1 Supplementary data for 2 Human health risk assessment of soil dioxin/furans contamination and dioxin-like 3 activity determined by ethoxyresorufin-O-deethylase bioassay 4 5 Yu Bon Man a, Ka Lai Chow b, Hong Sheng Wang c, Xiao Lin Sun d, Sheng Chun Wu e, 6 Zong Wei Cai f, Yuan Kang g, Hui Li h, Jin Shu Zheng f, John P. Giesy i, Ming Hung 7 Wong a,e,h * 8 9 a Consortium on Health, Environment, Education and Research (CHEER), and 10 Department of Science and Environmental Studies, Hong Kong Institute of Education, 11 Tai Po, Hong Kong, China 12 b 13 Kong, China 14 c 15 Sciences, Sun Yat-sen University, Guangzhou 510006, China 16 d 17 Yat-sen University, Guangzhou 510006, China. 18 e 19 University, Lin’an, Zhejiang 311300, China 20 f 21 University, Hong Kong, China 22 g 23 Laboratory of Theoretical Chemistry of Environment, Ministry of Education, Higher 24 Education Mega Center, Guangzhou 510006, China Department of Geography, Hong Kong Baptist University, Kowloon Tong, Hong Department of Microbial and Biochemical Pharmacy, School of Pharmaceutical Land Resources and Environment, School of Geography Science and Planning, Sun School of Environmental and Resource Sciences, Zhejiang Agriculture and Forestry Dioxin Analysis Laboratory, and Department of Chemistry, Hong Kong Baptist School of Chemistry & Environment, South China Normal University; Key 1 25 h 26 Guangdong, China 27 i 28 Hong Kong, China Department of Environmental Engineering, Jinan University, Guangzhou 510630, Department of Biology and Chemistry, City University of Hong Kong, Kowloon, 29 30 *Corresponding author: Ming Hung Wong - postal address: Consortium on Health, 31 Environment, Education and Research (CHEER) and Department of Science and 32 Environmental Studies , The Hong Kong Institute of Education, Tai Po, Hong Kong, 33 China; e-mail: minghwong@ied.edu.hk; tel.: (+852) 2948 8706 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 2 CsoilIngREF ED CF ) / RfD BW AT CsoilIngREF ED risk ingest CF SFO BW AT Non - cancer risk ingest ( 49 Cancer 50 Equation (A1) (US EPA, 1989) 51 Where 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85 Non-cancer riskingest Cancer riskingest Csoil IngR EF ED BW AT CF RfD SFO Non- Cancer risk via ingestion of soil particles Cancer risk via ingestion of soil particles Concentration of the pollutant in soil (mg/kg) Ingestion rate of soil (mg/day) Exposure frequency (days/yr) Exposure duration (years) Average body weight (kg) Averaging time (days) Conversion factor (1x10-6 kg/mg) Reference dose (mg/kg/day) Oral slope factor (mg/kg/day)-1 3 Non - cancer risk dermal ( 86 Cancer risk dermal BW AT Equation (A2) (US EPA, 1989) 88 Where Non-cancer riskdermal Cancer riskdermal SA AFsoil ABS GIABS BW AT Csoil SA AFsoil ABS EF ED 87 89 90 91 92 93 94 95 96 97 98 99 100 101 102 103 104 105 106 107 108 109 110 111 112 113 114 115 116 117 118 119 120 121 122 123 Csoil SA AFsoil ABS EF ED CF GIABS ) / Rfd CF SFO GIABS Non-Cancer risk via dermal contact of soil