Method for quantitative determination of uronic acid-containing polysaccharide
A technology for quantitative detection of uronic acid polysaccharides, which is applied to measurement devices, instruments, scientific instruments, etc., can solve the problems of non-specificity, large error, unsuitability for detection, and complicated operation of chemical color development methods, and achieves low reagent prices. , The effect of short time-consuming and simple operation steps
Active Publication Date: 2017-05-31
DALIAN POLYTECHNIC UNIVERSITY
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Problems solved by technology
[0003] At present, there are mainly chemical chromogenic methods for quantitative detection of uronic acid-containing polysaccharides, and high-performance liquid chromatography-liquid mass spectrometry for quantification of disaccharides produced after enzymatic hydrolysis of glycosaminoglycans, while chemical chromogenic methods No specificity, large error, high enzyme price, cumbersome operation, not suitable for routine detection
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Embodiment 1
[0053](1) Accurately weigh 10.0mg chondroitin sulfate (CS), 10.0mg hyaluronic acid (HA) and 20.0mg abalone gonadal polysaccharide AGSP, dissolve them with 1.3mol / L trifluoroacetic acid after mixing, and prepare a series of concentrations (AGSP : 0.04, 0.12, 0.2, 0.4, 0.8, 1.2, 1.6, 2.0mg / mL, CS, HA: 0.02, 0.06, 0.1, 0.4, 0.6, 0.8, 1.0mg / mL) mixed standard polysaccharide solution, each concentration Two parallel.
[0054] (2) Self-made sample: containing 10% hyaluronic acid, 20% chondroitin sulfate, and 70% starch.
[0055] (3) Prepare the polysaccharide solution of the sample to be tested: take 25mg of the self-made sample in a 10mL volumetric flask, dissolve it to volume with 1.3mol / L trifluoroacetic acid solution, take 1ml to be heated and hydrolyzed; accurately measure 0.50mL of the polysaccharide solution of the sample to be tested in Add 0.50mL mixed standard polysaccharide solution to the 1mL hydrolysis tube to make a spiked sample, seal it, and wait for heating and hyd...
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The invention provides a method for quantitative determination of uronic acid-containing polysaccharide. The method comprises the following specific steps of preparing a uronic acid-containing polysaccharide standard substance solution from trifluoroacetic acid, wherein the uronic acid-containing polysaccharide is the polysaccharide of which a main chain is formed by a hexuronic acid-hexosamine or hexuronic acid-hexose disaccharide repeated disaccharide fragment; carrying out acid hydrolysis and deacidification on a standard substance to obtain a to-be-tested sample acid hydrolysis residue; dissolving a standard sample acid hydrolysis residue, and carrying out analysis by using a high-performance liquid chromatography-tandem triple quadrupole mass spectrometer to obtain a multi-response monitoring chromatogram and a standard curve; processing and detecting the to-be-tested sample acid hydrolysis residue by adopting the same method; and identifying the variety of the uronic acid-containing polysaccharide and obtaining the content of the uronic acid-containing polysaccharide in the to-be-tested sample. The method is simple in operation step, low in reagent price and relatively short in elapsed time, and polysaccharide formed by repeated disaccharide fragments of uronic acid and hexosamine or hexose can be accurately quantified at the same time.
Description
technical field [0001] The invention relates to the technical field of detection of uronic acid-containing polysaccharides, in particular to a method for quantitative detection of uronic acid-containing polysaccharides. Background technique [0002] Uronic acid-containing polysaccharides widely exist in animals, including glycosaminoglycans and other non-glycosaminoglycans containing uronic acid. Glycosaminoglycans mainly refer to straight-chain polysaccharides composed of a disaccharide repeating unit composed of a hexaminoglycan and a hexuronic acid, such as hyaluronic acid and chondroitin sulfate. In some organisms such as shellfish, polysaccharides containing repeated disaccharide fragments connected by uronic acid and hexose, such as abalone gonad polysaccharide AGSP, have also been found. Polysaccharides containing uronic acid play an important role in physiological processes and have attracted widespread attention, some of which have been developed as medicines, heal...
Claims
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IPC IPC(8): G01N30/88G01N30/06G01N30/14
CPCG01N30/06G01N30/14G01N30/88G01N2030/062G01N2030/8836
Inventor 宋爽徐鑫温成荣艾春青鲁姣姣郭丽刘斌张豹孙晓玥
Owner DALIAN POLYTECHNIC UNIVERSITY
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