Background: The chemical composition of gallstones is essential to study aetiopathogenesis of gallstone disease. Objective: To determine the composition of gallstones in a Sudanese population. Patients and methods: It describes an extensive quantitative analysis of gallstones from patients presented with symptomatic gall stone disease and treated by cholecystectomy after the acceptance of the pre-given informed consent in Khartoum teaching hospital in the period between Jan 2010 and Dec 2010. Using a pretested questionnaire data collected from and analyzed statistically by SPSS computer program version 21. Results: Data are analyzed from 94 patients (six males and 88 females). Cholesterol stones showed a significantly higher cholesterol content than pigment stones (p = 0.0042), though not significantly higher than mixed stones. Their phospholipids content and inorganic phosphates were higher than in the other types of stones and oxalate content was significantly elevated in comparison with mixed stones (p = 0.0402). In mixed stones, the cholesterol, bile acids, and bilirubin were intermediate between cholesterol and pigment stones, whereas triglycerides were significantly more than pigment stones (p = 0.0007). Bilirubin (p = 0.0021) and bile acids (p = 0.0016) were significantly higher than cholesterol stones (p = 0.0001) and (p = 0.0001) respectively. However, they contained the lowest amounts of sodium, potassium, magnesium, and oxalate. In pigment stones, bilirubin was significantly higher (p = 0.0001) than both groups. Conclusion: Collaborations between surgeons, nutritionists, biochemists, and physicians should be stimulated in future studies to define the different types of gall stones in different areas in Sudan and the relevancy of such types with diets’ traditions.