Abstract: OBJECTIVE To establish a method for determination of fructose, glucose and xylitolum in carbohydrate and electrolyte injection by HPLC. METHODS A Waters Sugar Pak I column (300 mm×6.5 mm) was used with the mobile phase of water. Flow rate was 0.5 mL·min^-1. The tests were carried out by refractive index detector. RESULTS The resolution among fructose, glucose and xylitolum were good. The linear range of fructose was 20-100 μg(r=0.999 9), with average recovery of 99.53%, RSD=0.81%(n=9). The linear range of glucose was 40-200 μg (r=0.999 7), with average recovery of 99.75%, RSD=0.47%(n=9). The linear range of xylitolum was 10-50 μg (r=0.999 7), with average recovery of 99.78%, RSD=1.02%(n=9). CONCLUSION The established method is simple, sensitive and reproducible, and suitable for the quality control of multiple electrolytes and invert sugar injection.