Walnut green seedcase extractive, preparation method of extractive, and application of extractive as antioxidant
A technology of walnut green peel and an extraction method, which is applied in the directions of skin care preparations, antidote, cosmetics, etc., can solve the problems of complicated operation, high cost, and the activity of a single component is not necessarily the best.
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Embodiment 1
[0019] Embodiment 1: the preparation of extract
[0020] Weigh 60g of walnut green skin powder, add 900mL of anhydrous methanol, reflux and extract at 65°C for 3 times, each time for 2h, combine the extracts and concentrate under reduced pressure to form an extract to constant weight, and obtain 20.02g of methanol extract. Then, add acetone-water (1:1) until completely dissolved, extract with equal volumes of petroleum ether (60-90°C), chloroform, ethyl acetate, and n-butanol in sequence, and concentrate under reduced pressure to obtain petroleum ether extracts respectively , chloroform extract, ethyl acetate extract, n-butanol extract and raffinate were 1.38g, 0.81g, 1.45g, 4.33, 12.1g respectively.
Embodiment 2
[0021] Embodiment 2: extract free radical scavenging ability
[0022] The free radical scavenging ability of samples was determined by DPPH method. DPPH (1,1-diphenyl-2-trinitrophenylhydrazine (1,1-Diphenyl-2-picrylhydrazyl radical)) has a maximum absorption peak at 517nm. After DPPH captures an electron, the absorption at 517nm disappears, and the degree of discoloration is quantitatively related to the ability of antioxidants to scavenge free radicals. Based on this principle, the spectrophotometer is used to detect the change of absorbance value after the DPPH free radical reacts with the sample solution, which can test the ability of the sample to provide hydrogen atoms, scavenge free radicals and resist oxidation.
[0023] The extracts in Example 1 were dissolved in acetone-water (1:1) to different concentrations (100-500 μg / mL), and at the same time, the concentration was prepared with absolute ethanol to l×10 -4 mol / L DPPH solution, keep it away from light for later u...
Embodiment 3
[0033] Embodiment 3: extract reducing ability
[0034] The reducing ability of the extract was determined by the Prussian blue method. The principle of the Prussian blue reaction is that iron ions encounter ferrocyanide to produce blue ferric ferrocyanide precipitation, that is, 3K 4 Fe(CN) 6 +4FeCl 3 → Fe 4 [Fe(CN) 6 ] 3 ↓+12KCl. The reducing ability of the sample is quantitatively related to the precipitation of ferric ferrocyanide. Take 0.5mL of sample solutions with different concentrations (the preparation method is the same as in Example 1), add 0.2mol / L phosphate buffer solution of pH=6.6 and 1% K 3 [Fe(CN) 6 ] solution 2.5mL and mix well, the mixture was incubated at 50 ℃ for 20min, after adding 2.5mL10% trichloroacetic acid solution, after mixing, centrifuge at 3000rmp for 10min. Take 2.5mL supernatant, add 2.5mL distilled water and 1mL0.1% FeCl 3 , mix evenly, measure the absorbance at 700nm after 10min, the greater the absorbance value, the stronger the re...
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