邻苯二甲酸酯、双酚A在血液中的环境类激素效应研究
|
摘要
环境类激素是一类外源性干扰生物和人体正常内分泌机能的化学物质，也被称为内分泌干扰物质。它对生物体内分泌系统的复合干扰损害作用，主要包括对生物的生殖与发育系统的干扰或致畸影响，内分泌系统紊乱或免疫功能改变，致突变或致癌性影响，神经系统失调等现象。性激素作为类固醇激素，在体内的作用途径可分为血液—细胞质—细胞核三个环节，任何一个环节中类固醇激素的作用受到干扰，都将影响到类固醇激素在体内的正常调节作用。
目前，环境当中存在的能够证明或间接证明属于环境激素类的物质有70种。本文选取了两种已被确认的环境激素—邻苯二甲酸二丁酯(DBP)和双酚A(BPA)为研究对象，采用荧光光谱分析法，测定了DBP和BPA在水溶液中与雌激素(E_2)／雄激素(睾酮)和血清蛋白组分间的二元、三元体系结合常数，分析了二者在血液中引起的雌、雄激素平衡比例的变化。
荧光分析结果表明：DBP(BPA)血清蛋白组分和E_2(睾酮)之间能两两结合。当E_2、DBP和血清蛋白组分处于同一体系中，随着DBP浓度的增加，E_2与血清蛋白组分—DBP二元体系间的结合常数也逐渐增大，其最大值约为正常E_2与血清蛋白组分结合常数的3倍；在睾酮、DBP和血清蛋白组分的体系中，随着DBP浓度的增加，睾酮与血清蛋白组分—DBP二元体系的结合常数也呈现上升趋势，其最大值约为正常睾酮与血清蛋白组分结合常数的4倍。这表明DBP的存在可能不会与E_2和睾酮对血清蛋白组分进行竞争结合，但由于本身的结合特点，会减少血液中游离E_2和睾酮的含量，进而减少进入细胞内E_2和睾酮的量，同时DBP还会引起血液中雌、雄激素平衡比例的失调。雌、雄个体分组实验结果表明：DBP使雌性个体血液中游离E_2和睾酮的平衡比例降低，而对雄性个体作用不显著。
实验结果还表明：当E_2、BPA和血清蛋白组分处于同一体系中，随着BPA浓度的增加，E_2与血清蛋白组分—BPA二元体系间的结合常数也逐渐增加，其最大值高出正常E_2与血清蛋白组分结合常数约一个数量级；在睾酮、BPA和血清蛋白组分的体系中，随着BPA浓度的增加，睾酮与血清蛋白组分—BPA二元体系的结合常数明显上升，其最大值高出正常睾酮与血清蛋白组分结合常数近一个数量级。雌、雄个体分组实验结果表明：BPA使雄性个体血液中游离E_2和睾酮的平衡比例升高，而对雌性个体作用不显著。
本文研究结果表明：BPA在血液中的环境类激素效应明显强于DBP，而且二者在血液中对E_2和睾酮有着相似的干扰作用，它们会使血液中E_2和睾酮的结合量增大，从而限制血液内游离E_2和睾酮的浓度，减少进入细胞内E_2和睾酮的量，进而使细胞内E_2和睾酮的平衡受到影响。
In this dissertation, the combination of estradiol (E2), testosterone (T), dibutyl phathalate (DBP), bisphenol-A (BPA) and serum components had been determined in the binary and ternary systems by fluorescence method.
The results of fluorimetry showed that DBF (BPA), serum components and estradiol (testosterone) can combinate one another. In the E2-DBP-semm components ternary system, the combination constants of estradiol and binary system (DBP-serum components) would increase gradually with increasing the concentration of DBP, the maximum of combination constant is about three times of ordinary combination constant of binary system (E2-serum components). In the system of testosterone, DBP and serum components, with the increase of DBP concentration, the combination constants of testosterone and DBP-serum components showed a increase trend, the maximum can be four times of the combination constant of T-semm components. The result indicates that DBP may not compete with estradiol and testosterone for binding to serum components. On the contrary, because of the binding characteristic of DBP, it can decrease the free content of estradiol and testosterone in blood, and influence the balance of sex hormone in blood. The experiment showed that DBP make the balance proportion lower in female blood. But it has little effect on male.
In the E2-BPA-serum components ternary system, the maximum of combination constant is about ten times higher than ordinary combination constant of binary system (Ej-serum components). In the system of testosterone, BPA and serum components, with the increase of BPA concentration, the combination constants of testosterone and BPA-serum components would increase apparently, and the maximum can also be ten times higher than the combination constant of T-serum components. The result also indicates that BPA make the balance proportion higher in male blood. But it has little effect on female.
