Present study is aimed to explore the rapid and reliable method of analyzing the composition of concentric layers of stones by fourier transform infrared spectroscopy with Kidney Stone Library Software (KSLS-13). Total of 69 kidney stones recovered from kidney stone patients (33 males and 36 females, mean age ranges of males and females were 10.1 to 37.3 & 15.2 to 54.4 respectively) were analyzed by fourier transform infrared spectroscopy. Composition of central, middle, peripheral layers of each kidney stone and whole stone were analyzed in the research laboratory of Institute of Biochemistry. Spectra of layers of kidney stones were collected and compared with kidney stone library software (KSLS-13). Among 69 kidney stones, 25 (36.2%) were pure stones (23.2% pure Calcium oxalate mono hydrate (COM), 10% pure carbonate apatite (CA), 3% pure magnesium ammonium phosphate (MAP)) and 44 (63.7%) mixed stones. Among 69 kidney stones, most prevalent were pure calcium oxalate stones (23.2%) and calcium oxalate mono hydrate stones mixed with carbonate apatite and ammonium hydrogen urate (AHU) (18.8%). The IR bands were compared with KSLS-13 as well as with standards. Calcium oxalate mono hydrate, carbonate apatite and uric acid were significantly increased in middle layer, but ammonium hydrogen urate and calcium oxalate dihydrate (COD) were increased in periphery. Whereas, reverse was true for magnesium ammonium phosphate in central layer. In conclusion, KSLS-13 by FTIR is found to be the most rapid and reliable method to study composition of concentric layers of kidney stones.