Endocrine-disrupting chemicals, EDCs are of increasing interest from the viewpoint of health effects and human exposure as well as abnormality in wildlife reproductive systems. A nationwide survey of these EDCs was conducted to collect information on their existence in source and drinking waters in Japan. The analyzed 68 chemicals were as follows; 9 ethyl phthalates and similar compounds, 16 phenols, 8 styrenes, 2 organic tin compounds, 2 hormones, 26 pesticides and 5 other chemicals listed in SPEED '98 announced by the Environment Agency. Diethylhexyl phthalate (DEHP) was most frequently detected in 88% of 45 raw waters and 88% of 42 treated waters. Di-n-butyl phthalate (DBP) was found both in 26% of raw waters and in 12% of treated waters, as well. The maximum concentration of DEHP in treated water was 0.00026 mg/l, and that of DBP was 0.00018 mg/l. Bisphenol A (32%) and phenol (21%) were also common in raw waters, though they were hardly found in treated waters after chlorination. Among pesticides, pentachlorophenol (PCP) was found in 4 raw waters, with the maximum concentration of 0.00004 mg/l, and one treated water
Journal of Water and Environment Technology, Vol.2, No.1, 2004 - 17 - A NATIONWIDE SURVEY OF ENDOCRINE DISRUPTING CHEMICALS IN SOURCE AND DRINKING WATERS IN JAPAN S. Kunikane*, M. Ando**, T. Aizawa*, Y. Kanegaki*** * Department of Water Supply Engineering, National Institute of Public Health, 4-6-1 Shorokanedai, Minato-ku, Tokyo 108-8638 Japan ** Department of Environmental Chemistry, National Institute of Health Sciences, 1-18-1Kamiyoga, Setagaya-ku, Tokyo 158, Japan *** The Japan Water Research Center, 2-8-1 Toranomon, Minato-ku, Tokyo 105-0001, Japan ABSTRACT Endocrine-disrupting chemicals, EDCs are of increasing interest from the viewpoint of health effects and human exposure as well as abnormality in wildlife reproductive systems. A nationwide survey of these EDCs was conducted to collect information on their existence in source and drinking waters in Japan. The analyzed 68 chemicals were as follows; 9 ethyl phthalates and similar compounds, 16 phenols, 8 styrenes, 2 organic tin compounds, 2 hormones, 26 pesticides and 5 other chemicals listed in SPEED '98 announced by the Environment Agency. Diethylhexyl phthalate (DEHP) was most frequently detected in 88% of 45 raw waters and 88% of 42 treated waters. Di-n-butyl phthalate (DBP) was found both in 26% of raw waters and in 12% of treated waters, as well. The maximum concentration of DEHP in treated water was 0.00026 mg/l, and that of DBP was 0.00018 mg/l. Bisphenol A (32%) and phenol (21%) were also common in raw waters, though they were hardly found in treated waters after chlorination. Among pesticides, pentachlorophenol (PCP) was found in 4 raw waters, with the maximum concentration of 0.00004 mg/l, and one treated water. KEYWORDS Bisphenol; DEHP; drinking water; Endocrine disrupting chemicals; pesticides INTRODUCTION Synthetic chemicals having hormone-like effects increasingly draw scientific interests with respect to both wildlife and human health. These chemicals are suspected to cause abnormality in wildlife reproductive systems and similar symptoms eventually in human beings. They are called endocrine-disrupting chemicals, EDCs. Once they are released to the environment, they are widely spread over land, air and water, and some of them may be accumulated in plants and animals. In 1998, the Environment Agency announced its Strategy Plan for Endocrine Disrupters, SPEED '98 (also revised in 2000) including possible 67 endocrine disrupting chemicals (except metals). In order to clarify the occurrence of these EDCs and other chemicals in raw and treated waters for drinking water supply, the Ministry of Health and Welfare, Japan, conducted a nationwide survey of these chemicals and hormones originated from human beings and domestic animals, in cooperation with the Japan Water Supply Research Center from 1999 to 2000. MATERIALS AND METHODS Selection of sampling points The research group