Analysis of 17β-Estradiol Metabolites via Human CYP1B1 by HPLC-ECD / 中国药师

ABSTRACT

Objective:To study the metabolites of 17β-estradiol via human cytochrome enzyme CYP 1B1 and analyze the genera-tion rate of products by a high performance liquid chromatography coupled with electrochemical detector (HPLC-ECD) method.Meth-odsA Mightysil RP-18GP (250 mm ×3.0 mm, 5 μm) column was used at the temperature of 40 ℃.The electrochemical detector with E=+900 mV was applied, the mobile phase was 0.5％NaH2PO4(pH 3.0) and methanol (4555), the flow rate was 0.5 ml · min-1 , and the injection volume was 5μl.Results:The main metabolite was 4-OH-E2 accompanied with a little of 2-OH-E2.The average hydroxylation rate of 4-OH-E2 was about five times as much as that of 2-OH-E2 at the same concentration of estrogen E 2 me-tabolized via CYP1B1.Conclusion:Taken together, CYP1B1 catalyzed hydroxylation sites of 17-beta estradiol based on NADPH me-tabolism are maily 4.