Abstract: OBJECTIVE To establish a method for determining content of hypoxanthine in Muscular Amino Acids and Peptides and Nucleosides injection by ion used exchange chromatography of sulfonic cation exchange bonded silicone. METHODS The ion chromatographic column used exchange chromatography of sulfonic cation exchange bonded silicone were adopted, the mobile phase was water(pH adjusted to 2.5±0.1 with sulfuric acid) at a flow rate of 0.8 mL·min^-1, the column temperature was 30℃, the photo-diode array detector was used with the detection wavelength of 260 nm. RESULTS There was a good linear relationship of hypoxanthine in the concentration of 1-50 μg·mL^-1. The average recovery was 99.0%. CONCLUSION The method has the advantages of simple operation, stable chromatogram baseline, symmetrical principal component peaks, high resolution, good repeatability, durability and high accuracy. It can be used for the determination of hypoxanthine in Muscular Amino Acids and Peptides and Nucleosides injection