天津城市水、土环境中典型药物与个人护理品（PPCPs）分布及其复合雌激素效应研究
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摘要
药物与个人护理品(Pharmaceuticals and Personal Care Products,PPCPs)是一类与人们日常生活联系最为密切的新型污染物。本文选取在我国大量使用的氟喹诺酮类抗生素、磺胺类抗生素和四环素类抗生素为目标物质,对其在天津城市水、土环境中的分布状况进行了调查。同时选取了雌酮、17β-雌二醇、雌三醇和17α-乙炔基雌二醇这4种典型雌激素类物质,对其二元混合溶液的复合雌激素效应进行了研究。
本文建立并优化了基于液相色谱-质谱联用技术,可对水环境样品中氟喹诺酮类抗生素,磺胺类抗生素和四环素类抗生素同时进行检测的方法。该方法检出限为0.4-3.2 ng/L,目标抗生素加标回收率在60-105 %之间,相对标准偏差小于15 %。
应用上述方法对天津市市内河流、市内排污河、污水处理厂出水与再生水、饮用水水源地以及自来水中18种氟喹诺酮类抗生素、磺胺类抗生素、四环素类抗生素的分布情况进行了检测。检测结果表明:
在天津市内河流水样中,检测出13种目标抗生素,浓度范围在3.0-1080.0 ng/L。在天津市内排污河水样中,检测出9种目标抗生素,浓度范围在4.7-448.4 ng/L。在污水处理厂出水样品中,检测出12种目标抗生素,浓度范围在43.0-486.0 ng/L。在再生水样品中,检测出9种目标抗生素,浓度范围在4.0-311.0 ng/L。在饮用水水源地水样及自来水水样中均未检出目标抗生素。
此外,本文还建立了基于液相色谱-质谱联用技术,可对土壤环境样品中氟喹诺酮类抗生素,磺胺类抗生素和四环素类抗生素同时进行检测的方法。该方法检出限为0.1-1.9μg/kg,目标抗生素加标回收率在56-97 %之间,相对标准偏差小于20 %。
应用上述方法对天津市大沽排污河、北塘排污河,饮用水水源地于桥水库周边地区土壤中18种氟喹诺酮类抗生素、磺胺类抗生素、四环素类抗生素的分布情况进行了检测。检测结果表明:
在大沽排污河周边土壤样品中,检测出12种目标抗生素,浓度范围在1.4-114.8μg/kg。在北塘排污河周边土壤中,检测出9种目标抗生素,浓度范围在0.7-306.9μg/kg,在饮用水水源地周边土壤样品中,检测出全部18种目标抗生素,浓度范围在0.5-20343.2μg/kg。
本文对重组基因酵母检测方法的实验条件进行优化,确定了酵母与受试溶液最优反应时间为2 h,确定了酵母与O-NPG反应时间为30 min。
对人乳腺癌细胞(MCF-7)增殖检测方法的实验条件进行优化,确定以DMSO为溶剂时,可导致MCF-7细胞增殖效应显著降低,故应选取无酚红DMEM培养基为配制待测溶液时所用溶剂。确定培养基中的酚红、血清中内源性雌激素对MCF-7细胞有增殖效应,应选取无酚红DMEM培养基与经活性炭/葡聚糖处理的胎牛血清。确定使用无酚红DMEM培养基最优预培养时间为7 d。确定加入待测样品后最优培养时间为48 h。
应用优化后重组基因酵母方法和人乳腺癌细胞(MCF-7)增殖方法检测由雌酮、17β-雌二醇、雌三醇及17α-乙炔基雌二醇4种雌激素组成的二元混合体系的复合雌激素效应,并使用浓度加和模型(CA)与独立作用模型(IA)对二元混合体系的复合雌激素效应进行预测,结果证明CA模型更适用于重组基因酵母检测体系,IA模型更适用于人乳腺癌细胞(MCF-7)增殖检测体系。在重组基因酵母检测体系中,二元混合溶液表现出的复合雌激素效应比大于1的比例为19.0 %,在人乳腺癌细胞(MCF-7)增殖检测体系中,二元混合溶液表现出的复合雌激素效应比大于1的比例仅为6.2 %。
Pharmaceuticals and Personal Care Products as Pollutants (PPCPs) comprise a diverse collection of chemical substances, including any product used by individuals for personal health or cosmetic reasons or used by agribusiness to enhance growth or health of livestock. In this paper, fluoroquinolones, tetracyclines and sulfonamides were selected as traget compounds, considering that they are the the three most commonly used antibiotic groups in China. The combined estrogenic activity of binary mixtures of estrone, 17β-estradiol, estiol and 17α-ethynylestradiol at different equipotent concentrations were also tested.
The simultaneous analysis method for multiple classes of antibiotics in water samples via solid phase extraction and liquid chromatography-tandem mass spectrometry were established and optimized. The limit of detection of target antibiotics were at the range of 0.4-3.2 ng/L, the recoveries were between 60% and 105%, and the relative standard deviation was less than 15%.
