Determination of estradiol metabolites in human liver microsome by high performance liquid chromatography-electrochemistry detector

AIM: To constitute a method to determine the estradiol metabolites in human liver microsome in low concentration of estradiol. METHODS: Use high perfo rmance liquid chromatography after solvent extraction, evaporation, and rec onstitution to separate the metabolites and use a electrochemistry detector to detect the metabolites. RESULTS: With a mobile phase of acetic acid buf fer-acetonitrile (50:50, v/v, pH 4.5) at flow rate of 1.0 ml/min and a pote ntial of +0.7 V vs Ag/AgCl, all six composition were well separated and sat isfactorily detected. There are E-3, 16 alpha -OHE1, 2-OHE2, E-1, and two u nidentified composition. The minimum detectable amount is about 100 pg on c olumn. This method is sensitive enough to detect E-1 in a substrate concent ration of 1 mu mol/L. CONCLUSION: The method can be used to study the metab olism mechanism of estradiol in liver microsome.