Preliminary Study On Polysaccharide Of Salvia Miltiorrhiza

Polysaccharide, one class of natural polymers in Chinese herbs, is thebasic material to maintain normal operation of the lives, and has better regulation of theimmune system function and anti-aging effects. So, there is a tremendous potentialdevelopment in the herbal plant polysaccharides. Danshen, the dried root and rhizomeof Salvia miltiorrhiza Bunge, is a rare traditional Chinese medicine in our country.There are many reports on other chemical constituents of S. miltiorrhiza, but few onpolysaccharides.In the present paper, we studied the polysaccharides of S. miltiorrhiza, includingprocess of extraction, isolation, purification, and the contents of total polysaccharides indifferent parts of S. miltiorrhiza determined by phenol-sulfuric acid method, thecomposition and content of monosaccharides determined by HPLC analysis. The DNAdamage protecting activity of polysaccharides was also studied. The paper aimed tooffer data for studying polysaccharides of S. miltiorrhiza. The primary results were asfollows:(1) Using traditional method for extracting polysaccharides: ethanol degreasing,water extraction and alcohol precipitation, protein removing by Sevage method andrepeated freeze-thaw method, the small molecules impurities removing by dialysis, andthen polysaccharide obtained by freeze-dried.(2) Using phenol-sulfuric acid method to determine the polysaccharide content,the results indicated that there was a good linearity of glucose in the range of 0.01 to0.1mg. The regression equation was y=9.6584x-0.0395 (r=0.9992, n=6).Methodological experiments showed that the method had high precision and goodrepeatability and can be used as the detection means of polysaccharide in S.miltiorrhiza.(3) Comparing the total polysaccharide content. The results showed that thereexisted a distinct difference among different parts of S. miltiorrhiza, 18.66% in root,1.20% in stem, 1.14% in leaves, 4.59% in flowers and 0.75% in seeds. The distributionratio of total polysaccharide in the five parts are 24.88: 1.60: 1.52: 6.12: 1.(4) The online HPLC detection method for eight monosaccharides simultaneously was established. The chromatographic conditions were as follows: the wavelength of250nm; the column temperature of 40℃, the solvent flow 1.0mL/min; the solvent A:15% (V) acetonitrile +0.05mol/L phosphate buffer (KH₂PO₄-NaOH,pH6.9); the solventB: 40% (V) acetonitrile +0.05mol/L phosphate buffer (KH₂PO₄-NaOH,pH6.9); timegradient: 0→5→10→60 min; corresponding solvent B concentration gradient:10%→11.6%→14%→20%. Methodological experiments showed that the method hadhigh precision and good repeatability and can be used as the detection means of eightmonosaccharides.(5) Comparing monosaccharide components of the polysaccharide in differentparts of S. miltiorrhiza by HPLC analysis and accounting the content ofmonosaccharides, the results showed that, the same monosaccharide components, suchas Man, Rha, GlcUA, GalUA, Glc, Gal and Ara, existed in root, stem, leaf and flower.However, there was Xyl instead of Gal existed in seed. There were big differences incomponents and content of monosaccharides among different parts of the S.miltiorrhiza.(6) The effect of polysaccharide solution on protecting DNA damage was tested.The experiments determined the optimal concentration, and compared the DNA damageprotecting activity between different parts of S. miltiorrhiza polysaccharide. The resultsshowed the polysaccharides in root, stem and leaves had obvious activity.