Method for extracting polysaccharide from brandy distillation residual liquid

Abstract

The invention relates to the technical field of polysaccharide extraction, in particular to a method for extracting polysaccharide from brandy distillation raffinate, which comprises the following steps: mixing the brandy distillation raffinate with a precipitant, and performing precipitation treatment to obtain brandy distillation raffinate precipitate; wherein the precipitator is selected from glacial acetic acid; washing the brandy distillation raffinate precipitate with ethanol, methanol, a methanol / chloroform mixed solution, chloroform and acetone in sequence to obtain a washed precipitate; adding a Sevage reagent into the washed precipitate for deproteinization treatment to obtain a deproteinized crude polysaccharide solution; and dialyzing the deproteinized crude polysaccharide solution to obtain brandy distillation residual liquid polysaccharide. According to the method, the byproduct utilization rate of the brandy distillation residual liquid is increased, and the production cost is reduced, so that effective utilization of resources and environmental protection are realized.

Description

Technical Field

[0001] The invention relates to the technical field of polysaccharide extraction, and in particular to a method for extracting polysaccharide from brandy distillation residue. Background Art

[0002] A large amount of distillation residue is produced during the production of brandy. Although it is often regarded as a by-product of production, its potential application value cannot be ignored. These brandy distillation residues are rich in polyphenols, reducing sugars and flavonoids. These ingredients not only give brandy a unique nutritional composition and flavor characteristics, but also show broad application prospects in the fields of food, medicine and chemical industry. In the food industry, polysaccharides can be used as natural food additives to enhance the nutritional value and taste of products; in the field of medicine, the biological activity of polysaccharides provides the possibility for their application in drug research and development; and in the field of chemical industry, polysaccharides are ideal raw materials for the preparation of biomaterials such as bio-glue and biofuel. In particular, polysaccharides in brandy distillation residues, such as cellulose, hemicellulose and pectin, have significant nutritional and functional properties. Therefore, extracting and utilizing these polysaccharides can not only help to realize the resource utilization of brandy distillation residues, reduce environmental pollution and resource waste in the production process, but also significantly increase the economic added value of production by-products.

[0003] The production process of brandy produces a large amount of distillation residue, which, if discharged naturally, will seriously pollute the environment and cause waste of resources. However, the distillation residue contains rich nutrients such as polyphenols, reducing sugars and flavonoids, so the recycling of the distillation residue has become a key technical issue. Summary of the invention

[0004] In view of the shortcomings of the prior art, the present invention provides a method for extracting polysaccharides from brandy distillation residues. The present invention selectively precipitates polysaccharides using glacial acetic acid as a precipitant, and then sequentially washes with 96% ethanol, methanol, a methanol / chloroform mixture, chloroform and acetone to improve the purity and obtain a washed precipitate, and then performs a dialysis treatment after protein removal with a Sevage reagent to finally obtain brandy distillation residue polysaccharides. The method for extracting polysaccharides from brandy distillation residues not only improves the utilization rate of by-products of brandy distillation residues, but also reduces production costs, thereby achieving effective utilization of resources and environmental protection.

[0005] In order to achieve the above object, the present invention provides a method for extracting polysaccharides from brandy distillation residue, comprising the following steps:

[0006] S1. Mixing the brandy distillation residue with a precipitant and performing a precipitation treatment to obtain a brandy distillation residue precipitate; wherein the precipitant is selected from glacial acetic acid.

[0007] S2. Wash the brandy distillation residue precipitate with ethanol, methanol, methanol / chloroform mixed solution, chloroform and acetone in sequence to obtain a washed precipitate.

[0008] S3. Add Sevage reagent to the washed precipitate for deproteinization to obtain a deproteinized crude polysaccharide solution.

[0009] S4, dialyzing the deproteinized crude polysaccharide solution to obtain brandy distillation residue polysaccharide.

[0010] Preferably, the precipitant is glacial acetic acid, and the volume ratio of the brandy distillation residue to the glacial acetic acid is 1:4-6; wherein the volume fraction of the glacial acetic acid is 96%, and it contains 0.3% hydrochloric acid, and the pH of the glacial acetic acid is adjusted to 0.20 with hydrochloric acid.

[0011] Preferably, the specific operation method of the washing treatment is:

[0012] The precipitate of brandy distillation residue is washed with 96% ethanol solution by volume for 2h to 3h, and the obtained solid is ethanol washing, wherein the mass volume ratio of 96% ethanol solution by volume to the precipitate of brandy distillation residue is 2mL to 4mL:1g; the ethanol washing is washed with methanol for 2h to 3h, and the obtained solid is methanol washing; wherein the mass volume ratio of methanol to ethanol washing is 2mL to 4mL:1g; the methanol washing is washed with methanol / chloroform mixed solution for 2h to 3h, and the obtained solid is methanol / chloroform mixed solution washing, The volume ratio of methanol to chloroform in the methanol / chloroform mixture is 4:6; wherein the mass volume ratio of the methanol / chloroform mixture to the methanol washing is 2mL-4mL:1g; the methanol / chloroform mixture washing is washed with chloroform for 2h-3h, and the obtained solid is a chloroform washing; wherein the mass volume ratio of chloroform to the methanol / chloroform mixture washing is 2mL-4mL:1g; the chloroform washing is washed with acetone for 2h-3h, and the obtained solid is an acetone washing, wherein the mass volume ratio of acetone to chloroform washing is 2mL-3mL:1g.