particles Cancer risk via dermal contact of soil particles Surface area of the skin that contacts soil (cm2/day) Skin adherence factor for soil (mg/cm2) Dermal absorption factor (chemical specific) Gastrointestinal absorption factor 4 Non - cancer risk inhale 124 125 126 127 128 129 130 131 132 133 134 135 136 137 138 139 140 Cancer risk inhale Csoil EF ET ED PET AT * Csoil EF ET ED PET AT * Equation (A3) (US EPA, 2009) RfC IUR Where Non-cancer riskinhale Cancer riskinhale ET IUR AT* RfC PEF Non-cancer risk via inhalation of soil particles Cancer risk via inhalation of soil particles Exposure time (hours/day) Inhalation unit risk (mg/m3)-1 Averaging time (hours) RfC (mg/m3) Particle emission factor = 1.36 x 109 m3/kg PEF is the inhalation of pollutants that are adsorbed to respirable particles (PM10) and relates the concentration of a pollutant in soil with the concentration of respirable particles in the air, due to fugitive dust emissions from contaminated soils (US EPA, 2001). 141 142 143 144 145 146 147 148 149 150 151 152 153 5 154 Details and parameters used in the health risk assessment 155 Seventeen 2,3,7,8-substituted PCDD and PCDF congeners with their respective 2,3,7,8 – 156 TEFs were used to conduct non-cancer and cancer risks assessment using 157 TeCDD toxicity equivalence (Van den Berg, 2006) (Table S2) (supplementary data). 158 The parameters, reference doses and slope factors for estimating human non-cancer 159 and cancer risks are listed in Table S3 (supplementary data). The equations for 160 estimating the potential non-cancer risks of concerned pollutants via the exposure of 161 PCDD/Fs contaminated soils can be obtained from the ratio of estimated average daily 162 dose (ADD) (mg/kg/day) to a reference dose (RfD) = Hazard Quotient (HQ) (Equations 163 (A1) (A2) and (A3) (supplementary data). The reference dose (mg/kg/day) is an 164 estimation of maximum permissible risk on human populations through daily exposure 165 taking the vulnerable group (of children), during a lifetime into consideration. The HQ 166 ≤ 1 indicates no adverse health effects, whereas HQ > 1 indicates likely adverse health 167 effects (US EPA, 1989). The procedure for conducting a cancer risk assessment on 168 humans is similar to a non-cancer risk assessment. Instead of dividing the ADD by the 169 reference dose (RfD), the ADD (µg/kg/day) is multiplied by an slope factor (SF) 170 (mg/kg-day)-1 (US EPA, 1989) (Equations (A1) (A2) and (A3). Qualitative 171 descriptions of non-cancer and cancer risks are shown in Table S4 (supplementary 172 data). 