TWI805271B - Purification method of ganoderma cell wall composition - Google Patents

Abstract

A purification method of ganoderma cell wall composition includes following steps: (1) taking a residue of a ganoderma fruiting body went through extraction process for obtaining an extract thereof, wherein the extract contains terpenoids, sterols, polysaccharides, or a combination thereof, and the residue contains the cell wall of the fruiting body; (2) placing the residue into an aqueous percarbonate solution to form a mixture liquid, which is then reacted at a first temperature for decolorizing the residue, wherein the concentration of the aqueous percarbonate solution is 5 to 20% (w/v), and the first temperature is between 15 to 40℃; (3) after the decolorization process, raising the temperature of the mixture liquid to a second temperature for the mixture liquid to react, so as to digest and decompose the residue, wherein the second temperature is between 80 to 100℃; and (4) filtering the treated mixture liquid to obtain a purified product.

Description

Purification method of Ganoderma lucidum cell wall composition

The present invention relates to the related technology of Ganoderma lucidum, especially a method for purifying the cell wall composition of Ganoderma lucidum.

Ganoderma lucidum is a fungus with multiple components and is an important source of health food. Its fruiting body contains about 1~3% triterpenoid and sterol, and about 1~2% water-soluble polysaccharide (water -soluble polysaccharide), and the usual method of extracting Ganoderma lucidum is to use 65~95% alcohol as the first step of extraction to obtain the low-polarity small molecule triterpenes and sterols, and then use hot water for the second step of extraction to obtain The polar macromolecular polysaccharides are obtained, and the remaining Ganoderma lucidum residue can only be used as a soil conditioner and compost except for the excipient of the above-mentioned extract.

However, the above-mentioned ganoderma residues are mainly the cell walls of ganoderma fruiting bodies, and still contain many useful components including ultra-high molecular weight polysaccharides and chitin. Therefore, in order to effectively utilize these ganoderma residues, Taiwan Patent No. 442496 is It is disclosed that the Ganoderma lucidum residues were further purified to obtain the purified cell wall composition, that is, a complex of chitin and polysaccharides, which is a polyglucosamine fiber, and has N-acetylglucosamine (N-Acetylglucosamine) and (l→3) polydextrose ((l→3)-β-D-Glucan) The copolymerized structure, and the experimental results show that the polyglucamine fiber can be applied to skin wound dressings, cosmetic excipients, or pharmaceutical excipients.

According to Taiwan Patent No. 442496, the above-mentioned purification method is firstly treated with strong alkali (1N sodium hydroxide) to digest and decompose the remaining proteins, nucleic acids, and lipids of Ganoderma lucidum residue at a high temperature of 80~100°C, and take the filtrate After cleaning, the second stage of treatment is carried out with a decolorizing agent (0.1% hypochlorite) to remove the pigment of the Ganoderma lucidum residue at a high temperature of 80-100 ° C, and become a white branched pulp-like mycelial cell wall; but , this kind of digestion and decomposition process using strong alkali will produce a large amount of extremely dark alkaline waste liquid, which is not only difficult to handle, but also causes a great burden on the environment. There is a problem of residual chlorine in cleaning, and at the same time, the reaction conditions of hypochlorite are not easy to control under the condition of high temperature, which often causes excessive oxidation and makes the recovery rate poor.

In order to solve the problem of poor recovery (only about 8-12%) in Taiwan Patent No. 442496, Taiwan Patent No. I620570 changed the second stage of treatment to 25-45% hydrogen peroxide to remove pigment, although the recovery rate It can be increased to 25~35% (as shown in Figure 2 and paragraph 0030 of Taiwan Patent No. I620570), but there are still difficulties in over-oxidation and difficult control of reaction temperature. A little carelessness in the reaction will not only produce a large number of bubbles and overflow The risk of over-reaction equipment also causes a sharp drop in recovery rate, especially high-concentration hydrogen peroxide is a highly toxic, reactive, corrosive and oxidizing liquid, and there is a greater risk in large-scale operation and storage; moreover, with The waste liquid treated by this method still contains a large amount of unreacted hydrogen peroxide, and it cannot solve the problem of a large amount of strong alkali waste liquid produced in the first stage of treatment, so it still causes a great burden on waste liquid treatment.

In order to solve the above problems, the present invention provides a method for purifying ganoderma-like cell wall components, which uses percarbonate aqueous solution as a treatment agent for decolorization, digestion and decomposition at one time, replacing the prior art The technique is a two-stage treatment of sodium hydroxide and hypochlorite or hydrogen peroxide, which not only reduces the burden on waste liquid treatment, but also improves the recovery rate of purified products.

