Hydrophilic interaction and reverse-phase liquid chromatography combined with quadrupole-time-of-flight mass spectrometry for untargeted urine metabolomic profiling in rats during sodium aescinate injection-induced acute kidney injury

Acute kidney injury (AKI) induced by sodium aescinate injection is a significant clinical issue, but the mechanisms responsible for this nephrotoxicity are not fully understood. Conventional reversed-phase liquid chromatography-mass spectrometry (RPLC-MS) often fails to provide a comprehensive metabolite profile in urine due to its limitations in effectively analyzing metabolites with varying polarities. This study aims to address this issue by utilizing a combination of hydrophilic interaction liquid chromatography (HILIC) and reversed-phase liquid chromatography (RPLC) to investigate the urine metabolites in a rat model of sodium aescinate induced AKI and to explore potential metabolic pathways associated with this condition. A combination of HILIC and RPLC, equipped with quadrupole-time-of-flight mass spectrometry, was established for untargeted metabolomic profiling of urine samples from rats with sodium aescinate-induced AKI. Metabolomic profiling identified 61 differential metabolites that were significantly altered in the urine of AKI rats. Pathway enrichment analysis indicated that these metabolites were classified into 12 metabolic pathways. This study captures a broader spectrum of metabolites under HILIC and RPLC modes than a single mode, enhancing our understanding of the metabolic mechanisms involved in sodium aescinate-induced kidney injury.