The paper showed that the hormone mimic effect of BPA is stronger than the effect of DBF. Moreover, both of them have same disturb function on estradiol and testosterone in blood. They can make the binding content of estradiol and testosterone increase in blood. Then they restricted the free concentration of estradiol and testosterone in blood and reduce the content of estradiol and testosterone which entered the cell. Finally, the balance of estradiol and testosterone is affected in cell.
目前，环境当中存在的能够证明或间接证明属于环境激素类的物质有70种。本文选取了两种已被确认的环境激素—邻苯二甲酸二丁酯(DBP)和双酚A(BPA)为研究对象，采用荧光光谱分析法，测定了DBP和BPA在水溶液中与雌激素(E_2)／雄激素(睾酮)和血清蛋白组分间的二元、三元体系结合常数，分析了二者在血液中引起的雌、雄激素平衡比例的变化。
荧光分析结果表明：DBP(BPA)血清蛋白组分和E_2(睾酮)之间能两两结合。当E_2、DBP和血清蛋白组分处于同一体系中，随着DBP浓度的增加，E_2与血清蛋白组分—DBP二元体系间的结合常数也逐渐增大，其最大值约为正常E_2与血清蛋白组分结合常数的3倍；在睾酮、DBP和血清蛋白组分的体系中，随着DBP浓度的增加，睾酮与血清蛋白组分—DBP二元体系的结合常数也呈现上升趋势，其最大值约为正常睾酮与血清蛋白组分结合常数的4倍。这表明DBP的存在可能不会与E_2和睾酮对血清蛋白组分进行竞争结合，但由于本身的结合特点，会减少血液中游离E_2和睾酮的含量，进而减少进入细胞内E_2和睾酮的量，同时DBP还会引起血液中雌、雄激素平衡比例的失调。雌、雄个体分组实验结果表明：DBP使雌性个体血液中游离E_2和睾酮的平衡比例降低，而对雄性个体作用不显著。
实验结果还表明：当E_2、BPA和血清蛋白组分处于同一体系中，随着BPA浓度的增加，E_2与血清蛋白组分—BPA二元体系间的结合常数也逐渐增加，其最大值高出正常E_2与血清蛋白组分结合常数约一个数量级；在睾酮、BPA和血清蛋白组分的体系中，随着BPA浓度的增加，睾酮与血清蛋白组分—BPA二元体系的结合常数明显上升，其最大值高出正常睾酮与血清蛋白组分结合常数近一个数量级。雌、雄个体分组实验结果表明：BPA使雄性个体血液中游离E_2和睾酮的平衡比例升高，而对雌性个体作用不显著。
本文研究结果表明：BPA在血液中的环境类激素效应明显强于DBP，而且二者在血液中对E_2和睾酮有着相似的干扰作用，它们会使血液中E_2和睾酮的结合量增大，从而限制血液内游离E_2和睾酮的浓度，减少进入细胞内E_2和睾酮的量，进而使细胞内E_2和睾酮的平衡受到影响。
In this dissertation, the combination of estradiol (E2), testosterone (T), dibutyl phathalate (DBP), bisphenol-A (BPA) and serum components had been determined in the binary and ternary systems by fluorescence method.
The results of fluorimetry showed that DBF (BPA), serum components and estradiol (testosterone) can combinate one another. In the E2-DBP-semm components ternary system, the combination constants of estradiol and binary system (DBP-serum components) would increase gradually with increasing the concentration of DBP, the maximum of combination constant is about three times of ordinary combination constant of binary system (E2-serum components). In the system of testosterone, DBP and serum components, with the increase of DBP concentration, the combination constants of testosterone and DBP-serum components showed a increase trend, the maximum can be four times of the combination constant of T-semm components. The result indicates that DBP may not compete with estradiol and testosterone for binding to serum components. On the contrary, because of the binding characteristic of DBP, it can decrease the free content of estradiol and testosterone in blood, and influence the balance of sex hormone in blood. The experiment showed that DBP make the balance proportion lower in female blood. But it has little effect on male.
In the E2-BPA-serum components ternary system, the maximum of combination constant is about ten times higher than ordinary combination constant of binary system (Ej-serum components). In the system of testosterone, BPA and serum components, with the increase of BPA concentration, the combination constants of testosterone and BPA-serum components would increase apparently, and the maximum can also be ten times higher than the combination constant of T-serum components. The result also indicates that BPA make the balance proportion higher in male blood. But it has little effect on female.
The paper showed that the hormone mimic effect of BPA is stronger than the effect of DBF. Moreover, both of them have same disturb function on estradiol and testosterone in blood. They can make the binding content of estradiol and testosterone increase in blood. Then they restricted the free concentration of estradiol and testosterone in blood and reduce the content of estradiol and testosterone which entered the cell. Finally, the balance of estradiol and testosterone is affected in cell.
