2. . har . ci. :2 2 2 lop an . i J. . an S. Di ayu a . 1994. Co puter auto ate lo P calculations ase on an e ten e roup contri ution approach. J. Che . nf. Co put. Sci. 4: 1 roto, ., oreau, ., andyc e, C. . Eur. . ed. Che . : ne ovich J.P. a ton .P. noue .S. 19 9. ioaccu ulation an tissue istri ution of a uaternary a oniu Chiou, . . artition coefficients of organic co pounds in lipid ater syste s and correlations ith fish surfactant in three a uatic species. ull. Environ. Conta . o icol. 4 : 9 ioconcentration factors. En iron. ci. echnol : 62
2 1 - 512 - Copyright@United Nations, 2017. All rights reserved
angen erg . ., . . . . ei nen urg E. ori e 6 . On the usefulness and relia ility of existing s for C O P. 199 . Dayli ht Che ical nfor ation Syste s nf. Sys. nc. rvine Ca ris assess ent and priority setting. A and A in Environmental esearch , 6 CS EE (1999): D Scientific Co ittee for o icity an Ecoto icity an the Environ ent Opinion on revise oonen ., . indgren, . ansen . archer 6 . rediction of iodegrada ility fro che ical structure. n: proposal for a list of Priority su stances in the conte t of the ater fra e or irective (CO s Proce ure) prepare ei nen urg . . . . . Da ors y eds. . iodegrada ility rediction. lu er Acade ic u lishers y the rauenhofer nstitute er any . inal report opinion a opte at the 11th CS EE plenary eetin on th of Septe er 1999 2 . iodegradation and ioaccu ulation data on existing data ased on the C C apan. apan che ical industry, Ecology toxicology infor ation center. 0 0 Co otto R. . i erle R.A. S isher R.D. 19 9. ioconcentration an eta olis of linear al yl en enesulfonate y Daphni s an athea inno s. . . ar in R.A. i erle E s. A uatic o icolo y (AS 19 9) vol. AS ie el 2000 . ersonal co unication to OECD En iron ent Directorate, 20 arch 2000 S P yhol ., . . erg . ngersle 6 . Acti ated sludge iodegrada ility si ulation test. Danish E A, Connell D. . a er D. . 19 . Use of polyno ial e pressions to escri e the ioconcentration of hy ropho ic En iron ental eport o. che icals y fish. Ecoto icol. Environ. Saf. 1 : 4 yhol . . ngersle . inetic iodegradation tests ith lo test su stance concentrations: ha e flas test Connell D. . 199 . ioaccu ulation of eno iotic co poun s lori a: CRC Press nc. pp.1 1 ith surface ater and short ter rate easure ent in acti ated sludge. n: ales . . ed. . iodegradation inetics: eneration and use of data for regulatory decision a ing. ro the E AC Europe or shop. ort unlight. De ruijn J. usser . Seinen . er ens J. 19 9. Deter ination of octanol/ ater partition coefficients ith the epte er 6. pp. 0 . E AC Europe, russels “slow stirring” method. Environ. Toxicol. Chem. 8:499 1 yhol . . or ng . eport of ing test: hal e flas atch test ith surface ater or surface Devillers J. intein S. Do ine D. 199 . Co parison of C o els ase on lo P. Che osphere ( ):1 4 1 ater sedi ent suspensions. O CD 2 ater uality E aluation of the aero ic iodegrada ility of organic DoE 199 . ui ance on the a uatic to icity testin of ifficult su stance. Unites in o Depart ent of the co pounds at lo concentrations, O C C iodegrada ility Environ ent on on OECD . tructure Acti ity elationships for iodegradation. OECD En iron ent onographs o. 6 . aris Doucette .J. An ren A. . 19 . Correlation of octanol/ ater partition coefficients an total olecular surface area for hi hly hy ropho ic aro atic co poun s. Environ. Sci. echnol. 