This optimized analytical method were applied for the detection of fluoroquinolones, tetracyclines and sulfonamides in rivers, discharge river, wastewater treatment effluent, reclaimed water, drinking water resource and tap water.
Thirteen target antibiotics were detected in the river, with the concentration of 3.0-1080.0 ng/L. Nine target antibiotics were detected in the discharge river, with the concentration of 4.7-448.4 ng/L. In the wastewater treatment effluent, twelve target antibiotics were determined, of which the contents were at the range between 43.0 ng/L and 486.0 ng/L. Nine target antibiotics were detected in the reclaimed water, with the concentration of 4.0-311.0 ng/L. None of the target antibiotics were detected in drinking water resource and tap water.
Moreover, the simultaneous analysis method for multiple classes of antibiotics in soil samples via solid phase extraction and liquid chromatography-tandem mass spectrometry were established and optimized. The limit of detection of target antibiotics were at the range of 0.1-1.9μg/kg, the recoveries were between 56% and 97%, and the relative standard deviation was less than 20%.
This optimized analytical method were applied for the detection of fluoroquinolones, tetracyclines and sulfonamides in soil samples collected from the irrigation area of Dagu discharge river and Beitang discharge river. The soil samples collected from the farmland and orchard near the Yvqiao reservoir were also analysed.
Twelve target antibiotics were detected in the samples collected from the irrigation area of Dagu discharge river, the concentration were from 1.4μg/kg to114.8μg/kg. Nine target antibiotics were detected in the samples collected from the irrigation area of Beitang discharge river, with the concentration of 0.7-306.9μg/kg. All of the target antibiotics were detected in the samples collected from Yvqiao reservoir, with the concentration of 0.5-20343.2μg/kg.
The procedure of recombined yeast estrogen screen were optimized by adjusting the sample reaction time to 2 h and set the color reaction time as 30 min.
The parameters for cell proliferation assay using MCF-7 human breast cancer cells were optimized. The DMEM medium without phenol was selected as the solvent for analytical compounds. The fetal bovine serum treated by activated carbon / dextran was necessary for the proliferation test. The optimized pro-incubation time with DMEM medium without phenol was 7 d. The analytical reaction time was set as 48 h.
The optimized recombined yeast estrogen screen and cell proliferation assay were applied for the test of combined estrogenic activity of binary mixtures of estrone, 17β-estradiol, estiol and 17α-thynylestradiol at different equipotent concentrations. Concentration addition model and independent action model were used to predict the combined estrogenic activity of binary mixtures. The results demonstrated that the prediction of concentration addition model was less deviated in the recombined yeast estrogen screen than the prediction of independent action model. However, the independent action model was better than concentration addition model in the cell proliferation assay.
本文建立并优化了基于液相色谱-质谱联用技术,可对水环境样品中氟喹诺酮类抗生素,磺胺类抗生素和四环素类抗生素同时进行检测的方法。该方法检出限为0.4-3.2 ng/L,目标抗生素加标回收率在60-105 %之间,相对标准偏差小于15 %。
应用上述方法对天津市市内河流、市内排污河、污水处理厂出水与再生水、饮用水水源地以及自来水中18种氟喹诺酮类抗生素、磺胺类抗生素、四环素类抗生素的分布情况进行了检测。检测结果表明:
在天津市内河流水样中,检测出13种目标抗生素,浓度范围在3.0-1080.0 ng/L。在天津市内排污河水样中,检测出9种目标抗生素,浓度范围在4.7-448.4 ng/L。在污水处理厂出水样品中,检测出12种目标抗生素,浓度范围在43.0-486.0 ng/L。在再生水样品中,检测出9种目标抗生素,浓度范围在4.0-311.0 ng/L。在饮用水水源地水样及自来水水样中均未检出目标抗生素。
此外,本文还建立了基于液相色谱-质谱联用技术,可对土壤环境样品中氟喹诺酮类抗生素,磺胺类抗生素和四环素类抗生素同时进行检测的方法。该方法检出限为0.1-1.9μg/kg,目标抗生素加标回收率在56-97 %之间,相对标准偏差小于20 %。
应用上述方法对天津市大沽排污河、北塘排污河,饮用水水源地于桥水库周边地区土壤中18种氟喹诺酮类抗生素、磺胺类抗生素、四环素类抗生素的分布情况进行了检测。检测结果表明:
在大沽排污河周边土壤样品中,检测出12种目标抗生素,浓度范围在1.4-114.8μg/kg。在北塘排污河周边土壤中,检测出9种目标抗生素,浓度范围在0.7-306.9μg/kg,在饮用水水源地周边土壤样品中,检测出全部18种目标抗生素,浓度范围在0.5-20343.2μg/kg。
本文对重组基因酵母检测方法的实验条件进行优化,确定了酵母与受试溶液最优反应时间为2 h,确定了酵母与O-NPG反应时间为30 min。
对人乳腺癌细胞(MCF-7)增殖检测方法的实验条件进行优化,确定以DMSO为溶剂时,可导致MCF-7细胞增殖效应显著降低,故应选取无酚红DMEM培养基为配制待测溶液时所用溶剂。确定培养基中的酚红、血清中内源性雌激素对MCF-7细胞有增殖效应,应选取无酚红DMEM培养基与经活性炭/葡聚糖处理的胎牛血清。确定使用无酚红DMEM培养基最优预培养时间为7 d。确定加入待测样品后最优培养时间为48 h。
应用优化后重组基因酵母方法和人乳腺癌细胞(MCF-7)增殖方法检测由雌酮、17β-雌二醇、雌三醇及17α-乙炔基雌二醇4种雌激素组成的二元混合体系的复合雌激素效应,并使用浓度加和模型(CA)与独立作用模型(IA)对二元混合体系的复合雌激素效应进行预测,结果证明CA模型更适用于重组基因酵母检测体系,IA模型更适用于人乳腺癌细胞(MCF-7)增殖检测体系。在重组基因酵母检测体系中,二元混合溶液表现出的复合雌激素效应比大于1的比例为19.0 %,在人乳腺癌细胞(MCF-7)增殖检测体系中,二元混合溶液表现出的复合雌激素效应比大于1的比例仅为6.2 %。
