Milk from 24 Nili Ravi buffaloes were collected on fortnight basis from Tarir Dairy Farm, Dhanola on Sargodha Road, Faisalabad. Data obtained for each attribute was subjected to statistical analysis. Oxytocin administration to buffaloes milk resulted in increased pH, proteins, lactose and solid-not-fat and decreased in fat percent as compared to normal animals. Alkaline phosphotase, acid phosphotase, lactoperoxidase activity, and thiocyanate were high in oxytocin treated while lipase activity was low in buffaloes. Caprylic acid(C8), capric acid (C10), myristic acid (C14), palmitic (C16) were high while butyric acid (C4), caproic acid (C6), lauric acid (C12), steric acid (C18), oleic acid (C18:1), linoleic acid (C18:2) were significantly low in oxytocin treated buffalo. Vitamins (A, D, E and C) were high in oxytocin treated buffaloes as compared to control. Milk sodium, phosphorus and copper were high while low in potassium, iron and zinc in experimental group. From this study it is concluded that regular use of oxytocin injections do alter the milk composition which could have a promising effect on the milk value added products to effect the human health.