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*TESTOSTERONE
The Journal of Clinical Endocrinology & Metabolism Vol. 89, No. 2 534-543
Copyright © 2004 by The Endocrine Society

Special Feature

Measurement of Total Serum Testosterone in Adult Men: Comparison of Current Laboratory Methods Versus Liquid Chromatography-Tandem Mass Spectrometry

Christina Wang, Don H. Catlin, Laurence M. Demers, Borislav Starcevic and Ronald S. Swerdloff

Division of Endocrinology (C.W., R.S.S.), Department of Medicine, Harbor-UCLA Medical Center and Research and Education Institute, Torrance, California 90502; UCLA-Olympic Analytical Laboratory (D.H.C., B.S.), Los Angeles, California 90025; and Department of Pathology and Medicine (L.M.D.), Pennsylvania State University College of Medicine, H. S. Hershey Medical Center, Hershey, Pennsylvania 17033

Address all correspondence and requests for reprints to: Christina Wang, M.D., UCLA School of Medicine, General Clinical Research Center, Box 16, 1000 West Carson Street, Torrance, California 90502.

The diagnosis of male hypogonadism requires the demonstration of a low serum testosterone (T) level. We examined serum T levels in pedigreed samples taken from 62 eugonadal and 60 hypogonadal males by four commonly used automated immunoassay instruments (Roche Elecsys, Bayer Centaur, Ortho Vitros ECi and DPC Immulite 2000) and two manual immunoassay methods (DPC-RIA, a coated tube commercial kit, and HUMC-RIA, a research laboratory assay) and compared results with measurements performed by liquid chromatography-tandem mass spectrometry (LC-MSMS). Deming’s regression analyses comparing each of the test results with LC-MSMS showed slopes that were between 0.881 and 1.217. The interclass correlation coefficients were between 0.92 and 0.97 for all methods. Compared with the serum T concentrations measured by LC-MSMS, the DPC Immulite results were biased toward lower values (mean difference, -90 ± 9 ng/dl) whereas the Bayer Centaur data were biased toward higher values (mean difference, +99 ± 11 ng/dl) over a wide range of serum T levels. At low serum T concentrations (<100 ng/dl or 3.47 nmol/liter), HUMC-RIA overestimated serum T, Ortho Vitros ECi underestimated the serum T concentration, whereas the other two methods (DPC-RIA and Roche Elecsys) showed differences in both directions compared with LC-MSMS. Over 60% of the samples (with T levels within the adult male range) measured by most automated and manual methods were within ± 20% of those reported by LC-MSMS. These immunoassays are capable of distinguishing eugonadal from hypogonadal males if adult male reference ranges have been established in each individual laboratory. The lack of precision and accuracy, together with bias of the immunoassay methods at low serum T concentrations, suggests that the current methods cannot be used to accurately measure T in females or serum from prepubertal subjects.

This work was supported by the Core Endocrine Laboratory at Penn State-Hershey Medical Center; National Institutes of Health (NIH) Grant MO1 RR00543 to the GCRC at Harbor-UCLA Medical Center; NIH Grants RO1 CA 71053 and RO1 DK 61006 (to C.W., D.H.C., and R.S.S.); and United States Anti-Doping Agency (to D.H.C.). The samples were collected by the nurses of the Harbor-UCLA GCRC, supported by NIH Grant MO1 RR00425.

Abbreviations: CV, Coefficient of variation; GC, gas chromatograph; HRP, horseradish peroxidase; HUMC, Harbor-UCLA Research and Education Institute Endocrine Research Laboratory; LC-MSMS, liquid chromatography-tandem mass spectrometry; LOQ, limit of quantification; MS, mass spectrometry; T, testosterone.




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