CN107602721B - Method for extracting tremella polysaccharide from acid buffer solution - Google Patents
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Abstract
The invention provides a method for extracting tremella polysaccharide from an acidic buffer solution, which comprises the following steps: under the heating condition, leaching the tremella powder by adopting an acidic buffer solution to obtain a leaching solution; carrying out solid-liquid separation on the leaching solution to obtain a liquid component, wherein the liquid component contains tremella polysaccharide; and carrying out alcohol precipitation on the liquid components to obtain a precipitate which is the tremella polysaccharide. The extraction rate of the tremella polysaccharide can reach 31.57 percent, which is 3-4 times of that of the tremella polysaccharide extracted by the traditional water extraction method, and the extraction rate of the tremella polysaccharide is greatly improved.
Description
Technical Field
The invention belongs to the technical field of plant polysaccharide extraction, and particularly relates to a method for extracting tremella polysaccharide from an acidic buffer solution.
Background
Tremella, also known as Tremella, is a higher fungus, and has effects of nourishing yin, moistening lung, invigorating qi and regulating blood. Tremella polysaccharide is the main effective component of Tremella, and is an acidic heteropolysaccharide prepared from Tremella (Tremella fuciformis (berk.) berk)Tremella fuciformisBerk) sporophore, its main chain structure is composed of α - (1 → 3) glucoside-linked mannan, and its branched chain is composed of glucuronic acid and xylose, the tremella polysaccharide has the functions of regulating immunity, resisting thrombosis, prolonging blood coagulation time, reducing blood sugar and blood fat, inhibiting tumor, delaying senility and resisting radiation.
How to further improve the extraction rate of the tremella polysaccharide is the key to realizing the industrial production of the tremella polysaccharide. At present, the tremella polysaccharide can be extracted by a hot water extraction method, an ultrasonic-assisted extraction method, a microwave-assisted extraction method, an extrusion-assisted extraction method, an enzymatic extraction method, an alkaline extraction method and the like.
The cell wall of the tremella sporophore cell has a compact structure formed by combining double-layer protein and polysaccharide, so that the cell wall structure is difficult to damage by a hot water extraction method, and polysaccharide components of the tremella sporophore are fully released. If auxiliary instruments such as an ultrasonic generator, a microwave reaction system or a pressure cooker are added to extract the tremella polysaccharide, the cost of additional instruments is increased, and the energy consumption is high. Although the enzyme extraction method is mild, problems of increased viscosity of the extract, difficulty in filtration, increased impurities such as protein, and low cost of enzyme reagents are encountered in the extraction process.
Disclosure of Invention
In view of this, the invention aims to provide an extraction method of tremella polysaccharide, which has high extraction rate, low energy consumption and low cost.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a method for extracting tremella polysaccharide from an acidic buffer solution, which comprises the following steps:
1) under the heating condition, leaching the tremella powder by adopting an acidic buffer solution to obtain a leaching solution;
2) carrying out solid-liquid separation on the leaching solution to obtain a liquid component, wherein the liquid component contains tremella polysaccharide;
3) precipitating the liquid components with ethanol, and centrifuging to obtain precipitate as Tremella polysaccharide.
The acidic buffer solution in the step 1) is a citric acid-sodium citrate buffer solution, an acetic acid-sodium acetate buffer solution or a disodium hydrogen phosphate-citric acid buffer solution. The concentration of the citric acid-sodium citrate buffer solution is 0.1mol/L, the pH range is 3.0-6.6, and the citric acid-sodium citrate buffer solution is composed of citric acid and sodium citrate; the concentration of the acetic acid-sodium acetate buffer solution is 0.2mol/L, the pH range is 3.6-5.8, and the acetic acid-sodium acetate buffer solution consists of acetic acid and sodium acetate; the disodium hydrogen phosphate-citric acid buffer solution is prepared from 0.1mol/L citric acid solution and 0.2mol/L disodium hydrogen phosphate solution, and the pH range is 2.2-6.0.
The ratio of the material to the water extracted in the step 1) is 1: 30-1: 70 g/mL.
The heating mode in the step 1) is water bath heating, the water bath temperature is 80-100 ℃, and the water bath time is 2-6 hours.
And 2) performing solid-liquid separation in the step 2) by centrifugation at 7000-9000 rpm for 5-10 min.
