Separation and purification method of hyaluronic acid

The invention provides a separation and purification method of hyaluronic acid. The separation and purification method comprises the following steps: 1) dissolving freeze-dried powder of a crude hyaluronic acid product to obtain a hyaluronic acid solution; 2) loading the hyaluronic acid solution obtained in the step 1) into a chromatographic column filled with a UniGel medium for chromatography, and respectively carrying out loading and elution by taking pure water and inorganic salt as mobile phases; and 3) segmentally collecting the hyaluronic acid solution with the target peak value after chromatography and elution in the step 2), and analyzing a component solution meeting the requirements to obtain purified disaccharide, tetrasaccharide, hexasaccharide and octasaccharide. According to the separation and purification method of the hyaluronic acid, only one-step elution is needed, various saccharides in the hyaluronic acid can be separated through purification of the filler, no organic solvent is used durin