Measurement of sex steroids in murine blood and reproductive tissues by liquid chromatography-tandem mass spectrometry

Keely May Mcnamara, D. T. Harwood, U. Simanainen, K. A. Walters, M. Jimenez, D. J. Handelsman

Research output: Contribution to journalArticle

90 Citations (Scopus)

Abstract

Accurate measurement of sex steroids is essential to evaluate mouse models for human reproductive development and disorders. The recent advent of liquid chromatography-tandem mass spectrometry (LC-MS/MS) assays that match the sensitivity of steroid immunoassay could overcome problems arising from the limited specificity of steroid immunoassay. In this current study we validate a LC-MS/MS assay for the measurement of key sex steroids from murine serum and reproductive tissues. The assay gave excellent dilutional linearity (r2≥0.98) and reproducibility (CV≤10% of replicate samples) in serum and reproductive tissues with sensitive quantitation limits; testosterone (T; 2pg), dihydrotestosterone (DHT; 10pg), 5α-androstane-3α,17β-diol (3αDiol; 40pg), 5α-androstane-3β,17β-diol (3βDiol; 40pg), estradiol (E2; 0.5pg) and estrone (E1; 0.3pg). Using 0.1mL sample, T was the only consistently detectable steroid (detection limit 20pg/ml) in both male and female mouse serum. In the testis, T and DHT were quantifiable as were both diols at relatively high levels. Prostatic T levels were low and DHT was determined to be the most abundant androgen in this tissue. Uterine and ovarian levels of E2, E1 and T were measurable, with levels varying according to estrous cycle stage. Hence, we demonstrate that this LC-MS/MS method has the sensitivity, specificity and multi-analyte capability to offer accurate steroid profiling in mouse serum and reproductive tissues.

Original languageEnglish
Pages (from-to)611-618
Number of pages8
JournalJournal of Steroid Biochemistry and Molecular Biology
Volume121
Issue number3-5
DOIs
Publication statusPublished - 2010 Aug 1

Keywords

  • Immunoassay
  • LC-MS/MS
  • Mouse
  • Reproductive tissue
  • Serum
  • Steroids

ASJC Scopus subject areas

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

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