1 pa es 1 4 OECD (1994): “US EPA/EC Joint Project on the Evaluation of (Quantitative) Structure Activity Relationships.” OECD Doucette .J. An ren A. . 19 . Esti ation of octanol/ ater partition coefficients: evaluation of si etho s for En iron ent onograph o. . aris hi hly hy ropho ic aro atic co poun s. Che osphere 1 pa es 4 9 OECD . Detailed e ie aper on iodegrada ility esting. OECD En iron ental onograph o. . aris Driscoll S. . cElroy A.E. 199 . ioaccu ulation an eta olis of en o(a)pyrene in three species of polychaete OECD . uidance docu ent on direct phototransfor ation of che ical in ater. OECD D 2 . aris or s. Environ. o icol. Che . 1 ( ):14 1 141 OECD . ar oni ed integrated ha ard classification syste for hu an health and en iron ental effects of ECE OC 199 . he role of ioaccu ulation in environ ental ris assess ent: he a uatic environ ent an relate che ical su stances. aris. Docu ent E O O 200 6 foo e s russels el iu edersen ., . yle, . . ie el , . utt ann. . ander A. ede rand . En iron ental a ard Classification ECEOOC 199 . A uatic to icity testin of sparin ly solu le volatile an unsta le su stances. ECE OC ono raph – data collection and interpretation guide for su stances to e e aluated for classification as dangerous for the en iron ent. o. ECE OC russels ordic Council of inisters. 2nd edition. e a ord : , 66 pp European Co ission 199 . echnical ui ance Docu ent in support of Co ission Directive 9 /9 /EEC on Ris ch ar en ach . ., . . sch end D. . oden . En iron ental organic che istry st ed. ohn iley Assess ent for ne notifie su stances an Co ission Re ulation (EC) o 14 /94 on Ris Assess ent for E istin ons, nc. e or Su stances. russels co . . 2 . ate of iodegradation. n: y an . ., . . eehl D. . osen latt 2 : and oo of hose A. . Prottchet A. Crippen . . 19 . J. 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ne ovich . . no e . . 99 . The in l ence o sediment nd colloid l m teri l on the io v il ilit o tern r mmoni m s r ct nt. Ecotoxicol. Environ. et . : 4 ristensen . 99 . ioconcentr tion in ish: Com rison o C s derived rom EC nd T testing methods in l ence o rtic l te m tter to the io v il ilit o chemic ls. nish ter lit nstit te c . 98 . Correl tion o ioconcentr tion ctors. Environ. ci. Technol. : 4 8 cC rth . . imene . . 98 . ed ction in io v il ilit to l egills o ol c clic rom tic h droc r ons o nd to dissolved h mic m teri l. Environ. Toxicol. Chem. 4: c im . . oeden . . 98 . direct me s re o the t e e icienc o xeno iotic chemic l cross the gill o roo tro t alvelinus fontinalis nder normoxic nd h oxic conditions. Com . iochem. h siol. C: 4 e l n . . nd ow rd . . 99 . tom r gment Contri tion ethods or Estim ting ct nol ter rtition Coe icients. . h rm. ci. 84 8 iemel . . 99 . T rogr m ver . . nish Environment l rotection genc iemi . . s .C. eith . . r nw ld . Environ. Toxicol. Chem. :89 9 iimi . . 99 . ol ilit o org nic chemic ls in oct nol triolin nd cod liver oil nd rel tionshi s etween sol ilit nd rtition coe icients. t. es. : – EC 99 . lic tion o str ct re ctivit rel tionshi s to the estim tion o ro erties im ort nt in ex os re ssessment. EC Environment irector te. Environment onogr h o. EC 998. rmoni ed integr ted h rd cl ssi ic tion s stem or h m n he lth nd environment l e ects o chemic l s st nces. s endorsed the 8th oint meeting o the chemic ls committee nd the wor ing rt on chemic ls in ovem er 998 EC . id nce oc ment on tic Toxicit Testing o i ic lt st nces nd ixt res EC ris Cyprinus carpio erh i en . n der elde E. . o s . . .C. iem . . . n der teen . . . t inger . 