Study On Separation And Purification Of Polysaccharides From Portulaca Oleracea L. And Their Biological Activities

Portulaca oleracea L. is a kind of plants which can be used as vegetable and drag. Recently, general attention is paied to Portulaca oleracea L., because of its affluent nutrition , distinct pharmacologic action and the character as vegetable and drag. Investigations indicated that the Portulaca oleracea L. had ditissimus pharmacologic action as adjusting blood fat, anti-angiosclerosis, anti-tumor, anti-aging, and nor-blood sugar. At present, there were more researches of its flavonoids, but less researches about other active compounds.Polysaccharides(PS) has many biological activities. General attention is paied to PS's effections of anti-tumor and immunological regulation. Right now, there are some reports about the Portulaca oleracea L. polysaccharides, but there is no report about the genealogical research of the extraction, separation and purification of the Portulaca oleracea L. polysaccharides. In this article, the Portulaca oleracea L. was used as raw material, and to study and research the extraction, separation, purification and activities as immunological activity in vivo and anti-tumor, antioxygen activity out-of-body of the Portulaca oleracea L. polysaccharides. This research provided the foundation for the investigating and exploitating of anti-tumor function food and medicine. And it can provide a new path for the developing and utilizing of Portulaca oleracea L..The maine research contents and results as follows:1,To study the extraction technology of polysaccharides from Portulaca oleracea L.. The content of polysaccharides of Portulaca oleracea L. was determined by the method of phenol-sulphate acid. On the basis of single factors tests, the four factors including temperature, solid-liquid ratio, time and the extraction times on the extraction rate of the polysaccharides were studied by orthogonal experimental design. The results showed that temperature was an important factor for the extraction rate of the polysaccharides. The optimal extraction technology conditions were that the temperature was 100℃, the extraction time was 2h, the solid-liquid ratio was 1:15 and the extraction times were 4 times. The extraction rate, yield and content of polysaccharides were 94．26%, 15．50% and 22．60%, respectively.2,To study the technology of separating and purificating of polysaccharides from Portulaca oleracea L..(1) Three methods: macroporous resin adsorptive process,H₂O₂ method and active carbon adsorption of separating pigment were uesd on the Portulaca oleracea L. polysaccharides, and than compared these methods.The result was that the best method of separating pigment was macroporous adsorptive resins AB-8．(2) After decolored, study on the deproteinization experiment of the Portulaca oleracea L. polysaccharides. Compared there methods of TAC method,sevag method and TCA-n-butanol method. The best way was sevag method.(3) Purified the Portulaca oleracea L. polysaccharide with DEAE cellulose column chromatography. Obtain pure Portulaca oleracea L. polysaccharide: POPS₀,POPS₁,POPS₂,POPS₃ and POPS₄．The purify of POPS₀,POPS₁,POPS₂,POPS₃ and POPS₄ were 84．31%, 92．14%, 90．45%, 88．47% and 82．16%. To study the molecular weight of these holosaccharides by HPLC, the results are: POPS₀: 1．801×10⁴D;POPS₁:5．583×10⁴D;POPS₂:2．721×10⁵D;POPS₃:6．006×10⁵D;POPS₄: 9．443×10⁵D. To study the composition of the monosaccharide by the vapor phase chromatography.The results are: POPS₀ is composed of glucose and galactose, these molar ratio is 1．76:1; POPS₁ is composed of rhamnose, glucose and galactose, these molar ratio is 1．61:1．12:1．4; POPS₂ is composed of arabinose, xylose, mannitose, and galactose, these molar ratio is 1．34:1．84:1．52:0．15; POPS₃ is composed of xylose, glucose and galactose, these molar ratio is 1．62:1．22:1．48; POPS₄ is composed of arabinose, glucose and galactose, these molar ratio is 1．22:0．67:0．41．3,To study the immunological activities of crude polysaccharides(POP-1) and de-proteinization (POP-2) polysaccharides from Portulaca oleracea L.. The resules were as follows: in the dose of 100mg/kg, POP-1 could significantly enhance mice macrophage to swallow chicken erythrocytes(p<0．01) ; in the dose of 300mg/kg, POP-1 significantly enhance the content of haemolysin in mice body(p<0．001) ; in the dose of 100 mg/kg, 300mg/kg, POP-1 increases thymus index and spleen index in immunosuppression mice (P<0．05) . In the dose of 300mg/kg, POP-2 could enhance mice macrophage to swallow chicken erythrocytes (p<0．05 ); in the dose of 300mg/kg, POP-2 significantly enhance the content of haemolysin in mice body(p<0．001) ; in the dose of 100mg/kg,300mg/kg, POP-2 significantly increases thymus index and spleen index in immunosuppression mice (P<0．01or p<0．001) .4,To study the founction of POP and POPS₁,POPS₂,POPS₃,POPS₄ to inhibite the growth of SGC, AGS, and U937．The result is: these polysaccharides are all having the ability to inhibite the growth.. POPS₁ had the best effection on SGC, when its density is 160μg/mL, the inhibition ratio is 49．27%; POPS₂ had the best effection on AGS, when its density is 320μg/mL, the inhibition ratio is 26．33%; POPS₃ had the best effection on U937, when its density is 640μg/mL, the inhibition ratio is 66．74%.5,To study the antioxidize activity in vitro of the polysaccharides from Portulaca oleracea L. by orthohydroxybenzoic acid and DPPH. POPS₂ had the best effection. When its density is 960μg/mL, the clearance rate of hydroxide radical is 92．15%, and the clearance rate of DPPH radical is 80．52%.