Studies On Separation, Purification And Composition Of The Effective Polysaccharides In Atractylodes Macrocephala Koidz

The extraction,separation,purification and composition of active polysaccharides at pingjiang in Atractylodes macrocephala Koidz. (AMK)were investigated.Through the methodology of comparison, optimum technical was obtained for the first time to determine Crude Atractylodes macrocephala Koidz.polysaccharides(CAMP).The comparison showed phenol sulfuric-acid is more stable and accurate.The method was easy,stable and reliable.CAMP was determined as 41.77%.By studying the process of ethanol precipitation,decolorizing and deproteinized, an optimal polysaccharide purification method were established. It showed that a favorable yield of polysaccharide was obtained:precipitated by 80% ethanol;the deproteinization effect was optimal with TCA method;the optimal decoloration condition included as following:temperature 60℃,time 15 min, quantity of active carbon 1%and pH 4.5.After purified polysaccharides were separated by DEAE-52 column chromatography, and RAMP-1,RAMP-2,RAMP-3 were collected. RAMP-1 was further isolated by Sephadex G-100 column chromatography.HPLC-ELSD method was applied for the first time to determine weight-average molecular weight (Mw) of polysaccharides,Using standard dextran T5,T1O,T20,T40,T70 to establish standard curve.The experimental results showed that the method's linear range was Mw between 5000 and 70000.Standard curve equation was gained as lgMw =-0.303tR+3.4816,r=-0.9900.Mw of the sample polysaccharide was 32767.It is practical,reliable, accurate to determine Mw with HPLC-ELSD method.The structure and composition of RAMP were determined by IR,PC,GC,GC-MS and FGC/SAW. IR showed RAMP had typical absorption peaks of polysaccharides;RAMP was made up of D-Mannose, D-Glucose,D-Galactose,D-Arabinose,and the ration was 1:0.74:1.88: 1.07. FGC/SAW, as a new method for Polysaccharide Research,will have wide application prospects in pharmaceutical analysis.In recent years,Atractylodes macrocephala Koidz. polysaccharides (AMP) arouse more and more attention in enhancing immune function, anti-oxidation, the role of lowering blood sugar and so on.It is expected to be a great development in the value of health products and medicines. This research provides scientific reference for the further study and development of Atractylodes macrocephala Koidz.(AMK)as a natural plant resources in Hunan.