Canola oil has been used in the Pakistan for the treatment of various diseases and skin infections. Oil was extracted with n-hexane from the seeds of canola (Brassica napus L.) and was evaluated for free fatty acid value. Four microorganisms namely; Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas originals, and Klebsiella pneumonia, has known to cause some infections treatable with these oils were investigated. The results showed that all oil shown inhibitory effects against Klebsiella pneumoniae, Staphylococcus epidermidis, and Pseudomonas originals but no inhibitory effects was found against Staphylococcus aureus.