超高效液相色谱-串联质谱法测定运动营养食品中60种类固醇激素  

Determination of 60 steroid hormones in sports nutrition foods by ultra performance liquid chromatography-tandem mass spectrometry

作　　者：蔡小明 吴少明 余清 黄何何 郑秋萍 何孟杭 戴明 欧阳立群 CAI Xiaoming;WU Shaoming;YU Qing;HUANG Hehe;ZHENG Qiuping;HE Menghang;DAI Ming;OUYANG Liqun(Institute of Food Inspection,Fujian Inspection and Research Institute for Product Quality,National Quality Supervision and Testing Center for Processed Food,Fuzhou 350002,China)

机构地区：[1]福建省产品质量检验研究院食品检验研究所,国家加工食品质量检验检测中心,福建福州350002

出　　处：《色谱》2024年第12期1173-1188,共16页Chinese Journal of Chromatography

基　　金：国家市场监督管理总局科技计划项目(2021MK054);福建省市场监督管理局科技项目(FJMS2023014)~~。

摘　　要：基于QuEChERS,结合超高效液相色谱-串联质谱(UPLC-MS/MS)技术,建立了一种同时测定运动营养食品中60种类固醇激素的分析方法。样品以1.0%(v/v)甲酸乙腈提取,氯化钠进行分层,150 mg PSA和150 mg C18粉末净化,净化液经氮气吹至近干后用50%甲醇水溶液复溶,在Waters Acquity UPLC HSS T3分析柱上,以0.1%(v/v)甲酸水溶液(含10 mmol/L乙酸铵)-甲醇为流动相,电喷雾离子源(ESI)正离子模式下进行UPLC-MS/MS多反应监测(MRM)测定。在最优的仪器参数及前处理条件下进行方法验证,60种类固醇激素在1.0~100 ng/mL范围内具有良好的线性关系,相关系数(r)均大于0.99;方法的检出限(LOD)和定量限(LOQ)分别为0.2~0.8μg/kg和0.6~2.4μg/kg。以乳清蛋白粉(固体)、果胶蛋白肽(半固体)、运动营养液(液体)为空白基质,进行3个水平的加标回收试验(n=6),60种类固醇激素的平均回收率为73.7%~112.7%,相对标准偏差(RSD)为3.2%~10.1%。将新开发的方法应用于13批次实际样品的测定,在其中1份样品中检出禁用药物(勃地酮),含量为14.6μg/kg,说明运动营养食品中确有非法添加类固醇激素的现象,后续应加强对运动营养食品中该类固醇类激素的监控。该方法简单高效,灵敏度高,重复性好,适用于运动营养食品中类固醇激素的快速筛查和定量分析,为运动营养食品中类固醇激素的日常监督提供技术支撑。Recent societal developments and improving living standards have witnessed a growing enthusiasm for sport and fitness along with a boom in sports nutrition foods.However,rapid industrial development has led to product quality and safety issues becoming increasingly important.Steroid hormones exhibit physiological effects that include promoting protein synthesis,increasing muscle mass,and reducing body fat.Driven by vested interests,some unscrupulous traders illegally add steroid hormones to sports nutrition foods to improve sports performance and relieve muscle pain,which poses a potential threat to human health.Therefore,establishing a method for determining steroid hormones in sports nutrition foods is of great practical importance.In this study,an analytical method for the simultaneous determination of 60 steroid hormones in sports nutrition foods was developed using QuEChERS combined with ultra performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS).The MS parameters and chromatographic conditions were optimized for the 60 steroid hormones,and the extraction solvents and purification methods were systematically optimized to facilitate rapid screening and accurate quantitative analysis of these hormones in sports nutrition foods.Samples were extracted with 1.0%formic acid in methanol,partitioned with sodium chloride,and purified using 150 mg of PSA and 150 mg of C 18 powder.The clean-up solution was evaporated to near dryness under a stream of nitrogen and the residue was reconstituted with 50%methanol aqueous solution.The 60 target compounds were separated on a Waters Acquity UPLC HSS T 3 column(100 mm×2.1 mm,1.8μm)with 0.1%(v/v)formic acid aqueous solution(containing 10 mmol/L ammonium acetate)and methanol as the mobile phases.The analytes were determined by multiple reaction monitoring(MRM)in positive electrospray ionization mode(ESI+)and quantified using the external standard method.The method was validated under the optimized conditions,with the 60 steroid hormones found to exhibit good l

关 键 词：超高效液相色谱-串联质谱 QUECHERS 类固醇激素 运动营养食品 

分 类 号：O658[理学—分析化学]