Isotope-dilution TurboFlow-LC-MS/MS method for simultaneous quantification of ten steroid metabolites in serum
Isotope-dilution TurboFlow-LC-MS/MS method for simultaneous quantification of ten steroid metabolites in serum
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DOI：
10.1016/j.cca.2017.03.002
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发表时间：
2017-05-01
影响因子：
5
通讯作者：
Juul, Anders
中科院分区：
文献类型：
--
作者：
Soeborg, Tue;Frederiksen, Hanne;Juul, Anders
建立了同时定量血清中甾体代谢产物脱氢表雄酮硫酸酯（DHEAS）、孕酮、17-羟孕酮（17- ohp）、4-雄烯二酮（Adione）、皮质酮、11-脱氧皮质醇、皮质醇、可的松、睾酮(T)和3-硫酸雌酮（El-S）的同位素稀释TurboFlow-LC-MS/MS方法，并进行了验证。定量限、变异性（日内和日内）、分析范围和线性均可用于临床。此外，还评估了样品的稳定性，包括冻融循环的影响以及温度和储存时间的影响。该方法应用于391份10-18岁丹麦健康男孩的血清样本。所包括的类固醇代谢物的浓度范围为这一群体提出。将391份血清样品的DHEAS、17-0HP、Adione和Tin浓度与本实验室现有的LC-MS/MS方法进行比较。两种方法之间的一致性非常好。此外，新方法的灵敏度提高了，可以对许多低于现有方法loq的样品进行定量分析。因此，这两种仪器及其相关方法被验证为彼此可能的备份，我们认为这是高通量实验室定期分析临床样本的一个极其重要的问题。所包括的十种分析物可以同时分析，但也可能只包括一些用于特定诊断目的的分析物，这使得新方法成为临床实验室中非常有用的工具。(C) 2017 Elsevier B.V.版权所有
An isotope-dilution TurboFlow-LC-MS/MS method for simultaneous quantification of the ten steroid metabolites dehydroepiandrosterone sulfate (DHEAS), progesterone, 17 alpha-hydroxyprogesterone (17-OHP), Delta 4-androstenedione (Adione), corticosterone, 11-deoxycortisol, cortisol, cortisone, testosterone (T), and estrone 3-sulfate (El-S) in serum was developed and validated. Limits of quantification, variability (inter- and infra-day), analytical range and linearity were all found to be acceptable for clinical use. Furthermore, sample stability was evaluated including the influence of freeze-thaw cycles and the effects of temperature and storage time.The method was applied to 391 serum samples from healthy, Danish boys 10-18 years old. The concentration ranges of the included steroid metabolites for this population are presented.Concentrations of DHEAS, 17-0HP, Adione and Tin the 391 serum samples were furthermore compared to results obtained using an existing LC-MS/MS method in our laboratory. Excellent agreement was found between the methods. Furthermore, the improved sensitivity of the new method allowed for quantification of a number of samples found to be below the LOQs of the existing method. Thus, the two instruments and their associated methods were validated as possible back-ups for each other, which we consider an extremely important issue in high-throughput laboratories analyzing clinical samples on a regular basis.The ten analytes included can be analyzed simultaneously but it is also possible only to include some of these analytes for specific diagnostic purposes which make the new method an extremely useful tool in the clinical laboratory. (C) 2017 Elsevier B.V. All rights reserved.