A kind of arborvitae leaf polysaccharide with antiviral and immune-enhancing activities, its preparation method and application
A technology for enhancing immunity and oriental orientalis leaves is applied in the field of polysaccharides, which can solve the problems such as no reports of oriental orientalis leaf polysaccharides, and achieve the effects of good antiviral efficacy, high purity, and simple and reasonable preparation process.
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Embodiment 1
[0040] (1) Weigh 200g of Arborvitae leaves, dry at 40 °C for 8 h, and then pulverize them with a powder machine;
[0041] (2) Extract with hot water at 100 °C for 2 h, the ratio of solid to liquid is 1 : 40 g / mL, and the number of extraction times is 1 time; centrifuge at 4000 r / min for 10 min, collect the supernatant and spin at 65 °C Concentrate by evaporation under reduced pressure.
[0042] (3) Deproteinize the hot water extract by using the Sevage method. The preparation method of the Sevage reagent is to mix chloroform and n-butanol at a volume ratio of 4:1; Mix at a ratio of 1:1, shake with a shaker for 20 min, centrifuge at 4000 r / min for 10 min, and take out the upper layer of orientalis leaf polysaccharide solution; the deproteinization process is repeated 10 times.
[0043] (4) Add 2 times the volume of absolute ethanol to the crude polysaccharide solution after deproteinization, and let stand at 4 °C for 48 h; then centrifuge at 4000 r / min for 15 min, collect the ...
Embodiment 2
[0046] (1) Weigh 200g of Arborvitae leaves and dry them at 80 °C for 2 h, then pulverize them with a powder machine;
[0047] (2) Extract with hot water at 100 °C for 2 h, the ratio of solid to liquid is 1 : 20 g / mL, and the extraction times are 3 times; centrifuge at 4000 r / min for 10 min, collect the supernatant and spin at 65 °C Concentrate by evaporation under reduced pressure.
[0048] (3) Use the Sevage method to deproteinize the hot water extract. The preparation method of the Sevage reagent is to mix chloroform and n-butanol in a volume ratio of 4:1; Mix 1:1, shake with a shaker for 20 min, centrifuge at 4000 r / min for 10 min, and take out the polysaccharide solution of the upper layer of orientalis leaves; the deproteinization process is repeated 10 times.
[0049] (4) Add 5 times the volume of ethanol (the volume fraction of ethanol used is 65%) to the crude polysaccharide solution after deproteinization, let stand at 4 °C for 48 h; then centrifuge at 8000 r / min for...
Embodiment 3
[0052] (1) Weigh 200g of Arborvitae leaves and dry them at 60 °C for 5 h, then pulverize them with a powder machine;
[0053] (2) Extract in hot water at 90 °C for 4 h, the ratio of solid to liquid is 1 : 30 g / mL, and the number of extraction times is 2; centrifuge at 4000 r / min for 10 min, collect the supernatant and spin at 65 °C Concentrate by evaporation under reduced pressure.
[0054] (3) Use the Sevage method to deproteinize the hot water extract. The preparation method of the Sevage reagent is to mix chloroform and n-butanol in a volume ratio of 4:1; Mix 1:1, shake with a shaker for 20 min, centrifuge at 3000 r / min for 10 min, and take out the polysaccharide solution of the upper layer of orientalis leaves; the deproteinization process is repeated 11 times.
[0055] (4) Add 3 times the volume of ethanol (the volume fraction of ethanol used is 80%) to the crude polysaccharide solution after deproteinization, let stand at 4 °C for 48 h; then centrifuge at 4000 r / min for...
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