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Isotope-Dilution Mass Spectrometry for Quantification of Urinary Active Androgens Separated by Gas Chromatography

Author(s): Su Hyeon Lee | Man Ho Choi | Won-Yong Lee | Bong Chul Chung

Journal: Mass Spectrometry Letters
ISSN 2233-4203

Volume: 1;
Issue: 1;
Start page: 29;
Date: 2010;

Keywords: Androgen | Testosterone | Isotope-dilution | GC-MS | Drug efficacy

Cross reacting antibodies can cause an overestimation of the results of immunoassays. Therefore, alternative methodsare needed for the accurate quantification of steroids. Gas chromatography combined with isotope-dilution mass spectrometry(GC-IDMS) is developed to quantify urinary active androgens, testosterone, epitestosterone and dihydrotestosterone, which areclinically relevant androgens to both hair-loss and prostate diseases. The method devised involves enzymatic hydrolysis with β-glucuronidase, solid-phase extraction, liquid-liquid extraction using methyl tert-butyl ether and subsequent conversion to pentafluorophenyldimethylsilyl-trimethylsilyl (flophemesyl-TMS) derivatives for sensitive and selective analysis in selected-ionmonitoring mode. Flophemesyl-TMS derivatization not only eliminates matrix interference but also has a good peak resolutionwithin a 6 min-run. A selective and sensitive GC technique with flophemesyl-TMS derivatives also allows accurate quantitativeanalysis of three active androgens when combined with IDMS. The limit of quantification of the three analytes was
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