Twenty-four milk samples were collected form milking Murrah buffaloes of National Dairy Research Institute herd, irrespective of their parity, stage of lactation and body weight. Just after collection milk serum was prepared through TCA precipitation, filtration and centrifugation and preserved at 4°C for 30 days. MU content was estimated on day 0, 5, 10, 15, 20, 25 and 30 of preservation. The average MU concentration was highest at day 20 (57.16±1.61) and lowest at day 25 (55.48±1.61). But there was no significant effect of storage on milk urea concentration. On the other hand a total of 64 milk samples were collected from all four quarters of 16 different animals. The mean urea concentration (mg/dl) of milk from left front, left hind, right front and right hind were estimated as 64.77±4.14, 64.98±3.88, 66.22±4.06 and 65.23±3.90, respectively, for front and hind quarters. No significant differences were found among the quartets, between the front and hind quarters and between left and right quarters. Urea concentration in milk were also estimated from morning and evening milking. Urea concentration (mg/dl) in the morning milk (54. 13±0.62) was significantly (P<0.01) lower than the evening (56.40±0.60). The correlation between morning and evening milk urea concentration was 0.84 (P0.01). In conclusion, storage and quarter had no effect on milk urea concentration in Murrah buffaloes, but diurnal variation exist.