Journal
JOURNAL OF STEROID BIOCHEMISTRY AND MOLECULAR BIOLOGY
Volume 121, Issue 3-5, Pages 546-555Publisher
PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/j.jsbmb.2010.01.005
Keywords
Steroid conjugates; Catechol estrogens; Electron capture atmospheric pressure chemical ionization; Normal-phase HPLC
Funding
- National Institutes of Health [P30ES013587, R01CA90744]
- Prostate Cancer Foundation [R01CA091016]
- [P30ES13587]
- Grants-in-Aid for Scientific Research [22590130] Funding Source: KAKEN
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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 5 alpha-dihydrotestosterone (DHT)-17 beta-glucuronide, DHT-17 beta-sulfate, and tibolone-17 beta-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 (El)) and its metabolites as pentafluorobenzyl (PFB) derivatives in human plasma in the attomole range. The limit of detection for E1-PFB was 740 attomole 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, 16 alpha-hydroxy-E2, estrone (E1), 2-methoxy-E1, 4-methoxy-E1, and 16 alpha-hydroxy-E1 from 5 pg/mL to 2000 pg/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. (C) 2010 Elsevier Ltd. All rights reserved.
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