三重四极杆液质联用技术建立检测血清中胰岛素样生长因子1的定量方法研究  

Quantitation of IGF-Ⅰ in the Human Serum Using Liquid Chromatography Triple-quadrupole Mass Spectrometry

作　　者：河春姬[1,2] 申利[2] He Chunji;Shen Li(Beijing Sport University,Beijing 100084,China;China Anti-Doping Agency,Beijing 100029,China)

机构地区：[1]北京体育大学,北京100084 [2]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》2021年第7期549-555,共7页Chinese Journal of Sports Medicine

基　　金：国家体育总局科研项目(2014B128)。

摘　　要：目的:采用三重四极杆液质联用(LC-QQQ)技术建立检测血清中全链胰岛素样生长因子1(IGF-Ⅰ)的方法,并对该方法进行方法验证。方法:采用酸溶液游离人血清中IGF-Ⅰ并通过乙腈沉淀大分子复合物的方法提取血清中IGF-Ⅰ,应用高效液相色谱仪C18反向色谱柱进行分离提纯,通过电喷雾离子源带电的方式,用串接三重四极杆质谱技术对特定质荷比的离子对进行定量分析。结果:使用大鼠血清配制人IGF-Ⅰ标准曲线的线性范围为50~1200 ng/mL,r>0.99。对50 ng/mL、200 ng/mL、500 ng/mL三个浓度的血清样本进行重复性检测,相对标准偏差分别为5.39%、2.91%和3.62%,中间精密度相对标准偏差分别为14.00%、6.10%和6.11%。方法定量限可达到50 ng/mL。结论:LC-QQQ检测血清全链IGF-Ⅰ的方法符合世界反兴奋剂机构(WADA)相关文件关于方法验证的要求。Objective To establish a method for the detection of the intact insulin-like growth factor1(IGF-Ⅰ) using the liquid chromatography triple-quadrupole mass spectrometry(LC-QQQ) technique,and implement the validation.Method IGF-Ⅰ was isolated from human serum by acid solution and extracted by acetonitrile precipitation of macromolecular complex. Then it was separated and purified using a reversed-phase C18 high-performance liquid chromatography column. The ion pairs with specific mass-to-charge ratio were quantitatively analyzed by tandem electrospray ionisation triple-quadrupole mass spectrometry.Results The linear range of human IGF-Ⅰ standard curve prepared with rat serum was 50-1200 ng/mL,r>0.99. The relative standard deviation(RSD) of serum samples with the concentration of 50 ng/mL,200 ng/mL and 500 ng/mL were 5.39%,2.91% and 3.62%,respectively,and the RSD of intermediate precision were 14.00%,6.10% and 6.11% accordingly. The limit of quantitation was 50 ng/mL.Conclusion The LC-QQQ method established for the quantification of intact IGF-Ⅰ in human serum meets the criteria of the World Anti-Doping Agency(WADA).

关 键 词：兴奋剂检测 LC-QQQ IGF-Ⅰ 方法验证 

分 类 号：R446.1[医药卫生—诊断学] O657.63[医药卫生—临床医学]