稳定同位素iTRAQ标记/高效液相色谱-串联质谱法同时定量分析人体中42种氨基酸及典型病例  

Simultaneous Determination of 42 Amino Acids in Human Body by Stable Isotope Isobaric Tage for Relative and Absolute Quantitation Labeled-LC-MS/MS with Typical Example Cases

作　　者：李鹏飞[1] 王燕[1] 陶蓓蓓 王静[1] 罗静[1] 张绪得 安卓玲[1] 刘丽宏[1] 

机构地区：[1]第二炮兵总医院药学部,北京100088 [2]北京艾米诺医学研究有限公司,北京100088

出　　处：《分析化学》2012年第5期762-767,共6页Chinese Journal of Analytical Chemistry

摘　　要：建立稳定同位素iTRAQ标记/高效液相色谱-串联质谱法同时定量分析人体中42种氨基酸的方法。人生物样本经磺基水杨酸沉淀蛋白,稳定同位素iTRAQ-115衍生化后,加入iTRAQ-114同位素标记的氨基酸内标液进样,选用AAA-C18色谱柱,以水-乙腈(含有0.01%七氟丁酸、0.1%甲酸)为流动相,采用梯度洗脱进行分离,选用3200QTRAP型质谱仪的多重反应监测(MRM)扫描方式进行检测。同位素内标消除了系统误差,实现了氨基酸的定量分析,42种氨基酸及同分异构体均能基线分离。本方法快速、灵敏、专属性强、高通量,可用于临床氨基酸代谢疾病的诊疗和营养评估。To develop a LC-MS/MS method for the simultaneous determination of 42 amino acids in human body by stable isotope isobaric tage for relative and absolute quantitation（iTRAQ） labeled.After preciptitated by sulfosalicylic acid and labeled by iTRAQ reagent 115,the biological samples were added iTRAQ reagent 114-labeled amino acid as isotope internal standard,and separated on an AAA-C18 column using water and acetonitrile（both containning 0.01% heptafluorobutyric acid and 0.1% formic acid） as mobile phase by gradient elution.Detection was carried out by multiple reaction monitoring（MRM） on a 3200QTRAP LC-MS/MS system.Quantitation was completed by isotope internal standard which also could remove the systematic error.42 amino acids and isomers were chromatographic baseline separated.The assay was successful applied to clinical diagnose and treat.The method is a rapid,sensitive,selective and high-flux for the determination of amino acids metabolism disease and nutritional evaluation.

关 键 词：高效液相色谱-串联质谱 全谱氨基酸 高通量定量 稳定同位素标记 

分 类 号：O657.63[理学—分析化学] R96[理学—化学]