Objective:To investigate the relationship of statins (drug given to reduce serum levels of LDL-cholesterol) on vitamin D levels of Pakistani type 2 diabetes mellitus (DM) patients in a hospital in Karachi.Methods:In a cross-sectional survey, 312 consecutive patients with type 2 DM (219 males and 93 females, age 22-70 years) were recruited with informed consent. A questionnaire was administered to find out whether they were statin users or non-users. Serum was analyzed for concentrations of 25(OH) vitamin D [25(OH)D] and other related biomarkers such as serum cholesterol, triglycerides, HDL-cholesterol, LDL-cholesterol, phosphate and calcium using kit methods. Multiple Linear Regression was used to evaluate association of statin use with serum levels of vitamin D while adjusting for related covariates including duration of statin use, duration of type 2 DM and smoking.Results:Mean concentrations of serum cholesterol, and LDL-cholesterol were lower among statin users compared to statin non-users (P < 0.01), while HDL-cholesterol levels were higher (P<0.01). No relationship was observed between statin use and serum levels of vitamin D (P=0.768), when adjusted for age, gender, BMI, duration of type 2 DM, smoking, serum cholesterol and LDL-cholesterol. The adjusted regression coefficient (β) and standard error [SE(β)] for statin use duration were 0.012 (0.042), when serum levels of vitamin D was taken as an outcome.Conclusion:Lack of association was found between statin use and vitamin D levels in a hospital-based population of Pakistani patients with type 2 DM.