selected 45 surface and ground water treatment plants so as to inclusively clarify the current status of EDC contamination of raw and treated waters for drinking water supply all over Japan. As for Journal of Water and Environment Technology, Vol.2, No.1, 2004 - 18 - surface water, 22 large rivers were selected with respect to area distribution. As for groundwater, those for dinking water use were selected. Capacities of these water treatment plants in this study were mostly above 100,000 m 3 /day. Cautions were paid to include water treatment plants in any of major cities. These selected treatment plants are conducting periodical monitoring of monitoring items included in Japanese water quality standard. Sampling points were shown in Figure 1. Analyzed Chemicals Target chemicals are selected, with priority on EDCs selected in SPEED '98, domestic production amount and historical importance, with additional consideration of alkyl phenols. Ester phthalates listed in SPEED '98, individual alkyl phenols, despite those are expressed in total alkyl phenols in SPEED '98, individual styrenes, organic tin compounds, and hormones from human beings and livestocks were selected for further study. Pesticides of large production amount, that of prohibited production or prohibited use, and no-intended by-products were, included in the list as well. This research excluded such pesticides with no or least domestic production or those with no appropriate analytical method. Benzo (a) pylene and other normal industrial chemicals were also focused in this study. Dioxins were excluded from the scope of this paper. Sampling Dates The first samplings were conducted in summer, between July 27 and October 1, 1999 for raw waters and treated waters. The second samplings were between October 19 and December 1999 for raw water, so called winter. Samplings were prolonged when their turbidity was above 3 times higher than normal turbidity in raw water or their river flow was 2 times or higher than normal. Since the concentration of pesticides mainly depends upon their usage in the catchment area, timing of pesticide usage was also taken into consideration for sampling. Analytical Methods In total, 12 analytical methods were employed in this study, mainly depending upon chemical properties of the target substances. Analytical methods were designed to detect each group of chemicals by simultaneous detection methods with sufficient sensitivity. Analytical methods were summarized in Table 1. It basically follows 'Provisional Manual for EDCs Study' prepared by the Environment Agency, and it was revised for higher sensitivity in our research group. To minimize analytical variations between samples, samples were divided into 3 professional analytical centers and the same group of substances was analyzed in the same laboratory for all the samples. Quantitative detection limits, QDLs, were determined based on variances in each analytical method. The data detected below QDLs were expressed as "ND". Sufface water Groundwater Figure 1 45 Sampling Points (Raw water and treated water) Journal of Water and Environment Technology, Vol.2, No.1, 2004 - 19 - Basic water quality items, such as temperature, water temperature, turbidity, pH, KMnO 4 consumption, color, electric conductivity, alkalinity and residual chlorine (only for treated water), were also measured in each water treatment plant. Table 1 TARGET SUBSTANCES AND ANALYTICAL METHODS Group / substances Methods fundamentally employed 1. Volatile Chemicals Purge and trap GC/MS (DBCP, styrene monomer, n-butyl benzene) 2. Ester Phthalates 1000 times (x) concentration with hexane liquid-liquid extraction (LLE), GC/MS 3. Phenolic compounds Adjustment of low pH. 2000x hexane LLE, delivatized by BSTFA, GC/MS 4. Poly-brominated biphenyls 2000x hexane LLE, GC/MS-SIM 5. Styrenes and others 2500x hexane LLE, GC/MS 6. Pesticides 1000x with Solid phase extraction (SPE), GC/MS 7. Methomyl and carbaryl Direct injection to HPLC 8. Hydrophilic pesticides SPE or