[0013] Preferably, the mass volume ratio of the washed precipitate to the Sevage reagent is 5 g:1 mL to 2 mL.

[0014] Preferably, a rinsing treatment is performed before the deproteinization treatment, and the operation of the rinsing treatment is: ultrapure water is added to the washed precipitate, and then freeze-dried to obtain crude polysaccharides from brandy distillation residue; the purpose of adding ultrapure water is to dissolve and rinse the washed precipitate.

[0015] Preferably, the dialysis treatment tool is a dialysis bag, and the dialysis treatment conditions are: dialysis treatment at a molecular weight cutoff of 3000Da to 3500Da for 48h to 72h.

[0016] Preferably, the precipitation treatment is performed under the conditions of: standing at 4°C to 10°C for 20h to 24h.

[0017] Preferably, during the rinsing process, the mass volume ratio of the washed precipitate to ultrapure water is 1 g: 1 mL to 5 mL.

[0018] Preferably, the specific operation of the deproteinization treatment is:

[0019] Sevage reagent is added to the washed precipitate several times, and the supernatant is shaken and centrifuged until no protein layer appears, and then concentrated to obtain a deproteinized crude polysaccharide solution; wherein, the shaking treatment conditions are: shaking at an shaking speed of 30rpm / min to 35rpm / min for 1h to 2h; the centrifugal treatment conditions are: centrifuging at 5000r / min to 6000r / min for 10min.

[0020] Preferably, during the rinsing process, the freeze-drying conditions are: first freezing at -20°C to -80°C for 16h to 18h, and then freezing at -6°C to 10°C for 48h to 72h.

[0021] Preferably, the brandy distillation residue is also concentrated before use, and the operation of the concentration treatment is: concentrating the brandy distillation residue to 1 / 5 to 1 / 3 of the original volume at 45° C. to 55° C.

[0022] Compared with the prior art, the present invention has the following beneficial effects:

[0023] 1. The present invention provides a method for extracting polysaccharides from brandy distillation residue, comprising the steps of mixing the brandy distillation residue with a precipitant and performing a precipitation treatment to obtain a brandy distillation residue precipitate; wherein the precipitant is selected from glacial acetic acid; then, the brandy distillation residue precipitate is washed with ethanol, methanol, a methanol / chloroform mixed solution, chloroform and acetone in sequence to obtain a washed precipitate; then, Sevage reagent is added to the washed precipitate to perform a deproteinization treatment to obtain a deproteinized crude polysaccharide solution; and finally, the deproteinized crude polysaccharide solution is dialyzed to obtain brandy distillation residue polysaccharide.

[0024] Among them, glacial acetic acid is used as a precipitant for polysaccharide extraction, which has high selectivity for brandy distillation residue polysaccharides and usually does not significantly interact with other components such as proteins, which helps to improve the purity of brandy distillation residue polysaccharides. The method is simple to operate, efficient and environmentally friendly.

[0025] The precipitate of brandy distillation residue was washed with ethanol, methanol, methanol / chloroform mixture, chloroform and acetone in sequence. Among them, the precipitate of brandy distillation residue was washed with ethanol solution with a volume fraction of 96%, which can effectively remove the residual glacial acetic acid and eliminate the impurities in the brandy distillation residue dissolved by glacial acetic acid and the unprecipitated substances in the brandy distillation residue, thereby reducing the contamination of the brandy distillation residue precipitate by glacial acetic acid and improving the purity of the brandy distillation residue precipitate. Since some polysaccharides will precipitate in glacial acetic acid to form solid structures, such as dextran and pectin, and the ethanol solution with a volume fraction of 96% can partially dissolve these solid structures, it is helpful to wash the precipitate of brandy distillation residue more thoroughly; using methanol to wash the ethanol washings can effectively remove the ethanol solution with a volume fraction of 96%, and promote the drying of the brandy distillation residue; using methanol / chloroform mixture to wash the methanol washings can effectively dissolve a variety of compounds and biomolecules in the methanol washings, including lipids, proteins and some polysaccharides; using chloroform to wash the methanol / chloroform mixture washings can effectively remove the methanol The attachments on the surface of the alcohol / chloroform mixed liquid washings and the impurities dissolved therein help to improve the purity of the polysaccharides in the brandy distillation residue, while showing less interference with the structure of polysaccharides and proteins, which is beneficial to maintaining the original integrity and functionality of the polysaccharides and proteins; acetone is a polar organic solvent. Using acetone to clean the chloroform washings can not only effectively dissolve water-soluble compounds (such as ethanol), some organic matter (ketones, aldehydes and alcohols) and lipids, but also remove attachments on the surface and inside of the chloroform washings, such as inorganic salts, monosaccharides and oligosaccharides, thereby further improving the purity of the precipitate of the brandy distillation residue.