In order to achieve the above-mentioned purpose, the purification method of the ganoderma cell wall composition of the present invention comprises the following steps: (1) take a ganoderma fruiting body and undergo an extraction process to obtain the residue after the extract, wherein the extract contains terpenoids, solid Alcohol, or polysaccharide, or its combination, the residue contains the cell wall of the fruiting body; (2) put the residue into an aqueous percarbonate solution to form a mixed solution, and react at a first temperature for about 24 hours, to The residue is decolorized, wherein the concentration of the percarbonate aqueous solution is 5-20% (w/v), and the first temperature is between 15-40°C; (3) after the decolorization is completed, the mixed solution is The temperature is raised to a second temperature and reacted at the second temperature to digest and decompose the residue, wherein the second temperature is between 80 and 100°C; (4) filtering the treated mixture , to obtain a purified product.

Wherein, the percarbonate aqueous solution is sodium percarbonate aqueous solution (Sodium percarbonate, 2Na ₂ CO ₃ ‧3H ₂ O ₂ ).

Wherein, the extraction process of the step (1) is sequentially through an organic solvent extraction and a hot water extraction, through the organic solvent extraction to obtain an extract containing triterpenes and sterols, and through the hot water extraction to obtain a polysaccharide-containing Extract.

Wherein, the purified product is a complex of chitin and polysaccharide.

Wherein, the purified product is polyglucosamine fiber, and is a copolymerized structure of N-acetylglucosamine (N-Acetylglucosamine) and (1→3) polydextrose ((1→3)-β-D-Glucan).

Wherein, the purified product can be applied to skin wound dressings, cosmetic or skin care products excipients, or pharmaceutical excipients.

Based on the above, in step (2), the residue of the fruiting body of Ganoderma lucidum is treated with an aqueous solution of sodium percarbonate at room temperature. The aqueous solution of sodium percarbonate can slowly release hydrogen peroxide (H ₂ O ₂ ) to decolorize the residue , and add sodium percarbonate depending on the degree of decolorization. After decolorization is completed, the original reactant mixture is heated in step (3). Because sodium percarbonate is added with water, sodium carbonate can be released, and sufficient alkalinity is provided when the concentration is high and the temperature is raised, so it can be digested. Decomposition reaction, decomposes proteins, nucleic acids, and a small amount of lipids in Ganoderma lucidum residues to ensure that there are no allergens left.

Therefore, the present invention uses sodium percarbonate aqueous solution as the treatment agent that decolorizes and digests and decomposes once, and replaces the two-stage treatment that prior art is sodium hydroxide and hypochlorite or hydrogen peroxide, and because the decolorization reaction of the present invention Carried out at room temperature, the reaction is mild so as not to cause excessive oxidation, so the recovery rate of the purified product can be increased to about 50-60%. In addition, the purification method of the present invention only has a waste liquid with a lighter color once in the whole process, and this The waste liquid only contains sodium carbonate and a small amount of hydrogen peroxide, which can reduce the burden on waste liquid treatment; moreover, sodium percarbonate is white powdery particles, which not only has a low cost of use, but is also quite safe during storage and use.

S1~S4: steps

[FIG. 1] is a flow block diagram of the present invention.

In order to illustrate the central idea of the present invention expressed in the column of the above-mentioned summary of the invention, it is expressed in specific embodiments.

Please refer to Fig. 1, the present invention provides a method for purifying ganoderma cell wall composition, which comprises the following steps:

Step S1: Take a ganoderma lucidum fruiting body and go through an extraction process to obtain the residue after the extract. In some embodiments, the extraction process of the present invention is to firstly take a fruiting body of Ganoderma lucidum, break it and soak it in an organic solvent to perform organic solvent extraction to obtain an extract containing triterpenes and sterols, wherein , the organic solvent is methanol, ethanol, or propanol, preferably ethanol (alcohol); afterward, the residue after organic solvent extraction is cleaned, and soaked in hot water to obtain polysaccharide-containing The extract; finally, wash the residue after the reaction, and dry the residue at about 35~45°C.

