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J Steroid Biochem Mol Biol. 2016 Jan;155(Pt A):56-62. doi: 10.1016/j.jsbmb.2015.09.032. Epub 2015 Sep 28.

Assay reproducibility of serum androgen measurements using liquid chromatography-tandem mass spectrometry.

Author information

1
Division of Cancer Epidemiology and Genetics, National Cancer Institute, National Institutes of Health, Department of Health and Human Services, Bethesda, MD 20892, USA. Electronic address: britton.trabert@nih.gov.
2
Cancer Research Technology Program, Leidos Biomedical Research, Inc., Frederick National Laboratory for Cancer Research, Frederick, MD 21701, USA.
3
Division of Cancer Epidemiology and Genetics, National Cancer Institute, National Institutes of Health, Department of Health and Human Services, Bethesda, MD 20892, USA.
4
Pharmacogenomics Laboratory, Centre Hospitalier Universitaire de Québec (CHUQ) Research Center, Laval University, Québec, Canada.
5
Departments of Obstetrics and Gynecology, and Preventive Medicine, University of Southern California Keck School of Medicine, Los Angeles, CA 90033, USA.

Abstract

BACKGROUND:

Valid and precise measures of androgen concentrations are needed for etiologic studies of hormonally-related cancers. We developed a high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) method with two sample preparations to measure 11 androgens, including adrenal and gonadal androgenic precursors and their 5α-reduced metabolites.

METHODS:

Androgen levels were measured in serum from 20 healthy volunteers (5 men, 10 premenopausal women, 5 postmenopausal women). Two blinded, randomized aliquots per individual were assayed in each of three batches. A fourth batch of samples was measured at an external laboratory using comparable methodology to measure 9 of the 11 androgens. Coefficients of variation (CV) and intraclass correlation coefficients (ICC) were calculated from the individual components of variance. Comparability of 9 androgens across laboratories was assessed using Spearman ranked correlations, Deming regression and bias plots.

RESULTS:

The laboratory CVs were <5% and ICCs were uniformly high (>95%) for all androgens measured across sex/menopausal status groups. Spearman ranked correlations for 9 hormones measured in the comparison laboratory were high (>0.85), suggesting good agreement.

CONCLUSION:

Our high-performance LC-MS/MS assays of 11 androgens, including adrenal and gonadal androgenic precursors and their 5α-reduced metabolites demonstrated excellent laboratory reproducibility, and good comparability with an established method that measured 9 of the 11 hormones tested. The serum androgen metabolite assays are suitable for use in epidemiologic research.

KEYWORDS:

Endogenous hormones; Liquid chromatography tandem mass spectrometry; Metabolites; Reproducibility; Serum androgen assay

PMID:
26416142
PMCID:
PMC4663146
DOI:
10.1016/j.jsbmb.2015.09.032
[Indexed for MEDLINE]
Free PMC Article

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