Simultaneous measurement of testosterone, androstenedione and dehydroepiandrosterone (DHEA) in serum and plasma using Isotope-Dilution 2-Dimension Ultra High Performance Liquid-Chromatography Tandem Mass Spectrometry (ID-LC–MS/MS)

Comparison of 7 Published LC-MS/MS Methods for the Simultaneous Measurement of Testosterone, Androstenedione, and Dehydroepiandrosterone in Serum

“…One of the other methods in this study (method A) was found to agree indirectly with the reference method for testosterone (10 ). Because the testosterone methods investigated in this study agreed well, we believe that the investigated testosterone methods report values that are close to the true testosterone concentrations.…”

“…Others, however, reported much higher variation and less strong agreement between the investigated LC-MS/MS methods for testosterone (8,9 ). Data on the agreement of LC-MS/MS assays for adrenal steroids are even scarcer (10 ). Second, since the publication of Taieb et al…”

“…Several articles on LC-MS/MS methods for testosterone and other steroid hormones have recently been published (5 ). This development, as well as the urgent need for reliable assays, prompted the editorial board of the Journal of Clinical Endocrinology and Metabolism (JCEM) 10 to state that from 2015 on, "manuscripts reporting sex steroid assays as important endpoints must use MS-based assays," and "it is anticipated that this requirement [to use] MS-based assays will extend to adrenal steroids and vitamin D in the near future" (6 ). Although we agree with the requirement for reliable steroid assays and understand the superiority of LC-MS/MS compared with immunoassays, the JCEM statement raises several questions.…”

Inaccurate First-Generation Testosterone Assays Are Influenced by Sex Hormone–Binding Globulin Concentrations

“…This method is very accurate, sensitive, and well standardized, as was shown in previous studies (7)(8)(9). Intraassay variation in both the female and male concentration range was 4%.…”

“…In all serum samples, total testosterone was measured using an isotope dilution (ID)-LC-MS/MS method, as de- scribed earlier (7). In short, the serum samples underwent a liquid-liquid extraction using hexane ether (4:1) after addition of internal standards:…”