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Studys On Separation And Purification, Structure Analysis And Antioxidant Activity Of Abalone Viscera Ploysaccharide

Posted on:2014-10-26Degree:MasterType:Thesis
Country:ChinaCandidate:W W ZouFull Text:PDF
GTID:2311330485495167Subject:Food Science
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Abalone is very expensive and previous seafood, which is well known as "the champion of seafood", and it has a good reputation in the international market. Recent years, with the gradual increase of abalone production in our country, large numbers of abalone internal organs (20-30% of its weight) have been abandoned or turned into low-price products, which result in the waste of resources and environmental pollution. Our group has extracted crude polysaccharide from the abalone viscera at earlier stage, and verified its physiological activity. In this study, abalone viscera crude polysaccharides were isolated and purified for further analysis of its nature, structure and antioxidant activity.The crude polysaccharide, named AVP, was isolated from pre-treated abalone viscera by pulsed electric field (PEF) and ethanol precipitation, containing 60.39% of sugar. The yield of AVP was 3.0%. Comparing the effects of three methods of removing protein, Sevag method is the best according to the results. AVP was fractionated by stepwise elution from DEAE-52 anion-exchange chromatography and three fractions (AVP-1, AVP-2, AVP-3) were obtained. The yields of three fractions were 25.3%,39.7%,15.8% respectively. The three fractions are eluted by distilled water from Sephadex G-100 molecular sieve column and the elution curves are single symmetrical peak.The vitro antioxidant analysis showed that abalone viscera polysaccharide exhibited certain reducing power and the ability to scavenging free radicals, such as superoxide radical and hydroxyl radical, among which AVP has greater reducing abilities than the other three fractions. The scavenging activity of AVP-1 on superoxide radical and hydroxyl radical is the strongest. The scavenging activity of AVP-1 on superoxide radical was 50% or more at concentration of 10 mg/mL, and the scavenging activity of AVP-1 on hydroxyl radical was 100% at high concentrations.The H202-induced oxidative stress model of HepG2 cells was established to further investigate the antioxidant activity of AVP on the level of cells. The evaluations include the survival rate, the content of malondialdehyde (MDA) and superoxide dismutase (SOD) activity. In particular, at concentration of 60?g/mL, the survival rate of oxidative damage cell was 89.1% with the handling by AVP-1. The MDA content was significantly lower after handling by AVP. And for SOD, AVP-1 and AVP-2 show better performance than AVP-3 in increasing SOD activity.The chemical analysis showed that AVP-1 contained 93.3% of sugar,1.57% of protein,4.7% of sulfate; AVP-2 contained 87.6% of sugar,6.23% of protein,9.3% of sulfate; AVP-3 contained 90.4% of sugar,3.8% of protein,12.5% of sulfate. The molecular weights were 148252,75162 and 60954 respectively. IR analysis demonstrated that the backbone of AVP-1 and AVP-3 were ?-type glycosidic linkages, and the three fractions all contained pyranose. Monosaccharide composition analysis showed that AVP-1 was composed of L-Rhamnose, D-Xylose, D-Mannose, D-Glucose, D-Galactose, Glucuronic acid and Fucose; AVP-2 was composed of L-Rhamnose, D-Glucose, Glucuronic acid and Fucose, and AVP-3 was composed of L-Rhamnose, D-Glucose, D-Mannose, D-Galactose, Glucuronic acid.
Keywords/Search Tags:abalone viscera polysaccharide(AVP), Separation and purification, antioxidant activity, structural properties
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