Ultra‐performance liquid chromatography/tandem mass spectrometric determination of testosterone and its metabolites in in vitro samples
- 22 June 2006
- journal article
- research article
- Published by Wiley in Rapid Communications in Mass Spectrometry
- Vol. 20 (14), 2215-2221
- https://doi.org/10.1002/rcm.2580
Abstract
This paper describes the development and partial validation of a fast, sensitive and specific ultra‐performance liquid chromatography/tandem mass spectrometric (UPLC/MS/MS) method for the determination of testosterone (T) and its four metabolites, 6β‐OH‐T, 16α‐OH‐T, 16β‐OH‐T and 2α‐OH‐T, in in vitro samples. The analytical method involves direct dilution of samples with acetonitrile containing an internal standard, followed by separation of testosterone and the four metabolites on an Acquity UPLC™ C18 column and detected by selected reaction monitoring (SRM) in positive ionization mode using turbo ionspray ionization. The parent compound and its metabolites investigated were well separated (Rs >1.5) with a run time of 4 min under a gradient condition. The method was partially validated. The linear concentration range was 0.01 to 5 µM for all the compounds of interest. Inter‐assay mean bias and relative standard deviation (RSD) were in the range of −12% to 8% and 4.1% to 8.5%, respectively. Intra‐assay mean bias and RSD were in the range of −8.0% to 5.2% and 3.4% to 9.6%, respectively. The lower limit of quantitation for this assay was 0.01 µM. The differences in LC/MS performance were investigated by conducting a comparison of UPLC with another method previously optimized for HPLC‐based separation and quantification of testosterone and its metabolites. Copyright © 2006 John Wiley & Sons, Ltd.Keywords
This publication has 16 references indexed in Scilit:
- Orthogonal extraction/chromatography and UPLC, two powerful new techniques for bioanalytical quantitation of desloratadine and 3-hydroxydesloratadine at 25 pg/mLJournal of Pharmaceutical and Biomedical Analysis, 2006
- Improving LC–MS sensitivity through increases in chromatographic performance: Comparisons of UPLC–ES/MS/MS to HPLC–ES/MS/MSJournal of Chromatography B, 2005
- Determination of testosterone concentrations in rat plasma using liquid chromatography–atmospheric pressure chemical ionization mass spectrometry combined with ethyl oxime and acetyl ester derivatizationJournal of Chromatography B, 2005
- Testosterone Measurement by Isotope-Dilution Liquid Chromatography–Tandem Mass Spectrometry: Validation of a Method for Routine Clinical PracticeClinical Chemistry, 2005
- Increasing throughput and information content for in vitro drug metabolism experiments using ultra‐performance liquid chromatography coupled to a quadrupole time‐of‐flight mass spectrometerRapid Communications in Mass Spectrometry, 2005
- Peak capacity in gradient ultra performance liquid chromatography (UPLC)Journal of Pharmaceutical and Biomedical Analysis, 2005
- Determination of testosterone in saliva and blow of bottlenose dolphins (Tursiops truncatus) using liquid chromatography–mass spectrometryJournal of Chromatography B, 2005
- Simultaneous determination of steroid composition of human testicular fluid using liquid chromatography tandem mass spectrometrySteroids, 2004
- Quantitative analysis of eight testosterone metabolites using column switching and liquid chromatography/tandem mass spectrometryRapid Communications in Mass Spectrometry, 2004
- Measurement of Total Serum Testosterone in Adult Men: Comparison of Current Laboratory MethodsVersusLiquid Chromatography-Tandem Mass SpectrometryJournal of Clinical Endocrinology & Metabolism, 2004