引文
1. Tyler C R, Jobling S, Sumpter J E Endocrine disruption in wildlife : A critical review of the evidence. Critical Reviews/n Toxicology, 1998, 28 (4): 319-361.
2. Harries J E, Janbakhsh A, et al. Estrogenic potency of effluent from two sewage treatment works in the United Kingdom. Environ. Toxicol Chem., 1999,18(5): 932.
3．周庆祥，江桂斌．浅谈环境内分泌干扰物质．科技术语研究，2001，3(3)：12～14．
4．邓茂先，陈祥贵．环境内分泌干扰物研究进展．国外医学卫生学分册，2000，27(2)：65～68．
5. Workshop sponsored UN Environment programme, U S EPA, White House Office of Science and Technology Policy, et al. International workshop on endocrine disruptors, 1997, 1.
6. ICPS, OECD. International programme on chemical safety report of ICPS/OECD scoping meeting on endocrine disruptors (EDs). US Washington D. C, 1998,3.
7 出云逾明．威胁人类存亡的定时炸弹—环境荷尔蒙．深圳：海天出版社，1999．
8．任仁．环境激素的种类和污染途径．大学化学，200 1(5)：28～32．
9．井口泰泉．环境荷尔蒙问题的现状及其今后动向．98’中日环境测试技术与环境管理研讨会，1998．
10. Kelce W, Stone C, et al. Persistent DDT metabolite p, p'-DDE is a potent androgen receptor antagonist. Nature, 1996 (375): 581～585.
11. Stone R. Environmental estrogens stir debate Science, 1994 (265): 308～310.
12. Hutchinson T H, Brown R, Brugger K E, et al. Ecological risk assessment of endocrine disruptors. Environ. Health Perspect, 2000, 108 (11): 1007～1014.
13. Cheek A O, Vonier P M, et al. Environmental aignaling: A biological context for endocrine disruption. Environ. Health Perspect, 1998, 106 (Suppl. 1): 5～10.
14. Routledge E J, Sheahan D, et al. Identification of estrogenic chemicals in STW effluent. In vivo responses in trout and roach. Environ. Sei. Technol., 1998 (32): 1559～1565.
15. Paflak M. A testing deadline for endocrine disruptors. Environ. Sci. Technol., 1996 (30): 540A～544A.
16．郝明德，刘晓宏．环境激素的危害与预防方法．世界农业，1999(9)：42～43．
17．香山不二雄．环境激素问题研究现状．世界环境，1999(2)：34～35．
18. Allen Y, et al. The extent of oestrogenic contaminants in the UK estuarine and marine environments- further surveys of flounder. The Science of the Total Environment, 1999 (233): 5～20.
19. Key SW. New study highlights hazardson hormone disrupting chemicals. World Oisease (Weekly Plus), 1998 (9): 11～12.
20．小林秀幸．内分泌扰乱化学物质问题．环境管理，1998，34(11)：1～7．
21. Ternes TA, Stumpf M, et al. Behavior and occurrence of estrogens in municipal sewage treatment plants-Ⅰ. Investigations in Germany, Canada and Brazil. The Science of the Total Environment, 1999 (225): 81～90.
22．武木田薰，井上述．内分泌扰乱化学物质对生物体影响．环境管理，1998，34(11)：8～14．
23. Harries JE, et al. Estrogeuic potency of effluent from two sewage treatment works in the United Kingdom. Environmental Toxicology and Chemistry, 1999, 18 (5): 932～937.
24．王兰，蒋学之，顾祖维编著．环境与生殖．上海：上海医科大学出版社，1994．
25．高京生．环境激素．中国检验医学与临床，2001，2(3)：97～99．
26．夏星辉，杨居荣，许嘉琳．环境激素污染研究进展．上海环境科学，2001，20(2)：56～59．
27．邱东茹，吴振斌，贺铎．内分泌扰乱化学品对动植物的影响和作用机制．环境科学研究，2000，13(6)：52～55．
28．陈岚，王欣．环境激素及其对生物的危害．山西中医学院学报，2002，3(3)：45～46．
29．王毓秀，张利民，邹敏．化学农药与环境激素．农村生态环境，1999，15(4)：37～41．
30．周宜开．环境激素导致阴盛阳衰?中国环境报，1999(8)：31．
31．邓苹译．环境荷尔蒙为何会给人类带来危机．环境科学，1998(12)：26～29．
32．江田汉，杨晓明，赖玉平．环境激素问题浅析．中国环境管理，2001(1)：43～45．
33. John A. Mclachlan, Arnold S F, Klotz D M, et al. Synergistic Activations of Estrogen Receptor with Combinations of Environmental Chemicals. Science, 1996 (272): 1489-1492.
