Rapid Classification and Identification of Chemical Compounds and Semi-Quantitative Metabolism of Huangkui Capsules and the Protective Effects of Its Quercetin Derivatives against Tacrolimus-induced HK-cell Reduction

Abstract

The study aimed to establish an effective strategy for systematically characterizing and verifying compounds in Huangkui capsules (HKCs). An ultrahigh-performance liquid chromatography quadrupole time-of-flight mass spectrometry (MS) method was effectively established and utilized for the chemical compound characterization in HKC, with the support of MS-DIAL, MS-FINDER, and Global Natural Product Social Molecular Network. Multiple rat samples were analyzed after the oral administration. Metabolites were identified based on specific cleavage behaviors, and metabolic pathways were predicted. Semi-quantitative analysis of the metabolome profiles was conducted using post-data processing. High concentrations in vivo were investigated for their role in tacrolimus-induced death of HK-2 cells. In total, 129 compounds were identified in HKC, of which 74 were reported for the first time. In this study, we successfully identified and uncovered 19 prototypes and 123 metabolites from the biosamples. The concentrations of glucuronidation and methylation metabolites of quercetin were the highest in the kidney and intestinal tissues. In contrast, significant glucuronidation of quercetin metabolites was observed with high blood concentrations. Notably, quercetin glucuronidation and methylation metabolites protected HK-cell against tacrolimus-induced injury in a dose-dependent manner. This study successfully established a reliable and efficient strategy for comprehensive characterization of traditional Chinese medicine compounds. This strategy harnessed the power of various intelligent postprocessing technologies to provide a more thorough understanding of active components and their metabolic transformations in the body. These results suggest that quercetin metabolites should be evaluated for their protective capacity against kidney disease.