Identification of naringin metabolites mediated by human intestinal microbes with stable isotope-labeling method and UFLC-Q-TOF-MS/MS

•A total of 13 naringin metabolites mediated by human intestinal microbes were identified, among which 5 were reported for the first time.•Naringin underwent extensive phase I metabolism in human intestinal microbes.•Microbial metabolic profiles of naringin among different human individuals were diverse.•Stable isotope-labeling method was efficient to eliminate the endogenous interferences during metabolism research on pharmaceutical compounds. Widely presented in medicinal plants, naringin is one of the major flavanones with various pharmaceutical bioactivities. After oral administration, naringin predominantly undergoes metabolisms mediated by liver cytochrome P450 and gut microbes, while its human microbes-mediated metabolic profiling is still largely obscure due to the endogenous interferences, which makes it extremely difficult to analyze metabolites precisely. In this study, we aim of systematically investigating the biotransformation of naringin mediated by human intestinal microbes through applying stable isotope-labeling method. [2′,3′,5′,6′-D4]naringin was synthesized and incubated anaerobically with human gut microbes. A total of 13 microbial metabolites were detected and identified by UFLC-Q-TOF-MS/MS, among which 5 were reported for the first time. Furthermore, the proposed metabolic pathway revealed that naringin went through extensive phase I metabolism in human intestinal microbes. Of note, diverse metabolic profiles of naringin among human participants were obtained, which could be attributed to the distinct gut microbiota compositions of individuals.