34．曹信孚译．国外环境激素问题的研究动态．广州环境科学，2002，17(1)：46～48．
35. Colbom T, Dumaoski D, Meyers J P. Our Stolen Future, New York: Penguin Books, 1996.
36. Gillesby BE and Zacbarewski TR. Exoestrogens: mechanisms of action and strategies for identification and assessment. Environmental Toxicology and Chemistry, 1998, 17 (1): 3～14.
37. Scott Alex UK. Industry prepares for tough rules. Chemical Week, 1998, 160 (2): 22.
38．杜克久，储少岗等．用MCF-7细胞增殖法研究国产多氯联苯的类雌激素效应．环境科学学报，2000，20(2)：230～233．
39．赵丙顺，邹继超等．小鼠子宫增重法检测国产三氯杀虫螨醇的雌激素生物括性．环境科学学报，2000，20(2)：244～247．
40. Zaeharewski T. In vitro bioassays for assessing eslrogenic substances. Environmental Science and Technology, 1997, 31 (3): 613～623.
41. Bolger R, Wiese TE and Ervin K, et al. Rapid Screening of environmental chemicals for estrogen receptor binding capacity. Environmental Health Perspectives, 1998, 106 (9): 551～557.
42. Safe SH and Gaido K. Phytoestrogens and anthropogenic estrogenic compounds. Environmental Toxicology and Chemistry, 1998, 17 (1): 119～126.
43. Andersen HR, Andersson Anna-Maria, et al. Comparison of short-term estrogenicity tests for identification of hormone-disrupting chemicals. Environmental Health Perspectives, 1999, 107 (supplement 1): 89～100.
44．Olsson P,Westcrlund L and Teb SW, et al. 母体对雌激素暴露和PCB对斑马鱼(Danio rerio)不同生活阶段的影响．AMBIO，1999，28(1)：100～106．
45. Brown RJ, Conradi M and Depledge MH. Long-term exposure to 4-nonylphonel affects sexual differentiation and growth of amphlpod corphium volutator(Pallas,1766). The Science of the Total Environment, 1999 (233): 77～88.
46. Reitrer LW, Derosa C, et al. The U. S. Framework for research on endocrine disruptors and an analysis for research programs supported during fiscal year 1996. Environmental Health Perspectives, 1998, 106 (3): 105～117.
47. Karcher W. Recent Trends and developments m the EU in the environmental control and management of chemicals. Eeotoxicology and Environmental Safety, 1998 (40): 97～102.
48. Shi LM, Fang H and Tong W, et al. QSAR models using a large diverse set of estrogeus. J Chem InfCompol Sci., 2001 (41):168～195.
49. Kareher W, Hansen BG, et al. Predictions for existing chemicals- a multilateral QSAR project. SAR and QSAR Environmental Research, 1995 (3): 217～221.
50. Hermeas J, Balaz S, et al. Assessment of QSAR for predicting fate and effects of chemicals in the environment: an international European project. SAR and QSAR Environmental Research, 1995 (3): 223-236.
51. Clark LB and Rosen RT, et al. Determination of alkylphenol ethoxylates and their acetic and derivatives in drinking water by particle beam liquid chromatograph/mass spectrometry. Int J Anal. Chem., 1992 (47): 167～180.
52．齐文启，孙宗光．环境荷尔蒙物质及其检测分析．环境科学，1999，18(12)：531～533．
53．吴平谷，韩关根，王惠华等．饮用水中邻苯二甲酸酯类的调查．环境与健康杂志，1999，16(6)：338～339．
54．丁鹏，赵晓松等．酞酸酯类化合物(PAES)研究新进展．吉林农业大学学报，1999，21(3)：119～123,128．
55．叶常明．环境中的邻苯二甲酸酯．环境科学进展，1993，1(2)：36～45．
56. Klamsmeier RE. In: I. E. Liener (Ed). Microbial Biodeterioration. Academic Press, London, 1981: 431～471.
57．石万聪编．增塑剂．北京：化学工业出版社，1989：60．
58. Atlas E and Giam CS. Global transport of organic; pollutants: ambient concentration in the remote marine atmosphere. Science, 1981 (211): 163～165.
59．周文敏等编．环境优先污染物．北京：中国环境科学出版社．1989：48．
60．金相灿等编．有机化合物污染化学-有毒有机物污染化学．北京：清华大学出版社，1990：266～275．
61．庞金梅等．苯二甲酸酯的微生物降解与转化．环境科学，1993，18(3)：88～90．
62. Yin Mei-chin et al. Znvestigation on risk of phthalate ester in drink water and marked foods. Medicines Food Analysis, 1996, 4 (4): 313～318.
