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.