The ethanol precipitation in the step 3) is performed by using an ethanol solution, and the volume concentration of the ethanol solution is 85-99%; the volume ratio of the liquid components to the ethanol solution in the alcohol precipitation is 1: 3-5.
The alcohol precipitation time in the step 3) is 8-14 h.
The rotating speed during centrifugation in the step 3) is 8000-11000 rpm, and the centrifugation time is 5-10 min.
The invention has the beneficial effects that: according to the method for extracting the tremella polysaccharide, the acidic buffer solution is used as an extraction solvent to replace the traditional water extraction, and the tremella polysaccharide is obtained by performing alcohol precipitation after extraction. The principle that the solubility of the tremella polysaccharide in an acidic buffer solution is higher than that of the tremella polysaccharide in hot water is utilized. The extraction rate of the tremella polysaccharide by the method provided by the invention can reach 31.57%, the extraction rate of the method provided by the invention is 3-4 times of that of the traditional water extraction method, and the extraction rate of the tremella polysaccharide is greatly improved. In addition, the invention does not need other additional auxiliary instruments or reagents, such as ultrasonic waves or subcritical states, and has low energy consumption and low cost.
Detailed Description
The invention provides a method for extracting tremella polysaccharide from an acidic buffer solution, which comprises the following steps: 1) leaching the tremella powder by adopting an acid buffer solution under a water bath condition to obtain a leaching material liquid; the acidic buffer solution comprises a citric acid-sodium citrate buffer solution, an acetic acid-sodium acetate buffer solution and a disodium hydrogen phosphate-citric acid buffer solution; the temperature of the water bath is 80-100 ℃, the time of the water bath is 2-6 h, and the ratio of materials to water is 1: 30-1: 70 g/mL; 2) carrying out solid-liquid separation on the leaching solution to obtain a liquid component, wherein the liquid component comprises tremella polysaccharide; 3) precipitating the liquid components with ethanol, and centrifuging to obtain precipitate as Tremella polysaccharide.
In the invention, the source of the tremella powder is not limited, and the tremella powder can be obtained by drying commercially available tremella in the sun, crushing and sieving. The sun-drying method is conventional in the field, and particularly, the sun-drying method can be adopted for naturally drying in sunlight. The grinding is preferably carried out by adopting a Chinese herbal medicine grinder, and the screening mesh number is preferably 60-100 meshes, and more preferably 80 meshes; the granularity of the tremella powder is preferably 150-250 μm, and more preferably 200-220 μm.
After the tremella powder is obtained, the tremella powder is mixed with an acidic buffer solution to obtain an extraction feed liquid. The acidic buffer solution is a citric acid-sodium citrate buffer solution, an acetic acid-sodium acetate buffer solution or a disodium hydrogen phosphate-citric acid buffer solution; the concentration of the citric acid-sodium citrate buffer solution is 0.1mol/L, the pH range is 3.0-6.6, and the citric acid-sodium citrate buffer solution is composed of citric acid and sodium citrate; the concentration of the acetic acid-sodium acetate buffer solution is 0.2mol/L, the pH range is 3.6-5.8, and the acetic acid-sodium acetate buffer solution consists of acetic acid and sodium acetate; the disodium hydrogen phosphate-citric acid buffer solution is prepared from 0.1mol/L citric acid solution and 0.2mol/L disodium hydrogen phosphate solution, and the pH range is 2.2-6.0.
The preferred volume ratio of the mass of the tremella powder to the volume of the acidic buffer solution is 1: 30-70 g/mL; more preferably 1: 35-45 g/mL; most preferably 1:40 g/mL.
The invention heats the leaching liquid in water bath. The temperature of the water bath is 80-100 ℃, preferably 90-100 ℃, and more preferably 100 ℃; the water bath time is 2-6 h, preferably 4-5 h; most preferably 5 hours. The water bath is prepared by adopting water bath equipment which is conventional in the field, and has no other special limitation. In particular, a water bath can be adopted in the embodiment of the invention. In the water-bath heating process, the tremella polysaccharide in the tremella powder is extracted by the acidic buffer solution to obtain an extraction liquid, wherein the extraction liquid comprises tremella powder solid residues and tremella polysaccharide dissolved in a system.
