Abstract: Renal calculi collected from adult male/female patients of Multan, Pakistan were analyzed for major constituents, free amino acids and metallic contents. Composition of calculi (whole powder) was determined by chemical method using stone analysis kit. Thirteen distinct types of renal calculi were identified. Most of the calculi were compound in nature, however, some simple calculi containing calcium oxalate only were also found. Two dimensional descending paper chromatography was used to identify free amino acids in some prominent classes of renal calculi. Arginine, aspartinine, glycine, leucine, isoleucine and tryptophan were found in nearly all the calculi whereas glutamic acid, histidine, hydroxyproline, phenylalanine, threonine and tryosine occurred randomly. Analysis of major (Na, K, Ca and Mg) and trace (Fe, Zn, Cu, Ni, Pb and Mn) elements in the calculi was carried out by flame atomic absorption spectrometry. Elements such as Na, K, Ca, Mg, Fe and Zn have shown their presence in nearly all the calculi however Pb, Ni, Mn and Cu levels were below the limit of detection. This study has revealed useful information about the chemical nature of renal calculi from Multan region. It will be helpful in adopting preventive strategies to minimize stone formation and their reoccurrence.