Study On The Preliminary Separation And Activities Of Flavonoids From Walnut Leaves And Seedcases

This article studied the extraction of flavonoids from walnut leaves and seedcases, the optimum process condition with a macroporous resin to enrich the flavonoids and its resistance activity, then total flavonoids from walnut seedcases was separated preliminarily .The results are as follows:(1) We have got the standard curve on the measurement of the content of the flavonoids in walnut leaves and seedcases by spectrophotometer. The equation is Y=0.1335+0.0001X, the relative coefficient is 0.9996.(2) With the recovery rate of flavonoids as indexes, optimum process condition for separation and purification of flavonoids from walnut leaves and seedcases with macroporous resin was studied. By static and dynamic adsorption and desorption,LS303 type,D101 type macroporous resin is respectively the best for separation and purification of flavonoids from walnut leaves and seedcases among six types of macroporous adsorption resin chosen. The optimum process condition for walnut leaves flavonoids was the loaded amount of flavonoids vs dry resin: 1:8, the extract solution at a concentration about 4.67mg.mL^-1, pH of 6. After eluted with 4BV of distilled water and 4BV(bed volume, the same follow)of 70% ethanol, the yield of flavonoids was above 80% and product purity was above 70%.The optimum process condition for seedcases flavonoids was the loaded amount of flavonoids vs dry resin: 1:12, the extract solution at a concentration about 3.0075⁶.175mg.mL^-1, pH of 5. After eluted with 6BV of distilled water and 5BV of 70% ethanol, the yield of flavonoids was above 60% and product purity was above 80%.(3) Amplification experiment was studied with the optimum process condition for separation and purification of flavonoids from walnut leaves and seedcases. It shows that this purification has a better reproducibility. It gets a high yield, and only needs low cost, simple process, and safe solvents. It is suitable for mass production.(5) The inhibiting rate of walnut leaf and peel flavonoids (10mg.mL^-1) separated by macroporous resin on TMV infect to Nicotiana glutinosa is 77.52%and 79.8%. Apart from walnut leaves flavonoids has an effect on promoting Magna porthe grow, walnut leaves and seedcases flavonoids both have a weak inhibition on 5 kinds of pathogenic fungi. Compared with walnut leaves flavonoids, seedcase flavonoids has a higher inhibition rate on Venturia pyrina and Sclerotinia sclerotiorum, respectively 42.96%and 40.75%. Walnut leaves, seedcase flavonoids both have different range activity to 7 kinds of bacteria, walnut seedcase flavonoids have a stronger bacteriostasis effect on Staphylococcus aureus, Bacterium prodigious and Corynebacterium pekinense, leaves flavonoids has a weak inhibition on Proteus species and Bacillus subtilis. Different concentrations of walnut leaves, seedcase flavonoids both have different levels of inhibition on the 7 kinds of bacteria. No matter inhibiting the TMV or fungal and bacteria, compared to walnut leaves flavonoids, walnut seedcase flavonoids has a stronger effect.(6) Walnut seedcase flavonoids were chromatographic separated by using polyamide. It gets a total of 18 fractions, of which nine have the flavonoids characteristics response of color, the merger of 17²0 bottles repeated recrystallization gets a white needle-like crystal. Qualitative tests showed: The compound has the typical reaction of flavonoids. And purity tests showed it's pure.