The invention carries out solid-liquid separation on the extraction feed liquid and collects liquid components. In the present invention, the solid-liquid separation method is preferably centrifugation; the rotating speed of the centrifugation is preferably 7000-9000 rpm, and more preferably 8000 rpm; the centrifugation time is preferably 5-10 min, and more preferably 8 min. The solid-liquid separation of the invention is to remove the solid residue which is insoluble in the acid buffer solution in the tremella powder, so as to be beneficial to the subsequent alcohol precipitation of the tremella polysaccharide.
After the liquid component is obtained, carrying out alcohol precipitation on the liquid component to obtain alcohol precipitation feed liquid, wherein the alcohol precipitation feed liquid comprises precipitated tremella polysaccharide and an alcohol precipitation liquid phase, and the obtained precipitate is the tremella polysaccharide. In the present invention, the alcohol precipitation reagent is preferably an ethanol solution; the volume concentration of the ethanol solution is preferably 85-99%, and more preferably 95%; the volume ratio of the liquid component to the ethanol solution is preferably 1: 3-5, and more preferably 1: 4.
In the embodiment of the invention, the alcohol precipitation is specifically to mix the liquid component with an ethanol solution, and to perform precipitation by standing. The mixing method of the present invention is not particularly limited as long as mixing can be achieved. The time of alcohol precipitation in the invention is preferably 8-14 h, more preferably 11-13 h, and most preferably 12 h; according to the invention, the hydrogen bonds in the liquid components are destroyed by ethanol, so that the solubility of the tremella polysaccharide in the liquid components is reduced, and the polysaccharide is precipitated.
After the alcohol precipitation material liquid is obtained, the second solid-liquid separation is carried out on the alcohol precipitation material liquid to obtain the tremella polysaccharide. In the present invention, the second solid-liquid separation method is preferably centrifugation; the rotating speed of the centrifugation is preferably 8000-11000 rpm, more preferably 9500-10500 rpm, and most preferably 10000 rpm; the centrifugation time is preferably 5-10 min, and more preferably 8 min. The invention collects solid precipitate after the second solid-liquid separation, namely the extracted tremella polysaccharide.
The formula for calculating the extraction rate of the tremella polysaccharide is as follows:
in the invention, the tremella polysaccharide obtained by the extraction is dissolved in water to obtain the tremella polysaccharide water solution. And (3) measuring the polysaccharide content in the tremella polysaccharide aqueous solution by using a phenol-sulfuric acid method, and calculating the extraction rate of the tremella polysaccharide.
The extraction method of tremella polysaccharide provided by the present invention is described in detail with reference to the following examples, but these should not be construed as limiting the scope of the present invention.
Example 1
Drying commercially available tremella in the sun, grinding and sieving with a 90-mesh sieve. Adding 40ml of citric acid-sodium citrate buffer solution with pH of 3.0 into 1g of tremella powder, placing the tremella powder in hot water at 100 ℃ for water bath extraction, after 5h, centrifuging the extraction liquid at 8000rpm for 8min, removing residues, adding 4 times volume of 90% ethanol solution into the supernatant, precipitating for 12h, centrifuging at 10000rpm for 8min, and removing the supernatant to obtain tremella polysaccharide precipitate.
The control experiment was performed by replacing the citric acid-sodium citrate buffer with water as the extraction solvent, as described above.
Dissolving the obtained tremella polysaccharide precipitate in 20ml of water completely, measuring the polysaccharide content in the solution to be 15.79mg/ml by a phenol-sulfuric acid method, calculating the mass of the polysaccharide in the polysaccharide precipitate to be 315.8mg, wherein the polysaccharide extraction rate is 31.58%, and the extraction rate of the tremella polysaccharide extracted by taking water as an extraction solvent is 7.55% compared with the extraction rate of the tremella polysaccharide extracted by taking water as an extraction solvent, and is improved by 318.3%.
Example 2
Drying commercially available Tremella fuciformis in the sun, pulverizing, and sieving with 80 mesh sieve. Adding 40ml of citric acid-sodium citrate buffer solution with pH of 3.0 into 1g of tremella powder, placing the tremella powder in hot water at 100 ℃ for water bath extraction, centrifuging the extraction liquid at 7500rpm for 5min after 4h, removing residues, adding 4.5 times volume of 95% ethanol solution into the supernatant, precipitating for 11h, centrifuging at 11000rpm for 6min, and removing the supernatant to obtain tremella polysaccharide precipitate.
The control experiment was performed by replacing the citric acid-sodium citrate buffer with water as the extraction solvent, as described above.
