Repeated use of exogenous oxytocin in newly calved buffalo has made buffalo breeder anxious about its reproductive and productive performances. Moreover, misconceptions about its harmful effect on public health are increasing day by day during postpartum period when animal has been already in stress. To know effect of oxytocin injection on cervix, uterus and milk composition during postpartum interval in Nili-Ravi buffalo, a study was conducted at Buffalo Research Institute Pattoki, Pakistan from September to October. For this purpose, 25 animals were randomly divided into three groups: group-1 (Control; n=8), group-2 (Low dose 10IU; n=8) and group-3 (High dose 30IU; n=9) while one animal excluded at milk sampling from group-3 due to unavoidable circumstances. Oxytocin was administered to each buffalo twice daily @10IU and 30IU into group-2 and group-3 respectively, whereas, saline into group-1 within one week of post calving for 48 days. Ultrasonography was performed twice a week to monitor involution changes while milk composition analysis was done once a week. Results depicted that initially, anechoic lumen filled areas with echogenic border in cervix and uterus was found but at involution, cervix and uterus became moderately hyperechoic without any fluid filled areas. Moreover, there was non-significant effect (P>0.05) of treatment on cervix, uterine body, non-gravid and gravid horn at involution among three groups when these attained non pregnant size. Regarding progesterone, it remained non-significant (P>0.05) throughout postpartum period among treatments. Furthermore, milk composition results showed that fat, protein, lactose, freezing point, SNF and solids were significantly higher (P<0.05) in group-2 and group-3 as compared to group-1 except density and pH which remained non-significant (P>0.05) in all groups. On the basis of result, it may be concluded that oxytocin had no effect on uterine involution and progesterone; however, it had some role to affect the normal composition of milk in postpartum involution interval.