A high performance liquid chromatographic method for the determination of glycerol (material) , acetic acid(by-product), 1,3-propanediol(production), and ethanol(by-product) in fermentation broth from glycerol to 1,3-propanediol has been developed.Differential refractometric detector, column oven, column of stationary phase ofPL Hi-Plex H ligand exchange resin (5μrm, 300 mm× 7.7 mm I.D.), and mobile phase of 0.005 mol/L H2SO4 water solution were used.The flow rate of mobile phase was 0.6 mL/min and the column temperature was 65 ℃.The calibration graphs for these compounds were linear with correlation coefficients.at >0.9997(n=8) over the ranges of 0 mg/L ~3874 mg/L、 0 mg/L ~1418 mg/L、 0 mg/L ~5209.0 mg/L, and 0 mg/L ~3570 mg/L.The limits of detection.(signal-to-noise ratio=2) for them were in the range of 2.1 mg,/L~ 6.5mg/L.The relative standard deviations (RSD, n=6) were in the range of 0.13%-0.59%, and the average recovery rates were in the range of 99.5%-100.2%.The method was p roved to be effective way to determine the compounds in the fermentation broth.