Step S2: Put the residue after step S1 into an aqueous percarbonate solution to form a mixed solution, and react at a first temperature to decolorize the residue. In some embodiments, the percarbonate aqueous solution is sodium percarbonate aqueous solution, and its concentration is 5 ~ 20% (w/v), preferably 7.5 ~ 15% (w/v); or preferably, over The concentration of the aqueous carbonate solution is 5, 7.5, 10, 12.5, 15, 17.5, or 20% (w/v); or preferably, the concentration of the aqueous percarbonate solution is 7.5 or 15% (w/v). In some embodiments, the first temperature is between 15~40°C, preferably 20~37°C; or preferably, the first temperature is 15, 17.5, 20, 22.5, 25, 27.5, 30, 32.5 , 35, or 37°C. Therefore, step S2 is to treat the residue of Ganoderma lucidum fruiting bodies with sodium percarbonate aqueous solution at room temperature. The sodium percarbonate aqueous solution can slowly release hydrogen peroxide to decolorize the residue, and add sodium percarbonate depending on the degree of decolorization.

Step S3: After the decolorization is completed, the temperature of the mixture is raised to a second temperature, and reacted at the second temperature to digest and decompose the residue. In some embodiments, the second temperature is between 80-100°C, preferably 85-90°C; or preferably, the second temperature is 80, 82.5, 85, 87.5, 90, 92.5, 95, 97.5 , or 100°C. Therefore, step S3 is to raise the temperature of the original reactant mixed solution. Because sodium percarbonate can release sodium carbonate after adding water, it also provides sufficient alkalinity when the concentration is high and the temperature is raised, so the digestion and decomposition reaction can be carried out, and the ganoderma lucidum can be decomposed. Proteins, nucleic acids, and a small amount of lipids in the residue to ensure that there are no allergens left.

Step S4: Filter the treated mixture to obtain a purified product. In some embodiments, the purified product of the present invention is a complex of chitin and polysaccharide, which is a polyglucose Amine fiber, and has a copolymer structure of N-acetylglucosamine (N-Acetylglucosamine) and (l→3) polydextrose ((l→3)-β-D-Glucan).

In some embodiments, the purified product can be washed and added to deionized water to form a suspension, and then filter the suspension with filter paper to form a film on the filter paper, which can be applied to Skin wound dressings, or cosmetic or cosmetic excipients. In some embodiments, the purified product can be processed after cleaning, such as freeze-drying, to be used as an excipient for pharmaceutical tablets, which can improve the physical properties of the powder during the tableting process.

Thereby, the purification method of the present invention uses sodium percarbonate aqueous solution as the treatment agent that decolorizes and digests and decomposes once, replaces the two-stage treatment of sodium hydroxide and hypochlorite or hydrogen peroxide in the prior art, and due to decolorization The step S2 of the reaction is carried out at room temperature, and the reaction is mild so as not to cause excessive oxidation, so the recovery rate of the purified product can be increased to about 50-60%.

In addition, the purification method of the present invention only uses sodium percarbonate aqueous solution, and there is only one waste liquid with a lighter color in the whole process, and the waste liquid only contains sodium carbonate and a small amount of hydrogen peroxide, so the burden on waste liquid treatment can be reduced.

Specific embodiments of the present invention are described below:

Example 1: Take a ganoderma lucidum fruiting body and extract triterpenes, sterols, and polysaccharides with alcohol and hot water to extract residues, wherein the residues are mainly composed of macromolecules such as cell walls and proteins, nucleic acids, and melanin (melanin). ). The weight of the residue is 100 equal parts (moisture content 12.44%), add 1000 volume equal parts of 15% (w/v) sodium percarbonate solution, and place it at a room temperature of 20~37 ° C, react for about 24 hours, percarbonate Sodium aqueous solution slowly releases hydrogen peroxide and decolorizes the residue of Ganoderma lucidum; after that, the temperature of the original reactant is raised to 85-90°C and maintained for 1-2 hours to decompose the protein, nucleic acid, and a small amount of lipid in the residue of Ganoderma lucidum. After cooling to room temperature, filter to obtain a purified product, wash the purified product with water until the pH value is neutral, and detect the concentration of hydrogen peroxide with hydrogen peroxide test paper until it is not detected. The wet weight of the purified product recovered was 185.45 equal parts, the water content was 74.0%, and the recovery rate was 55.06%.

