中国药房2012，Vol.23Issue(25)：2318-2320,3.DOI:10.6039/j.issn.1001-0408.2012.25.06
以睾酮为探针的HPLC法测定CYP3A1酶活性的方法学研究
Methodology Research on Determination of CYP3A1 Activity by HPLC with Testosterone as Probe
摘要
Abstract
OBJECTIVE: To develop a method for the determination of CYP3A1 activities in rats with testosterone as probe. METHODS: HPLC method was adopted using midazolam as internal standard, and samples were extracted by ethyl-acetate, evaporated and re-dissolved. The method validation of the determination of 6-β-hydroxytestosterone (6-β-OHT) concentration had been examined. The determination was performed on Hypersil BDS C18 column with mobile phase consisted of methanol-Na2HPO4 (pH= 8.25) (60:40) at the flow rate of 1 mL·min-1. Detection wavelength was set at 254 nm. RESULTS: The linear range of 6-β-OHT was 1.25—100μg·mL-1 (r=0.999 8). Limit of quantitation was 1.25 μg·mL-1. Extraction recoveries were 83.9%~87.4% (RSD≤ 8.82% ). Relative recoveries were 100.4% ~103.4% (RSD≤7.05% ). The RSD of intra-day and inter-day were less than 10%. Sample were stable at room temperature for 4 hours. CONCLUSION: The method is rapid, simple and repeatable. It's applicable to determination of testosterone and 6-^-OHT in vitro. Also it's applicable for the evaluation of CYP3A1 activities in vitro.关键词
高效液相色谱法/睾酮/6-β羟基睾酮/咪达唑仑/肝微粒体/方法学Key words
HPLC/ Testosterone/ 6-β-hydroxytestosterone/ Midazolam/ Liver microsomes/ Methodology分类
医药卫生引用本文复制引用
周望,邓多,周文菁,刘韬,黄红兵..以睾酮为探针的HPLC法测定CYP3A1酶活性的方法学研究[J].中国药房,2012,23(25):2318-2320,3.基金项目
2009年广东省医学科研基金项目(B2009090) （B2009090）