63．赵振华．我国环境中酞酸酯污染的研究．环境科学丛刊，1986，7(5)：50～56．
64. Sehiedek Thomas. Impact of plasticizers (phthalicacid esters) on soil and groundwater quality. IAHS Publ, 1995 (225): 149～156.
65. Nagini S and Selvam S. Phthalate ester plasticizers- A review. Trends Life Sci., 1994, 9 (1): 9～16.
66. Wei wende. A Complete Collection of Organic Raw Materials in Chemical Industry. Vol.Ⅲ, Beijing: Chemical Industry Press, 1990.
67．刘程，江小梅等．表面活性剂应用大全．北京：北京工业大学出版社，1992：113．
68．王跃志．化工进展，2000，19(4)：965～967．
69. Olea N, Pulgar R, et al. Environ. Health Perspect., 1996 (104): 298～305.
70. Shintani H J. Liq. Chromatogr., 1995, 18 (3): 613～626.
71. Ben-Jonathan N, Steinmetz R. Trends Endocrinol. Metab., 1998, 9 (3): 124～128.
72. Brotons J A, Oles-Serraao M F, et al. Environ. Health Perspect., 1995 (103): 608～612.
73. Soto A M, Justicia H, et al. Environ. Health Perspect., 1991 (92): 167.
74. Krishnan A V, Stathis P, et al. Endocrinology, 1993 (132): 2279～2286.
75. Bitman J, Cecil H C, et al. Science, 1969 (162): 371～372.
76. Sonnenschein C, Soto Am. Journal of Steroid Biochemistry and Molecular Biology, 1998, 65 (1-6): 143～150.
77. Jobling S. Pure and Appl. Chem., 1998,70 (9): 1805～1827.
78. Thiele B, Guenther K, et al. Chem. Rev., 1997, 97 (8): 3247～3272.
79. Murray, H.E., Ray, L.E., et al. Chemosphere, 1981 (10): 1327.
80. Gledhill, W.E., Kaley, R.C., et al. Environ. Sci. Technol., 1980 (14): 301.
81. Kurane, R., Suzaki, T., et al. Aqric. Biol. Chore.,1980 (44): 529.
82．赵振华．酞酸酯对人和环境潜在危害的研究概况．环境化学，1991，10(3)：64～68．
83. Mol H G J, Sunarto S, et al. Chromatogr. A, 2000 (879): 97～112.
84. Van der Kraak G. Pure and Appl. Chem., 1998, 70 (9): 1785～1794.
85．赵美萍，李元宗等．酚类环境雌激素的分析研究进展．分析化学，2003，31(1)：103～109．
86．靳翠红，赵剑等．双酚A对雄性小鼠生殖系统的影响．中国医科大学学报，2002，31(2)：123～124．
87. Staples C A, Dora P B, et al. Bisphenol A concentrations in receiving water near US manufacturing and processing facilities. Chemosphere, 2000 (40): 521～525.
88. Belfroid A C, et al. Analysis and occurrence of estrogenic hormones and their glucuronides in surface water and waste water in the Netherlands. The Science of the Total Environment, 1999 (225): 101～108.
89. Desbrow C and Routledge E J, et al. Identifieation of estrogenic chemicals in STW effluent. Ⅰ Chemical fractionation and in vitro biological screening. Environmental Science and Technology, 1998, 32 (11): 1549～1557.
90. Del Olmo M, Zafra A, et al. Int. J. Environ. Anal. Chem., 1998, 69 (1): 99～110.
91. Del Olmo M, Zafra A, et al. Talanta, 2000, 50 (6): 1141～1148.
92. Seifert M, Brenner-Weib G, et al. Fresenius J. Anal. Chem., 1999 (363): 767～770.
93．旌拉德 W T，奥马利B W[美]著．激素作用与分子内分泌学的实验方法手册．北京：科学出版社，1988：1～3．
94．郭栋生，袁小英，吕晓军等．万家寨引黄工程水源地水质调查分析．环境化学，2002，21(3)：271～275．
95．杨频，杨曼曼，杨斌盛．科学通报，1993，38(22)：2051～2054．
96．颜承农，上官云凤等．双嘧达莫与牛血清白蛋白结合热力学特征的荧光光谱法研究．光
谱学与光谱分析，2003，23(3)：543～546．
97．张立伟，杨频等．丹参铜ⅡA等中草药有效成分与白蛋白结合反应的光谱研究．光谱学与光谱分析，2001，21(5)：694～696．
98．杨曼曼，杨频，张立伟．荧光法研究咖啡酸类药物与白蛋白的作用．科学通报，1994，39(1)：31～35．
99．易平贵，商志才．丝裂霉素C、阿霉素与过氧化氢酶的相互作用．化学通报，2003(7)：496～499．
100．袁小英，陈亮等．荧光法研究环丙沙星-铜(Ⅱ)-DNA的三元络合物，光谱学与光谱分析，1999，19(4)：572～574．
2. Harries J E, Janbakhsh A, et al. Estrogenic potency of effluent from two sewage treatment works in the United Kingdom. Environ. Toxicol Chem., 1999,18(5): 932.