Dissolving the obtained tremella polysaccharide precipitate in 20ml of water completely, measuring the polysaccharide content in the solution to be 14.35mg/ml by a phenol-sulfuric acid method, calculating the mass of the polysaccharide in the polysaccharide precipitate to be 287.0mg, wherein the polysaccharide extraction rate is 28.70%, and the extraction rate of the tremella polysaccharide extracted by taking water as an extraction solvent is 7.35% compared with the extraction rate of the tremella polysaccharide extracted by taking water as an extraction solvent, and is improved by 290.5%.
Example 3
Drying commercially available Tremella fuciformis in the sun, pulverizing, and sieving with 80 mesh sieve. Adding 60ml of citric acid-sodium citrate buffer solution with pH of 3.0 into 1g of tremella powder, placing the tremella powder in hot water at 80 ℃ for water bath extraction, centrifuging the extraction liquid at 9000rpm for 7min after 5h, removing residues, adding four times of ethanol solution with volume concentration of 90% into the supernatant, precipitating for 10h, centrifuging at 10000rpm for 10min, and removing the supernatant to obtain tremella polysaccharide precipitate.
The control experiment was performed by replacing the citric acid-sodium citrate buffer with water as the extraction solvent, as described above.
Dissolving the obtained tremella polysaccharide precipitate in 20ml of water completely, measuring the polysaccharide content in the solution to be 5.10mg/ml by a phenol-sulfuric acid method, calculating the mass of the polysaccharide in the polysaccharide precipitate to be 102.0mg, wherein the polysaccharide extraction rate is 10.20%, and the extraction rate of the tremella polysaccharide extracted by taking water as an extraction solvent is 7.25% compared with the extraction rate of the tremella polysaccharide extracted by taking water as an extraction solvent, and is increased by 40.69%.
Example 4
Drying commercially available Tremella fuciformis in the sun, pulverizing, and sieving with 70 mesh sieve. Adding 30ml of citric acid-sodium citrate buffer solution with pH of 3.0 into 1g of tremella powder, placing the tremella powder in hot water at 90 ℃ for water bath extraction, centrifuging the extraction liquid at 7000rpm for 7min after 4h, removing residues, adding four times volume of 90% ethanol solution into the supernatant, precipitating for 9h, centrifuging at 11000rpm for 5min, and removing the supernatant to obtain tremella polysaccharide precipitate.
The control experiment was performed by replacing the citric acid-sodium citrate buffer with water as the extraction solvent, as described above.
Dissolving the obtained tremella polysaccharide precipitate in 20ml of water, measuring the polysaccharide content in the solution to be 8.16mg/ml by a phenol-sulfuric acid method, calculating the mass of the polysaccharide in the polysaccharide precipitate to be 162.2mg, wherein the polysaccharide extraction rate is 16.32%, and the extraction rate of the tremella polysaccharide extracted by taking contrast water as an extraction solvent is 6.22%, which is improved by 162.4%.
Example 5
Drying commercially available Tremella fuciformis in the sun, pulverizing, and sieving with 60 mesh sieve. Adding 60ml of citric acid-sodium citrate buffer solution with pH of 3.0 into 1g of tremella powder, placing the tremella powder in hot water at 100 ℃ for water bath extraction, after 3h, centrifuging the extraction liquid at 8000rpm for 6min, removing residues, adding ethanol solution with volume concentration of 99% into the supernatant, precipitating for 10h, centrifuging at 9000rpm for 9min, and removing the supernatant to obtain tremella polysaccharide precipitate.
The control experiment was performed by replacing the citric acid-sodium citrate buffer with water as the extraction solvent, as described above.
The obtained tremella polysaccharide precipitate is completely dissolved in 20ml of water, the content of the polysaccharide in the solution is measured to be 13.34mg/ml by a phenol-sulfuric acid method, the mass of the polysaccharide in the polysaccharide precipitate is calculated to be 266.8mg, the extraction rate of the polysaccharide is calculated to be 26.68%, and the extraction rate of the tremella polysaccharide by taking contrast water as an extraction solvent is calculated to be 5.62%, which is improved by 374.7%.
