Abstract: A method for simultaneous determination of lactic acid and acetic acid in jiaosu solution by high performance liquid chromatography (HPLC) had been established. This article focused on the establishment of the simultaneous determination method of lactic acid and acetic acid in jiaosu solution by high performance liquid chromatography through the optimization of the detection wavelength, mobile phase and its concentration and pH. The results showed that the optimal Chromatographic conditions are as follows:Chromatographic column (SinoChrom ODS-BP 4.6 mm×150 mm, 5 μm), 0.01 mol/L KH₂PO₄ solution (pH2.88) as mobile phase, flow rate of 0.8 mL/min, wavelength of 215 nm, injection volume of 20 μL, column temperature of 25℃. The method could be used for simultaneous determination of lactic acid and acetic acid in jiaosu solutions. The method has the advantages of fast detection, wide linear detection range of lactic acid (0~2100 μg/mL) and acetic acid (0~1800 μg/mL), good precision and repeatability (the RSD of parallel determination of lactic acid and acetic acid were 0.99% and 1.21%, respectively) and good recoveries (92.31%~102.20%).