Penthiopyrad (PEN) is a novel fungicide as succinate dehydrogenase inhibitor (SDHI) with chiral characteristics, and it is crucial to investigate its enantioselective residue behavior given its extensive application. This study aimed to comprehensively evaluated the storage stability, deposition, dissipation, accumulation, and dietary risk associated with PEN enantiomers and its metabolite 1-methyl-3-trifluoromethyl-1 H-pyrazole-4-carboxamide (PAM) in apple and strawberry. Two enantiomers of PEN accumulated primarily on leaves after spraying, whose concentrations were much higher than those observed in the apple and strawberry fruits, respectively. S-(+)-PEN was found to be preferentially degraded in apples (EF: 0.5113–0.5925), resulting in a relative enrichment of its antipode. Multiple sprays of PEN led to the accumulation of both of its enantiomers, with average residue accumulation value (RAaverage) of 1:2.06:1.11 for the R-(−)-PEN and 1:2.06:1.09 for the S-(+)-PEN in strawberries, respectively. The terminal residues of PAM were lower than the limit of quantification (LOQ), and that of total PEN in both fruits were below the maximum residue limit (MRL) established by China. Furthermore, the acute risk quotient (RQa) of target pesticide in apples were 0.821 %–2.105 %. The investigation provided a theoretical foundation and scientific guidance for applying PEN at the enantiomeric level in fruits. Display omitted •The penthiopyrad (PEN) enantiomers are mainly distributed on leaves 2 h after foliar spraying.•PEN exhibits enantioselective degradation in apples with S-(+)-PEN preferential degradation.•Repeated spraying of PEN could lead to residual accumulation in strawberries.•The acute dietary exposure risk quotient of PEN in apples was comparably acceptable.