Development and validation of a sensitive liquid chromatography-tandem mass spectrometry assay to simultaneously measure androgens and estrogens in serum without derivatization
Development and validation of a sensitive liquid chromatography-tandem mass spectrometry assay to simultaneously measure androgens and estrogens in serum without derivatization
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DOI：
10.1016/j.cca.2009.09.003
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发表时间：
2009-11-03
影响因子：
5
通讯作者：
Handelsman, David J.
中科院分区：
文献类型：
--
作者：
Harwood, D. Tim;Handelsman, David J.
背景：免疫分析法被广泛用于定量生物样品中的类固醇激素。然而，它们缺乏特异性，特别是在低水平上。方法：建立了稳定同位素稀释LC-MS/MS测定血清中睾酮（T）、双氢睾酮（DHT）、雌二醇（E2）和雌酮（E1）的方法。样品制备涉及用含有氘代内标的己烷：乙酸乙酯（3：2）进行液-液萃取（LLE）。结果：使用200 μ L血清，定量限分别为0.3 pg（1.5 pg/mL）E（1），0.5 pg（2.5 pg/mL）E（2），2 pg（10 pg/mL）T和10 pg（50 pg/mL）DHT。准确度（93-110%）和精密度（中位数4%，所有
Background: Immunoassays are widely used to quantify steroid hormones in biological samples. However, they lack specificity, especially at low levels. This study aimed to develop a sensitive LC-MS/MS method to measure serum androgens and estrogens without derivatization within a single run.Methods: A stable-isotope dilution LC-MS/MS method was established using atmospheric pressure photoionization to quantify testosterone (T), dihydrotestosterone (DHT), estradiol (E2) and estrone (E1) from serum. Sample preparation involved liquid-liquid extraction (LLE) with hexane:ethyl acetate (3:2) containing deuterated internal standards. Accuracy was assessed by spiked recovery of serum pools, and imprecision by quality controls.Results: Using 200 mu L serum, limits of quantification were 0.3 pg (1.5 pg/mL) E(1), 0.5 pg (2.5 pg/mL) E(2), 2 pg (10 pg/mL) T and 10 pg (50 pg/mL) DHT. Accuracy (93-110%) and precision (median 4%, all