98 . el tionshi etween ioconcentr tion in ish nd steric ctors o h dro ho ic chemic ls. Chemos here 4: 8 89 erh i en . 98 . ioconcentr tion o h dro ho ic chemic ls in ish. n: oston T. . rd . eds tic Toxicolog nd Environment l te: inth ol me T T 9 . meric n ociet or Testing nd teri ls e erence or hil del hi 4 erh i en . chr . . 98 . el tionshi etween eo s ox gen concentr tion nd t e nd elimin tion r tes d ring ioconcentr tion o h dro ho ic chemic ls in ish. Environ. Toxicol. Chemos here : 4 erh i en . i m .T. . . 99 . io cc m l tion nd iotr ns orm tion o ol chlorin ted di en o dioxins nd di en o r ns in ish. Environ. Toxicol. Chem. 9: 8 hydrophobic organic chemicals with the “slow stirring method,” Environ. Toxicol. edersen . T le . iemel . . ttm nn . nder . nd ede r nd . 99 . Environment l rd Cl ssi ic tion – d t collection nd inter ret tion g ide nd edition . Tem ord 99 : 8 etersen . . ristensen . 998. io cc m l tion o li o hilic s st nces in ish e rl li e st ges. Environ. Toxicol. Exploring A Chem. : 8 9 uantitative Treatments of olute solvent nteractions e er . . de ort . . 9 9. The h dro ho ic r gment l const nt: n extension to d t oint set. E r. . Theoretical and Computational Chemistry ol. ed. Chem. – Chim. Ther. 4:4 9 488 o erts . . 989. tic toxicit o line r l l en ene s l hon tes – n l sis. Comm nic ciones resent d s l s orn d s del Comite Es nol de l etergenci 989 4 . lso in .E. T rner . . Engl nd T. . ch lt nd . . w eds. 88. roc. Third ntern tion l or sho on lit tive tr ct re ctivit el tionshi s in Environment l Toxicolog 988 noxville Tennessee . 9 98. v il le rom the tion l Technic l n orm tion ervice e t. o Commerce ring ield chr . . erh i en . 99 . el tionshi etween io v il ilit nd h dro ho icit : red ction o the t e o org nic chemic ls ish d e to the sor tion o rticles. Environ. Toxicol. Chem. 9: 4 Trends harmacol. ci hi o cette . o s C. ndren . c . 988. h sic l chemic l ro erties o chlorin ted di en o dioxins. Environ. ci. Technol. : ges 8 Trends harmacol. ci i m .T. . . v n der inde . 99 . i e de endent ioconcentr tion inetics o h dro ho ic org nic chemic ls in ish sed on di sive m ss tr ns er nd llometric rel tionshi s. Environ. ci. Technol. 9: 9 i m .T. . . rt . erh i en . 99 . The in l ence o tem er t re on the t e r te const nts o Environ. Toxicol. Chem h dro ho ic com o nds determined the isol ted er sed gill o r in ow tro t Oncorhynchus my iss . t. Toxicol. : 4
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ne ovich . . no e . . 99 . The in l ence o sediment nd colloid l m teri l on the io v il ilit o tern r mmoni m s r ct nt. Ecotoxicol. Environ. et . : 4 ristensen . 99 . ioconcentr tion in ish: Com rison o C s derived rom EC nd T testing methods in l ence o rtic l te m tter to the io v il ilit o chemic ls. nish ter lit nstit te c . 98 . Correl tion o ioconcentr tion ctors. Environ. ci. Technol. : 4 8 cC rth . . imene . . 98 . ed ction in io v il ilit to l egills o ol c clic rom tic h droc r ons o nd to dissolved h mic m teri l. Environ. Toxicol. Chem. 4: c im . . oeden . . 98 . direct me s re o the t e e icienc o xeno iotic chemic l cross the gill o roo tro t alvelinus fontinalis nder normoxic nd h oxic conditions. Com . iochem. h siol. C: 4 e l n . . nd ow rd . . 