<p class="abstract"><strong>Background:</strong> The biochemical constituents of gallstones predict its etiopathogenesis. Hence, this study is conducted to investigate the biochemical constituents of gallstones from indigenous Nigerian patients.</p><p class="abstract"><strong>Methods:</strong> It was a retrospective study of the biochemical constituents of post-operative gallstones from 48 adult patients of the University of Port Harcourt Teaching Hospital, Nigeria. Records on age, sex, and concentrations of gallstone biochemical constituents (total cholesterol, triglyceride, total bilirubin, and calcium) from 1<sup>st</sup> January 2007 to December 2016 were extrapolated from laboratory records and analyzed using SPSS version 15. </p><p class="abstract"><strong>Results:</strong> The gallstones were of more females than the males (70.8% versus 29.2%; p=0.004). The mean age of the study cohorts was 46.88±7.96 with age range 31–64 years. No sex difference in age was observed, however, females were younger (males 50.39±8.31 versus females 45.47±7.52; p=0.056) with a lower age range (males 40–64 versus females 31- 63) than the males. The gallstone comprised of cholesterol stones (64.7%), mixed stones (22.9%), and pigment stones (12.5%). Though the females dominated among the three types of gallstones, the predominant stone type in both sexes remained cholesterol stones. Cholesterol lipid was the main constituent of the gallstones with no sex difference (p=0.227).</p><p class="abstract"><strong>Conclusions:</strong> Cholesterol gallstones are the most common type of gallstones in this study and the cholesterol lipid is the most prevalent constituent of these gallstones. The females presented with more of the gallstones disease than the males.</p>