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Extraction, Purification And Antioxidant, Heart Protection Activities Of Polysaccharides From Trichosanthes Kirilowii Maxim.

Posted on:2017-11-23Degree:MasterType:Thesis
Country:ChinaCandidate:X X WangFull Text:PDF
GTID:2311330485457290Subject:Food Science
Abstract/Summary:PDF Full Text Request
Trichosanthes kirilowii Maxim.(TKM), perennial, scandent herb, was one of the Cucurbitaceae Trichosanthes, which was a kind of famous and native drug of Shandong province, has many pharmaceutical properties, such as clearing heat, expelling phlegm, removing stasis and clearing bowel aperients and so on. Trichosanthes kirilowii Maxim. polysaccharide(TKMP) is one of the important fractions of TKM, numerous studies have shown that polysaccharides exhibit a tempting prospects in anti-oxidation, anti-tumor, hypoglycaemic and immunnoregulatory and so on, but according to the literatures at home and abroad, researches on TKMP were so little. In the paper, extraction, viscosity, purification, antioxidant and heart protection activities of TKMP had been studied. This paper will provide theoretical basis for TKMP’s further study. The main results are as follows: 1. The determination of optimum extraction process of TKMPBased on single factor test, the optimum extraction conditions of UPE of TKMP were ensured by the method of orthogonal test. The yield of polysaccharide was the highest to 19.11% at the conditions of 100 MPa of extraction pressure, 3 min of extraction time and 1:40(g/mL) raw materials to water ratio, 0.32% lower than HRE, 1.00% higher than UAE, but the extraction time of UPE is only 3 min which is 1/10 and 1/20 of HRE and UAE, respectively. 2. Study on viscosity properties of TKMPThe effects of polysaccharide concentration, temperature, pH value and different metallic ions on viscosity of TKMP extracted by UPE were investigated through comparing with TKMP extracted by HRE in this study. The results showed that the polysaccharide was a kind of low viscosity polysaccharides, all those factors could affect its viscosity to different extent. The higher the solution concentration, the greater the viscosity; the higher the solution temperature, the smaller the viscosity; and alkalinity or acidity and metallic ions have less impact on its viscosity, indicating the nature of the polysaccharide is relatively stable. The viscosity of TKMP extracted by UPE was lower than TKMP extracted by HRE 3. Purification and characterization of the primary structure of TKMPThe crude TKMP was deproteinized through the Sevage method, then, purified by DEAE-52 cellulose column chromatography stepwise eluted with 0, 0.05, 0.1 and 0.2 M Na Cl solutions, four components were got, called TKMP-1, TKMP-2, TKMP-3, TKMP-4 respectively, the highest content component TKMP-1 was purified by Sephadex G-75 column chromatography for further separation and purification, and eluted with distilled water, two components was got, called TKMP-1-1, TKMP-1-2, The two fractions were identified by high-performance gel-permeation chromatography(HPGPC), TKMP-1-2 was a homogeneous, stable and pure component, having a molecular weight(Mw) of 12 KDa. The analysis of GC-MS showed that the purified fraction was composed of L-arabinose and D-mannose, and molar ratio was 1:2.72. TKMP-1-2 is a typical polysaccharide compounds, contained furan ring which had α-glycoside linkage and β-glycoside linkage two kinds of link methods. 4. Antioxidant activities in vitro of TKMPThe in vitro antioxidant activities of TKMP were investigated by measuring its scavenging ability on DPPH·, O2-·, ·OH radical, H2O2 and reducing power assay. The results showed that all the free radical scavenging rates had concentration-dependent, the IC50 values of DPPH·, O2-·, ·OH radical and H2O2 were 0.63, 1.48, 4.04, 1.03 mg/mL respectively. Reducing power of TKMP was gradually improved with the increase of the concentration, when the polysaccharide concentration was 0.5mg/m L, the reducing power was maximum. Purified fraction TKMP-1-2 had better scavenging impacts on DPPH· and O2-· radical, but the effect is not as good as crude TKMP. 5. Heart toxicity of zebrafish induced by terfenadine protective activities of TKMPThe experiment was set as low-dose group(1 μg/m L), middle-dose group(10 μg/mL) and high-dose group(100 μg/mL), the effects of TKMP and TKMP-1-2 on zebrafishes heart-rate reduction induced by terfenadine were analyzed. The r esults showed that the low-dose group could significantly improve the zebrafishes’ heart rate, however, zebrafishes of high-dose group died.
Keywords/Search Tags:Trichosanthes kirilowii Maxim.polysaccharide, ultrahigh pressure, viscosity, purification, antioxidant, heart protection
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