甲基屈他雄酮在人体内的代谢产物研究  

Analysis of methasterone in human urine by gas chromatography-mass spectrometry

作　　者：景晶[1] 王杉[1] 刘欣[1] 杨声[1] 邓静[1] 张玉梅[1] 杨志勇[1] 张力思[1] 张亦农[1] JING Jing;WANG Shan;LIU Xin;YANG Sheng;DENG Jing;ZHANG Yu-mei;YANG Zhi-yong;ZHANG Li-si;ZHANG Yi-nong(National Anti-doping Laboratory,China Anti-doping Agency,Beijing 100029,China)

机构地区：[1]国家体育总局反兴奋剂中心,北京100029

出　　处：《药物分析杂志》2020年第4期644-650,共7页Chinese Journal of Pharmaceutical Analysis

摘　　要：目的:研究口服甲基屈他雄酮(methasterone)后人尿中代谢产物的情况。方法:招募健康志愿者口服甲基屈他雄酮100 mg,收集服药前空白尿样,服药后72 h内全部尿样,以及第4至20天晨尿。尿样经过酶解、液-液萃取、衍生化等前处理步骤,采用GC/MSD-MS和GC-MS/MS检测尿样中甲基屈他雄酮及代谢产物,并研究其在人尿中的代谢情况。色谱-质谱条件:HP-1毛细管色谱柱(25 m×0.2 mm×0.11μm),柱温为程序升温(起始温度177℃,以3℃·min^-1的速率升至245℃,然后以17℃·min^-1的速率升至320℃,保持2.5 min),载气为高纯氦气,进样口温度280℃,柱压138 kPa,分流比10∶1,进样量2μL;质谱离子源为EI源,源温度230℃,电子能量70 eV,采集模式为Scan(m/z 40~700)和MRM。结果:研究结果表明,口服甲基屈他雄酮在人尿中的以原型甲基屈他雄酮及C3位羟基化还原产物为主,这2种物质是目前兴奋剂检测领域检测甲基屈他雄酮摄入的目标物质。结论:通过对口服甲基屈他雄酮在人尿中代谢产物的研究,发现可作为辅助检测甲基屈他雄酮摄入的代谢产物M2~M4,通过对多种代谢物信息综合研判,可降低假阳性风险,提高兴奋剂检测的可靠性、科学性。Objective:To investigate the metabolic profile of methasterone in human urine after oral administration of methasterone.Methods:A male volunteer orally took a single dose of 100 mg methasterone substance.Urine blank was collected before administration,and then all urine excreted within 72 hours after administration were collected.From day 4 to 20,the morning urines were also collected.The urine samples were pre-treated using enzymatic hydrolysis,liquid-liquid extraction and derivation and then examined by GC/MSD-MS and GCMS/MS to detect methasterone and its metabolites and investigate metabolic profile of methasterone.Analytical parameters:a HP-1 cross-linked capillary column(25 m×0.2 mm×0.11μm)was adopted,the initial column temperature was set at 177℃ and then increased at a rate of 3℃·min^-1 to 245℃ and continue to increase to 320℃ at a rate of 17℃·min^-1 to and then holding for 2.5 min;the transfer line temperature was maintained at 280℃,and the column pressure was 138 kPa;the split ratio was 10∶1,with 2μL injection volume;the electronsource temperature was 230℃ with electron energy of 70 eV;Data were obtained in Scan(m/z 40 to 700)and multiple reaction monitoring(MRM)mode.Results:This study found that methasterone and the C3 hydroxylated reduzate were the major metabolites of methasterone in human urine,which were now the target compounds to determine methasterone abuse in doping detection.Conclusion:The findings of this study suggested that M2-M4 metabolites could be as the auxiliary markers to enhance the reliability of methasterone determination and reduce the possibility of false positive results.

关 键 词：甲基屈他雄酮 代谢产物 蛋白同化类固醇 甾体激素 硅烷化衍生物 气相色谱-质谱 

分 类 号：R917[医药卫生—药物分析学]