LLE, HPLC 9. MCPP and 2,4,5-T pH 3.5, SPE, methyl delivertization, hexane LLE, GC/MS 10. Hormones 2000x or 10000x SPE, LC/MS 11. Acephate 200x SPE, LC/MS-SIM 12. Amitrole 50x SPE, HPLC RESULTS AND DISCUSSION Occurrence of EDCs in raw water Table 2 shows the results of the survey for EDCs in raw and treated waters. Among 37 target EDCs, 3 ester phthalates and 10 phenols were detected. No styrenes, organic tin compounds, or hormones were detected from raw water. Out of 13 detected chemicals, diethylhexyl phthalate (DEHP) were detected with very high occurrences (79 samples, 88%). Bisphenol A (29 samples), di-n-butyl phthalate (DBP, 21 samples) and phenol (19 samples) were also common. Other substances were detected in only limited number of samples. The maximum concentration of DEHP was 0.00053 mg/l, bisphenol A was 0.00023 mg/l and DBP was 0.00065 mg/l. Seasonal difference of phenol was remarkable. It was detected only in one sample out of 45 samples in summer, however, detected in 18 samples out of 45 samples in winter. This is presumably attributable to bio- or self-degradation effects of phenol in the environment and difference of river flow affecting the dilution rate. Table 3 summarized the results from pesticides and other synthetic chemicals. Malathion, methomyl, and benomyl were detected. In spite of the prohibited production and usage for long time, hexachlorocychloro- hexane (HCH) and pentachlorophenol (PCP) was detected in raw waters even at very low concentration. It is notable that PCP was still detected in several raw waters, despite that; its registration had been eliminated since 1990. It demonstrates the potential use of PCP, impurities in other agricultural chemicals with PCP or high resistance in environment. Benzofenon was detected in three raw waters as well. It is utilized as a material to synthesize pharmaceuticals, but information is limited on its toxicity and environmental behavior. Suspected by-products from pesticides listed in Table 3 were not detected. No clear interrelation was found between detected chemicals and sampling area or size of water treatment plants. Almost all detected cases were those of surface water except that DEHP was universally found in ground water. Occurrence of EDCs in treated waters Journal of Water and Environment Technology, Vol.2, No.1, 2004 - 20 - In treated water samples, only diethylhexyl phthalate (DEHP) were detected with very high occurrences (37 samples, 88%), as shown in Table 2. Dibutyl phthalate (DBP), diethyl adipate (DEHA), phenol and 2-sec-butyl phenol were quantitatively detected in treated water, in 5, 2, and 1 of the samples, respectively. The maximum concentration of DEHP was 0.00026 mg/l, and that of DBP was 0.00018 mg/l. The concentrations of DEHP was much lower than 0.008 mg/l, the WHO guideline value of DEHP for drinking water quality, and 0.06 mg/l, the guideline value for monitoring items in Japanese drinking water standard, though these guideline values were derived from conventional chronic toxicity studies. Among phenolic compounds, only phenol and 2-sec-butyl phenol were detected respectively in one sample. HCH and PCP were detected respectively in one treated water sample at very low concentration, 0.00001 mg/l Table 2 EDCs in raw and treated waters by simultaneous measuring methods mg/l mg/l Summer Winter Total mg/l 1 Diethylhexyl Phthalate, DEHP 0.00005 ND ～ 0.00053 42/45 37/45 79/90 ND ～ 0.00026 37/42 2 Diethyl Phthalate 0.00005 ND 0/45 0/45 0/90 ND 0/42 3 Di-n-buthyl phthalate, DBP 0.00005 ND ～ 0.00065 13/45 8/45 23/90 ND ～ 0.00018 5/42 4 Buthylbenzyl Phthalate 0.00005 ND 0/45 0/45 0/90 ND 0/42 5 Di-2-ethylhexyl adipate, DEHA 0.00001 ND ～ 0.00002 1/45 0/45 1/90 ND ～ 0.00002 2/42 6 Dicyclohexyl phthalate 0.00005 ND 0/45 0/45 0/90 ND 0/42 7 Di-n-propyl phthalate 0.00005 ND 0/45 0/45 0/90 ND 0/42 8 Dipentyl phthalate 0.00005 ND 0/45 0/45 0/90 ND 0/42 9 Di-n-hexyl phthalate 0.00005 ND 0/45 0/45 0/90 ND 0/42 10 Bis-phenol A 0.00001 ND ～ 0.00023 14/45 15/45 29/90 ND 0/42 11 