Ethyl acetate extraction procedure and isocratic high-performance liquid chromatographic assay for testosterone metabolites in cell microsomes

Journal of Chromatography. B, Biomedical Sciences and Applications
M R BaltesM Hanocq

Abstract

An isocratic reversed-phase high-performance liquid chromatography (HPLC) method was developed and validated for separation of testosterone and its main metabolites over the nominal range 20 to 40 microg/ml and 280 to 4600 ng/ml, respectively. Mobile phase composition (phosphate buffer-methanol-acetonitrile, 50:38.5:11.5) was optimised by studying the influence of numerous chromatographic parameters. The most critical one was the ratio CH3CN/CH3OH. Good recoveries (around 90% for all compounds) and an improved specificity were assessed by a double ethyl acetate extraction of biological samples. According to the performance criteria tested, the method could be applied to enzymatic inhibition and induction in vitro studies.

References

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Citations

Dec 17, 2002·Journal of Pharmacological and Toxicological Methods·J L FayerK J Ruterbories
Feb 1, 2008·Toxicological Sciences : an Official Journal of the Society of Toxicology·Jing-Ying HuangTzuu-Huei Ueng
Sep 16, 2011·Pakistan Journal of Biological Sciences : PJBS·Wanna Sirisangtrakul, Bungorn Sripanidkulchai
Oct 23, 2013·African Journal of Traditional, Complementary, and Alternative Medicines : AJTCAM·Wanna Sirisangtragul, Bungorn Sripanidkulchai
Mar 10, 2015·Yakugaku zasshi : Journal of the Pharmaceutical Society of Japan·Isamu MurataIkuo Kanamoto
Jan 5, 2002·Journal of Chromatography. a·K ShimadaT Higashi

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