Background: Vitamin D deciency is prevalent among obese individuals. Some authors have postulated that Vitamin D is sequestered in the excess adipose tissue, leading to less bioavailability, whereas others suggest that low Serum 25-hydroxy Vitamin D may be a result of volumetric dilution of vitamin D in the large adipose stores. Aim: To study and compare the Vitamin D levels in Obese and Non-obese diabetic female with obese and Non-obese nondiabetic females. Method & Materials: The present study was conducted in the Department of Medicine, MLN Medical College & SRN Hospital, Prayagraj to assess Vitamin D levels of diabetic and non-diabetic obese and non-obese females. A total of 115 diabetic female patients attending Clinic fullling the inclusion criteria and giving consent were included as Cases and 115 female non-diabetic patients without active or chronic problems attending different departments were included in the study. Serum samples of all the participants were collected for estimation of 25-hydroxy vitamin D (25-OH vitamin D). Quantitative determination of 25- hydroxy vitamin D (25- OH vitamin D) in human serum is done by chemiluminescent microparticle immunoassay (CMIA). Result: Majority of Controls had Normal Vitamin D levels (52.2%) , on the other hand only 1.7% cases had normal Vitamin D levels. Vast majority of Cases (98.3%) had Vitamin D deciency. Higher percentage of Cases were found to have mild and severe vitamin D deciency as compared to that in controls. [Mild Vitamin D deciency -12.2% vs. 11.3% in cases and controls respectively and severe Vitamin D deciency - 86.1% vs. 36.5% in cases and controls respectively] . Difference in vitamin D status of Cases and Controls was found to be signicant statistically. Serum Vitamin D levels of Controls (29.18±20.31 ng/ml) was found to be signicantly higher as compared to Cases (9.06±6.03 ng/ml). Conclusion- Mean serum vitamin D levels were signicantly lower in Diabetic females as compared to that in Non-Diabetic females.Obesity was associated with a signicantly lower vitamin D levels in both Diabetic females as well as Non-Diabetic females.