Aim: To determine the serum Iron Levels in Patients of Gall Bladder Stone Disease. Methods: This Prospective study which was carried in the Department of General surgery, Government medical College, and Hospital, Bettiah, Bihar, India for 14 months, Patients diagnosed with gallstone disease admitted in various surgical wards and confirmed by ultrasonography, both male and female patients. Serum ferritins, serum cholesterol, serum iron along with other routine investigations were estimated in all 50 patients. Patients with gallstones were diagnosed with iron deficiency anemia based on hemoglobin levels, serum iron, and serum ferritin levels. Adult male with hemoglobin level <13 and adult female with <12 are diagnosed as anaemic. Gallstone patients were divided into anaemic and non-anaemic groups and compared with each other. Results: Mean age of subjects was 41.27±13.9 years, range was 20–72 years and median age was 42 years. Majority of subjects were in the age group 20–30 years. In this study majority, i.e. 80% were females and 20% were males. Among females mean serum iron in subjects with iron deficiency was 20.7 ± 6.8 and among males was 43.2 ± 5.2. Among females mean serum iron in subjects without iron deficiency was 85.6 ± 21.5 and among males was 102.7 ± 23.2. Most of the patients with gallstone have serum iron level less than normal, and are anaemic. This association between low serum iron with anemia and gallstone disease was statistically significant in our study, P value of 0.002. Conclusion: The low body store of serum iron is a risk factor for cholelithiasis in females and serum iron, serum ferritin may be used as marker of iron store so that low serum iron status could be diagnosed at early stage.