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Extraction method of lycium ruthenicum polysaccharide and application of lycium ruthenicum polysaccharide

The technology of a black fruit wolfberry and an extraction method is applied in the directions of medical preparations containing active ingredients, pharmaceutical formulations, organic active ingredients, etc., which can solve the problems of low extraction efficiency, severe extraction conditions, and lower activation energy, and achieves the promotion of cell apoptosis. Death, the method is simple, the effect of shortening the time

Active Publication Date: 2020-02-04
黑龙江邦超生物科技有限公司
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AI Technical Summary

Problems solved by technology

The acid-base extraction method needs to be neutralized in the later stage, and the extraction conditions are relatively severe, which is more destructive to the sugar chain, so it is rarely used at present; the extraction time of the microwave method should not be long, and the power should not be too high, otherwise it will lead to excessive evaporation of water and dissolution of polysaccharides At the same time, this method reduces the activation energy of some reactions, so that new forces are formed between polysaccharide molecules and between polysaccharides and other molecules to prevent the dissolution of polysaccharide molecules; the enzymatic extraction reaction is mild, and the disadvantages are cellulase and The application of pectinase is difficult to control, and it is often easy to obtain a large amount of oligosaccharides, and the oligosaccharides with anti-tumor biological activity usually contain very little
Although the above extraction methods have their own advantages and disadvantages, the overall extraction efficiency is low, the polysaccharide is lost in the refining process, and the application of Lycium barbarum polysaccharide in anti-tumor is less, so the present invention provides a polysaccharide of black fruit Extraction method and application in antitumor

Method used

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  • Extraction method of lycium ruthenicum polysaccharide and application of lycium ruthenicum polysaccharide
  • Extraction method of lycium ruthenicum polysaccharide and application of lycium ruthenicum polysaccharide
  • Extraction method of lycium ruthenicum polysaccharide and application of lycium ruthenicum polysaccharide

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Embodiment 1 A kind of extraction method of Lycium barbarum polysaccharide

[0024] Include the following steps:

[0025] (1) After washing the black wolfberry, dry it at 40°C, dehydrate, pulverize, pass through a 300-mesh sieve, and dry to obtain the black wolfberry powder;

[0026] (2) Add Lycium barbarum powder into distilled water, stir evenly, extract at 70° C. for 3 hours, and filter to obtain the extraction solution of Lycium barbarum polysaccharide; The liquid ratio is 1:25g / mL;

[0027] (3) adding ethanol to the extraction solution of Lycium barbarum polysaccharide in step (2), after the alcohol meter detects that the alcohol content is 90%, carry out suction filtration, place the obtained filter cake in an oven and dry at 60° C. for 30 minutes, Obtain polysaccharide powder;

[0028] (4) After adding 50 mL of DMSO to 5 g of polysaccharide powder obtained in step (3), dissolve it, stir it magnetically for 12 hours, add 50 mL of distilled water, continue to st...

experiment example 1

[0030] The drawing of experimental example 1 glucose standard curve:

[0031] (1) Preparation of anthrone-ethyl acetate solution

[0032] Weigh 1g of anthrone into a 50mL volumetric flask, slowly add ethyl acetate to volume, shake well to dissolve, and store in a dark place for later use.

[0033] (2) Determine the maximum absorption wavelength:

[0034] Weigh 0.1g of glucose powder with an analytical balance and place in a 100mL volumetric flask to constant volume, prepare a glucose standard solution with a concentration of 1mg / mL, and pipette 0, 0.1, 0.2, 0.3, 0.4, 0.5mL of glucose standard solutions to 10mL, the final concentration of glucose is 0μg / mL, 10μg / mL, 20μg / mL, 30μg / mL, 40μg / mL, 50μg / mL respectively, take 2mL of the glucose solution of the above concentration and add 0.5mL of anthrone-ethyl acetate, 5mL concentrated sulfuric acid solution, shake well, scan at a wavelength of 500-800nm, and find that the absorption value at 620nm is the largest, so this wavelengt...

Embodiment 2

[0037] Embodiment 2 Different solid-liquid ratios influence polysaccharide extraction yield

[0038] Weigh 5 portions of 10g black wolfberry powder and add different volumes of aqueous solutions to make the solid-liquid ratio (g / mL) 1:10, 1:15, 1:20, 1:25, 1:30. Stir the glass rod to fully soak it, extract at 90°C for 2 hours, filter it with gauze, filter it with a Buchner funnel, take the filtrate, and add anthrone (C 14 h 10 O)-Ethyl acetate (CH 3 COOC 2 h 5 ) solution (0.5mL), concentrated sulfuric acid (H 2 SO 4 ) solution (5mL), put it into boiling water for 1 min, take it out immediately and cool it at room temperature, use distilled water as a blank control, measure the absorbance at 620nm with a UV-visible spectrometer and adjust the concentration so that the absorbance value is between 0.2-0.7, compare with the standard curve , to obtain the concentration of Lycium barbarum polysaccharide, and the results are shown in Table 1.

[0039] Table 1 Effect of solid-l...

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Abstract

The invention uses lycium ruthenicum as a raw material to extract a polysaccharide, and uses an anthrone-ethyl acetate-concentrated sulfuric acid solution to determine the content of the polysaccharide. The fruits of lycium ruthenicum are subjected to crushing, soaking, heating, filtering, suction filtration and other treatment to obtain a solution containing the lycium ruthenicum polysaccharide;the extracted lycium ruthenicum polysaccharide is subjected to water extraction and alcohol precipitation, and suction filtration, a filter cake is oven-dried in an oven, a polysaccharide powder is taken out, DMSO is added for dissolution, stirring is performed for 12 h under a magnetic stirrer, distilled water is added, and suction filtration is performed; a precipitate is washed with anhydrous ethanol, acetone, and diethyl ether in order, and drying is performed to obtain a lycium ruthenicum polysaccharide refined product; and the lycium ruthenicum polysaccharide refined product is subjectedto macro-porous resin decolorization and Sevage method deproteinization to obtain the lycium ruthenicum polysaccharide. The yield of the polysaccharide is greatly improved by using the DMSO method for refining, the lycium ruthenicum polysaccharide is applied to K562 cells, cell apoptosis can be promoted by changing an expression amount of relevant apoptotic proteins in the mitochondrial pathway,and the lycium ruthenicum polysaccharide has the anti-leukemia tumor effect.

Description

technical field [0001] The invention belongs to the technical field of polysaccharide extraction, and in particular relates to a method for extracting polysaccharide from Lycium barbarum and its application. Background technique [0002] Lycium ruthenicum Murr. is a perennial shrub belonging to Solanaceae and Lycium genus. Divided into wild and cultivated products, the anthocyanin content of wild products is higher than that of cultivated products. The main production areas in my country are Qinghai, Xinjiang and Gansu, and the Qaidam Basin in Qinghai is the best. This area has a long sunshine time and a large temperature difference between day and night, and a large area of ​​black wolfberry bushes can be formed on the desert saline soil. This experiment was taken from the wild wolfberry black fruit in Qaidam Basin, Qinghai. The most prominent feature of Lycium barbarum is its high anthocyanin content, and there are many studies, but polysaccharides are the main material ...

Claims

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Application Information

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IPC IPC(8): C08B37/00A61K31/715A61P35/00
CPCA61K31/715A61P35/00C08B37/0003C08B37/006
Inventor 芦静波张现峰李燕子姚欣宇
Owner 黑龙江邦超生物科技有限公司
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