拟干酪乳杆菌胞内代谢产物的GC-MS分析方法的建立

李晓雪; 田锡炜; 王永红; 储炬; 张嗣良; 刘玉伟

doi:10.13386/j.issn1002-0306.2013.18.024

拟干酪乳杆菌胞内代谢产物的GC-MS分析方法的建立

Metabolites research of Lactobacillus paracasei with gas chromatography-mass spectrometry

摘要

摘要: 用气相色谱-质谱联用（GC-MS）技术建立了拟干酪乳杆菌发酵过程中胞内代谢物的代谢组学研究方法。考察了不同溶解剂（吡啶、四氢呋喃、二甲基甲酰胺）和衍生化试剂（N-甲基叔丁基二甲基硅基三氟乙酰胺（MTBDSTFA）、N,O-双（三甲基硅烷基）三氟乙酰+三甲基氯硅烷（BSTFA+TMCS）、N-甲基-N-三甲基硅烷基三氟乙酰胺（MSTFA））,对质谱峰的信号强度和个数的影响,确定使用吡啶作为溶解剂、BSTFA+TMCS作为衍生剂的两步衍生法效果最佳。以拟干酪乳杆菌为分析对象,共鉴定出38种胞内中间代谢物。方法学考察结果表明,该方法具有良好的精密度、稳定性和重现性。本方法已成功地应用于拟干酪乳杆菌发酵过程研究,并且发现了发酵过程中可能的生物标志物。

Abstract: A reliable metabolomics method for investigating intracellular metabolites in Lactobacillus paracasei by gas chromatography-mass spectrometry was developed.The effect of different solvents (pyridine, tetrahydrofuran and dimethylformamide) , and derived reagents (MTBDSTFA, BSTFA+TMCS and MSTFA) , for peak signal intensity and numbers were investigated.The best condition with pyridine as solvent and BSTFA+ TMCS as derived reagent was determined.Total 38 kinds of intracellular metabolites were identified in the samples of Lactobacillus paracasei.The method had been successfully employed in fermentation process study.Several biomarkers had been detected and identified.