99 . tom r gment Contri tion ethods or Estim ting ct nol ter rtition Coe icients. . h rm. ci. 84 8 iemel . . 99 . T rogr m ver . . nish Environment l rotection genc iemi . . s .C. eith . . r nw ld . Environ. Toxicol. Chem. :89 9 iimi . . 99 . ol ilit o org nic chemic ls in oct nol triolin nd cod liver oil nd rel tionshi s etween sol ilit nd rtition coe icients. t. es. : – EC 99 . lic tion o str ct re ctivit rel tionshi s to the estim tion o ro erties im ort nt in ex os re ssessment. EC Environment irector te. Environment onogr h o. EC 998. rmoni ed integr ted h rd cl ssi ic tion s stem or h m n he lth nd environment l e ects o chemic l s st nces. s endorsed the 8th oint meeting o the chemic ls committee nd the wor ing rt on chemic ls in ovem er 998 EC . id nce oc ment on tic Toxicit Testing o i ic lt st nces nd ixt res EC ris Cyprinus carpio erh i en . n der elde E. . o s . . .C. iem . . . n der teen . . . t inger . 98 . el tionshi etween ioconcentr tion in ish nd steric ctors o h dro ho ic chemic ls. Chemos here 4: 8 89 erh i en . 98 . ioconcentr tion o h dro ho ic chemic ls in ish. n: oston T. . rd . eds tic Toxicolog nd Environment l te: inth ol me T T 9 . meric n ociet or Testing nd teri ls e erence or hil del hi 4 erh i en . chr . . 98 . el tionshi etween eo s ox gen concentr tion nd t e nd elimin tion r tes d ring ioconcentr tion o h dro ho ic chemic ls in ish. Environ. Toxicol. Chemos here : 4 erh i en . i m .T. . . 99 . io cc m l tion nd iotr ns orm tion o ol chlorin ted di en o dioxins nd di en o r ns in ish. Environ. Toxicol. Chem. 9: 8 hydrophobic organic chemicals with the “slow stirring method,” Environ. Toxicol. edersen . T le . iemel . . ttm nn . nder . nd ede r nd . 99 . Environment l rd Cl ssi ic tion – d t collection nd inter ret tion g ide nd edition . Tem ord 99 : 8 etersen . . ristensen . 998. io cc m l tion o li o hilic s st nces in ish e rl li e st ges. Environ. Toxicol. Exploring A Chem. : 8 9 uantitative Treatments of olute solvent nteractions e er . . de ort . . 9 9. The h dro ho ic r gment l const nt: n extension to d t oint set. E r. . Theoretical and Computational Chemistry ol. ed. Chem. – Chim. Ther. 4:4 9 488 o erts . . 989. tic toxicit o line r l l en ene s l hon tes – n l sis. Comm nic ciones resent d s l s orn d s del Comite Es nol de l etergenci 989 4 . lso in .E. T rner . . Engl nd T. . ch lt nd . . w eds. 88. roc. Third ntern tion l or sho on lit tive tr ct re ctivit el tionshi s in Environment l Toxicolog 988 noxville Tennessee . 9 98. v il le rom the tion l Technic l n orm tion ervice e t. o Commerce ring ield chr . . erh i en . 99 . el tionshi etween io v il ilit nd h dro ho icit : red ction o the t e o org nic chemic ls ish d e to the sor tion o rticles. Environ. Toxicol. Chem. 9: 4 Trends harmacol. ci hi o cette . o s C. ndren . c . 988. h sic l chemic l ro erties o chlorin ted di en o dioxins. Environ. ci. Technol. : ges 8 Trends harmacol. ci i m .T. . . v n der inde . 99 . i e de endent ioconcentr tion inetics o h dro ho ic org nic chemic ls in ish sed on di sive m ss tr ns er nd llometric rel tionshi s. Environ. ci. Technol. 9: 9 i m .T. . . rt . erh i en . 99 . The in l ence o tem er t re on the t e r te const nts o Environ. Toxicol. Chem h dro ho ic com o nds determined the isol ted er sed gill o r in ow tro t Oncorhynchus my iss . t. Toxicol. : 4
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