Flavonoids Extraction, Purification, Identification And Anti-oxidative Activity Research Of LAOLING Jujube Leaves

LaoLing thread jujube, is famous for its origin shandong Texas LaoLing county,has a long cultivation history. Many studies have shown that flavonoids withantioxidant and anti-inflammatory, and treatment of cardiovascular diseases andother health care function. Taking LaoLing jujube leaf as the raw material, by usingethanol reflux extraction of flavonoids in the leaves and stems of jujube, extractionprocess was optimized by using an orthogonal experiment, using a macroporousresin purification jujube leaves flavonoids, liquid chromatography-massspectrometry identification jujube leaf flavonoids, and research on antioxidantactivity of total flavonoids of jujube leaf, the experimental results are as follows:Using ethanol refluxing extraction to the extraction technology of flavonoids injujube leaf single factor experiment, through the orthogonal method to get the besttechnological conditions as follows:50%ethanol concentration, solid-liquid ratio1:10, reflux time1.0h, extraction temperature70℃. Under this condition, thecontent of total flavonoids of14.3151mg/g.Will get the crude extract of jujube leaf flavonoids of macroporous resinpurification, through static experiments of macroporous resin, selected from the7kinds of macroporous resin D101macroporous resin in the separation andpurification of flavonoids. Through macroporous resin in the static and dynamicadsorption and adsorption experiments to determine the best technology ofpurification of jujube leaf flavonoids: jujube leaves flavonoids onthe sample amountto40ml, pH value of sample liquid on5, select the volume fraction of70%ethanolelution, elution volume of200mL.At the high performance liquid chromatography separation chromatographicconditions of jujube leaf flavonoids, mobile phase of acetonitrile-water was finallychosen. By liquid chromatography-mass spectrometry coupled analysis of jujubeleaf flavonoids of monomer, got the7kinds of compounds, separation by massspectrogram, reference related literature, the basic five flavonoids can be determined,they as follow: the wild baicalin, rutin, trifoliate-orange immature fruit glycosides,naringin, hesperidin.To investigate the in vitro scavenging free radicals of jujube leaf flavonoidsability and zebra fish in vivo antioxidant activity, the experimental results show that jujube leaf flavonoids on DPPH free radical, hydroxyl free radical, superoxide anionhas good scavenging effect, by restoring force experiment, found jujube leafflavonoids of reducing power compared with the same concentration of Vc, thedifference is not big, that jujube leaf flavonoids have good antioxidant activityability. In zebra fish antioxidant activity in vivo experiments, through microscopicobservation, the results show that the jujube leaf flavonoids could partially restoreskin fluorescence point on the fluorescent zebra fish, explain jujube leaf flavonoidshas good antioxidant activity.