Several studies demonstrate the effectiveness of eye drops Platelet Rich Plasma (E-PRP) as a promoter of the healing of different ophthalmological disorders in human, due to the high concentration of growth factors as EGF, PDGF-BB and Fibronectin which represent the key to obtain the desired regenerative effects on the ocular surface. There is different protocol for PRP eye drops preparation for use in human corneal disorders. This study was to evaluate an easy and affordable technique for preparation of canine autologous E-PRP according to standard measure used before for human corneal disorders to be used in canine corneal ulcers in comparison with control group, all treatments were applied four times daily. Twelve healthy dogs were divided into two groups first group included eight dogs which received autologous E-PRP compared with second group which included four dogs and received saline eye drops as control. Complete blood count is performed to whole blood and E-PRP to validate last one content of platelets which must equal more 1.5 those in whole blood. E-PRP contain platelets 2.41-fold than those in whole blood with one step centrifugation technique. Autologous canine E-PRP mean healing time was 3.62±0.23 days while with control group mean healing time was 6.75±0.30 days. In conclusion, As the E-PRP is now widely documented in human ophthalmology, it had been proven as a great result in veterinary practice when prepare autologous E-PRP by single step centrifugation to be used for canine corneal wound and healing rate was better than control group by significant difference. which make it novel option for treatment for canine corneal ulcer.