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多囊卵巢综合征相关雄性激素的液相色谱-串联质谱检测
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作者单位
曹正,刘颖,丛宇婷,卢一凡,董莹,刘京瑞,唐国栋,翟燕红 1.首都医科大学附属北京妇产医院检验科2.上海爱博才思分析仪器贸易有限公司3.北京市海淀区妇幼保健院产前诊断中心 
基金项目:国家自然科学基金青年科学基金项目(81702057)
中文摘要:建立了液相色谱-串联质谱(LC-MS/MS)检测血清中硫酸脱氢表雄酮(DHEAS)、雄烯二酮(A4)、睾酮(T)、17-羟基孕酮(17-OHP)、双氢睾酮(DHT)5种激素的分析方法。血清样本经蛋白沉淀后,采用固相萃取,经Agela Venusil MP C18色谱柱(3.0 mm×50 mm,3 μm)分离,以含0.1%甲酸的甲醇和含0.02%甲酸的水为流动相进行梯度洗脱。使用正-负离子多反应监测模式进行数据采集。结果表明,5种激素在各自质量浓度范围内线性关系良好(r2>0.995),回收率为96.0%~105%,各激素的批内精密度和批间精密度均小于10%,DHEAS、A4、T、17-OHP、DHT的定量下限(LOQ)分别为5.00、0.05、0.05、0.025、0.025 ng/mL。该方法快速、准确、灵敏,适用于临床对多囊卵巢综合征(PCOS)患者血清雄性激素水平的检测。
中文关键词:多囊卵巢综合征  液相色谱-串联质谱法(LC-MS/MS)  硫酸脱氢表雄酮  雄烯二酮  睾酮  17-羟基孕酮  双氢睾酮
 
Determination of Androgens Related to Polycystic Ovary Syndrome by Liquid Chromatography-Tandem Mass Spectrometry
Abstract:A liquid chromatography-tandem mass spectrometric(LC-MS/MS) method was established for the determination of five androgens,ie. dehydroepiandrosterone sulfate(DHEAS),androstenedione(A4),testosterone(T),17-hydroxyprogesterone(17-OHP) and dihydrotestosterone(DHT),in serum.The serum samples were extracted by solid phase extraction after protein precipitation,then separated on an Agela Venusil MP C18 column(3.0 mm×50 mm,3 μm) by gradient elution using methanol containing 0.1% formic acid and water with 0.02% formic acid as mobile phases,finally detected with electrospray ionization in positive or negative mode under multiple reaction monitoring(MRM) mode.As a result,there were good linear relationships for five androgens in their respective mass concentration ranges with correlation coefficients(r2) more than 0.995,and the recoveries ranged from 96.0% to 105%.The intra- and inter-assay precision values for all androgens were less than 10%.The limits of quantitation for DHEAS,A4,T,17-OHP and DHT were 5.00,0.05,0.05,0.025,0.025 ng/mL,respectively.The method is rapid,accurate and sensitive,and could satisfy the requirements for clinical determination of serum androgens in patients with polycystic ovary syndrome(PCOS).
Key Words:polycystic ovary syndrome(PCOS)  liquid chromatography-tandem mass spectrometry(LC-MS/MS)  dehydroepiandrosterone sulfate(DHEAS)  androstenedione(A4)  testosterone(T)  17 hydroxyprogesterone(17 OHP)  dihydrotestosterone(DHT)
引用本文:曹正,刘颖,丛宇婷,卢一凡,董莹,刘京瑞,唐国栋,翟燕红.多囊卵巢综合征相关雄性激素的液相色谱-串联质谱检测[J].分析测试学报,2020,39(1):146-151.
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