Example 6
Drying commercially available Tremella fuciformis in the sun, pulverizing, and sieving with 70 mesh sieve. Adding 50ml of citric acid-sodium citrate buffer solution with pH of 3.0 into 1g of tremella powder, placing the tremella powder in hot water at 90 ℃ for water bath extraction, after 6h, centrifuging the extraction liquid at 9000rpm for 8min, removing residues, adding four times volume of 95% ethanol solution into the supernatant, precipitating for 8h, centrifuging at 10000rpm for 10min, and removing the supernatant to obtain tremella polysaccharide precipitate.
The control experiment was performed by replacing the citric acid-sodium citrate buffer with water as the extraction solvent, as described above.
The obtained tremella polysaccharide precipitate is completely dissolved in 20ml of water, the content of the polysaccharide in the solution is measured to be 12.88mg/ml by a phenol-sulfuric acid method, the mass of the polysaccharide in the polysaccharide precipitate is calculated to be 257.6mg, the extraction rate of the polysaccharide is calculated to be 25.76%, and the extraction rate of the tremella polysaccharide extracted by taking contrast water as an extraction solvent is calculated to be 7.95%, which is improved by 224.0%.
Example 7
Drying commercially available Tremella fuciformis in the sun, pulverizing, and sieving with 80 mesh sieve. Adding 50ml of citric acid-sodium citrate buffer solution with pH of 3.0 into 1g of tremella powder, placing the tremella powder in hot water at 90 ℃ for water bath leaching, centrifuging the leaching material liquid at 7000rpm for 5min after 2h, removing residues, adding 3 times volume of 99% ethanol solution into the supernatant, precipitating for 12h, centrifuging at 11000rpm for 7min, and removing the supernatant to obtain tremella polysaccharide precipitate.
The control experiment was performed by replacing the citric acid-sodium citrate buffer with water as the extraction solvent, as described above.
And completely dissolving the obtained tremella polysaccharide precipitate in 20ml of water, measuring the polysaccharide content in the solution to be 4.72mg/ml by a phenol-sulfuric acid method, calculating the mass of the polysaccharide in the polysaccharide precipitate to be 94.4mg, wherein the polysaccharide extraction rate is 9.44%, and the extraction rate of the tremella polysaccharide extracted by taking the reference water as an extraction solvent is 3.67%, which is improved by 157.2%.
Example 8
Drying commercially available Tremella fuciformis in the sun, pulverizing, and sieving with 100 mesh sieve. Adding 70ml of citric acid-sodium citrate buffer solution with pH of 3.0 into 1g of tremella powder, placing the tremella powder in hot water at 90 ℃ for water bath extraction, centrifuging the extraction liquid for 7min at 8000rpm after 4h, removing residues, adding ethanol solution with volume concentration of 99% 4 times of the volume of the supernatant, precipitating for 8h, centrifuging for 7min at 11000rpm, removing the supernatant, and obtaining tremella polysaccharide precipitate.
The control experiment was performed by replacing the citric acid-sodium citrate buffer with water as the extraction solvent, as described above.
The obtained tremella polysaccharide precipitate is completely dissolved in 20ml of water, the content of the polysaccharide in the solution is measured to be 12.19mg/ml by a phenol-sulfuric acid method, the mass of the polysaccharide in the polysaccharide precipitate is calculated to be 243.8mg, the extraction rate of the polysaccharide is calculated to be 24.38%, the extraction rate of the tremella polysaccharide by taking the contrast water as an extraction solvent is calculated to be 7.32%, and the extraction rate is improved by 233.1%.
Example 9
Drying commercially available Tremella fuciformis in the sun, pulverizing, and sieving with 80 mesh sieve. Adding 50ml of disodium hydrogen phosphate-citric acid buffer solution with the pH value of 2.2 into 1g of tremella powder, putting the tremella powder into hot water with the temperature of 90 ℃ for water bath extraction, centrifuging the extraction liquid for 8min at the rotating speed of 9000rpm after 4h, removing residues, adding 5 times volume of 90% ethanol solution into the supernatant, precipitating for 14h, centrifuging for 5min at the rotating speed of 11000rpm, and removing the supernatant to obtain tremella polysaccharide precipitate.
The control experiment was performed by replacing the disodium hydrogen phosphate-citric acid buffer with water as the extraction solvent, as described above.
