To establish a carbazole method for the simultaneous determination of sodium hyaluronate (SH) and chondroitin sulfate (CS).By regulating acidity or alkalinity and ionic strength with sodium acetate, CS was selectively precipitated with cetylpyridinium chloride (CPC), whereas SH was not, so SH and CS were separatedThe sample pretreatment conditions were as follows: measuring 10 mL mixture of SH and CS accurately, adding 1.6 g sodium acetate and 10 mL CPC of 12.5 mmol/L, standing for 1 h after stirring thoroughly, then, the solution was filtered with 20 μm filter membrane and the filtrate was used to determine the content of SH, the precipitates on the filter membrane were resolved with 1.4 mol/L NaCl solution by ultrasonic treatment for the determination of CS.The average recoveries of SH and CS were 101.2% and 99.4% with RSD of 1.05% and 0.96%, resp.This method can be used for the simultaneous determination of SH and CS without interference with each other.