ISOCRATIC HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHIC METHOD FOR THE SEPARATION OF TESTOSTERONE METABOLITES

被引：42

作者：

SANWALD, P ^{[1
]}

BLANKSON, EA ^{[1
]}

DULERY, BD ^{[1
]}

SCHOUN, J ^{[1
]}

HUEBERT, ND ^{[1
]}

DOW, J ^{[1
]}

机构：

[1] MARION MERRELL DOW INC, DEPT PHARMACOKINET & DRUG METAB, F-67080 STRASBOURG, FRANCE

来源：

JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES | 1995年 / 672卷 / 02期

关键词：

D O I：

10.1016/0378-4347(95)00216-6

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

An isocratic reversed-phase high-performance liquid chromatographic (HPLC) method using an Ultrasphere IP column has been developed for the determination of testosterone and its metabolites after incubation of 4-C-14-labelled or unlabelled testosterone with rat liver microsomes. Compounds were eluted with methanol-water-tetrahydrofuran (35:55:10, v/v, pH 4.0) and detected by ultraviolet (UV) absorption at 245 nm. UV or on-line radioactivity detection can be used although, due to differences in detector cell volumes, peak resolution is slightly better with UV detection. Selectivity was validated by collecting HPLC peaks and verifying their identity by gas chromatography-mass spectrometry after derivatization by N,O-bis(trimethylsilyl)trifluoroacetamide chlorosilane. A three-day validation was performed to determine the linearity, repeatability, reproducibility and accuracy of the method, using corticosterone as internal standard. The method is applicable to the measurement of cytochrome P-450 isoenzyme activities in rat liver.

引用

页码：207 / 215

页数：9

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