An LC-MS/MS analysis for seven sex hormones in serum
An LC-MS/MS analysis for seven sex hormones in serum
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血清中七种性激素的 LC-MS/MS 分析
DOI：
10.1016/j.jpba.2018.09.014
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发表时间：
2019-01-05
影响因子：
3.4
通讯作者：
Li, Yan
中科院分区：
文献类型：
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作者：
Yuan, Teng-Fei;Le, Juan;Li, Yan
背景：全面的血清性激素谱对于监测许多相关疾病的发生和发展至关重要。然而，现有的多类性激素检测方法缺乏标准参考物质（SRM）认证，样品消耗量大。为了改进现有的液相色谱-串联质谱联用技术，（LC-MS/MS）方法侧重于SRM认证和最小化血清消耗，用于同时定量包括雌激素在内的7种主流血清性激素（雌酮E1、雌二醇E2和雌三醇E3）、雄激素（睾酮T、雄烯二酮AD、脱氢表雄酮DHEA）和孕激素方法：为了实现一批分析，设计并仔细优化了一种简单的策略。示意性地，首先用同位素标记的内标物加标血清。然后，用甲基叔丁基醚进行液-液萃取。在氮气下干燥后，引入丹磺酰氯用于衍生化。结果：E1、E2和E3的最低定量浓度为0.005ng/mL，T、P和AD的最低定量浓度为0.01ng/mL，DHEA的最低定量浓度为0.25ng/mL。批间和批内CV均小于11.8%。通过干扰加标试验证明选择性令人满意。通过系统SRM验证，观察到的平均偏倚为-5.4%至4.7%，这表明方法具有良好的可靠性。我们发现化学发光免疫法（CLIA）检测结果与现有方法相比存在显著的正偏倚，这促使我们重新考虑我们先前关于男性冠状动脉粥样硬化性疾病患者性激素调节的研究结果。结论：建立了一种多类别血清性激素谱分析的LC-MS/MS方法，该方法具有SRM认证和最小化血清用量的优点。利用这一可靠的方法，对以往基于CLIA的男性冠状动脉粥样硬化患者性激素调节的研究结果进行了修正和完善。(C)2018 Elsevier B. V.版权所有。
Background: Comprehensive serum sex hormone profiling is essential for monitoring the occurrence and development of many related diseases. However, the current methods for multi-class sex hormone detection were always lack of Standard Reference Material (SRM) certification and suffered from large sample consumption. For improvement, we developed a new liquid chromatography-tandem mass spectrometry (LC-MS/MS) method focused on SRM certification and minimization of serum consumption for simultaneous quantification of seven mainstream serum sex hormones including estrogens (estrone E1, estradiol E2 and estriol E3), androgens (testosterone T, androstenedione AD, dehydroepiandrosterone DHEA) and progestogens (progesterone P).Methods: To achieve one-batch analysis, a straightforward strategy was designed and carefully optimized. Schematically, serum was firstly spiked with isotope-labeled internal standards. Then, liquid-liquid extraction was performed with methyl tert-butyl ether. After drying under nitrogen, dansyl chloride was introduced for derivatization. Finally, the mixture was submitted to LC-MS/MS for quantification.Results: The limit of quantification was 0.005 ng/mL for E1, E2 and E3, 0.01 ng/mL for T, P and AD, 0.25 ng/mL for DHEA. Inter- and intra-assay CVs were less than 11.8%. The selectivity was proved satisfactory by interference spiking tests. With systematical SRM validation, the mean bias of -5.4 to 4.7% was observed, which indicated excellent method reliability. We found significant positive bias in chemiluminescence immunoassay (CLIA) detection comparing with current method, which promoted us to reconsider our previous results on sex hormone regulation in male patients with coronary atherosclerotic disease. After redetecting the related samples, modified and improved conclusions were proposed.Conclusions: A LC-MS/MS method for multi-class serum sex hormone profiling was developed with SRM certification and minimized serum consumption. Taking advantages of such reliable method, the previous CLIA-based research findings on sex hormone regulation in male patients with coronary atherosclerosis were modified and improved after redetecting the same sample-pool. (C) 2018 Elsevier B.V. All rights reserved.