Analysis Of The Chemical Constituents And Study Of The Antioxidative Activities In Bleeding Sap Of Xinjiang Grape Tree

Objective:1. Comparatively analyse of the chemical constituents and the antioxidativeactivities of different kinds Bleeding sap of Xinjiang grape tree from different areas.2.Using vitro antioxidant tests as active evaluation tools,bleeding sap of Xinjiang kashimunage grape tree was enriched by HPD400macroporous resin whose the antioxidantactivity of extract was analysed by HPLC, preparative HPLC. Methods:1.The preliminaryanalysis of the chemical constituents in Bleeding sap of Xinjiang Grape tree had beendetected with systematic pre-experimental method.2.Using gallic acid as the standard, theoptimized conditions for determining content of phenol Bleeding sap of Xinjiang GrapeTree by Folin-Ciocalteu Colorimetry.3. Derivated by phenylisothiocyanate （PITC） anddetermined17kinds of amino acids from9varieties by RP-HPLC method.4.The mineralelements of samples were determined by ICP-OES. The principal components analysisand cluster analysis using SPSS17.0software were applied to evaluate characteristicelements.5.Using VC as the standard, Bleeding sap of Xinjiang Grape tree’s reductionability,·OH, DPPH·,O₂^-·,NO₂^-·scavenging capability was detemin ed.6.Bleeding sap ofXinjiang kashi munage Grape tree was purified by HPD400macroporous resin.60%ethanol eluted fractions from HPD400macroporous resin were analyzed by HPLC.Thecondition for determining content of resveratrol by HPLC.The condition for determiningcontent of gallic acid, catechin,caffeic acid, epicatechin, coumaric acid,ferulicacid.7.60%ethanol eluted fractions was further purified by pre-HPLC,and9compoundsobtained were determined by kit about·OH scavenging capability. Results:1.Bleeding sapof Grape tree may contain amino acids, reteins, carbohydrates, glycosides, saponin. maynot contain flavonoids and alkaloids or the concentration of low-less than the detectionlimit.2.Using gallic acid as the standard,there is a good linearity between content ofphenol and absor-bance in the range of0-6mg/L. The relative standard deviations ofstability, reproducibility, precision, and spikerecovery rate of this method were all satisfactory.3.There were11^-16kinds of free amino acids in Bleeding sap of XinjiangGrape Tree,of which,7kinds were human essential amino acids.4.After principalcomponent analysis of all samples, three principal factors were gained. The cumulativecontribution rate of three principal factors to total variances was92.49%and the variancecontribution of the first factor was60.92%, indicating Ca, Mg, PandS were thecharacteristic elements of bleeding sap of grape tree. After a cluster analysis of all samples,samples could be clustered into two groups and the elemental distribution characteristicswere related to the habitats and varieties of bleeding sap of grape tree.5.The bleeding sapof grape tree have scavenging activity:·OH> DPPH·>O2^-·, and have some scavengingability for nitrite.6.Reseratrol was enriched form34.53μg· L^-1to344.65μg· L^-1by60%ethanol eluted fractions from HPD400macroporous resin.Content of gallic acid in allsamples from different area were different. Gallic acid was enriched form1.03μg·mL^-1to1.44μg·mL^-1by60%ethanol eluted fractions from HPD400macroporous resin. But theother five kinds of monomers phenolic were part found.7. The range of nine compoundshydroxyl radical scavenging rate was48.89%-70.71%,VC(0.2mg·mL^-1)was57.40%.Conclusion:1.The chemical composition of Bleeding sap of Grape tree were analyzedfirstly which provided a theoretical reference for the development of resources.2.There aredifferences in the region of different species and in the same species of different area forpolyphenol content.The cause of the difference of the polyphenol content have a certainrelationship with the geographical environment except species.3.There was significantdifference in amino acid contents among eleven varieties,But,all kinds of amino acidcontents in9samples is similar to the distribution and composition.4.Mineral elements inbleeding sap of grape tree were related to its cumulation and habitats.5.The bleeding sapof Grape tree have strong scavenging activity on·OH but low toO2^-·.6.Reseratrol was goodenriched by HPD400macroporous resin.Reseratrol,galic acid and other phenoliccompounds may be one of antioxidative substances from Bleeding sap from Grapetree.7.The chemical composition of Bleeding sap of Grape tree were analyzed firstly,andnine pure compounds had a certain antioxidant activity.This method can be used to purifythe antioxidant components from bleeding sap of grape.