BEHAVIOUR OF UV FILTERS, UV BLOCKERS AND PHARMACEUTICALS IN HIGH RATE ALGAL PONDS TREATING URBAN WASTEWATER

Lucas Vassalle1,2, Adrià Sunyer Caldú3, M. Silvia Díaz-Cruz3,Larissa Terumi Arashiro1, Ivet Ferrer1, Marianna Garfí1, Mª Jesús García-Galán1

1 GEMMA – Group of Environmental Engineering and Microbiology, Department of Civil and Environmental Engineering, Universitat Politècnica de Catalunya·BarcelonaTech, c/Jordi Girona 1-3, Building D1, 08034, Barcelona, Spain

2 Department of Sanitary and Environmental Engineering, Universidade Federal de Minas Gerais (UFMG), Av. Antônio Carlos, 6627, Belo Horizonte, MG, Brazil

3 ENFOCHEM-Water, Environmental and Food Chemistry Unit, Department of Environmental Chemistry, Institute of Environmental Assessment and Water Research (IDAEA-CSIC), c/Jordi Girona 18-26, 08034, Barcelona, Spain. Table S1. Target PPCPs, classified by their chromatographic retention time, and their optimized UPLC–QqLIT-MS/MS parameters by positive ionization mode.

Solubility Molecular (25ºC) Log Family Abbreviation Compound CAS no. Formula pKa Molecular Structure weight Kow ( mg L-1)

Formatted: Font: 7 pt

BZT 1H-benzotriazole 95-14-7 C6H5N3 119.120 20000 1.23 8.2

MeBZT 5-methyl-1-H-benzotriazole 136-85-6 C7H7N3 133.150 5 1.89 8.66

Benzotriazoles 5,6-Dimethyl-1H-benzotriazole DMBZT 4184-79-6 C8H11N3O 165.192 914.2 2.26 - monohydrate

2-(2'-Hydroxy-5'- UVP 2440-22-4 C13H11N3O 225.25 - - - methylphenyl)benzotriazol

Benzophenone-1 BP1 131-56-6 C13H10O3 214.220 240 3.17 7.1 2,4-Dihhydroxybenzophenone*

Benzophenone-2 BP2 2,2’,4,4’- 131-55-5 C13H10O5 246.053 98.42 2.78 - Tetrahydroxybenzophenone* Benzophenones

Benzophenone-3/Oxybenzone BP3 2-Hydroxy-4- 131-57-7 C14H12O3 228.240 0.21 3.79 - methoxybenzophenone*

4HB 4-hydroxybenzophenone 1137-42-4 C13H10O2 198.220 405.8 3.02 -

4DHB 4,4 -dihydroxybenzophenone 611-99-4 C13H10O3 214.220 1905 2.55 -

2,2’-Dihydroxy-4- DHMB 131-53-3 C14H12O4 244.074 52.73 3.82 - methoxybenzophenone

Enzacamene Camphor 4MBC 3-(4-Methylbenzilidene) 36861-47-9 C18H22O 254.400 0.1966 4.95 - derivatives camphor*

Cinnamate EHMC Ethylhexyl methoxycinnamate 5466-77-3 C18H26O3 290.400 0.1548 5.8 - derivatives

Benzocaine EtPABA 94-09-7 C9H11NO2 165.189 1671 1.86 - Ethyl 4-aminobenzoate p-aminobenzoic acid derivatives Padimate-O2-Ethylhexyl-4- ODPABA 21245-02-3 C17H27NO2 277.400 0.1977 6.15 - (dimethylamino)benzoate

