Shibata Yasuhiro, Arai Seiji, Honma Seijiro
Department of Urology, Gunma University Graduate School of Medicine, Maebashi, Japan.
Department of Urology, Gunma University Graduate School of Medicine, Maebashi, Japan.
J Steroid Biochem Mol Biol. 2015 Jan;145:193-9. doi: 10.1016/j.jsbmb.2014.04.002. Epub 2014 May 2.
A reliable and sensitive method for analyzing steroids using liquid chromatography tandem mass spectrometry (LC-MS/MS) is required for research concerning dehydroepiandrosterone (DHEA), which plays a central role in steroid hormone biosynthesis and metabolism. Furthermore, after the first proposal of the concept of intracrine DHEA, stable isotope tracer analysis, which is useful for structural recognition as well as determination of steroids, has been required to evaluate physiological action and hormone biosynthesis/metabolism in target organs. We describe sample processing and analysis methods for simultaneous quantification of multiple hormones, including DHEA, in serum, saliva and tissue using LC-MS/MS. A derivatization technique compatible with each functional group for measuring 3β-hydroxy-5-enes, such as DHEA and 5α/5β-steroids, with high sensitivity and specificity is also described. Finally, we describe a newly developed method for intracrine research using stable isotope-labeled (13)C-steroid substrates with tracer analysis of their metabolites by LC-MS/MS.This article is part of a Special Issue entitled'Essential role of DHEA'.
对于在类固醇激素生物合成和代谢中起核心作用的脱氢表雄酮（DHEA）的研究，需要一种可靠且灵敏的液相色谱串联质谱法（LC-MS/MS）来分析类固醇。此外，在首次提出内分泌性DHEA的概念后，为了评估靶器官中的生理作用以及激素生物合成/代谢，需要进行稳定同位素示踪分析，这对于类固醇的结构识别和测定很有用。我们描述了使用LC-MS/MS同时定量血清、唾液和组织中包括DHEA在内的多种激素的样品处理和分析方法。还描述了一种与每个官能团兼容的衍生化技术，用于高灵敏度和特异性地测量3β-羟基-5-烯类物质，如DHEA和5α/5β-类固醇。最后，我们描述了一种新开发的用于内分泌研究的方法，该方法使用稳定同位素标记的（13）C-类固醇底物，并通过LC-MS/MS对其代谢物进行示踪分析。本文是名为“DHEA的重要作用”的特刊的一部分。