Dissolving the obtained tremella polysaccharide precipitate in 20ml of water, measuring the polysaccharide content in the solution to be 12.45mg/ml by a phenol-sulfuric acid method, calculating the mass of the polysaccharide in the polysaccharide precipitate to be 249.0mg, wherein the polysaccharide extraction rate is 24.90%, and the extraction rate of the tremella polysaccharide extracted by taking the reference water as the extraction solvent is 7.31%, which is improved by 240.6%.
Example 10
Drying commercially available tremella in the sun, grinding and sieving with a 90-mesh sieve. Adding 50ml of acetic acid-sodium acetate buffer solution with pH of 3.6 into 1g of tremella powder, placing the tremella powder in hot water at 90 ℃ for water bath extraction, centrifuging the extraction liquid at 7000rpm for 10min after 4h, removing residues, adding ethanol solution with volume concentration of 95% in 4 times of volume into the supernatant, precipitating for 10h, centrifuging at 10000rpm for 10min, removing the supernatant, and obtaining tremella polysaccharide precipitate.
The control experiment was performed by replacing the acetic acid-sodium acetate buffer with water as the extraction solvent, as described above.
The obtained tremella polysaccharide precipitate is completely dissolved in 20ml of water, the content of the polysaccharide in the solution is measured to be 6.13mg/ml by a phenol-sulfuric acid method, the mass of the polysaccharide in the polysaccharide precipitate is calculated to be 122.6mg, the extraction rate of the polysaccharide is calculated to be 12.26%, the extraction rate of the tremella polysaccharide by taking the contrast water as an extraction solvent is calculated to be 7.31%, and the extraction rate is improved by 67.72%.
Example 11
Drying commercially available Tremella fuciformis in the sun, pulverizing, and sieving with 80 mesh sieve. Adding 50ml of acetic acid-sodium acetate buffer solution with pH of 5.8 into 1g of tremella powder, placing the tremella powder in hot water at 90 ℃ for water bath extraction, after 4h, centrifuging the extraction liquid at 8000rpm for 8min, removing residues, adding 3 times volume of 95% ethanol solution into the supernatant, precipitating for 12h, centrifuging at 10000rpm for 8min, and removing the supernatant to obtain tremella polysaccharide precipitate.
The control experiment was performed by replacing the acetic acid-sodium acetate buffer with water as the extraction solvent, as described above.
The obtained tremella polysaccharide precipitate is completely dissolved in 20ml of water, the content of the polysaccharide in the solution is 9.20mg/ml through a phenol-sulfuric acid method, the mass of the polysaccharide in the polysaccharide precipitate is calculated to be 184.0mg, the extraction rate of the polysaccharide is calculated to be 18.40%, the extraction rate of the tremella polysaccharide by taking the contrast water as an extraction solvent is calculated to be 7.31%, and the extraction rate is improved by 151.7%.
Example 12
Drying commercially available Tremella fuciformis in the sun, pulverizing, and sieving with 70 mesh sieve. Adding 50ml of citric acid-sodium citrate buffer solution with pH of 5.0 into 1g of tremella powder, placing the tremella powder in hot water at 90 ℃ for water bath extraction, centrifuging the extraction liquid for 5min at 8000rpm after 4h, removing residues, adding ethanol solution with volume concentration of 95% in four times of volume into the supernatant, precipitating for 12h, centrifuging for 5min at 10000rpm, and removing the supernatant to obtain tremella polysaccharide precipitate.
The control experiment was performed by replacing the citric acid-sodium citrate buffer with water as the extraction solvent, as described above.
The obtained tremella polysaccharide precipitate is completely dissolved in 20ml of water, the content of the polysaccharide in the solution is measured to be 6.98mg/ml by a phenol-sulfuric acid method, the mass of the polysaccharide in the polysaccharide precipitate is calculated to be 139.6mg, the extraction rate of the polysaccharide is calculated to be 13.96%, and the extraction rate of the tremella polysaccharide by taking contrast water as an extraction solvent is calculated to be 7.31%, which is improved by 90.97%.
Example 13
Drying commercially available tremella in the sun, grinding and sieving with a 90-mesh sieve. Adding 50ml of citric acid-sodium citrate buffer solution with pH of 6.0 into 1g of tremella powder, placing the tremella powder in hot water at 90 ℃ for water bath extraction, centrifuging the extraction liquid at 8500rpm for 6min after 4h, removing residues, adding ethanol solution with volume concentration of 99% in a volume which is three times that of the tremella powder into the supernatant, precipitating for 10h, centrifuging at 11000rpm for 8min, and removing the supernatant to obtain tremella polysaccharide precipitate.
