高效液相色谱-串联质谱法分析大肠杆菌代谢组中样品提取方法的比较  

Comparison of Extraction Methods for E. coli Metabolome Analysis Using Liquid Chromatography Tandem Mass Spectrometry

作　　者：梅辉[1,2] 戴军[1] 刘文卫[3] 凌霞[3] 朱鹏飞[3] 赵志军[1] 

机构地区：[1]江南大学食品科学与技术国家重点实验室,无锡214122 [2]江南大学食品学院,无锡214122 [3]无锡市疾病预防控制中心,无锡214002

出　　处：《分析化学》2011年第4期534-539,共6页Chinese Journal of Analytical Chemistry

基　　金：江南大学食品科学与技术国家重点实验室自由探索课题基金(No.SKLF-TS-200803)资助项目

摘　　要：比较了基于液相色谱和串联质谱联用技术（LC-MS/MS）的E.coli代谢组分析的5种不同样品提取方法。在对样品进行淬灭和洗涤后,分别使用冷甲醇法、热乙醇法、甲醇/氯仿法、热甲醇法和高氯酸法对样品平行提取3次,每个样品重复进样3次。结果显示,冷甲醇法所得到的相对提取率最高,且重复性好（RSD〈5%）。从相对提取率、重复性等方面综合考察5种提取方法,从优到劣依次为：冷甲醇法〉甲醇/氯仿法〉高氯酸法〉热乙醇法〉热甲醇法。此外,以冷甲醇法的提取溶剂配制系列不同浓度的代谢物标样测定的结果表明,本方法的代谢物检测线性较好,线性范围较宽,提取溶剂的基体效应对代谢物的定量分析影响较小。本方法的检出限为0.05~0.36 mmol/L。Five extraction method for E. coli metabolome analysis using liquid chromatography tandem mass spectrometry （LC-MS/MS） were compared. After being quenched and washed, three replications of the same sample were extracted using the following solvent system： cold methanol, hot ethanol, methanol/chloroform, perchloric acid, and hot methanol, each sample injection repeated 3 times. The results showed that the cold methanol extraction method obtained the highest rate of relative recovery with the best reproducibility （RSD〈5%）. Considering relative yield and reproducibility, the five extraction starting from favorably to poor were： cold methanol〉ethanol/chloroform 〉perehlorie acid〉hot ethanol〉 hot methanol. In addition, targeted metabolites series of different concentrations were added to the cold methanol solvent, and the results indicated that the linearity of 24 metabolites was good with the linear ranges wide, and the extraction media have little matrix effect on the quantification of metabotites. The limit of detection （LOD） was 0.05--0. 36 μmol/L.

关 键 词：代谢组分析 液-质联用 提取方法 大肠杆菌 

分 类 号：Q93-3[生物学—微生物学]