摘要
建立了一种快速、准确的同时测定大肠杆菌(E.coli)24种胞内代谢中间产物的液相色谱-串联质谱分析方法。样品经过淬灭、洗涤和提取后得到胞内代谢物,高速冷冻离心去除杂质。经Waters Atlantis HILIC silica色谱柱分离,以乙腈-10 mmol/L乙酸铵水溶液为流动相进行梯度洗脱,流速为250μL/min。电喷雾负离子(ESI-)模式电离,多反映监测模式(MRM)检测,基质标准溶液法进行定量。结果表明:24种代谢物在0.10～100μmol/L范围内呈良好的线性关系,相关系数均大于0.99;24种代谢物的检出限(S/N>3)为0.05～0.34μmol/L,3个添加水平的回收率为83.7%～108.1%,相对标准偏差(RSD)均小于10%。该方法具有简便、灵敏度高、定性准确等特点。
An high performance liquid chromatography-tandem mass spectrometry （HPLC-MS/MS） method was established for the determination of 24 intracellular metabolites in E. coli. After quenching, washing and extraction, the samples were centrifuged to remove impurities,and then separated on a Waters Atlantis HILIC silica column. The mobile phases were acetonitrile and water （containing 10 mmol/L ammonium acetate ） at a flow rate of 250 μL/min. The electrospray was operated in the negative ionization mode and the 24 metabolites were identified by multiple reaction monitoring （MRM） mode. The method of matrix-matched standard solution was adopted as the quantitative method. The calibration curves showed good linearity within the concentrations of 0. 10 - 100 μmol/L with the correlation coefficients r 〉 0.99. The limits of detection of the 24 metabolites were 0.05 - 0. 34 μmol/L. The recoveries of the 24 metabolites were 83.7% - 108.1% at the 3 spiked levels. The relative standard deviations were less than 10%. This method is simple, sensitive and accurate in the determination of 24 metabolites.
出处
《食品与发酵工业》
CAS
CSCD
北大核心
2011年第5期151-156,共6页
Food and Fermentation Industries
基金
江南大学食品科学与技术国家重点实验室自由探索资助课题(SKLF-TS-200803)
关键词
高效液相色谱-串联质谱
大肠杆菌
胞内中间代谢产物
代谢组学
high performance liquid chromatography-tandem mass spectrometry（HPLC-MS/MS） , E. coli,intracellular intermediate metabolites, metabolomics