173 In this study, the body weight (BD) of 60 kg for adults and 15 kg for children has 174 been chosen to reflect the current local situation (Lee et al., 1994). Ingestion rate (IngR) 175 of 100 and 200 mg/day were chosen for adults and children (Calabrese et al., 1987). In 176 addition, exposure duration (ED) of 25 years and 6 years for the adults and children 177 were also chosen (US EPA, 1991, 1989), with exposure frequency (EF) assumed to be 6 178 313 days/year, whereby workers are working all year round except for 1 day of holiday 179 per week and children are playing outdoors in the rural areas. Furthermore, exposure 180 time (ET) of 8 hours/day for continuous chronic exposure via the inhalation pathway 181 (US EPA, 2009) was chosen for adult workers and ET (3 hours/ day) was also assumed 182 for children to play outdoors in the rural areas. Surface area of skin that contacts the soil 183 (SA) was assumed to be 3300 and 2800 cm2/day for adult worker and children, 184 respectively (US EPA, 2013a). A life time (70 years) was chosen to calculate the 185 average time (AT) exposure for carcinogenic chemical exposure (US EPA, 1989). 186 187 188 189 190 191 192 193 194 195 196 197 198 199 200 201 7 202 203 A-D Y= +D B 204 205 1 + (X/C) Equation B (Luo et al., 2009; Qiao et al., 2006). 206 207 208 where Y is the EROD response, X is the inducer dose, A is the maximum EROD 209 response, B is the slope coefficient, C is the EC50, and D is the minimum EROD 210 response. EC50 is the concentration of TCDD eliciting 50% of the maximal inducible 211 EROD activity. 212 213 214 215 216 217 218 219 220 221 222 223 224 8 4 Log10 TEQbio (pg/g) 3 3 R² = 0.5908 2 2 1 0 1 2 3 4 5 Log10 TEQcal (pg/g) 225 226 Figure S1 Correlation of EROD assay or biologically derived TeCDD concentrations 227 (pg/g) (TEQbio) and chemically derived toxic equivalent concentrations (pg/g) of 228 dioxin-like chemicals (TEQcal (sum of 2,3,7,8-TeCDD toxic equivalent concentrations 229 (TEQPCDD/F) and dioxin-like PAHs toxic equivalent concentrations (TEQPAH)) in the 230 soil samples of 12 different types of land use. 9 100 TEQPCDD/F TEQPAH 99 98 97 96 W ) BS CD ) (O (D W EW BS O EW PS CW RW M CS (S ) Ab EW O A 95 F Percentage of TEQ PCDD/F and TEQPAH to theTEQcal 231 Different types of land use 232 233 Figure S2 Percentage of TEQPCDD/F and TEQPAH to the total TEQcal in different types 234 of land use 235 236 Note: OF = organic farm, A = agricultural, Ab = abandoned agricultural, EW (S) = 237 e-waste storage, CS = container storage, MRW = metal recycling workshop, CW = 238 constructions waste, PS = petrol station, OBS = open burning site, EW (DW) = 239 e-waste dismantling workshop, EW (OBS) = e-waste open burning site, CDW = car 240 dismantling workshop and TEQcal = sum of 2,3,7,8-TeCDD toxic equivalent 241 concentrations (TEQPCDD/F) and dioxin-like PAHs toxic equivalent concentrations 242 (TEQPAH)) 10 243 Table S1 Maximum concentrations of 16 individuals and total PAHs (μg/kg) amongst each 12 different types of land use (Man et al., 2013a). Sampling sites OF A Ab EW (S) CS MRW CW PS EW (OBS) OBS EW (DW) CDW Nap 207 313 364 203 317 11.6 159 7.61 312 21210 65 72.2 Any 6.81 9.16 14.6 3.57 2.72 19.9 7.2 3.06 70.1 39 37.2 26.6 Ane 1.7 1.96 4.25 0.787 2.44 1.11 0.999 0.707 14.8 38 3.13 12.1 Fle 9.74 20.9 35.2 10.5 17.5 3.7 9.35 0.942 134 179 10.3 20.8 Phe 56.3 101 148 50.7 81.6 55.8 229 12.2 607 80.9 