Clarithromycin 81103-11-9 C38H69NO13 747.953 0.33 3.16 8.99

Macrolide

Tylosin 1401-69-0 C46H77NO17 916.100 5 1.63 7.73

Flumequine 42835-25-6 C14H12FNO3 261.248 308.4 2.6 6.5

Fluoroquinolone

Ofloxacin 82419-36-1 C18H20FN3O4 361.367 6.76E+05 -2 2.1

Quinolone Oxolinic acid 14698-29-4 C13H11NO5 261.23 8007 0.94 8.77

Sulfabenzamide 127-71-9 C13H12N2O3S 276.311 1316 1.3 4.32

Sulfadimethoxine 122-11-2 C12H14N4O4S 310.329 433.1 1.63 6.91

Sulfonamides

Sulfamerazine 127-79-7 C11H12N4O2S 264.304 202 0.14 7.12

Sulfamethizole 144-82-1 C9H10N4O2S2 270.331 1050 0.54 2.1 / 5.3

Sulfamethoxazole 723-46-6 C10H11N3O3S 253.278 610 0.89 9.1

Sulfanitran 122-16-7 C14H13N3O5S 335.335 36.57 2.26 7.44

Sulfapyridine 144-83-2 C11H11N3O2S 249.289 268 0.35 8.43

Sulfathiazole 72-14-0 C9H9N3O2S2 255.317 373 0.05 7.2

Sulfisomidin 515-64-0 C12H14N4O2S 278.330 1620 -0.33 6.12

Dihydrofolate Trimethoprim 738-70-5 C14H18N4O3 290.318 400 0.91 7.2 reductases

Ketoprofen 22071-15-4 C16H14O3 254.281 51 3.12 4.45

Analgesic/ Mefenamic acid 61-68-7 C15H15NO2 241.285 20 (30ºC) 5.12 4.2 anti-inflammatories

Naproxen 22204-53-1 C14H14O3 230.259 15.9 3.18 4.15

Lipid regulators Gemfibrozil 25812-30-0 C15H22O3 250.333 4964 4.77 4.5

B-blocking agent Atenolol 29122-68-7 C14H22N2O3 266.336 685.2 0.16 9.6

Stimulants Caffeine 58-08-2 C8H10N4O2 194.191 21600 -0.07 14

Experimental data obtained from Chemspider database and the US Environmental Protection Agency’s EPISUITETM Table S2. Solar irradiation and air temperatures registered during the days of the experiment.

Temperature Date Irradiance (MJ/m2) (ºC)

10/07 23.3 24.9

11/07 26.6 25.5

12/07 28.9 26

13/07 25.3 25.6

14/07 7.7 24.6

15/07 27 25.9

16/07 28.1 25.8

17/07 28.3 25.8

18/07 27.1 26.2

19/07 25.3 25.9

20/07 24.6 25.1

.

Antibiotics Lipid regulators Benzothiazoles

1000

) -1

100 45%

100% 100%

Concentration (ng L 10

100% Influent Effluent 1

4DHB Flumequina Gemfibrozil Sulfabenzamidesulfamethoxazole

Figure S1. Concentrations of the targeted compounds detected with frequencies of detection (F%) ≤ 17% (1 samples out of 6) in the HRAP

Table S3. Chromatographic retention time and optimized MS/MS transitions for the target analytes

DP CE CXP DP CE CxP COMPOUND RT MRM1 MRM2 Internal standard (V) (eV) (eV) (V) (eV) (eV)

BZT 4.6 120>65 56 31 4 120>92 56 25 16 MeBZT 5.3 134>79 46 29 10 134>95 46 35 14 Benzotriazoles BZT-d4 DMBZT 5.8 148>77 56 39 2 148>93 56 27 6 UVP 10.8 226>120 46 25 10 226>107 46 19 18 BP1 7.7 215>137 40 27 10 215>105 40 29 6 BP2 6.6 247>137 46 25 8 247>109 46 45 8 BP3 9.2 229>151 40 25 12 229>105 40 27 16 Benzophenones BP3-d5 4HB 7.1 199>121 40 25 8 199>105 40 27 8 4DHB 5.9 215>121 45 27 8 215>93 45 45 6 DHMB 8.2 245>121 43 29 8 245>151 43 27 12 Camphor derivatives 4MBC 10.8 255>105 61 45 6 255>212 61 29 14

Cinnamate EHMC 11.9 291>179 51 19 4 291>161 51 25 10 4MBC-d4 derivatives p-aminobenzoic EtPABA 6.5 166>138 41 20 10 166>120 41 25 28 acid derivatives ODPABA 12.3 278>166 86 43 42 278>151 86 27 40 Clarithromycin 7.2 749>158 76 39 10 749>83 76 84 5 Clarithromycin-N- Macrolide Tylosin 5.74 917>174 106 53 10 917>773 106 46 12 methyl-d3 Quinolone Oxolinic acid 5.8 262>244 36 25 12 262>216 36 41 16 Flumequine-13C Flumequine 6.7 262>244 51 29 14 262>202 51 45 10 Fluoroquinolone Sulfamethazine-d4 Ofloxacin 4.5 362>318 56 29 18 362>261 56 41 18 Sulfabenzamide 5.8 277>156 56 17 10 257>92 56 41 6 Sulfadimethoxine 6.1 311>156 76 31 8 311>92 76 31 6

