Simultaneous measurement of the circulating testosterone (TS) and dehydroepiandrosterone sulfate (DHEAS) is deemed to be helpful for the assessment of men's health.Liquid chromatography/electrospray ionization-tandem mass spectrometry (LC/ESI-MS/MS) is the most reliable methodology for this purpose, however, it has room for improvement in analysis throughput.In this study, a quadruplicate of the Girard reagents was used to develop an LC/ESI-MS/MS method capable of quantifying TS and DHEAS in four different serum samples in a single run. The four serum samples were separately pretreated, derivatized with one of four Girard reagents, then combined.The LC/ESI-MS/MS analysis of the combined sample provided the androgen concentrations of four serum samples in parallel. The method had practical measuring ranges, in which good precision and accuracy, and negligible matrix effects were verified. The speed-up capability of the developed method was evaluated through the analysis of 10 batches of serum samples (total 40 samples); the method saved a 60% post-pretreatment analysis time compared to the non-derivatization method for 40 samples.
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