Influence Factors And Quality Control Of Environmental Estrogen Bioassay Based On Gene Recombined Yeast

It is paid much attention that environmental estrogen could interfere humanendocrine system．Based on demanding of water environmental estrogen management，we compared different analyzing methods and finally got this testing method，environmental estrogen effect analysis，which is on basis of recombinant geneyeast．It is adapt to actual water application and has close relationship with humanhealth．In order to improve reliability of testing result and be applied in actualcomplex water environmental estrogen analysis，a lot of experimental data has beenworked out，to a nalysis the effect of characteristics of water samples on thedetermination results，optimize testing condition and add suitable quality controlprocess during systemic analysis.The result shows that：（1）Recovery and proliferation of recombinant gene yeast could be cultured inSD/-Trp/-Leu auxotrophic medium and shaking in30℃．Sample could be exposedafter66hours’ culture．We could mix sample and yeast mixture in20~35℃waterbath for3~6hours．It could release β-Galactosidase by organic solvent treatment，react60minutes in35℃water batch，then420nm colorimetric analysis after0.5hours; It should be test more than one diluted water sample in reality．（2）The index such as DO，chroma and turbidity has small influence on testingresult．Due to affecting growth of yeast or gene convey，others in water sample，suchas micro，pH and some toxic in water has some influence on analyzing result．Theyneed to be treated further.（3）Setting up quality control could eliminate abnormal data．In order to avoid falsenegative result，we could test activity of recombinant gene yeast; When environmentalestrogen concentration is between0.025nmol/L~0.25nmol/L(cal． By E2)，normalized relative effect and environmental estrogen concentration is in linearpositive correlation．Linear gradient should be controlled on4.76±1.35; Besides，blank should be controlled on0.030±0.015; Relative effect value of positive referencesample(0.1nmol/L E2) should be better on49%±15%; Relative deviation of twoparallel samples should be controlled within30%（4） Main reason of environmental estrogen bioassay uncertainty is standard curvefitting and repeatability of sample analysis; Second reason is influence of differentenvironment or operators，sample collection process.