高效液相色谱串联质谱法同时定量测定人血清中脱氢表雄酮、睾酮及雄酮  

HPLC-MS/MS(ESI) simultaneous determination of dehydroepiandrosterone,testosterone and androsterone in human serum

作　　者：黄河花[1] 刘东阳[1] 胡蓓[1] 江骥[1] 

机构地区：[1]中国协和医科大学北京协和医院临床药理中心,北京100730

出　　处：《药物分析杂志》2012年第2期210-216,共7页Chinese Journal of Pharmaceutical Analysis

摘　　要：目的:建立一种快速、灵敏地同时测定人血清中脱氢表雄酮、睾酮及雄酮的高效液相色谱串联质谱(电喷雾电离模式)定量方法。方法:血清样品选用甲基叔丁基醚(MTBE)萃取后,取上清液用盐酸羟胺对3种激素进行柱前衍生化后进样测定。色谱柱为Waters YMC ODS-AQ(2.0 mm×150 mm,3.0μm),流动相为70%乙腈和30%醋酸(0.02%)溶液,流速为0.2 mL·min-1,柱温为50℃。本文采用电喷雾离子源进行正离子离子化,使用MRM检测模式选择性监测脱氢表雄酮(m/z 304.2/253.2)、睾酮(m/z 304.2/124.0)、雄酮(m/z 306.2/255.2)及内标炔诺酮(m/z 314.1/124.2)。结果:脱氢表雄酮、睾酮与雄酮的最低定量限(LLOQ)分别为0.10,0.05,0.10 ng·mL-1;标准曲线相关系数(r)均大于0.99;3种激素的日间及日内方法回收率均在±115%之内,日内及日间精密度(RSD)均小于15%。结论:实现在ESI电离模式下,用HPLC-MS/MS同时测定脱氢表雄酮、睾酮与雄酮3种甾体激素;本方法具有良好的特异性、灵敏度、准确度及精密度,能满足人血清样品定量分析需要。Objective：To establish a rapid and sensitive method for simultaneous quantitative determination of dehydroepiandrosterone （DHEA）, testosterone （T） and androsterone （ADT） using high performance liquid chroma- tographytandem mass spectrometry （HPLC -MS/MS） equipped with electrospray （ESI） ionization interface. Methods： Dehydroepiandrosterone, testosterone, androsterone, and internal standard （norethisterone） were extracted from human serum using MTBE （methylterbutylethylene） followed by treatment with hydroxylamine hydrochloride to produce derivatives before determination. Analytes were separated on Waters YMC ODS - AQ （2.0 mm × 150 mm,3.0 μm） with the mobile phase of aeetonitrile and 0. 02% acetic acid （70： 30）. The flow rate was 0.2 mL · min- 1, and the column temperature was 50℃, resulting the well separation of chromatography peaks. The electros- pray ionization method and MRM detection mode with positive ion were adopted to selectively detect DHEA （m/z 304. 20/253.20） ,T （ m/z 304.2/124.0 ）, ADT （ m/z 306.2/255.2 ） and internal standard norethisteron （ m/z 314. 1/124.2）. Results：The results indicated the lower limits of quantitation of DHEA,T and ADT were 0. 10, 0. 05 and 0. 10 ng · mL - 1, respectively. All of correlation coefficients of standard curves （r） were higher than 0.99 ; the intra day/inter day method recoveries and precisions （RSD） in determining these three hormones were within 115 % and less than 15 %, respectively. Conclusion：Our method provided an efficient determination method for the detection of three kinds of hormones （DHEA ,T and ADT） in human serum using HPLC -MS/MS with ESI interface. This method is specific, sensitive, accurate and precise, satisfying the quntifieaiton requirements for human serum samples.

关 键 词：液相色谱串联质谱 电喷雾 睾酮 雄酮 脱氢表雄酮 肟衍生化 定量分析 兴奋剂监测 

分 类 号：R917[医药卫生—药物分析学]