基于液相色谱-质谱技术的肾脏代谢组学分析方法研究

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

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。

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 100-1000 Da and a mass resolution of 120000.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.