3．周庆祥，江桂斌．浅谈环境内分泌干扰物质．科技术语研究，2001，3(3)：12～14．
4．邓茂先，陈祥贵．环境内分泌干扰物研究进展．国外医学卫生学分册，2000，27(2)：65～68．
5. Workshop sponsored UN Environment programme, U S EPA, White House Office of Science and Technology Policy, et al. International workshop on endocrine disruptors, 1997, 1.
6. ICPS, OECD. International programme on chemical safety report of ICPS/OECD scoping meeting on endocrine disruptors (EDs). US Washington D. C, 1998,3.
7 出云逾明．威胁人类存亡的定时炸弹—环境荷尔蒙．深圳：海天出版社，1999．
8．任仁．环境激素的种类和污染途径．大学化学，200 1(5)：28～32．
9．井口泰泉．环境荷尔蒙问题的现状及其今后动向．98’中日环境测试技术与环境管理研讨会，1998．
10. Kelce W, Stone C, et al. Persistent DDT metabolite p, p'-DDE is a potent androgen receptor antagonist. Nature, 1996 (375): 581～585.
11. Stone R. Environmental estrogens stir debate Science, 1994 (265): 308～310.
12. Hutchinson T H, Brown R, Brugger K E, et al. Ecological risk assessment of endocrine disruptors. Environ. Health Perspect, 2000, 108 (11): 1007～1014.
13. Cheek A O, Vonier P M, et al. Environmental aignaling: A biological context for endocrine disruption. Environ. Health Perspect, 1998, 106 (Suppl. 1): 5～10.
14. Routledge E J, Sheahan D, et al. Identification of estrogenic chemicals in STW effluent. In vivo responses in trout and roach. Environ. Sei. Technol., 1998 (32): 1559～1565.
15. Paflak M. A testing deadline for endocrine disruptors. Environ. Sci. Technol., 1996 (30): 540A～544A.
16．郝明德，刘晓宏．环境激素的危害与预防方法．世界农业，1999(9)：42～43．
17．香山不二雄．环境激素问题研究现状．世界环境，1999(2)：34～35．
18. Allen Y, et al. The extent of oestrogenic contaminants in the UK estuarine and marine environments- further surveys of flounder. The Science of the Total Environment, 1999 (233): 5～20.
19. Key SW. New study highlights hazardson hormone disrupting chemicals. World Oisease (Weekly Plus), 1998 (9): 11～12.
20．小林秀幸．内分泌扰乱化学物质问题．环境管理，1998，34(11)：1～7．
21. Ternes TA, Stumpf M, et al. Behavior and occurrence of estrogens in municipal sewage treatment plants-Ⅰ. Investigations in Germany, Canada and Brazil. The Science of the Total Environment, 1999 (225): 81～90.
22．武木田薰，井上述．内分泌扰乱化学物质对生物体影响．环境管理，1998，34(11)：8～14．
23. Harries JE, et al. Estrogeuic potency of effluent from two sewage treatment works in the United Kingdom. Environmental Toxicology and Chemistry, 1999, 18 (5): 932～937.
24．王兰，蒋学之，顾祖维编著．环境与生殖．上海：上海医科大学出版社，1994．
25．高京生．环境激素．中国检验医学与临床，2001，2(3)：97～99．
26．夏星辉，杨居荣，许嘉琳．环境激素污染研究进展．上海环境科学，2001，20(2)：56～59．
27．邱东茹，吴振斌，贺铎．内分泌扰乱化学品对动植物的影响和作用机制．环境科学研究，2000，13(6)：52～55．
28．陈岚，王欣．环境激素及其对生物的危害．山西中医学院学报，2002，3(3)：45～46．
29．王毓秀，张利民，邹敏．化学农药与环境激素．农村生态环境，1999，15(4)：37～41．
30．周宜开．环境激素导致阴盛阳衰?中国环境报，1999(8)：31．
31．邓苹译．环境荷尔蒙为何会给人类带来危机．环境科学，1998(12)：26～29．
32．江田汉，杨晓明，赖玉平．环境激素问题浅析．中国环境管理，2001(1)：43～45．
33. John A. Mclachlan, Arnold S F, Klotz D M, et al. Synergistic Activations of Estrogen Receptor with Combinations of Environmental Chemicals. Science, 1996 (272): 1489-1492.