Pharmaceuticals and Personal Care Products as Pollutants (PPCPs) comprise a diverse collection of chemical substances, including any product used by individuals for personal health or cosmetic reasons or used by agribusiness to enhance growth or health of livestock. In this paper, fluoroquinolones, tetracyclines and sulfonamides were selected as traget compounds, considering that they are the the three most commonly used antibiotic groups in China. The combined estrogenic activity of binary mixtures of estrone, 17β-estradiol, estiol and 17α-ethynylestradiol at different equipotent concentrations were also tested.
The simultaneous analysis method for multiple classes of antibiotics in water samples via solid phase extraction and liquid chromatography-tandem mass spectrometry were established and optimized. The limit of detection of target antibiotics were at the range of 0.4-3.2 ng/L, the recoveries were between 60% and 105%, and the relative standard deviation was less than 15%.
This optimized analytical method were applied for the detection of fluoroquinolones, tetracyclines and sulfonamides in rivers, discharge river, wastewater treatment effluent, reclaimed water, drinking water resource and tap water.
Thirteen target antibiotics were detected in the river, with the concentration of 3.0-1080.0 ng/L. Nine target antibiotics were detected in the discharge river, with the concentration of 4.7-448.4 ng/L. In the wastewater treatment effluent, twelve target antibiotics were determined, of which the contents were at the range between 43.0 ng/L and 486.0 ng/L. Nine target antibiotics were detected in the reclaimed water, with the concentration of 4.0-311.0 ng/L. None of the target antibiotics were detected in drinking water resource and tap water.
Moreover, the simultaneous analysis method for multiple classes of antibiotics in soil samples via solid phase extraction and liquid chromatography-tandem mass spectrometry were established and optimized. The limit of detection of target antibiotics were at the range of 0.1-1.9μg/kg, the recoveries were between 56% and 97%, and the relative standard deviation was less than 20%.
This optimized analytical method were applied for the detection of fluoroquinolones, tetracyclines and sulfonamides in soil samples collected from the irrigation area of Dagu discharge river and Beitang discharge river. The soil samples collected from the farmland and orchard near the Yvqiao reservoir were also analysed.
Twelve target antibiotics were detected in the samples collected from the irrigation area of Dagu discharge river, the concentration were from 1.4μg/kg to114.8μg/kg. Nine target antibiotics were detected in the samples collected from the irrigation area of Beitang discharge river, with the concentration of 0.7-306.9μg/kg. All of the target antibiotics were detected in the samples collected from Yvqiao reservoir, with the concentration of 0.5-20343.2μg/kg.
The procedure of recombined yeast estrogen screen were optimized by adjusting the sample reaction time to 2 h and set the color reaction time as 30 min.
The parameters for cell proliferation assay using MCF-7 human breast cancer cells were optimized. The DMEM medium without phenol was selected as the solvent for analytical compounds. The fetal bovine serum treated by activated carbon / dextran was necessary for the proliferation test. The optimized pro-incubation time with DMEM medium without phenol was 7 d. The analytical reaction time was set as 48 h.
The optimized recombined yeast estrogen screen and cell proliferation assay were applied for the test of combined estrogenic activity of binary mixtures of estrone, 17β-estradiol, estiol and 17α-thynylestradiol at different equipotent concentrations. Concentration addition model and independent action model were used to predict the combined estrogenic activity of binary mixtures. The results demonstrated that the prediction of concentration addition model was less deviated in the recombined yeast estrogen screen than the prediction of independent action model. However, the independent action model was better than concentration addition model in the cell proliferation assay.
引文
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