基于液质联用的玉米大斑病菌代谢组的方法优化

Optimization of Metabolomics Analysis Method of Setosphaeria turcica Based on LC-MS

为了对玉米大斑病菌菌丝的代谢组进行分析,通过正交试验优化液质联用检测的提取条件,并对其化合物进行定性分析。结果表明,优化后的提取条件为体积分数80%的甲醇4℃超声提取30 min,对UPLC-Triple TOF/MS洗脱条件优化后菌丝提取代谢物总离子色谱图峰形良好,分布相对均匀。共得到12323个质谱峰,比对数据库后发现,正离子模式下可鉴定化合物的数量优于负离子模式,且样品间相对标准偏差小于20%的化合物占89.607%以上。鉴定得到脂类代谢、氨基酸代谢和核苷酸代谢相关化合物及呋喃等杂环化合物,进一步提供的信息可用于玉米大斑病菌非靶向代谢组学的研究。

In this study, UPLC-MC was used to analyze the metabolome of S. turcica, and the extraction and detection conditions were optimized by orthogonal experiments. The results showed that the extraction condition was80% methanol and ultrasonic extraction at 4℃ for 30 min, and the total ion chromatogram detected by UPLC-Triple TOF/MS had good peak shape and uniform distribution. A total of 12,323 features in were obtained, and the number of compounds identified in the positive ion mode is more than that in the negative ion mode. Compounds with the relative standard deviation less than 20% accounted for more than 89.607%. The compounds related to lipid metabolism, amino acid metabolism and nucleotide metabolism, and heterocyclic compounds such as furan, can be identified and used for untargeted metabolomics studies of S. turcica.