Example 2: The residue after extracting triterpenes, sterols, and polysaccharides from a Ganoderma lucidum fruiting body with alcohol and hot water. Wherein, the weight of residue is 100 equal parts (moisture content 12.50%), add in the 7.5% (w/v) sodium percarbonate solution of 1000 volume equal parts, and be placed in room temperature 20～37 ℃, react about 24 hours, The sodium percarbonate aqueous solution slowly released hydrogen peroxide, and decolorized the ganoderma lucidum residue. Since the decolorization was not completed, 75 equal parts by volume of sodium percarbonate was added, and left for about 24 hours to complete the decolorization; after that, the original reactant Raise the temperature to 85-90°C and maintain it for 1-2 hours to decompose the protein, nucleic acid and a small amount of lipid in the ganoderma residue. After cooling to room temperature, filter to obtain a purified product, wash the purified product with water until the pH value is neutral, and detect the concentration of hydrogen peroxide with hydrogen peroxide test paper until it is not detected. The wet weight of the purified product recovered was 374.03 equal parts, the water content was 87.50%, and the recovery rate was 53.43%.

Example 3: The residue after extracting triterpenes, sterols, and polysaccharides from a fruiting body of Ganoderma recinaceum with alcohol and hot water. Wherein, the weight of residue is 100 equal parts (moisture content 11.87%), add in the 15% (w/v) sodium percarbonate solution of 1000 volume equal parts, and be placed in room temperature 20～37 ℃, react about 24 hours, The aqueous solution of sodium percarbonate slowly releases hydrogen peroxide, and decolorizes the residue of Ganoderma lucidum; after that, the temperature of the original reactant is raised to 85-90°C and maintained for 1-2 hours to decompose the protein, nucleic acid, and a small amount of the residue of Ganoderma lucidum Lipid. After cooling to room temperature, filter to obtain a purified product, wash the purified product with water until the pH value is neutral, and detect the concentration of hydrogen peroxide with hydrogen peroxide test paper until it is not detected. The wet weight of the purified product recovered was 244.75 equal parts, the water content was 81.03%, and the recovery rate was 52.68%.

The above-mentioned embodiments are only used to illustrate the present invention, and are not intended to limit the scope of the present invention. All modifications or changes that do not violate the spirit of the present invention belong to the intended protection category of the present invention.

S1~S4: steps

Claims (10)

A method for purifying ganoderma cell wall components, comprising the following steps: (1) taking a fruiting body of ganoderma lucidum and going through an extraction process to obtain the residue after the extract, wherein the extract contains terpenoids, sterols, or polysaccharides, or Its combination, the residue contains the cell wall of the fruiting body; (2) put the residue into a percarbonate aqueous solution to form a mixed solution, and react at a first temperature for about 24 hours to decolorize the residue, Wherein, the concentration of the percarbonate aqueous solution is 5-20% (w/v), and the first temperature is between 15-40°C; (3) after the decolorization is completed, the temperature of the mixed solution is raised to a first Two temperatures, and react at the second temperature to digest and decompose the residue, wherein the second temperature is between 80-100°C; and (4) filtering the treated mixed solution to obtain a purified product. The method for purifying the ganoderma lucidum cell wall composition as described in claim 1, wherein the percarbonate aqueous solution is an aqueous sodium percarbonate solution. The method for purifying ganoderma-like cell wall composition according to claim 1 or 2, wherein the concentration of the percarbonate aqueous solution is 7.5-15% (w/v). The method for purifying ganoderma-like cell wall composition as described in claim 1 or 2, wherein the concentration of the aqueous percarbonate solution is 7.5% (w/v) or 15% (w/v). The method for purifying ganoderma lucidum cell wall composition according to claim 1, wherein the first temperature is 20-37°C. The method for purifying Ganoderma lucidum cell wall composition according to claim 1, wherein the second temperature is 85-90°C. The method for purifying ganoderma-like cell wall composition as described in Claim 1, wherein, the extraction process of the step (1) is sequentially carried out through an organic solvent extraction and a hot water extraction, and through the organic solvent extraction, a compound containing triterpenes and The extract solution of sterol, and then obtain the extract solution containing polysaccharide through the hot water extraction. The method for purifying ganoderma-like cell wall composition as described in Claim 1, wherein the purified product is a complex of chitin and polysaccharide. Purification method of Ganoderma lucidum cell wall composition as described in claim item 8, wherein, the purified product is polyglucosamine fiber, and is N-ethylglucosamine (N-Acetylglucosamine) and (1→3) polyglucose ( (l→3)-β-D-Glucan) copolymer structure. The method for purifying the ganoderma-like cell wall composition according to any one of claims 1, 8 or 9, wherein the purified product can be applied to skin wound dressings, cosmetic or skin care product excipients, or pharmaceutical excipients .

Images