The present study is to investigate the composition of gallstones from human population. Gallstone samples removed Laparoscopically were washed, dried and analyzed for composition by Fourier Transform Infrared (FTIR) spectrometry and by Qualitative analysis to identify various bio-chemical components. Results based on FTIR spectrometry and Qualitative analysis suggests that cholesterol and bile pigment either with or without calcium content are the most predominant components in gallstones. Chemical analysis of gallstone using FTIR spectroscopic technique and Qualitative method often clues to the pathogenesis of the gallstone disease. Results obtained using FTIR Spectra is well correlated with that of result obtained by Qualitative analysis.