Background Citrus reticulata is one of the most cultivated fruit with great benefits for humans in the world. Citrus reticulata peel has several biological activities within them hypoglycemic, hepatoprotective, antimicrobial and antioxidant. The present study emphasizes on the in vivo wound healing and in vitro antimicrobial and antioxidant activities of Citrus reticulata peel extract. Methods Forty albino mice (23–29 g) of either sex were divided into four groups. The test groups were treated with experimental ointment (0.5% and 10% of methanolic extract), negative control were treated with Vaseline and the positive control were treated with silver sulfadiazine. Burn wounds were induced on dorsal area of mice bodies. Wound area measurement was carried out every day during 22 days. Biochemical screening was performed to identify possible compounds. Antioxidant and antimicrobial activities were also determined. Results Significant wound healing activity was observed with topical application of Citrus reticulata peel extract. Wound area reduction at day 16 of treatment was 100% for both treated groups (0.5% and 10%) when compared to positive and negative control it was 100% and 98.32% respectively at day 22. Furthermore higher rate of wound contraction (100% on 16th day) was observed for both treated groups. The result of biochemical screening showed that C. reticulata peel is characterized by highest amount of total polyphenols (13.19 mg/g), flavonoids (4.07 mg/g), vitamin C (13.20 mg/g), carotenoids (0.032 mg/g) and lowest content of macronutrients (Proteins: 0.40%, reducing sugars: 7.21%; lipids: 1.5%). Additionally C. reticulata peel exhibited remarkable antioxidant activity using DPPH and phosphomolybdate methods as well as the extract possess antimicrobial effect against pathogen bactria. Conclusion The findings from this research indicated that Citrus reticulata peel extract is effective in inhibiting the growth of pathogen bacteria and could be of therapeutic potentials for wound healing.