基于液相色谱-质谱技术的肾脏代谢组学分析方法研究

建立了一种基于高效液相色谱-超高分辨质谱技术的肾脏代谢组学分析方法。肾脏组织于液氮中研磨成粉后,采用甲基叔丁基醚/甲醇/水溶剂体系进行提取,分别得到强极性部分(下层)和弱极性部分(上层)提取物,依次采用HILIC亲水色谱柱和反相C18色谱柱进行梯度洗脱分离后,进行高分辨质谱分析。采用电喷雾离子源(ESI),正、负离子模式检测,扫描方式为全扫描,扫描范围为100~1000 Da,质量分辨率为120000。结果表明,该方法灵敏度高,专属性强,稳定性良好,可同时获取肾脏组织中的强极性和弱极性代谢物信息,可为肾脏疾病和药物肾毒性生物标志物的发现提供一种新的方法。

Research on a Kidney Metabolomics Analysis Method Based on Liquid Chromatography-Mass Spectrometry

A metabolomics method based on high performance liquid chromatography-ultrahigh resolution mass spectrometry was developed for the analysis of metabolites in kidney tissues.In order to achieve this goal,rat kidney tissues were ground into powder in liquid nitrogen,and extracted with a methyl tert butyl ether/methanol/water solvent system to obtain high polarity(lower layer) and low polarity(upper layer) extracts,respectively.Then,the high polarity and low polarity extracts were subjected to gradient elution separation on a hydrophilic HILIC column and a reversed phase C18 column,respectively,and subsequently detected by ultrahigh resolution mass spectrometry(Orbitrap Fusion Lumos).The mass spectrometry analysis was performed using an electrospray ionization(ESI) source operated in positive and negative ion mode with a scanning range of 100-1 000 Da and a mass resolution of 120 000.Results showed that with high sensitivity,high specificity and good stability,the proposed method could be used to obtain the comprehensive information of high polarity and low polarity metabolites in kidney tissues,providing a novel approach discovering new biomarkers for kidney disease and drug nephrotoxicity.