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Extraction method and application of gastrodia elata oligosaccharide GEP-2

A technology of GEP-2 and extraction method, which is applied in the direction of oligosaccharide-containing food ingredients, applications, and functions of food ingredients, etc., can solve the problems of large molecular weight and difficult to use, and achieve the effect of improving the dissolution state and increasing the differentiation rate

Active Publication Date: 2022-03-04
KUNMING INST OF BOTANY - CHINESE ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Gastrodia elata is rich in polysaccharides. The main type of polysaccharide skeleton reported in Gastrodia elata is α-(1-4)-pyrano-D-glucose. Previous studies have found that the molecular weight of the above-mentioned type of polysaccharides in Gastrodia elata is 1.54×10 3 kDa or 28.8kDa, it has the defect that the molecular weight is relatively large and difficult to use, and the molecular weight is 1.54×10 3 The kDa polysaccharide is actually a starchy component, and the aqueous solution tends to form a suspension, which is difficult to dissolve in water for use.

Method used

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  • Extraction method and application of gastrodia elata oligosaccharide GEP-2
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  • Extraction method and application of gastrodia elata oligosaccharide GEP-2

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Experimental program
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Effect test

Embodiment 1

[0056] Extraction, separation, purification and structure identification of GEP-1 and GEP-2

[0057] 1. Extraction, separation and purification of GEP-1 and GEP-2

[0058] Get 7.2 kg of fresh tubers of Gastrodia elata, cut into small pieces, add 95% ethanol aqueous solution for extraction three times, filter to obtain the filter residue, place the filter residue in a ventilated place under room temperature, and ventilate until the filter residue has no alcohol smell. Obtain defatted Gastrodia elata;

[0059] Extract the defatted gastrodia elata with boiling water, mix the defatted gastrodia elata with 20L of distilled water, boil and extract for 2 hours, and extract twice. Filtrate after the extraction, combine the two filtrates, and concentrate the combined filtrate to 7-10 L under reduced pressure at 65° C. to obtain a crude concentrated extract.

[0060] Add 95% ethanol aqueous solution to the crude concentrated extract, stir slowly and quickly to make the final concentra...

Embodiment 2

[0070] Take 100 mg of GEP-1 and GEP-2 prepared in Example 1 and dissolve them in 10 ml of pure water respectively, observe and record the dissolution state. Solubility of GEP-1 and GEP-2 as Figure 4 shown. Figure 4 The GEP-2 aqueous solution on the left is obviously a clear aqueous solution, while the GEP-1 aqueous solution on the right is obviously turbid and suspended, indicating that GEP-2 has good solubility.

[0071] Take 10 μL each of GEP-1 and GEP-2, and use a transmission electron microscope (TEM) to observe the morphology of polysaccharides with a large difference in molecular weight between the two. The form of GEP-1 is as Figure 5 As shown, it presents an irregular round or triangular shape similar to starch granules; the morphology of GEP-2 is as follows Figure 6 As shown, it presents a network-like morphology, indicating that it has the function of improving the dissolution state of the extract.

Embodiment 3

[0073] Neuroprotective activity assay

[0074] 1. PC12 is cultured in the medium of 1640+10%HS+5%FBS+100U / mL double antibody, the temperature of the incubator is 37℃, 5%CO 2 ;

[0075] 2. When the PC12 cells grow to an appropriate number, take the PC12 cells, digest with trypsin, and make a cell suspension;

[0076] 3. Aspirate the PC12 cell suspension into a 15mL centrifuge tube for centrifugation (1000rpm, 5min);

[0077] 4. After centrifugation, the centrifuge tube is disinfected with alcohol and taken into the ultra-clean bench, and the supernatant is poured into the waste liquid tank to leave the sediment;

[0078] 5. Add 5 mL of new complete medium to the pellet, pipette ten times to disperse the cells as much as possible, but not too hard, to obtain a dispersed cell suspension;

[0079] 6. Take 0.2ml of dispersed cell suspension, add it to the cell counting tube, add 0.8ml of PBS, mix well, and count;

[0080] 7. Adjust the concentration of PC12 cells to 5×10 4 cel...

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Abstract

The invention belongs to the technical field of medicines and biology, and particularly relates to an extraction method and application of gastrodia elata oligosaccharide GEP-2. The invention provides an extraction method of gastrodia elata oligosaccharide GEP-2. The gastrodia elata oligosaccharide GEP-2 provided by the invention has a similar net-shaped structure for preparing a dissolution assisting platform, and the dissolution state of an extract can be remarkably improved. Meanwhile, the gastrodia elata oligosaccharide GEP-2 also has excellent activity similar to nerve growth factors (NGF), and can induce nerve cell differentiation and promote pheochromocytoma (PC12) differentiation. Tests prove that the gastrodia elata oligosaccharide GEP-2 provided by the invention can effectively improve the differentiation rate of nerve cells.

Description

technical field [0001] The invention belongs to the field of medicine and biotechnology, and in particular relates to an extraction method and application of Gastrodia elata oligosaccharide GEP-2. Background technique [0002] The traditional Chinese medicine Gastrodia elata Bl. is the dry tuber of Gastrodia elata Bl., a plant belonging to the Orhidaceae family (Orhidaceae). The traditional Chinese medicine Gastrodia elata mainly treats internal liver wind, convulsions, convulsions, dizziness, headache, numbness of limbs, limb paralysis, rheumatic arthralgia, etc. It is a traditional Chinese medicine in my country. Research on the chemical constituents of Gastrodia can be traced back to the middle of the 19th century. At present, it can be known from literature reports that the main chemical components of Gastrodia genus are phenolic components and their derivatives, polysaccharides, polypeptides, etc., a total of more than 100 species. The above ingredients have shown a v...

Claims

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Application Information

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IPC IPC(8): C08B37/00A61P25/00A23L33/125A23L33/21
CPCC08B37/0003C08B37/00A61P25/00A23L33/125A23L33/21A23V2002/00A23V2200/322A23V2250/28
Inventor 胡江苗杨柳
Owner KUNMING INST OF BOTANY - CHINESE ACAD OF SCI
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