Study On Extraction, Separation, Structure Modification And Activity Of Polysaccharides From Panax Japonicus C.A.Mey.

This article has in-depth and systematically studied polysaccharides fromPanax japonicus C.A. Meyer, optimized the extraction of polysaccharide, obtainedfive kinds of polysaccharides, compared them and unrated polysaccharide (Z) ofchemical composition and structure differences, appraised their relatedpharmacological activities in vitro and vivo, carried on the research ofpolysaccharides modification, obtained the best synthesis processes of sulfatedpolysaccharide and polysaccharide iron compound. The main research contents areas follows:1. Response surface method (RSM) was used to optimize the extractionconditions of polysaccharides from the rhizomes of Panax japonicus C.A. Meyer.RSM analysis indicated that: the fitting equation confidence level is high, thefitting degree is good and the “lack of fit” is unremarkable. The optimal conditionfor the yield of polysaccharide was extraction time3.74h, extraction temperature93.90C and ratio of water to raw material40.22.2. Savage method used to remove the protein of Panax japonicuspolysaccharide. Graded it by alcohol and obtained40%,50%,60%,75%and90%of five kinds of polysaccharides which respectively named Z1, Z2, Z3, Z4and Z5.Polysaccharide content was measured by Phenol-Sulfuric acid method. Proteincontent was determined by the method of Bradford. The content of uronic acid wasdetermined by m-hydroxydiphenyl. Results showed that: the Z1was the highestpolysaccharide content about98.01%; Z2uronic acid content was the highest,about27.71%; Z contained a little protein and the rest of the sample was notdetected protein. Fourier transform infrared spectra indicated obvious characteristicpeaks of polysaccharide, the presence of uronic acids and α-pyranose of the glucoseresidue.3. Through TLC to qualitatively analyze monosaccharide. Gas chromatographymethod used to analyze monosaccharide quantitative. Compared HP-5, DB-17andDB-35MS three kinds of columns, analyzed the methods of sugar acetatederivative and sugar nitrile acetate derivative. The sugar acetate derivative and DB-35MS column were used to GC analyze. Result showed that the samples weremainly composed by arabinose, glucose and galactose. The molar ratio ofmonosaccharide composition in Z, Z1, Z2, Z3, Z4and Z5(Ara: Gal: Glc) were 0.49:1:5.799,0.505:1:13.170,0.344:1:6.197,0.319:1:1.311,0.436:1:1.007,2.252:1:2.824.4. Polysaccharide was sulfated with the method of chlorosulfonicacid-pyridine and product sulfur substitution degree is up to0.7801. Ultraviolet andinfrared spectra showed that the product contained-SO3absorption peaks. Theproduct still maintained the polysaccharide material nature. Polysaccharide ironcompound was obtained by the ferric chloride method. The iron core in product wasformed by trivalent iron which polymerized by oxygen bridge and hydroxyl bridgeand β-FeOOH iron core structure made the formation of the aggregation state.5. Pharmacological activities of Panax japonicus polysaccharides wereresearched. The antioxidant activities of Panax japonicus polysaccharides wereinvestigated including scavenging activity of hydrogen peroxide, DPPH and freeradicals of superoxide anion in vitro. The results of antioxidant activity exhibitedthem had a good potential for antioxidant. Antioxidant capacities between thesamples are quite different and Z5had the best activity on a certain concentration.At the same concentration, samples had the strongest capability to scavenge DPPHradical. When Z5concentration was10mg/ml, it reached90%efficiency toscavenge DPPH. Models of anti-inflammatory reaction were investigated bydimethyl benzene induced ear edema and celiac capillary’s permeabilityexperiments. Writhing test and hot-plate were used to examine analgesic effects.Panax japonicus polysaccharides effectively prolonged the pain threshold, reducedthe number of writhing, inhibited the ear swelling and decreased the capillarypermeability.