2,4-Dichlorophenol 0.00001 ND ～ 0.00003 0/45 1/45 1/90 ND 0/42 12 Phenol 0.00001 ND ～ 0.00005 1/45 18/45 19/90 ND ～ 0.00001 1/42 13 4-Ethylphenol 0.00001 ND ～ 0.00021 0/45 1/45 1/90 ND 0/42 14 2-tert-Buthylphenol 0.00001 ND ～ 0.00002 1/45 0/45 1/90 ND 0/42 15 2-sec-Buthylphenol 0.00001 ND ～ 0.00002 2/45 0/45 2/90 ND ～ 0.00002 1/42 16 3-tert-Buthylphenol 0.00001 ND 0/45 0/45 0/90 ND 0/42 17 4-tert-Buthylphenol 0.00001 ND ～ 0.00002 0/45 1/45 1/90 ND 0/42 18 4-sec-Buthylphenol 0.00001 ND 0/45 0/45 0/90 ND 0/42 19 4-Octyl phenol 0.00001 ND 0/45 0/45 0/90 ND 0/42 20 4-tert-Octyl phenol 0.00001 ND ～ 0.00001 1/45 1/45 2/90 ND 0/42 21 Nonyl phenol 0.0001 ND 0/45 0/45 0/90 ND 0/42 22 4-n-Nonyl phenol 0.00001 ND 0/45 0/45 0/90 ND 0/42 23 2-Hydroxybiphenyl 0.00001 ND ～ 0.00001 1/45 0/45 1/90 ND 0/42 24 3-hydroxybiphenyl 0.00001 ND 0/45 0/45 0/90 ND 0/42 25 4-hydroxybiphenyl 0.00001 ND ～ 0.00001 0/45 1/45 1/90 ND 0/42 26 Octachlorostyrene 0.00003 ND 0/45 0/45 0/90 ND 0/42 27 1,3-Diphenylpropane 0.00001 ND 0/45 0/45 0/90 ND 0/42 28 cis-1,2-Diphenylcyclobutane 0.00001 ND 0/45 0/45 0/90 ND 0/42 29 trans-1,2-diphenylcyclobutane 0.00001 ND 0/45 0/45 0/90 ND 0/42 30 2,4-Diphenyl-1-butene 0.00001 ND 0/45 0/45 0/90 ND 0/42 31 2,4,6-Triphenyl-1-hexene 0.00001 ND 0/45 0/45 0/90 ND 0/42 32 1e-Phenyl-4e （ 1'-Phenylethyl ） tetralin 0.00001 ND 0/45 0/45 0/90 ND 0/42 33 Stylene monomer 0.00001 ND 0/45 0/45 0/90 ND 0/42 34 Tributyltin 0.000005 ND 0/45 0/45 0/90 ND 0/42 35 Triphenyltin 0.000001 ND 0/45 0/45 0/90 ND 0/42 36 Ethinyl estradiol 0.000002 ND 0/45 0/45 0/90 ND 0/42 37 17β-Estradiol 0.000002 ND 0/45 0/45 0/90 ND 0/42 Note1: ND means 'not detected above Quantitative Detection Limits, QDLs'. Diethylestel phthalate is also included in monitoring items in drinking-water quality standard. Stylenes Note2: Detected/ Measured Substance Esteyl PhthalatesPhenols Hormone Organo tins No. QDLs Raw water Treated water Range detected Detected ／ Measured Range detected Journal of Water and Environment Technology, Vol.2, No.1, 2004 - 21 - for both compounds. The detected concentration is sufficiently lower than 0.002 mg/l for gamma-HCH and 0.009 mg/l for PCP with respect to the WHO guidelines for drinking water quality. Origin of these chemicals The most frequently detected compound, DEHP, is mainly used as plasticizer for vinyl chloride products. Total production of DEHP is above 300 kt in 1997. It hardly self-degrades in water environment, however, quick bio-degradation is expected in aerobic condition. DBP and DEHA are also used as a morphing chemical for vinyl chloride and plastics. It was reported that DEHP and DBP were contained and eluted from epoxy resins used for coating of water treatment facilities and distribution pipes (Kunikane, 1999), which indicates possibilities of contamination to occur during treatment. Since ester phthalates are also frequently contained in air and commercial products for daily use, it is difficult to conclude the source of contamination. Bisphenol is prevalently used in polycarbonate resins, epoxy resins, phenol resins, polyester, oxidation prohibitor and stabilizing agent for vinyl chloride. It was detected in high frequency in raw water, however, not found in that form in treated water. Since it has hydrophobic property with log Pow, 3.3-3.8, sort of Table 3 EDC Pesticides in raw water and treated water mg/l mg/l First Second Total mg/l 38 Benzoepin/Endosulfan 0.00005 ND 0/45 0/45 0/90 ND 0/42 39 Malathion 0.00001 ND ～ 0.00003 1/45 0/45 1/90 ND 0/42 40 Methomyl 0.00001 ND ～ 0.00013 3/45 0/45 3/90 ND ～ 0.00001 1/42 41 Benomyl (as MBC) 0.0001 ND ～ 0.0002 0/45 2/45 2/90 ND 0/42 42 Carbaryl 0.00001 ND 0/45 0/45 0/90 ND 0/42 43 Alachlor 0.00001 ND 0/45 0/45 0/90 ND 0/42 44 Trifluralin 0.00005 ND 0/45 0/45 0/90 ND 0/42 45 Hexachlorobenzene 0.00005 ND 0/45 0/45 0/90 ND 0/42 46 o,p'-DDT, p,p'-DDT 0.00005 ND 0/45 0/45 0/90 ND 0/42 47 Ardrin 0.00005 ND 0/45 0/45 0/90 ND 0/42 48 Endrin 0.00005 ND 0/45 0/45 0/90 ND 0/42 49 Dieldrin 0.00005 ND 0/45 0/45 0/90 ND 