水环境中雌激素的酶联免疫快速生物筛选技术研究

采用酶联免疫(ELISA)检测技术,对来自长江口及污水处理厂的水样分析典型雌激素雌二醇(E2)的浓度,将检测结果与化学方法检测数据进行比较验证,以阐明ELISA应用于水环境雌激素快速筛选的可行性。结果表明:ELISA的检测限可低至0.5 ng/L,试验的变异系数<30%,具有较高的灵敏性和良好的重复性;ELISA检测长江口水样中E2浓度范围≤0.7 ng/L,污水处理厂水样为2.5～11.5 ng/L,与液相色谱串联质谱(LC-MS/MS)的检测数据进行比较分析,其相关系数R~2为0.9147,表明2种方法具有良好的相关性;ELISA检测所需的水样量比LC-MS/MS更少,使用96微孔板可同时对43个样品进行检测,缩短了多个样品的分析时间,具有快速简便、经济适用的特点。因此,ELISA作为1种快速、简便、灵敏的雌激素高通量筛选技术,可为环境保护部门开展水环境雌激素物质调查和水体雌激素快速诊断等工作提供技术支持。

A Study on Rapid Biological Screening of Estrogens in Aquatic Environment by Enzyme-Linked Immunosorbent Assay

There has been more and more public concern about estrogens distributed extensively in aquatic environment as they may bring potential threat to human health and ecological safety due to their endocrine disrupting activities.The study aimed to determine contents of a typical estrogen 17β-estradiol(E2) in water samples collected from the Yangtze estuary and wastewater treatment plants by using enzyme-linked immunosorbent assay(ELISA),and then to verify the results by being compared with the measurements of a chemical method.It has shown that the lowest detection limit of ELISA could reach 0.5 ng/L and the coefficient of variance for the tests was below 30%indicating its high sensitivity and repeatability.E2 concentrations detected by ELISA were not greater than 0.7 ng/L in the water samples taken from the Yangtze estuary,and in the range of 2.5 ~ 11.5 ng/L in the samples from wastewater treatment plants,whilst they had a good correlation (R2 = 0.9147) with the results obtained by means of LC-MS/MS.On comparison,ELISA required less volume of water samples and took much less time for performing 43 sample analysis simultaneously by using 96-well microplates.It revealed that as a rapid,convenient and economical method and a highly throughput screening technique,ELISA could provide technical supports for environmental authorities to investigate and quickly examine estrogens in natural aquatic environment.