Introduction: Cholelithiasis is a common abdominal entity which results in increased hospital admissions. About 10-12% of adults have the incidence of developing gallstones. The prevalence of common bile duct stones in patients with gallstones varies from 8 to16%. Pure cholesterol stones are not frequent, and they account for about less than 10% of all stones. Gallstones are generally classified into pure cholesterol stones, black or brown pigmented stones and/or mixed stones. Materials and Methodology: The study population was basically divided into two groups. One was study group with 210 patients reported with gall stones disease and fulfilling all our inclusion criteria. The second one was control group with 55 patients without any gallstone disease admitted in surgery ward. Serum iron and ferritin contents of both groups was analysed and were compared with each other. Inclusion criteria followed in control group were age group of 15 – 70 years not suffering from gall stone disease which was confirmed by ultrasonography and for study group All patients with cholelithiasis which were confirmed by ultrasonography with age group of 15 - 70 years. Detailed history was taken from all the patients who were suffering from gall stone disease. Routine investigations were included such as ultrasound of abdomen was done in all the patients. 4 mL venous blood sample was taken for the evaluation of serum iron and serum ferritin level.Results: There were more females in cases group 175 (84.5%) than control group 20 (48%) and males in case were (35) 15.5% and (30) 48% in control. The mean of age in case group was 41.7812.81 and in control group was 40.0212.94. There was low prevalence of gallstones in young population and highest prevalence of gallstones in 30-39 years. In this study in males, the normal male reference value supplied with the kit was 60-160 µg/dl. Serum iron levels were low in 42.85% of cases and 23.3% of controls. Serum iron was normal in 57.14% of cases and in 76.6% of controls. The mean serum iron among cases 66.1228.35 and control 66.2319.53. P value was >0.05 which is not-significance. Serum iron level in females was low in 24% of cases and 35% of controls. Serum iron was normal in 74.28% cases and 65% of controls. The mean serum iron in female cases 58.3226.32 and control was 69.2719.66. P value was <0.05 which is significant statistically. Conclusion: From this study, it was concluded that a low storage of serum iron in the body is a potential risk factor for cholelithiasis in females. Serum iron, serum ferritin may be used as markers of iron store so that low serum iron status could be diagnosed at a much earlier stage.