Introduction: Diabetic nephropathy is an established risk factor for several cardiovascular diseases and the leading cause of chronic kidney disease in patients. Vitamin D promotes calcium metabolism and normal mineralization and formation of bone. Vitamin D has a great role in cell growth, proliferation, differentiation and apoptosis. It has important functions on neuromuscular and immune system. Aims: To evaluate the status of vitamin D3 level in diabetic nephropathy patients and to find out any sort of relation of vitamin D3 level with diabetic nephropathy. Materials and Methods: This observational, cross-sectional, case-control study was conducted in Hi-Tech Medical College & Hospital, in an urban area of Bhubaneswar, Odisha, over a period of two year (January 2013 to December 2014). Serum vitamin D3 levels were estimated in adult 200 cases of diabetic nephropathy as well as 50 age-sex matched normal individuals of same age group in the OPD of the Department of Medicine and Nephrology. Results: Serum vitamin D3 level is remarkably decreased in diabetic nephropathy patients in comparison with that of the control group. And it is observed that in the older age group of diabetic nephropathy patients, the serum vitamin D3 levels are drastically reduced in contrast to that of the same age group of controls. It is also revealed that vitamin D3 levels are remarkably decreased with the progression of diabetic nephropathy. Conclusion: From this study, we can draw a conclusion that the assessment of serum vitamin D3 in the cases of diabetic nephropathy is of prime importance and should start the treatment of vitamin D3 insufficiency and deficiency accordingly as soon as possible with the treatment of diabetic nephropathy.