34．曹信孚译．国外环境激素问题的研究动态．广州环境科学，2002，17(1)：46～48．
35. Colbom T, Dumaoski D, Meyers J P. Our Stolen Future, New York: Penguin Books, 1996.
36. Gillesby BE and Zacbarewski TR. Exoestrogens: mechanisms of action and strategies for identification and assessment. Environmental Toxicology and Chemistry, 1998, 17 (1): 3～14.
37. Scott Alex UK. Industry prepares for tough rules. Chemical Week, 1998, 160 (2): 22.
38．杜克久，储少岗等．用MCF-7细胞增殖法研究国产多氯联苯的类雌激素效应．环境科学学报，2000，20(2)：230～233．
39．赵丙顺，邹继超等．小鼠子宫增重法检测国产三氯杀虫螨醇的雌激素生物括性．环境科学学报，2000，20(2)：244～247．
40. Zaeharewski T. In vitro bioassays for assessing eslrogenic substances. Environmental Science and Technology, 1997, 31 (3): 613～623.
41. Bolger R, Wiese TE and Ervin K, et al. Rapid Screening of environmental chemicals for estrogen receptor binding capacity. Environmental Health Perspectives, 1998, 106 (9): 551～557.
42. Safe SH and Gaido K. Phytoestrogens and anthropogenic estrogenic compounds. Environmental Toxicology and Chemistry, 1998, 17 (1): 119～126.
43. Andersen HR, Andersson Anna-Maria, et al. Comparison of short-term estrogenicity tests for identification of hormone-disrupting chemicals. Environmental Health Perspectives, 1999, 107 (supplement 1): 89～100.
44．Olsson P,Westcrlund L and Teb SW, et al. 母体对雌激素暴露和PCB对斑马鱼(Danio rerio)不同生活阶段的影响．AMBIO，1999，28(1)：100～106．
45. Brown RJ, Conradi M and Depledge MH. Long-term exposure to 4-nonylphonel affects sexual differentiation and growth of amphlpod corphium volutator(Pallas,1766). The Science of the Total Environment, 1999 (233): 77～88.
46. Reitrer LW, Derosa C, et al. The U. S. Framework for research on endocrine disruptors and an analysis for research programs supported during fiscal year 1996. Environmental Health Perspectives, 1998, 106 (3): 105～117.
47. Karcher W. Recent Trends and developments m the EU in the environmental control and management of chemicals. Eeotoxicology and Environmental Safety, 1998 (40): 97～102.
48. Shi LM, Fang H and Tong W, et al. QSAR models using a large diverse set of estrogeus. J Chem InfCompol Sci., 2001 (41):168～195.
49. Kareher W, Hansen BG, et al. Predictions for existing chemicals- a multilateral QSAR project. SAR and QSAR Environmental Research, 1995 (3): 217～221.
50. Hermeas J, Balaz S, et al. Assessment of QSAR for predicting fate and effects of chemicals in the environment: an international European project. SAR and QSAR Environmental Research, 1995 (3): 223-236.
51. Clark LB and Rosen RT, et al. Determination of alkylphenol ethoxylates and their acetic and derivatives in drinking water by particle beam liquid chromatograph/mass spectrometry. Int J Anal. Chem., 1992 (47): 167～180.
52．齐文启，孙宗光．环境荷尔蒙物质及其检测分析．环境科学，1999，18(12)：531～533．
53．吴平谷，韩关根，王惠华等．饮用水中邻苯二甲酸酯类的调查．环境与健康杂志，1999，16(6)：338～339．
54．丁鹏，赵晓松等．酞酸酯类化合物(PAES)研究新进展．吉林农业大学学报，1999，21(3)：119～123,128．
55．叶常明．环境中的邻苯二甲酸酯．环境科学进展，1993，1(2)：36～45．
56. Klamsmeier RE. In: I. E. Liener (Ed). Microbial Biodeterioration. Academic Press, London, 1981: 431～471.
57．石万聪编．增塑剂．北京：化学工业出版社，1989：60．
58. Atlas E and Giam CS. Global transport of organic; pollutants: ambient concentration in the remote marine atmosphere. Science, 1981 (211): 163～165.
59．周文敏等编．环境优先污染物．北京：中国环境科学出版社．1989：48．
60．金相灿等编．有机化合物污染化学-有毒有机物污染化学．北京：清华大学出版社，1990：266～275．
61．庞金梅等．苯二甲酸酯的微生物降解与转化．环境科学，1993，18(3)：88～90．
62. Yin Mei-chin et al. Znvestigation on risk of phthalate ester in drink water and marked foods. Medicines Food Analysis, 1996, 4 (4): 313～318.
63．赵振华．我国环境中酞酸酯污染的研究．环境科学丛刊，1986，7(5)：50～56．
64. Sehiedek Thomas. Impact of plasticizers (phthalicacid esters) on soil and groundwater quality. IAHS Publ, 1995 (225): 149～156.
