Simultaneous Measurement of Serum Testosterone and Dihydrotestosterone by Liquid Chromatography-Tandem Mass Spectrometry

Background: Recent reports have described inherent problems with androgen immunoassays compared with mass spectrometry analyses. Methods: We developed a method for measuring serum testosterone (T) and 5α-dihydrotestosterone (DHT) simultaneously via liquid–liquid extraction followed by liquid chromat...

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Vydáno v:	Clinical chemistry (Baltimore, Md.) Ročník 54; číslo 11; s. 1855 - 1863
Hlavní autoři:	Shiraishi, Steve, Lee, Paul W. N, Leung, Andrew, Goh, Victor H. H, Swerdloff, Ronald S, Wang, Christina
Médium:	Journal Article
Jazyk:	angličtina
Vydáno:	Washington, DC Am Assoc Clin Chem 01.11.2008 American Association for Clinical Chemistry Oxford University Press
Témata:	Adult Analytical, structural and metabolic biochemistry Androgens Aqueous solutions Biological and medical sciences Biomedical research Calibration Chromatography, Liquid - methods Dihydrotestosterone - blood Female Females Fundamental and applied biological sciences. Psychology Humans Immunoassays Investigative techniques, diagnostic techniques (general aspects) Ionization Liquid chromatography Male Mass spectrometry Medical sciences Methods Middle Aged Physical examinations Radioimmunoassay Reference Values Research centers Sensitivity and Specificity Spectrometry, Mass, Electrospray Ionization Tandem Mass Spectrometry - methods Testosterone Testosterone - blood Validation studies Testosterone Dihydrotestosterone Androgen Mass spectrometry MS/MS Clinical biology Simultaneous measurement Serum Biochemistry Liquid chromatography Testicular hormone Molecular biology Sex steroid hormone
ISSN:	0009-9147, 1530-8561, 1530-8561
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Shrnutí:	Background: Recent reports have described inherent problems with androgen immunoassays compared with mass spectrometry analyses. Methods: We developed a method for measuring serum testosterone (T) and 5α-dihydrotestosterone (DHT) simultaneously via liquid–liquid extraction followed by liquid chromatography–tandem mass spectrometry (LC-MS/MS) with positive-mode electrospray ionization. Results: The DHT and T calibrators showed a linear response from 0.069 nmol/L to 34.4 nmol/L and 69.3 nmol/L, respectively. T interference in the DHT assay and vice versa were negligible. Within- and between-run imprecision values were <5% for both analytes. Percent recoveries of T and DHT spiked into samples at concentrations spanning the calibration curve were 100%–113% and 98%–107%, respectively. The lower limit of quantification was 0.069 nmol/L for both steroids. Serum T concentrations measured by LC-MS/MS were different from those obtained by RIA, especially at lower T concentrations. Serum DHT concentrations measured by LC-MS/MS were markedly lower than those generated by RIA because of the nonselectivity of the RIA without chromatography. The reference intervals (mean ± 2 SDs) determined for T and DHT were 9.2–33.7 nmol/L and 0.47–2.65 nmol/L, respectively, for 113 healthy adult men and 0.33–2.02 nmol/L and 0.09–0.91 nmol/L, respectively, for 133 healthy premenopausal women. Conclusions: We have developed and validated a selective and precise method for simultaneous measurements of serum T and DHT that can be adopted for routine measurements of these androgens in health and disease in men and women.
Bibliografie:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 14 content type line 23 ObjectType-Undefined-3
ISSN:	0009-9147 1530-8561 1530-8561
DOI:	10.1373/clinchem.2008.103846