Institute of Endocrinology, Národni 8, Prague, 11694, Czech Republic; University of Chemistry and Technology, Technická 6, Prague, 16628, Czech Republic.
Institute of Endocrinology, Národni 8, Prague, 11694, Czech Republic.
J Chromatogr B Analyt Technol Biomed Life Sci. 2022 Jun 30;1201-1202:123294. doi: 10.1016/j.jchromb.2022.123294. Epub 2022 May 20.
A development of robust and rapid method with simple sample preparation for the analysis of steroids of C-, C-, C- families is of interest of many research groups. Here we present a novel LC-MS/MS method for the simultaneous quantification of 32 steroid hormones in human plasma. Twenty-two of them were analyzed directly without the need for derivatization, while ten were derivatized with 2-fluoro-1-methylpyridinium p-toluenesulfonate. The steroids were separated on a C18 column with a gradient elution consisting of methanol and water with the addition of 0.1% formic acid. The mass spectrometer was operated in positive ESI mode. Validation demonstrated that the method was applicable for the quantitative analysis of two C- steroids (estrone, estradiol), nineteen C- steroids (testosterone, epitestosterone, dihydrotestosterone, 11-ketodihydrotestosterone, 11β-hydroxyandrostenedione, 11β-hydroxytestosterone, 11-ketotestosterone, dehydroepiandrosterone, 7α-hydroxydehydroepiandrosterone, 7β-hydroxydehydroepiandrosterone, 7-ketodehydroepiandrosterone, androsterone, epiandrosterone, androstenedione, androstenediol, 5α-androstane-3α,17β-diol, 5α-androstane-3β,17β-diol, 5β-androstane-3α,17β-diol, 5β-androstane-3β,17β-diol), and eleven C- steroids (cortisol, 21-deoxycortisol, 11-deoxycortisol, cortisone, corticosterone, 11-deoxycorticosterone, pregnenolone, 17-hydroxypregnenolone, progesterone, 17-hydroxyprogesterone, 5α-dihydroprogesterone). The lower limits of quantification are appropriate for analyses in both physiological and various pathophysiological conditions. The accuracy, intra- and inter-day precision values as well as stability tests were in accordance with FDA Guidelines. The method will be a useful tool in investigating the mechanisms of steroid-related diseases and will serve as a steppingstone for the development of other methods for steroid analyses in various biological matrices such as prostate tissue, cerebrospinal fluid, urine, seminal fluid, and saliva.
建立一种可同时分析 32 种人血浆甾体激素的灵敏、快速且样品处理简单的分析方法，这是许多研究小组感兴趣的课题。本文报道了一种新的 LC-MS/MS 方法，可直接分析其中 22 种甾体激素，而另外 10 种甾体激素则需要用 2-氟-1-甲基吡啶对甲苯磺酸盐衍生化。这些甾体激素在 C18 柱上进行梯度洗脱，流动相由甲醇和水组成，其中添加了 0.1%甲酸。质谱仪采用正电喷雾模式进行检测。方法学验证结果表明，该方法适用于两种 C-甾体激素（雌酮、雌二醇）、19 种 C-甾体激素（睾酮、表睾酮、二氢睾酮、11-酮基二氢睾酮、11β-羟基雄烯二酮、11β-羟基睾酮、11-酮基睾酮、脱氢表雄酮、7α-羟基脱氢表雄酮、7β-羟基脱氢表雄酮、7-酮基脱氢表雄酮、雄酮、表雄酮、雄烯二酮、雄烯二醇、5α-雄烷-3α,17β-二醇、5α-雄烷-3β,17β-二醇、5β-雄烷-3α,17β-二醇、5β-雄烷-3β,17β-二醇）和 11 种 C-甾体激素（皮质醇、21-脱氧皮质醇、11-脱氧皮质醇、皮质酮、皮质醇、11-脱氧皮质酮、孕烯醇酮、17-羟孕烯醇酮、孕酮、17-羟基孕酮、5α-二氢孕酮）的定量分析。定量下限适用于生理和各种病理生理条件下的分析。准确度、日内和日间精密度以及稳定性测试均符合 FDA 指南。该方法将成为研究甾体激素相关疾病机制的有用工具，并为开发其他生物基质（如前列腺组织、脑脊液、尿液、精液和唾液）中的甾体激素分析方法奠定基础。
