A compound fruit drink (CFD) is a functional beverage containing fruits, Chinese herbal medicine, and prebiotic fructose. Previous studies have shown the effect of a CFD on alleviating constipation and its impact on gut microbiota. However, a comprehensive analysis has not been reported in regard to the serum metabolism of CFDs. This study established a mouse constipation model, using loperamide hydrochloride. Herein, based on UHPLC–QTOF/MS analysis, 93 differential metabolites (mainly including phosphoglycerides and amino acid derivatives) among the groups of mice were identified. After CFD treatment, the content of phosphatidylethanolamine, amino acid derivatives (including N-Acetyl-L-aspartate, L-Norleucine, and cis-4-Hydroxy-D-proline), and fumarate increased, while that of esters decreased. Pathway enrichment analysis revealed that the CFD mitigated constipation by modulating nine metabolic pathways, which encompass glycerophospholipid metabolism, the tricarboxylic acid (TCA) cycle, pyruvate metabolism, and tyrosine metabolism. Notably, the glycerophospholipid metabolic pathway was identified as the most pertinent. Collectively, the results provide new ideas for developing functional foods that nourish the intestines and relieve constipation.
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