液相色谱-离子淌度差分质谱法测定人血清中睾酮及雄烯二酮的浓度  

Determination of testosterone and androstenedione in human serum by high performance liquid chromatography tandem differential mobility spectrometry method

作　　者：周从亚 徐慧玉 杨平 杨丽[1] 赵荣生[1] 翟所迪[1] 张现化[1] ZHOU Cong-ya;XU Hui-yu;YANG Ping;YANG Li;ZHAO Rong-sheng;ZHAI Suo-di;ZHANG Xian-hua(Department of Pharmacy,Peking University Third Hospital,Beijing 100191,China;Reproductive Center,Peking University Third Hospital,Beijing 100191,China)

机构地区：[1]北京大学第三医院药剂科,北京100191 [2]北京大学第三医院生殖中心,北京100191

出　　处：《中国临床药理学杂志》2020年第12期1701-1704,共4页The Chinese Journal of Clinical Pharmacology

基　　金：北京大学第三医院临床重点基金资助项目(BYSY2018022)。

摘　　要：目的建立一种能够灵敏、准确地测定人血清中睾酮及雄烯二酮的液相色谱-离子淌度差分质谱(HPLC-DMS-MS/MS)方法。方法空白血清采用活性炭吸附法处理去除待测物。用叔丁基甲醚液液萃取处理,进HPLC-DMS-MS/MS分析,色谱柱:Poroshell SB-C18(150×2.1 mm,2.7μm)。流动相为5 mM甲酸铵水溶液(含0.1%甲酸)-乙腈(含0.1%甲酸及5%水),梯度洗脱,流速为0.2 mL·min^-1,柱温:40℃,进样量:10μL,电喷雾电离,用多反应检测模式,定量分析离子对分别为m/z 289.2→97.0(睾酮),m/z 287.2→97.1(雄烯二酮),m/z:292.2→97.1(内标睾酮-D3)。结果睾酮的标准曲线方程为y=2.39x+8.34×10^-3(r=0.9933),线性范围为0.05~20 ng·mL^-1,雄烯二酮的标准曲线方程为y=1.52x+4.21×10^-3(r=0.9936),线性范围均为0.05~20 ng·mL^-1,睾酮和雄烯二酮的批内、批间精密度均在4.17%~13.61%,准确度均在93.33%~112.13%,平均提取回收率在64.20%~80.67%,稳定性好。结论本方法处理简便、灵敏度高,专属性强,适用于雄激素水平极低患者血清中睾酮及雄烯二酮的浓度检测。Objective To develop a method for the determining the concentration of testosterone and androstenedione simultaneously in human serum.Methods Blank matrix was obtained by purification of serum with activated carbon.The clinical serum samples were treated by liquid-liquid extraction with methyltertbutylether(MTBE),and then were determined by high performance liquid chromatography tandem differential mobility spectrometry(HPLC-DMS/MS/MS).The column was Poroshell SB-C18(150×2.1 mm,2.7μm).The flow rate was 0.2 mL·min^-1,the column temperature was 40℃,the injection volume was 10μL,and the electrospray ionization source was used.The multi-reaction monitoring mode was used to quantitatively analyze the ionization pairs as 289.2→97.0(Testosterone),m/z:287.2→97.1(Androstenedione)and m/z:292.2→97.1(Testosterone-D3).Results The standard curve of testosterone was y=2.39x+8.34×10^-3(r=0.9933)with a linear range of 0.05-20 ng·mL^-1.The standard curve of androstenedione was y=1.52x+4.21×10^-3(r=0.9936)with a linear range of 0.05-20 ng·mL^-1.The inter-and intra-assay precisions were between 4.17%and 13.61%,the accuracies were between 93.33%and 112.13%.The average extraction recoveries of the two analytes were between 64.20%and 80.67%,and the stability was good.Conclusion The method was simple,sensitive as well as specific,suitable for monitoring the concentration of testosterone and androstenedione in human serum.

关 键 词：睾酮 雄烯二酮 液相色谱-离子淌度差分质谱 

分 类 号：R97[医药卫生—药品]