65. Nagini S and Selvam S. Phthalate ester plasticizers- A review. Trends Life Sci., 1994, 9 (1): 9～16.
66. Wei wende. A Complete Collection of Organic Raw Materials in Chemical Industry. Vol.Ⅲ, Beijing: Chemical Industry Press, 1990.
67．刘程，江小梅等．表面活性剂应用大全．北京：北京工业大学出版社，1992：113．
68．王跃志．化工进展，2000，19(4)：965～967．
69. Olea N, Pulgar R, et al. Environ. Health Perspect., 1996 (104): 298～305.
70. Shintani H J. Liq. Chromatogr., 1995, 18 (3): 613～626.
71. Ben-Jonathan N, Steinmetz R. Trends Endocrinol. Metab., 1998, 9 (3): 124～128.
72. Brotons J A, Oles-Serraao M F, et al. Environ. Health Perspect., 1995 (103): 608～612.
73. Soto A M, Justicia H, et al. Environ. Health Perspect., 1991 (92): 167.
74. Krishnan A V, Stathis P, et al. Endocrinology, 1993 (132): 2279～2286.
75. Bitman J, Cecil H C, et al. Science, 1969 (162): 371～372.
76. Sonnenschein C, Soto Am. Journal of Steroid Biochemistry and Molecular Biology, 1998, 65 (1-6): 143～150.
77. Jobling S. Pure and Appl. Chem., 1998,70 (9): 1805～1827.
78. Thiele B, Guenther K, et al. Chem. Rev., 1997, 97 (8): 3247～3272.
79. Murray, H.E., Ray, L.E., et al. Chemosphere, 1981 (10): 1327.
80. Gledhill, W.E., Kaley, R.C., et al. Environ. Sci. Technol., 1980 (14): 301.
81. Kurane, R., Suzaki, T., et al. Aqric. Biol. Chore.,1980 (44): 529.
82．赵振华．酞酸酯对人和环境潜在危害的研究概况．环境化学，1991，10(3)：64～68．
83. Mol H G J, Sunarto S, et al. Chromatogr. A, 2000 (879): 97～112.
84. Van der Kraak G. Pure and Appl. Chem., 1998, 70 (9): 1785～1794.
85．赵美萍，李元宗等．酚类环境雌激素的分析研究进展．分析化学，2003，31(1)：103～109．
86．靳翠红，赵剑等．双酚A对雄性小鼠生殖系统的影响．中国医科大学学报，2002，31(2)：123～124．
87. Staples C A, Dora P B, et al. Bisphenol A concentrations in receiving water near US manufacturing and processing facilities. Chemosphere, 2000 (40): 521～525.
88. Belfroid A C, et al. Analysis and occurrence of estrogenic hormones and their glucuronides in surface water and waste water in the Netherlands. The Science of the Total Environment, 1999 (225): 101～108.
89. Desbrow C and Routledge E J, et al. Identifieation of estrogenic chemicals in STW effluent. Ⅰ Chemical fractionation and in vitro biological screening. Environmental Science and Technology, 1998, 32 (11): 1549～1557.
90. Del Olmo M, Zafra A, et al. Int. J. Environ. Anal. Chem., 1998, 69 (1): 99～110.
91. Del Olmo M, Zafra A, et al. Talanta, 2000, 50 (6): 1141～1148.
92. Seifert M, Brenner-Weib G, et al. Fresenius J. Anal. Chem., 1999 (363): 767～770.
93．旌拉德 W T，奥马利B W[美]著．激素作用与分子内分泌学的实验方法手册．北京：科学出版社，1988：1～3．
94．郭栋生，袁小英，吕晓军等．万家寨引黄工程水源地水质调查分析．环境化学，2002，21(3)：271～275．
95．杨频，杨曼曼，杨斌盛．科学通报，1993，38(22)：2051～2054．
96．颜承农，上官云凤等．双嘧达莫与牛血清白蛋白结合热力学特征的荧光光谱法研究．光
谱学与光谱分析，2003，23(3)：543～546．
97．张立伟，杨频等．丹参铜ⅡA等中草药有效成分与白蛋白结合反应的光谱研究．光谱学与光谱分析，2001，21(5)：694～696．
98．杨曼曼，杨频，张立伟．荧光法研究咖啡酸类药物与白蛋白的作用．科学通报，1994，39(1)：31～35．
99．易平贵，商志才．丝裂霉素C、阿霉素与过氧化氢酶的相互作用．化学通报，2003(7)：496～499．
100．袁小英，陈亮等．荧光法研究环丙沙星-铜(Ⅱ)-DNA的三元络合物，光谱学与光谱分析，1999，19(4)：572～574．