Sulfonamides Sulfamerazine 4.4 265>156 61 27 8 265>92 61 47 6 Sulfamethazine-d4 Sulfamethizole 4.4 271>156 56 25 10 271>108 56 27 10 Sulfamethoxazole 5.6 254>108 56 27 10 254>156 56 25 10 Sulfanitran 6.6 336>156 66 17 12 336>198 66 29 14 Sulfapyridine 4.2 250>156 51 28 12 250>92 51 31 6 Sulfathiazole 4.1 256>156 40 25 14 256>92 40 25 10 Sulfisomidin 4.6 279>124 76 33 8 279>186 76 23 14 Dihydrofolate Trimethoprim 3.9 291>230 76 33 20 291>261 76 35 12 Trimethoprim-d3 reductases Ketoprofen 7.3 255>105 66 33 18 255>209 66 25 10 BP3-d5 Analgesic/ Mefenamic acid 9.2 242>224 46 23 22 242>209 46 41 14 Mefenamic acid-d3 antiinflammatories Naproxen 7.4 231>185 56 17 14 231>115 56 77 10 Flumequine-13C

Lipid regulators Gemfibrozil 9.2 251>233 31 7 14 251>129 31 19 12 BP3-d5

B-blocking agent Atenolol 3.1 267>116 46 31 8 267>133 46 39 22 Trimethoprim-d3

Stimulants Caffeine 4 195>109.960 56 31 8 195>138 56 29 10 Trimethoprim-d3

BZT-d4 4.5 124>96 81 25 6

BP3-d5 9.2 234>151 36 27 12

4MBC-d4 10.8 259>216 76 27 14 Flumequine-13C 6.7 265>247 51 31 20

Sulfamethazine-d4 4.6 283>96 26 35 5 Internal standards Trimethoprim-d3 3.9 294>230 72 35 10

Gemfibrozil-d6 6.7 257.416>239 26 9 14

Mefenamic acid-d3 5.4 245>227.4 66 17 8

Caffeine-d3 4 198>140 71 25 8

Atenolol-d7 3.1 274>107 51 69 4

Table S4. Linearity and method limits of quantification (LOQ), given in ng L-1, for the targeted analytes.

Urban Family Compound Linearity r2 HRAPeff sewage BZT 1-700 0.9939 29.59 39.91 5-MeBZT 5-700 0.9978 9.01 31.44 Benzotriazoles DMBZT 30-500 0.9985 13.99 20.50 UVP 3-700 0.9932 282.02 209.00 BP1 30-700 0.9988 15.18 11.70 BP2 10-700 0.9982 - - BP3 1-700 0.9928 27.83 27.91 Benzophenones 4HB 5-700 0.9989 8.05 - 4DHB 3-700 0.9907 16.51 20.09 DHMB 10-700 0.9957 - - Camphor 4MBC 30-700 0.997 3.76 3.68 derivatives Cinnamate EHMC 5-700 0.9958 48.58 0.02 derivatives p-aminobenzoic EtPABA 50-700 0.9919 - - acid derivatives Clarithromycin 30-700 0.9989 - 8.00 Macrolide Tylosin 50-700 0.9957 - - Flumequine 30-700 0.9955 5.63 9.21 Fluoroquinolone Ofloxacin 30-700 0.9997 613.78 1036.76 Quinolone Oxolinic acid 1-700 0.9926 - - Sulfabenzamide 1-700 0.9919 8.91 - Sulfadimethoxine 3-700 0.9926 11.04 1.15 Sulfaguanidine 30-700 0.9958 - - Sulfamerazine 10-700 0.9961 - 7.69 Sulfamethizole 30-700 0.9927 17.50 13.41 Sulfamethoxazole 3-700 0.9929 18.98 2.25 Sulfonamides Sulfamethoxypyridazine 1-700 0.9958 - 7.71 Sulfanitran 3-700 0.9956 - - Sulfapyridine 1-700 0.9981 6.71 7.27 Sulfaquinoxaline 3-700 0.9979 9.23 - Sulfathiazole 10-700 0.9968 10.29 30.38 Sulfisomidin 5-700 0.9968 - - Dihydrofolate Trimethoprim 3-700 0.9931 11.83 16.41 reductases Ketoprofen 10-700 0.9955 113.15 67.78 Analgesic/ Mefenamic acid 1-700 0.9959 - - antiinflammatories Naproxen 10-700 0.9946 80.29 90.21 Lipid regulators Gemfibrozil 30-700 0.9878 30 765.63 B-blocking agent Atenolol 30-700 0.9983 80.02 8.89 Stimulants Caffeine 50-700 0.9944 291.00 183.53