Extraction purification and composition analysis of Folium hibisci mutabilis polysaccharides
Objective To investigate the extraction process of Folium hibisci mutabilis polysaccharides,and to analyze,isolate and purify the monosaccharide composition. Methods The enzymatic extraction process of Folium hibisci mutabilis polysaccharides was optimized by star point response surface method,and Folium hibisci mutabilis polysaccharides deproteinization was deproteinized by Sevag method and trichloroacetic acid method to screen the optimal deproteinization method. The deproteinized Folium hibisci mutabilis polysaccharides was separated and purified by DEAE-52 cellulose column and Sephadex G-100 glucan gel column. The monosaccharide components of Folium hibisci mutabilis polysaccharides and the purified product were analyzed by 1-phenyl-3-methyl-5-pyrazolone (PMP) precolumn derivatization-high performance liquid chromatography. Results The optimal extraction process of Folium hibisci mutabilis polysaccharides was as follows:3 hours of extraction,the liquid to material ratio of 70∶1,the temperature of 40 ℃,the enzyme dosage of 1.3％,and the yield was 56.42％. The best method for Folium hibisci mutabilis polysaccharides deproteinization was trichloroacetic acid. Folium hibisci mutabilis polysaccharides was mainly composed of six monosaccharides:mannose,rhamnose,glucose,galactose,xylose and fucose,the order of its content was glucose>galactose>xylose>mannose>fucose≈rhamnose,and the total polysaccharide content reached the highest in September. The isolated and purified Folium hibisci mutabilis polysaccharides Ⅰ contained glucose and galactose with a molar ratio of 9.06∶1,Folium hibisci mutabilis polysaccharides Ⅱα contained mannose,rhamnose,galactose,xylose and fucose with a molar ratio of 0.23∶0.15∶1∶0.45∶0.21,and Folium hibisci mutabilis polysaccharides Ⅱb contained only a small amount of fucosse. Conclusion The optimal extraction process of Folium hibisci mutabilis polysaccharides is stable and feasible,which can provide a theoretical basis for its production. The PMP precolumn derivatization-high performance liquid chromatography method is used to determine the composition of Folium hibisci mutabilis polysaccharides monosaccharides with good stability,accuracy and repeatability,which can be used as a quality control method for Folium hibisci mutabilis polysaccharides. The purification process of Folium hibisci mutabilis polysaccharides is stable and feasible,and can successfully purify homogeneous polysaccharides.
Folium hibisci mutabilisPolysaccharideExtraction processResponsive surface designIsolation and purificationDeproteinizationMonosaccharide compositionPrecolumn derivatization
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