Serum steroid profiling by isotopic dilution-liquid chromatography-mass spectrometry: Comparison with current immunoassays and reference intervals in healthy adults
Serum steroid profiling by isotopic dilution-liquid chromatography-mass spectrometry: Comparison with current immunoassays and reference intervals in healthy adults
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DOI：
10.1016/j.steroids.2010.11.005
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发表时间：
2011-02-01
期刊：
影响因子：
2.7
通讯作者：
Pagotto, Uberto
中科院分区：
文献类型：
--
作者：
Fanelli, Flaminia;Belluomo, Ilaria;Pagotto, Uberto
背景资料：同时，快速和可靠的测量广泛的类固醇面板是一个强大的工具，以解开生理和病理激素状态。目前临床实验室主要采用高通量免疫测定法，但由于交叉反应性和基质干扰，这些方法缺乏特异性。我们建立了一种同位素稀释-液相色谱-串联质谱（ID-LC-MS/MS）方法，用于同时测定血清中皮质醇、皮质酮、11脱氧皮质醇、雄烯二酮、脱氧皮质酮（DOC）、睾酮、17 OH孕酮、脱氢表雄酮（DHEA）和孕酮，并与我们实验室采用的常规免疫分析方法进行了比较。我们还建立了成人参考区间在416名健康subjects.Methods：0.9毫升的血清加标标记的内标（IS）和C18柱提取。将洗脱液注射到二维LC系统中，在灌注柱中纯化，并在21分钟梯度运行期间在C8柱上分离。结果：与ID-LC-MS/MS法比较，只有ElecsysE 170测定男性皮质醇、睾酮和> 1 ng/ml的孕酮的结果与ID-LC-MS/MS法一致，ElecsysE 170测定女性睾酮和孕酮的结果与ID-LC-MS/MS法一致
Background: The simultaneous, rapid and reliable measurement of a wide steroid panel is a powerful tool to unravel physiological and pathological hormone status. Clinical laboratories are currently dominated by high-throughput immunoassays, but these methods lack specificity due to cross-reactivity and matrix interferences. We developed and validated an isotopic dilution-liquid chromatography-tandem mass spectrometry (ID-LC-MS/MS) method for the simultaneous measurement of cortisol, corticosterone, 11deoxycortisol, androstenedione, deoxycorticosterone (DOC), testosterone, 17OHprogesterone, dehydroepiandrosterone (DHEA) and progesterone in serum, and compared it to routine immunoassays employed in our laboratory. We also established adult reference intervals in 416 healthy subjects.Methods: 0.9 ml of serum were spiked with labelled internal standards (IS) and extracted on C18 cartridges. Eluate was injected into a two-dimensional LC-system, purified in a perfusion column and separated on a C8 column during a 21 min gradient run. Analytes were revealed by atmospheric pressure chemical ionization (APCI) followed by multiple reaction monitoring (MRM) analysis.Results: Of the four immunoassays compared with the ID-LC-MS/MS method, only the results of ElecsysE170 for cortisol, testosterone in males and progesterone > 1 ng/ml were in agreement with ID-LC-MS/MS. ElecsysE170 for testosterone in females and progesterone