Separation And Analysis Of Polysaccharide From Prunella Vulgaris Linn

Prunella vulgaris Linn is the dry spike of Prunella vulgaris Linn which belongs to self-heal category, labiate .Its name backs from the saying"the grass will die away after the Summer Solstice".Now, research show the Prunella vulgaris Linn polysaccharides has many functions such as antivirus ,antibacterial,lower down blood pressure,antiinflammation, reducing blood fat, anticancer, restrating immunity and so on. The Prunella vulgaris Linn polysaccharides has gotten more and more attention on clinic value and exploitation value, just for it is a kind of potential resource.In this paper, we used Prunella vulgaris Linn as raw material, based on the research actuality of that, than did a series of work on Prunella vulgaris Linn .Firstly, polysaccharide from Prunella vulgaris Linn was extracted with water at 90℃. It was fractionated by anion-exchange chromatography and gel filtration chromatography. The results showed that weight average molecular weight of the major portion PLS3 was 8.3×10-5 Da by HPGPC. The primary structure of PLS3 was determined from its IR spectrum.Secondly, traditional phenol-vitriol method was amended to be more effective .In this paper, the manner of adding vitriol and the displaying temperature after adding vitriol effect on sensitive and absorbency Mixed standard samples were prepared based on the monosaccharide compositions to detect the quality of Prunella vulgaris Linn polysaccharide and PLS3.The result were 81.62%and 85.33% and the relative standard error were 0.56%and 0.34% respectively.Amended method effectively minished error and enhanced sensitive.Thirdly, an ion-pair reverse phase high performance liquid chromatographic (RP-HPLC)method for the simultaneous determination of carbohydrate and uronic acids was presented .p-Aminobenzoic acid(p-AMBA) was used for pre-column derivatization of the analytes ,enabling flurouescence (λex=313nm,λem=358nm)or photometric detection ( 303nm ) . Reaction condition such as reaction temperature and reaction time was optimized.Chromatographic column suitable for derivatization was selected. Effects of mobile phase compositions such as ion pairs and their concentrations and pH on the retention behaviors and separation results of 10 monosaccharides and 2 uronic were examzined. Derivatives of frucose, galactose, glucose, mannnose, xylose, arabose, ribose, N-acetyl-glucosamine, galacturonic acid, fucose, glucuronic acid , and rhamnose were separated on Atlantis dC18 column with hydrophilic end capping within 45 min ,applying sodium cyanoborohydride (TBAHSO4) as the ion pair reagent . The concentration detection limits was between 3.38 and 176×10-8 mol/L for fluorescence detection and between 2.55 and 13.4×10-7mol/L for UV detection. A good linearity was achieved in the concentration from 40 to143 mg/mL(R2>0.99). The changes of peak areas of derivative in 12-51 hours after derivatization were studied,and the RSD was from 2.5% to 3.9%. The described method has been applied for the determination of mono-/disaccharides and uronic acid in Prunella vulgaris Linn polysaccharide and spirulina polysaccharide after dissolved in trifluoroacetic acid solution (2 mol/L).And the result was nearly identical to that of ion chromatography and gas chromatography.After above research, we got better method to detect monosaccharide compositions of Prunella vulgaris Linn polysaccharide, discuss the detection and spectroscopy manner of Prunella vulgaris Linn polysaccharide.And these will benefit to food and medicine quality standard, offer useful gist to analysis on Prunella vulgaris Linn. And we look forward to making beneficial contribute to the deep exploitation on Prunella vulgaris Linn resource.