A comparative study on serum biochemical constituents was carried out at different physiological stages in females of Jersey crossbred cattle. A total of 72 healthy Jersey crossbred cattle (n=72), aged 3-6 years with a good body condition score (5-6 out of 10) were selected and they were divided into five groups, each group comprised of twelve (n=12) animals, Group 1: calves, Group 2: heifers, Group 3: pregnant lactating, Group 4: non-pregnant lactating, Group 5: pregnant dry and Group 6: non-pregnant dry Jersey crossbred cows. Blood samples were collected and analysed for biochemical profiles. Serum calcium and inorganic phosphorus were higher in calves followed by heifers. Cows in lactating groups had significantly lower calcium and higher phosphorus than those in dry groups. Cows in the pregnant group had significantly higher serum magnesium, potassium and chloride than those in pregnant lactating groups followed by cows in the pregnant dry group. However, serum sodium was higher in the pregnant dry group. Similarly, total protein, globulin and total cholesterol were higher in pregnant lactating group followed by comparable results observed in non-pregnant lactating and pregnant dry groups. Glucose was higher in female calves and blood urea nitrogen (BUN) and creatinine were higher in lactating groups. Therefore, variations in serum biochemical markers in Indian Jersey crossbred cows during different physiological stages were useful to frame the dietary and management strategies to enhance health, productivity and reproduction performances.