Liquid chromatography-mass spectrometry (LC-MS) of steroid hormone metabolites and its applications.

Abstract	Advances in liquid chromatography-mass spectrometry (LC-MS) can be used to measure steroid hormone metabolites in vitro and in vivo. We find that LC-electrospray ionization (ESI)-MS using a LCQ ion trap mass spectrometer in the negative ion mode can be used to monitor the product profile that results from 5alpha-dihydrotestosterone (DHT)-17beta-glucuronide, DHT-17beta-sulfate, and tibolone-17beta-sulfate reduction catalyzed by human members of the aldo-keto reductase (AKR) 1C subfamily and assign kinetic constants to these reactions. We also developed a stable isotope dilution LC-electron capture atmospheric pressure chemical ionization (ECAPCI)-MS method for the quantitative analysis of estrone (E1) and its metabolites as pentafluorobenzyl (PFB) derivatives in human plasma in the attomole range. The limit of detection for E1-PFB was 740attomole on column. Separations can be performed using normal-phase LC because ionization takes place in the gas phase rather than in solution. This permits efficient separation of the regioisomeric 2- and 4-methoxy-E1. The method was validated for the simultaneous analysis of plasma E2 and its metabolites: 2-methoxy-E2, 4-methoxy-E2, 16alpha-hydroxy-E2, estrone (E1), 2-methoxy-E1, 4-methoxy-EI, and 16alpha-hydroxy-E1 from 5pg/mL to 2000pg/mL. Our LC-MS methods have sufficient sensitivity to detect steroid hormone levels in prostate and breast tumors and should aid their molecular diagnosis and treatment.
Authors	Trevor M Penning, Seon-Hwa Lee, Yi Jin, Alejandro Gutierrez, Ian A Blair
Journal	The Journal of steroid biochemistry and molecular biology (J Steroid Biochem Mol Biol) Vol. 121 Issue 3-5 Pg. 546-55 (Aug 2010) ISSN: 1879-1220 [Electronic] England
PMID	20083198 (Publication Type: Journal Article, Research Support, N.I.H., Extramural)
Copyright	Copyright 2010 Elsevier Ltd. All rights reserved.
Chemical References	Hormones Steroids
Topics	Chromatography, High Pressure Liquid (methods) Hormones (blood) Humans Kinetics Reproducibility of Results Spectrometry, Mass, Electrospray Ionization (methods) Steroids (blood)

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