Biophytum sensitivum (L.) DC (family: Oxalidaceae) has been used in the Indian indigenous system of medicine, Ayurveda, for the treatment of various health aliments including renal calculi. The present study was undertaken to investigate the anti-urolithiatic activity of standardized methanolic extract of whole plant of B. sensitivum (MBS) in rats. Urolithiasis was induced by surgical implantations of zinc disc in the urinary bladders of rats. Upon postsurgical recovery, different doses of MBS (viz., 100, 200 and 400 mg/kg body weight) were administered to zinc disc-implanted rats for the period of 7 days by the oral route. Anti-urolithiatic activity was evaluated by measuring various dimensions of stones and estimating levels of various biomarkers in serum and urine samples. A significant decrease in urinary output was observed in the disc-implanted animals, which was prevented by the MBS treatment. Supplementation with MBS caused significant improvement in glomerular filtration rate and protein excretion. The elevated levels of serum creatinine, uric acid and blood urea nitrogen were also prevented by the MBS treatment. The MBS treatment showed reduced formation of deposition around the implanted zinc disc. The higher dose of MBS (400 mg/kg) found more effective. These results indicate that the administration of MBS significantly prevents the growth of urinary stones. The possible mechanism underlying this effect is mediated collectively through diuretic, antioxidant and anti-inflammatory effects of the plant. The results concluded that the methanolic extract of whole plant of B. sensitivum possessed significant anti-urolithiatic activity.