气相色谱-三重四极杆质谱法测定大肠杆菌中有机酸的含量

Determination of Organic Acids in E.coli by Gas Chromatography-triple Quadrupole Mass Spectrometry

建立了气相色谱-三重四极杆质谱联用法(GC-MS/MS)同时测定大肠杆菌三羧酸(TCA)循环中6种有机酸的方法。样品经细胞破壁处理、固相萃取(SPE)富集净化,由硅烷化试剂N-(特丁基二甲基硅烷)-N-甲基三氟乙酰胺(MTBSTFA)衍生化后,利用GC-MS/MS多反应监测模式测定,外标法定量。6种有机酸在20~10000μg/L范围内线性良好,相关系数均大于0.99,加标回收率为82.3%~100.4%,相对标准偏差为2.3%~4.5%,方法检出限为2.0~18μg/L。方法受基质影响小,重现性好,适合于复杂生物基质样本中有机酸含量的测定。

A method has been developed to determine the six organic acids in E.coli tricarboxylic acid cycle(TCA cycle)by gas chromatography-triple quadrupole mass spectrometry(GC-MS/MS).The sample was purified by cell disruption and solid-phase extraction(SPE),derivatized with the silylation reagent N-(tert-butyldimethylsilyl)-N-methyltrifluoroacetamide(MTBSTFA),and then analyzed by GC-MS/MS(MRM)mode,extraterritorial method for quantification.The linear correlation coefficients(R^2)of the six organic acids is greater than 0.99 in the linear range of 20~10000μg/L,the recovery was 82.3%~100.4%,the relative standard deviation(RSD)is 2.3%~4.5%,and the detection limit(LOD)of the method was 2.0~18μg/L.The method is less affected by the matrix and has good reproducibility,and is suitable for determining the content of organic acids in complex biological matrix samples.