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Introduction: The cholesterol stones are formed by super-saturation of the bile with cholesterol. Iron deficiency alters activity of several hepatic enzymes, leading to increased cholesterol saturation of bile in gall bladder thus promoting cholesterol crystallisation. Iron has a role in gallstone pathogenesis and ferritin is the most specific marker for iron levels in the body, but there are less studies which depict correlation between serum iron, serum ferritin and bile cholesterol. Aim: To study the correlation between serum iron, serum ferritin and bile cholesterol level in gallstone patients. Materials and Methods: The observational study was conducted in the Department of Surgery. The study population was 45 patients with gallstone disease. Serum iron, Serum ferritin and Bile cholesterol contents were analysed. Serum cholesterol levels were estimated using Folch method and estimation of bile cholesterol was done by Enzopak kit.Statistical analysis was done by using descriptive and inferential statistics using chi square test and software used in the analysis were SPSS 24.0 version and p<0.05 was considered as level of significance. Results: A positive but negligible correlation between serum iron and serum ferritin was observed (Pearson’s correlation coefficient of 0.247). Negative correlation between serum iron vs serum cholesterol and serum ferritin vs serum cholesterol was observed. Conclusion: It can be concluded that a low value of serum iron and serum ferritin is a risk factor for cholelithiasis, whereas, if the value of bile cholesterol increases then there is a high probability of cholelithiasis.