Application of smoke condensate derived from an indirect heating of jennet feces (Sargin) had been recommended by Iranian ancient scientists as a therapeutic agent. The present study was done to evaluate the healing effects of Guajol® ointment on burn wound in rat. The Guajol® ointment was prepared from the smoke condensate of Sargin samples. Wistar Rats (n = 50) were randomized into six groups including normal saline, silver sulfadiazine and 1.25%, 2.50%, 5.00% and 10.00% concentrations of Guajol® ointment. Under general anesthesia, dorsum of the rats were shaved and burn wounds were created using hot plate. Area of wounds and percent of healing were measured. Normal saline had the highest area of wound, followed by 1.25% Guajol® and silver-sulfadiazine groups. The group treated with 5.00% Guajol® showed the highest percent of healing. Percent of healing in NS, SSD and 5.00% Guajol® ointment groups on day 21 were 38.47%, 75.00% and 98.51%, respectively. Microscopic examination of wounds sections of rats treated with 5.00% Guajol® showed more collagen fibers and fibroblasts cells on day 7. Wounds of 5.00% Guajol® treated group was covered with healthy epithelial and epidermis tissues and hair follicles on day 21. This was the first report of using Sargin to heal the burn wound of rat. Further studies are recommended for investigation of the other effects of Guajol® ointment and its possible application in medicine.