基于受体作用理论的类雌激素效应测试方法

以17β雌二醇为标准参考物质,对基于受体作用理论的环境激素分析方法实验条件进行了重构和优化,建立了环境类雌激素分析方法。结果表明:用营养缺陷型培养基30℃恒温振荡66 h培养时间进行重组基因酵母的复苏与增殖,20℃～35℃水浴样品与酵母混合液培养3～6 h暴露,优先选用机溶剂处理法释放β-半乳糖苷酶,35℃恒温水浴进行酶活测试反应,采用反应终止后2.5 h 420 nm波长进行分析测试,同时实际样品的分析,加测不少于一个的稀释水样效应值,可使方法更加科学高效、准确度高,应用于各类实际样品分析。

TESTING METHOD OF ESTROGENIC EFFECT BASED ON THE RECEPTOR EFFECTS THEORY

The environmental hormones with accumulation and potential risk were concerned recently because of the inherent toxicity. The testing methods of the environmental hormones were studied by several research. Chemical methods were the major testing methods to determine the known environmental hormones during the past few years. In this paper we established and optimized a biological testing method based on the receptor effects theory. The experimental parameters of environmental hormone effects testing based on the receptor effects theory were optimized, including recovery and proliferation of recombinant gene yeast, exposure time and temperature of estrogenic compounds, the testing method of β- galactosidase, the optimum testing temperature, colorimetric time and colorimetric wavelength, statistical method by ORIGIN, and quantitative method, to analyze the estrogenic effect, in which 17β-estradiol was used as the standard reference materials. The results indicated that the method was more efficiently and accurately to determine the actual samples when recovery and proliferation of recombinant gene yeast was cultured in SD/-Trp/-Leu auxotrophic medium and shaked in 30℃ after 66 h when the yeast initiated the logarithmic growth. We could mix sample and yeast mixture in 20℃-35℃ water bath for 3-6 hours to improve the sensitivity.The β- Galactosidase should be released by organic solvent treatment in 35℃ water batch for 60 min, then 420 nm colorimetric analysis after 2.5 hours. The dose-effect curves of the environmental hormones fitted by ORIGIN were best when the concentration were between 0.025 nmol·L^-1 to 0.25 nmol·L^-1. In addition, we suggested that the dilution sample should be added in actual tests to increase the accuracy of the methods.