0/42 50 Heptachlor 0.00005 ND 0/45 0/45 0/90 ND 0/42 51 trans, cis-Chlordane 0.00005 ND 0/45 0/45 0/90 ND 0/42 52 Methoxychlor 0.00005 ND 0/45 0/45 0/90 ND 0/42 53 Hexachlorocychlorohexane, HCH 0.00001 ND ～ 0.00002 1/45 0/45 1/90 ND ～ 0.00001 1/42 54 Amitrole 0.00005 ND 0/45 0/45 0/90 ND 0/42 55 2,4,5-Trichlorophenoxyacetic acid 0.00005 ND 0/45 0/45 0/90 ND 0/42 56 1,2-Dibromo-3-chloropropane 0.00001 ND 0/45 0/45 0/90 ND 0/42 57 Nitrofen 0.00005 ND 0/45 0/45 0/90 ND 0/42 58 Pentachlorophenol, PCP 0.00001 ND ～ 0.00004 2/45 2/45 4/90 ND ～ 0.00001 1/42 59 tarns-Nonachlor 0.00005 ND 0/45 0/45 0/90 ND 0/42 60 p,p'-DDE, o,p'-DDE 0.000001 ND 0/45 0/45 0/90 ND 0/42 61 p,p'-DDD, o,p'-DDD 0.00001 ND 0/45 0/45 0/90 ND 0/42 62 Heptachlor epoxide 0.00005 ND 0/45 0/45 0/90 ND 0/42 63 Oxychlordane 0.00005 ND 0/45 0/45 0/90 ND 0/42 64 Benzo(a)pylene 0.00001 ND 0/45 0/45 0/90 ND 0/42 65 Polybrominated biphenyls 0.00001 ND 0/45 0/45 0/90 ND 0/42 66 4-Nitroroluene 0.00001 ND 0/45 0/45 0/90 ND 0/42 67 n-Butylbenzene 0.00001 ND 0/45 0/45 0/90 ND 0/42 68 Benzophenone 0.00001 ND ～ 0.00002 1/45 2/45 3/90 ND 0/42 Note1: "ND" means 'not detected above Quantitative Detection Limits, QDLs'. No-intended By-product Other organics Substance No. Major Pesticides Pestices of Prohibited Commercial Use or Manufacuring QDLs Raw water Treated water Range detected Detected ／ Measured Range detected Detected/ Measured Journal of Water and Environment Technology, Vol.2, No.1, 2004 - 22 - adsorption may occur in water treatment process, though, another possibility is shown in a part of our study (Hu et al, 2000). Bisphenol is very reactive with hypochlorite to form chlorinated bisphenols. Because chlorination process is obligatorily adopted for disinfection at the end of water treatment process in Japan, these reactive chemicals are supposed to yield chlorinated by-products. In addition, behavior of these chemicals in distribution network and water supply devices are important, especially in the existence of residual chlorine, because phenolic compounds are highly reactive with chlorine to form other chlorinated compounds which have endocrine affecting potential. CONCLUSION From our nationwide survey of raw and treated waters for drinking water supply, 19 chemicals were detected in raw waters and 8 chemicals were detected in treated waters out of 68 suspected EDCs. Ester phthalates, phenolic compounds, pesticides were detected in raw waters. Although ester phthalates were frequently found in treated waters, other chemicals were detected in only limited samples at low concentration. Among treated water samples, diethylhexyl phthalate (DEHP) were detected with very high occurrences (37 samples, 88%). Since the Ministry of Environment has been conducting risk assessment of DEHP and some other EDCs from 2000 (Ueda, 2000), risk evaluation of these chemicals should be conducted in a few years. The results of this research can be obtained via Internet; http://www.mizudb.or.jp/index.html. Acknowledgements The authors express special thanks to the drinking water supplies participated in this research. REFERENCES Hu J., Ohkubo S., Aizawa T. (2000). Evaluation on endocrine disrupting activity of by-products of phenolic chemicals. Proceeding of Japan Society on Water Environment the 34 th Annual Conference, 573, Kyoto, Japan. Kunikane S. (1999). Research on exposure of EDCs via drinking water. Grant-in-aid Research Report, the Ministry of Health and Wealfare, Japan. Ueda H. (2000). Current strategies against Environmental Endocrine Disrupters by the Ministry of Environment of Japan, International Symposium on Environmental Endocrine Disruptors 2000, pp.22-23, Yokohama, Japan. . 2- tert-Buthylphenol 0.00001 ND ～ 0.000 02 1/45 0/45 1/90 ND 0/ 42 15 2- sec-Buthylphenol 0.00001 ND ～ 0.000 02 2/45 0/45 2/ 90 ND ～ 0.000 02 1/ 42 16 3-tert-Buthylphenol 0.00001. 0/45 0/90 ND 0/ 42 20 4-tert-Octyl phenol 0.00001 ND ～ 0.00001 1/45 1/45 2/ 90 ND 0/ 42 21 Nonyl phenol 0.0001 ND 0/45 0/45 0/90 ND 0/ 42 22 4-n-Nonyl phenol