17β-雌二醇经人CYP1B1代谢产物研究

Analysis of 17β-Estradiol Metabolites via Human CYP1B1 by HPLC-ECD

目的:采用高效液相色谱与电化学检测器联用(HPLC-ECD)法研究17β-雌二醇经人细胞色素酶CYP1B1亚酶的代谢产物及各产物的生成速率。方法:色谱柱Mightysil RP-18GP(250 mm×3.0 mm,5μm)、柱温40℃,电化学检测器E=+900m V,流动相采用0.5%Na H_2PO_4(p H 3.0)和甲醇混合溶液(45∶55),流速0.5 ml﹒min^(-1),进样量为5μl。结果:HPLC-ECD法检测17β-雌二醇经CYP1B1代谢产物为4-OH-E_2,伴有少量2-OH-E_2。同浓度雌激素E_2经CYP1B1代谢生成4-OH-E_2的平均速率是2-OH-E_2的约5倍。结论:CYP1B1催化17β-雌二醇基于NADPH代谢的羟基化位点主要是4位。

Objective： To study the metabolites of 17β-estradiol via human cytochrome enzyme CYP! B1 and analyze the genera- tion rate of products by a high performance liquid chromatography coupled with electrochemical detector （HPLC-ECD） method, neth- otis： A Mightysil RP-18GP （250 mm×3.0mm,5μm） 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 （45： 55）, the flow rate was 0.5 ml mm-1 , and the injection volume was 5 μl. 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 E2 me- tabolized via CYP1B1. Conclusion： Taken together, CYP1 B1 catalyzed hydroxylation sites of 17-beta estradiol based on NADPH me- tabolism are really 4.