Quantitative detection method of polysaccharide containing uronic acid

Abstract

The invention discloses a quantitative detection method of polysaccharide containing uronic acid. The method comprises the following steps: preparing a standard polysaccharide solution and a sample solution to be tested; hydrolyzing polysaccharide in the standard solution and the sample solution to be tested so as to obtain hydrolyzed residue; derivatizing the residue with 1-phenyl-3-methyl-5-pyrazolone to prepare a sugar derivative; carrying out chromatographic analysis on the sugar derivative by the utilization of a high performance liquid chromatograph and a triple quadrupole linear ion trap mass spectrometer; determining which kind of polysaccharide containing uronic acid the sample to be tested is according to the chromatogram; and preparing a standard curve of chromatographic peak area and polysaccharide concentration according to the chromatogram of the standard polysaccharide disaccharide derivative obtained by high performance liquid chromatography-mass spectrum, and calculating content of polysaccharide containing uronic acid in the sample to be tested by the utilization of peak area of the sample to be tested. According to the method provided by the invention, quantitative detection of polysaccharides can be accurately carried out, and effective means is provided for quality control of products such as medicines, health product and the like of the polysaccharides.

Description

technical field [0001] The invention belongs to the technical field of uronic acid polysaccharide detection, in particular to a method for quantitative detection of uronic acid polysaccharides. Background technique [0002] The repeating disaccharide in the structure is a polysaccharide composed of an uronic acid and a hexosamine or hexose, which widely exists in nature. For example, heparin, chondroitin sulfate, and hyaluronic acid in glycosaminoglycans are all composed of repeating disaccharide fragments linked by uronic acid and hexosamine. In addition, polysaccharides containing repeated disaccharide fragments formed by linking uronic acid and hexose have also been found in some shellfish and other organisms, such as abalone gonad polysaccharide AGSP, etc. Due to their significant functional activity, some uronic acid-containing polysaccharides have been developed as medicines and health products. For example, chondroitin sulfate can effectively prevent arthritis, and ...

AI Technical Summary

Problems solved by technology

However, these chemical chromogenic methods lack specificity. For samples containing several polysaccharides of this type at the same time, only their total amount can be measured, and the content of each polysaccharide cannot be determined.

It has been reported in the literature that the disaccharides produced by the enzymatic hydrolysis of glycosaminoglycans were quantified by high performance liquid chromatography-liquid mass spectrometry, thereby quantifying various glycosaminoglycans, but the price of the degrading enzyme of glycosaminoglycanase are very expensive, and there is no commercial degrading enzyme for polysaccharides containing uronic acid-hexose disaccharide fragments, so this method is difficult to be widely used

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