气相色谱-燃烧-同位素比质谱在检测尿样中外源性睾酮及其代谢物中的应用  

Determination of the C/C Ratios of Testosterone and Its Main Metabolites by Gas Chromatographycombustion-isotope Ratio Mass Spectrometry

作　　者：温超 王静竹[1] 朱天硕 刘欣[1] 王杉[1] 张亦农[1] Wen Chao;Wang Jingzhu;Zhu Tianshuo;Liu Xin;Wang Shan;Zhang Yinong(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：《中国运动医学杂志》2020年第9期725-731,共7页Chinese Journal of Sports Medicine

摘　　要：目的:建立一套气相色谱-燃烧-同位素比质谱(GC-C-IRMS)检测方法,对尿样中的睾酮及其主要代谢物的来源进行确认。方法:按照世界反兴奋剂机构(WADA)的检测要求,建立GC-C-IRMS检测方法,并对该方法的最低检测浓度、稳定性、重现性、线性、不确定度、人群值等进行分析。结果:本研究建立的GC-C-IRMS检测方法的检测浓度线性范围为10~13000 ng/mL,在取用15 mL尿样时,对尿样中含量较低的睾酮、5α-雄烷-3α,17β-二醇、5β-雄烷-3α,17β-二醇、孕烷二醇的最低检测浓度为10 ng/mL,对尿样中雄酮、本胆烷醇酮等浓度较高的物质的最低检测浓度为100 ng/mL。本方法对各分析物的不确定度小于1‰,人群值检测结果满足MeanΔERC-TC±2SD<3‰,符合WADA检测标准。结论:本研究建立的GC-C-IRMS检测方法取用15 mL尿样即可对10 ng/mL的睾酮及其主要代谢产物进行分析。相比国际常用的二次液相分离与化学衍生化结合的检测方法,本文中的方法只需一次液相分离,无需衍生化,分析时间短,效率高,可用于兴奋剂常规检测及大型赛事检测。Objective To develop an efficient and stable gas chromatography-combustion-isotope ratio mass spectrometry(GC-C-IRMS) procedure to determine the origin of testosterone(T) in urine samples.Method According to the cretirion of the World Anti-Doping Agency(WADA),the procedure was established,and the lowest detection concentration,stability,repeatability,linearity,uncertainty,and population reference were analyzed. Results The linearity range of the procedure studies was from 10 to13000 ng/mL. The lowest detection concentration was 10 ng/mL for T,5α-Androstane-3α,17β-diol(5α-diol),5β-Androstane-3α,17β-diol(5β-diol) and pregnanediol(PD),but 100 ng/mL for An and Etio which were of high concentration in urine samples. The uncertainty of the procedure was less than 1‰,and the population reference met the WADA criterion(MeanΔERC-TC±2 SD<3‰). Conclusion The results obtained meet the criteria of WADA’s validation requirement. The running time of whole confirmation procedure is within 36 h for one batch of analysis. Only one HPLC purification procedure is required to isolate the target compounds and no chemical derivatization is involved. The present method can be used as routine anti-doping analysis for laboratory as well as for large-scale sports events.

关 键 词：气相色谱-燃烧-同位素比质谱 兴奋剂检测 液相色谱 睾酮 样品纯化 

分 类 号：R446.12[医药卫生—诊断学]