超高效液相色谱-高分辨质谱快速筛选贴膏剂中170种兴奋剂  

Fast screening of 170 doping substances in pastes by ultra-high performance liquid chromatography high resolution mass spectrometry method

作　　者：费巧玲 景晶[1] 陆江海[1] 王小兵[1] 徐友宣[1] Fei Qiaoling;Jing Jing;Lu Jianghai;Wang Xiaobing;Xu Youxuan(China Anti-Doping Agency,General Administration of Sport,Beijing 10029,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》2024年第10期815-823,共9页Chinese Journal of Sports Medicine

摘　　要：目的:建立超高效液相色谱-高分辨质谱(UPLC-HRMS)快速筛选贴膏剂中兴奋剂的检测方法,以加强药源性兴奋剂风险防控。方法:贴膏剂样品除去外衬后,分别采用叔丁基甲醚和无水乙醇超声提取,合并提取液后氮气流吹干,采用甲醇溶解残留物,过滤后采用赛默飞Vanquish-Q Exactive Plus超高效液相色谱-高分辨质谱仪检测。采用安捷伦ZORBAX Eclipse C18(2.1×100 mm,1.8μm)色谱柱分离,柱温为40℃,进样体积为3μL,以10 mmol/L甲酸铵(含0.1%甲酸,pH 3.5)和乙腈为流动相进行梯度洗脱,分别在电喷雾正、负离子模式下,以全扫描/数据依赖的二级质谱(Full MS/dd-MS2)扫描模式检测。结果:该方法具有良好的选择性,样品经前处理后在48 h内检测具有稳定性,92种兴奋剂检出限约为3 ng/g,31种兴奋剂检出限约为6 ng/g,47种兴奋剂检出限约为30 ng/g。采用该方法检测10例贴膏剂样品,检出4例含士的宁,3例含去甲乌药碱,3例含麻黄碱。结论:本研究建立了超高效液相色谱-高分辨质谱(UPLC-HRMS)快速筛选贴膏剂中9个类别共170种兴奋剂的检测方法,该方法简单、灵敏、可靠,适用于贴膏剂中兴奋剂的快速筛选。Objective To develop a detection method with ultra-high performance liquid chromatography-high resolution mass spectrometry(UPLC-HRMS)for fast screening of doping substances in pastes,so as to strengthen the drug-borne doping risk prevention system.Methods The paste samples were removed the covering film and extracted by using tert-butyl methyl ether and anhydrous ethanol,respectively.The extracts were combined and blown dry under nitrogen flow at 40℃,and methanol was used to dissolve the residues,which were then filtered and analyzed with the Thermo Vaniquish Q Exactive Plus.Then,3μL of the solution was injected into and separated by the Agilent ZORBAXEclipse C18 column(2.1×100 mm,1.8μm),and the gradient elution was performed with 10 mM ammonium formate(containing 0.1%formic acid,pH3.5)and acetonitrile as mobile phase.The 170 kinds of doping substances were detected in positive ion electrospray(ESI+)and negative ion electrospray(ESI-)respectively under Full Mass/data-dependent mass spectrometry(Full MS/ddMS2)mode.Results The results showed that the method had good selectivity and samples were stable within 48 h after pretreatment.The detection limit(LOD)of 92 doping substances were about 3 ng/g,while that of another 31 and 47 ones was about 6 ng/g and 30 ng/g,respectively.In this study,among 10 paste samples,4 were tested positive for strychnine,and 3 were positive for higenamine and ephedrine,respectively.Conclusion A fast screening method for 170 kinds of doping substances in 9 categories is developed by using ultra-high performance liquid chromatography with high resolution mass spectrometry in this study.Such method proves to be simple,sensitive and reliable,which is suitable for fast screening of doping substances in pastes.

关 键 词：贴膏剂 兴奋剂 超高效液相色谱–高分辨质谱(UPLC–HRMS) 快速筛选 

分 类 号：R286.0[医药卫生—中药学]