Milk has been an issue of public health concern due to unhealthy fats. This study investigated changes in milk fatty acids (FA) in relation to blood metabolites with advancing age and lactation stage in dairy buffaloes. A total of 12 dairy buffaloes were studied for six weeks in Peshawar. The serum glucose concentration averaged 59.02 mg/dl and triglyceride were 54.63 mg/dl. Glucose and triglyceride concentration was a bit high in older animals. Saturated FAs (SFAs) were averaged 70.41 g/100g. C14:1, C16:1 and C18:3 were significantly (P