The fruits of Rhus tripartita had recently attracted great attention due to its notable therapeutic effects. Their potential effects are attributed to the richness of diverse classes of secondary metabolites. The objective of this research was to access the determination of chemical composition and antioxidant properties of R. tripartita fruit extracts and its partitioned fractions (absolute petroleum ether, 70% aqueous ethanol, absolute ethyl acetate and water). LC-ESI-MS/MS and FTIR-ATR were used to assess the potential of R. tripartita fruits as a source of health-promoting constituents. A total of 38 phenolics, including flavones, flavonols, flavanones, organic acids, hydroxycinnamic acids and hydroxybenzoic acids, 26 of them were reported for the first time in R. tripartita. The main compounds were apigenin7-O-glucoside, apigenin and p-coumaric acid. The FTIR-ATR analysis results revealed the presence of characteristic functional groups such as − OH, C-O, − C = C and C-H of phenolic compounds, carboxylic acids, carbohydrates, lipophilic components and proteins in R. tripartita fruits. Furthermore, the ethyl acetate fraction showed the highest level of phenolic contents and strong antioxidant activities. The present study recommends R. tripartita fruits as source of natural antioxidants which can be used as bioactive ingredient for functional foods and nutraceuticals.