231 191 Ant 3.42 10.8 17.7 3.36 5.21 14.9 7.42 3.58 104 12.8 62 22.4 16 individuals PAHs Fla Pyr BaA Chr 40.3 36.6 6.18 8 73.9 43.8 13.4 24.8 64.4 42.2 11.6 19.1 35.3 21 5.29 9.4 52.9 26.3 9.26 12 121 55.4 61.7 75.9 395 151 15 20.6 29.7 17.1 19.6 24.1 163 7.56 3.64 3.19 25.9 42.7 6.24 30 571 948 521 532 116 2214 96.4 7095 BbkF 0.5 38.7 17.2 0.54 5.55 70.2 59.7 19 3.93 187 1581 3841 BaP 6.43 23.2 14.7 6.6 10.5 83.9 30.9 27.7 2.81 228 695 80546 IcdP 11.7 28.1 16.9 7.51 10.3 178 45.5 55.6 N.D. N.D. 680 1696 DahA 1.68 5.2 3.75 0.713 2.8 52.2 9 15.1 N.D. N.D. 149 819 BghiP 15.6 32.5 20.9 8.61 16.4 115 65.3 43.1 N.D. N.D. 622 715 Total PAHs 412 741 794 366 572 919 1205 279 1426 22078 6708 97484 244 245 246 247 248 249 250 251 252 Note: OF = organic farm, A = agricultural, Ab = abandoned agricultural, EW (S) = e-waste storage, CS = container storage, MRW = metal recycling workshop, CW = constructions waste, PS = petrol station, OBS = open burning site, EW (DW) = e-waste dismantling workshop, EW (OBS) = e-waste open burning site, CDW = car dismantling workshop, Nap = naphthalene, Any = acenaphthylene, Ane = acenaphthene (Ane), Fle = fluorene, Phe = phenanthrene, Ant = anthracene, Fla = fluoranthene, Pyr = pyrene, BaA = benz(a)anthracene, Chr = chrysene, BaP = benzo(a)pyrene, BbF = benzo(b)fluoranthene, BkF = (BkF), IcdP = indeno(1,2,3-cd)pyrene, DahA = dibenz(a,h)anthracene, BghiP = benzo(g,h,i)perylene (BghiP) and N.D. = not detected 11 253 254 Table S2 Concentrations of chemically derived PCDD/Fs (pg/g and pgWHO-TEQ/g) in 12 different soils and soil quality guidelines of the Netherlands, Canada and Japan Congeners WHO-TEF OF PCDD congeners 2,3,7,8-TeCDD 1 0.434 1,2,3,7,8-PeCDD 1 1.3 1,2,3,4,7,8-HxCDD 0.1 1.47 1,2,3,6,7,8-HxCDD 0.1 3.64 1,2,3,7,8,9-HxCDD 0.1 3.26 1,2,3,4,6,7,8-HpCDD 0.1 78.2 OCDD 0.0003 974 ΣPCDDs-WHO TEQ 10.7 PCDD congeners 2,3,7,8-TeCDF 0.1 3.31 1,2,3,7,8-PeCDF 0.03 2.71 2,3,4,7,8-PeCDF 0.3 3.62 1,2,3,4,7,8-HxCDF 0.1 4.75 1,2,3,6,7,8-HxCDF 0.1 4.54 2,3,4,6,7,8-HxCDF 0.1 4.99 1,2,3,7,8,9-HxCDF 0.1 0.417 1,2,3,4,6,7,8-HpCDF 0.01 29 1,2,3,4,7,8,9-HpCDF 0.01 2.22 OCDF 0.0003 34 ΣPCDFs-WHO TEQ 3.29 ΣPCDD/Fs-WHO TEQ/ TEQPCDD/F 14 The New Dutch List (Dutch Indicative level - serious contamination) (VROM, 2000) Canadian Environmental Quality Guidelines (Agricultural and industrial ) (CCME, 2007) Environmental Quality Standard (MOE, 2003) 255 256 257 258 Sampling sites MRW CW A Ab EW (S) CS 0.288 0.363 0.463 1.3 2.01 40.9 1130 5.46 0.927 4.24 8.14 25.9 26.1 811 9450 95.1 0.4 0.915 1.26 3.01 4.79 60.9 1140 8.65 0.14 0.363 0.463 1.46 2.1 32.3 1230 4.5 0.14 1.14 1.24 3.65 2.51 73.6 1600 9.86 0.583 0.15 0.228 0.705 0.55 0.801 0.119 3.2 0.34 6.03 0.386 5.84 6.68 7.76 12.5 21.2 18.6 26.7 5.11 106 7.14 65.8 13 108 1 4 1000 1.22 0.63 1.15 0.45 1.03 0.532 1.33 0.705 1.44 0.653 1.33 0.705 0.37 0.119 5.81 3.45 0.34 0.34 4.99 5.17 0.975 0.494 9.63 5 WHO-TEQ WHO-TEQ WHO-TEQ 2.27 1.99 5.14 5.89 4.91 8.59 0.405 62.9 2.37 26.8 4.47 14.3 PS OBS EW (DW) EW (OBS) CDW 0.78 2.39 2.23 7.88 4.41 786 9030 85.9 0.14 0.363 0.154 0.3 0.613 11.1 439 1.85 23700 1210 1.75 4.1 4.94 38.2 1290 24915 15.8 3.55 2.53 6.01 4.8 161 3190 37.7 199 838 717 1380 1130 7150 10900 2078 4.36 7.1 4.32 187 81.9 871 1450 126 3.55 3.05 5.59 5.01 5.26 6.63 0.303 44.8 3.42 84.4 4.35 90.3 0.544 0.45 0.554 0.892 0.708 0.728 0.0397 3.11 0.34 3.02 0.506 2.36 24.4 1.77 10.8 2.66 1.2 5.05 0.119 6.04 0.34 5.33 6.7 24922 10.1 6.36 9.47 10.8 9.28 12.8 0.66 63.8 6.51 103 8.13 45.9 1150 1340 2370 2420 2330 3160 844 8860 870 2810 1840 3918 5.65 5.82 11.8 13.4 12.4 18.9 2.57 94.1 9.99 55.1 10.1 136 OF = organic farm, A = agricultural, Ab = abandoned agricultural, EW (S) = e-waste storage, CS = container storage, MRW = metal recycling workshop, CW = constructions waste, PS = petrol station, OBS = open burning site, EW (DW) = e-waste dismantling workshop, EW (OBS) = e-waste open burning site, CDW = car dismantling workshop, HpCDD = hetachlorodibenzodioxin, HpCDF = heptachlorodibenzofuaran, HxCDF = hexachlorodibenzofuaran, HxCDD= hexahetachlorodibenzodioxin, OCDD = octachlorodibenzodioxin, OCDF = octachlorodibenzofuaran, TeCDD = tetrahetachlorodibenzodioxin and TeCDF= tetrachlorodibenzofuaran 12 259 260 Table S3 Parameters for estimating human non-cancer and cancer risks for 2,3,7,8–TeCDD via the exposure pathway of ingestion, dermal contact and inhalation, and dermal absorption factor (ABS) of 2,3,7,8–TeCDD Exposure factors Ingestion rate, IngR (mg/day) Exposed skin area, SA (cm2) Skin adherence factor, AFsoil (mg/cm2) Exposure frequency, EF (days/yr) Exposure duration, ED (yr) Exposure time, ET (hours/day) Body weight, BW (kg) Averaging time, AT (ATncA) (days) = (25 yr × 365 days/yr) Averaging time, AT (ATncC) (days) = (6 yr × 365 days/yr) Averaging time, AT* (ATncA*) (hours) = (25 yr × 365 days/yr × 24 hours/day) Averaging time, AT *( ATncC*) (hours) = (6 yr × 365 days/yr × 24 hours/day) Averaging time, AT (ATcAC) (days) = (70 yr × 365 days/yr) Averaging time, AT (ATcAC*) (hours) = (70 yr × 365 days/yr × 24 hours/day) Reference dose (RfD), slope factors (SFs) and dermal absorption factor (ABS) of 2, 3, 7, 8 – TeCDD Non-cancer risk via the exposure pathway of ingestion (RfDo (mg/kg-day) ) Non-cancer risk via the exposure pathway of dermal contact (RfDo × GIABS (mg/kg-day) ) Non-cancer risk via the exposure pathway of inhalation (RfCi (mg/m3)) Cancer risk via the exposure pathway of ingestion (SFO (mg/kg-day)-1 ) Cancer risk via the exposure pathway of dermal contact (SFO × GIABS (mg/kg-day)-1 ) Cancer risk via the exposure pathway of inhalation (IUR (mg/m3)-1) Dermal absorption factor (ABS) Adult 100 3300 0.2 313 25 8 60 9,125 / 213,000 / 25,550 613,200 Child 200 2800 0.2 313 6 3 15 / 2,190 / 52,560 25,550 613,200 7.00E-10 7.00E-10 4.00E-08 1.30E+05 1.30E+05 3.80E-02 0.03 7.00E-10 7.00E-10 4.00E-08 1.30E+05 1.30E+05 3.80E-02 0.03 13 261 262 263 264 265 Note: IngR was taken from Calabrese et al. 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