Objective <p>The purpose of this study was to preliminarily predict the chemical composition of urinary calculi using ultrasound in vivo.</p> Methods <p>The data of 267 urinary calculi patients were analyzed retrospectively, including non-contrast computed tomography (NCCT) and ultrasound imaging data before the intervention and post-interventional chemical composition. The Hounsfield unit (HU) value of calculi, the grayscale value of calculi and the grayscale value of posterior acoustic shadow of calculi on ultrasound were measured and analyzed statistically.</p> Results <p>The chemical composition analysis indicated that there were four types of mixed calculi; the main components were whewellite calculi, weddellite calculi, carbonate apatite calculi and anhydrous uric acid calculi. The HU value could distinguish between calcium-containing calculi and anhydrous uric acid calculi, with a cut-off value of 644.00, a sensitivity of 88.00% and a specificity of 95.04%, and <i>P</i> &lt; 0.001. The grayscale value of calculi on ultrasound could distinguish between calcium-containing calculi and anhydrous uric acid calculi with a cut-off value of 200.29, a sensitivity of 38.84% and a specificity of 96.00%, <i>P</i> &lt; 0.001. The grayscale value of the posterior acoustic shadow of calculi on ultrasound could distinguish between carbonate apatite calculi and anhydrous uric acid calculi with a cut-off value of 31.48, a sensitivity of 58.33% and a specificity of 84.00%, and <i>P</i> = 0.011.</p> Conclusion <p>Ultrasound can preliminarily distinguish the chemical composition of urinary calculi and provide certain information for clinicians to choose treatment plans.</p>