The present project was designed to study the prevalence of Vitamin D, calcium and phosphate in healthy females in Lahore. In this study 88 blood samples of healthy females were used to assay the concentration of Vitamin D, calcium and phosphate. ELISA was performed for the assessment of Vitamin D in the laboratory of LCWU. The calcium and phosphate were measured by using commercially available kits on chemistry analyzer. Statistically data was analyzed by using SPSS (version 13.0). The mean value of Vitamin D was 42.76 nmol ± 2.05. The mean of calcium was 8.78 mg/dl ± 0.06 and the mean of phosphate was 3.07 mg/dl ± 0.07 among the study group. To see the influence of lifestyle on the serum levels of vitamin D in healthy females, the data was divided into different lifestyle factors. On the basis of the age the data was stratified into two groups. Deficiency of vitamin D was also found in 23.86% females of study population. 24 (27.27%) young females showed the deficiency of calcium and in 21 (23.86%) healthy females the deficiency of phosphate was observed. Vitamin D was significantly and positively correlated with calcium and phosphate (r= 0.622 and r= 0.656 respectively).