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

Trevor M. Penning, Seon Hwa Lee, Yi Jin, Alejandro Gutierrez, Ian A. Blair

Research output: Contribution to journalArticlepeer-review

62 Citations (Scopus)

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 5α-dihydrotestosterone (DHT)-17β-glucuronide, DHT-17β-sulfate, and tibolone-17β-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 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, 16α-hydroxy-E2, estrone (E1), 2-methoxy-E1, 4-methoxy-EI, and 16α-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.

Original languageEnglish
Pages (from-to)546-555
Number of pages10
JournalJournal of Steroid Biochemistry and Molecular Biology
Volume121
Issue number3-5
DOIs
Publication statusPublished - 2010 Aug
Externally publishedYes

Keywords

  • Catechol estrogens
  • Electron capture atmospheric pressure chemical ionization
  • Normal-phase HPLC
  • Steroid conjugates

ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism
  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Endocrinology
  • Clinical Biochemistry
  • Cell Biology

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