LC-MS/MS检测癫痫患者神经递质类氨基酸

Detection of Amino Acids Neurotransmitter in Epilepsy by LC-MS/MS

目的:建立液相色谱串联质谱同位素内标法检测神经递质类氨基酸并用于癫痫患者临床评价。方法:选用AAA-C18柱色谱柱,以乙腈水(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,血浆样品用iTRAQ-115衍生化试剂处理后,加入iTRAQ-114衍生化的氨基酸内标并进样,选用3200QTRAP型质谱仪的多重反应监测(MRM)扫描方式进行检测。疾病组与健康组的统计采用t检验和主成份分析。结果:疾病组和健康组氨基酸测定结果显示:Trp、GABA两组间没有显著性差异(P>0.05),Arg、Gly、Ser、Tau、Asp、Glu、EtN、两组间有显著性差异(P<0.05),通过PCA分析显示,疾病组与健康组之间差异明显,Asp、Glu、Ser等是引起差异的主要氨基酸。结论:试验方法灵敏、专属性强,并初步的用于癫痫患者体内氨基酸评价。

Obiective： To develop a LC-MS/MS method with amino acids isotope as internal standard,to detect amino acids neu-rotransmitter in the Epileptic for clinical evaluation.Methods： After derivatizated by iTRAQ-115,plasma samples were added internal standard which had been derivatizated by iTRAQ-114.The separation was carried out on an AAA-C18 column using acetonitrile and wa-ter（both containing 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.T-test and PCA analysis used as the statistics methods.Results： There was no significant difference for Trp and GABA and significant difference for Arg,Gly,Ser,Tau,Asp,Glu,EtN between disease group and health group.There was significant difference between disease group and health group due to Asp,Glu,Ser by PCA analysis.Conclusion： The method is sensitive and selective and applied to detect amino acids neurotransmitter in the Epileptic for clinical evaluation.