The control experiment was performed by replacing the citric acid-sodium citrate buffer with water as the extraction solvent, as described above.
Dissolving the obtained tremella polysaccharide precipitate in 20ml of water, measuring the polysaccharide content in the solution to be 8.50mg/ml by a phenol-sulfuric acid method, calculating the mass of the polysaccharide in the polysaccharide precipitate to be 170.0mg, wherein the polysaccharide extraction rate is 17.0%, and the extraction rate of the tremella polysaccharide extracted by taking the reference water as an extraction solvent is 7.31%, which is improved by 132.6%.
Example 14
Drying commercially available tremella in the sun, grinding and sieving with a 90-mesh sieve. Adding 50ml of disodium hydrogen phosphate-citric acid buffer solution with the pH value of 3.0 into 1g of tremella powder, putting the tremella powder into hot water at the temperature of 90 ℃ for water bath leaching, centrifuging the leaching liquid for 6min at the rotating speed of 8500rpm after 4h, removing residues, adding ethanol solution with the volume concentration of 99% in a volume which is three times that of the tremella powder into the supernate, precipitating for 10h, centrifuging for 8min at the rotating speed of 11000rpm, and removing the supernate to obtain tremella polysaccharide precipitate.
The control experiment was performed by replacing the disodium hydrogen phosphate-citric acid buffer with water as the extraction solvent, as described above.
Dissolving the obtained tremella polysaccharide precipitate in 20ml of water, measuring the polysaccharide content in the solution to be 12.27mg/ml by a phenol-sulfuric acid method, calculating the mass of the polysaccharide in the polysaccharide precipitate to be 245.3 mg, wherein the polysaccharide extraction rate is 24.53%, and the extraction rate of the tremella polysaccharide extracted by taking the reference water as the extraction solvent is 7.31%, which is improved by 235.6%.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Claims (1)
1. The method for extracting tremella polysaccharide from the acidic buffer solution is characterized by comprising the following steps of:
1) under the heating condition, leaching the tremella powder by adopting an acidic buffer solution to obtain a leaching solution;
2) carrying out solid-liquid separation on the leaching solution to obtain a liquid component, wherein the liquid component contains tremella polysaccharide;
3) precipitating the liquid components with ethanol, and centrifuging to obtain precipitate as Tremella polysaccharide;
the acidic buffer solution in the step 1) is a citric acid-sodium citrate buffer solution, the concentration of the citric acid-sodium citrate buffer solution is 0.1mol/L, and the pH value is 3.0-6.6;
the ratio of the material to water extracted in the step 1) is 1: 30-1: 70 g/mL;
the heating mode in the step 1) is water bath heating, the water bath temperature is 90-100 ℃, and the water bath time is 2-6 hours;
the solid-liquid separation mode in the step 2) is centrifugation, the rotation speed of the centrifugation is 7000-9000 rpm, and the centrifugation time is 5-10 min;
the ethanol precipitation in the step 3) is performed by using an ethanol solution, and the volume concentration of the ethanol solution is 85-99%; the volume ratio of the liquid components to the ethanol solution during alcohol precipitation is 1: 3-1: 5;
the alcohol precipitation time in the step 3) is 8-14 h;
the rotating speed during centrifugation in the step 3) is 8000-11000 rpm, and the centrifugation time is 5-10 min.
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| CN102219932B (en) * | 2011-05-06 | 2013-06-12 | 齐齐哈尔大学 | Preparation method of edible soup bases packaging film containing tremellan |
| CN102796204A (en) * | 2012-08-27 | 2012-11-28 | 福建中烟工业有限责任公司 | Extraction method of tremella polysaccharide and application of tremella polysaccharide in tobaccos |
| CN103923222B (en) * | 2014-04-29 | 2016-04-13 | 中华全国供销合作总社昆明食用菌研究所 | A kind of low viscosity golden fungus polysaccharides extraction and separation method |
| CN105685961A (en) * | 2016-02-18 | 2016-06-22 | 重庆医药高等专科学校 | Method of preparing instant Tremella powder through microwave-assisted compound enzyme process |
| CN106119229A (en) * | 2016-08-25 | 2016-11-16 | 重庆市中药研究院 | The method separating cellulase from waste tremella cultivating material |
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