[0026] Sevage reagent can effectively precipitate and remove proteins from the crude polysaccharide solution of brandy distillation residue. It has high selectivity for proteins, simple operation, mild conditions, and will not damage the structure of polysaccharides, thereby ensuring the biological activity and functional integrity of polysaccharides.

[0027] 2. The present invention freeze-dries the mixture of the washed precipitate and ultrapure water, thereby not only efficiently removing 95% to 99% of the water without damaging the original structure and biological activity of the washed precipitate, but also effectively avoiding the risk of degradation of heat-sensitive components caused by traditional heat treatment. In addition, the polysaccharide of the brandy distillation residue obtained by the present invention is loose and porous, in a sponge-like state, and can be redissolved by adding water, which is easy to dissolve and use.

[0028] 3. The present invention provides a method for extracting polysaccharides from brandy residue, which is not only simple to operate, low in production cost and high in yield, but also has high purity of polysaccharides in the brandy distillation residue. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1This is a diagram of the precipitate after acetone washing and centrifugation in Example 1 of the present invention.

[0030] Figure 2 This is a diagram of crude polysaccharides from brandy distillation residue according to Example 1 of the present invention.

[0031] Figure 3 This is a diagram of the deproteinized crude polysaccharide solution of Example 1 of the present invention.

[0032] Figure 4 This is a diagram of the dialysis bag of Example 1 of the present invention dialyzed in ultrapure water for 72 hours.

[0033] Figure 5 This is a picture of the polysaccharide sample of brandy distillation residue obtained by freeze drying in Example 1 of the present invention.

[0034] Figure 6 This is the infrared spectrum of polysaccharide in brandy distillation residue in Example 1 of the present invention. DETAILED DESCRIPTION

[0035] The technical solution of the present invention will be clearly and completely described below in combination with the data in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0036] It should be noted that the professional terms used in the present invention are only for the purpose of describing specific embodiments and are not intended to limit the scope of protection of the present invention. Unless otherwise specified, the various raw materials, reagents, instruments and equipment used in the following embodiments of the present invention can be purchased from the market or prepared by existing methods. Among them, the Sevage reagent used in the present invention is a mixture of chloroform and n-butanol, and the volume ratio of chloroform to n-butanol is 4:1; the brandy distillation residue is provided by Nongken Nuanquan Winery.

[0037] When producing brandy using traditional technology, in order to obtain a higher alcohol content, alcohol is usually separated from these polysaccharides during the distillation process, causing these polysaccharides to dissolve in the distillation residue, resulting in a waste of raw materials.

[0038] In view of the problems existing in the prior art, the present invention provides a method for extracting polysaccharides from brandy distillation residue, comprising the following steps: mixing the brandy distillation residue with a precipitant and performing a precipitation treatment to obtain a brandy distillation residue precipitate; wherein the precipitant is selected from glacial acetic acid; washing the brandy distillation residue precipitate with ethanol, methanol, a methanol / chloroform mixed solution, chloroform and acetone in sequence to obtain a washed precipitate; adding a Sevage reagent to the washed precipitate to perform a deproteinization treatment to obtain a deproteinized crude polysaccharide solution; and dialyzing the deproteinized crude polysaccharide solution to obtain brandy distillation residue polysaccharide.

[0039] The present invention utilizes the dissolution characteristics of different solvents to remove impurities, retain and purify polysaccharide components. First, glacial acetic acid is used as a precipitant to selectively precipitate polysaccharides to obtain brandy distillation residue precipitates; then ethanol, methanol, methanol / chloroform mixed solution, chloroform, and acetone are used to wash in sequence to improve the purity, and then Sevage reagent is added to remove protein and then dialyzed to finally obtain high-purity brandy distillation residue polysaccharides. The present invention not only improves the utilization rate of by-products of brandy distillation residues, but also reduces production costs, thereby achieving effective resource utilization and environmental protection.

[0040] In order to enable those skilled in the art to more clearly understand the technical solution of the present invention, the technical solution of the present invention will be described in detail below in conjunction with specific embodiments.

[0041] Example 1

[0042] A method for extracting polysaccharides from brandy distillation residue comprises the following steps:

[0043] S1. Take 500 mL of brandy distillation residue and concentrate it under reduced pressure on a rotary evaporator at 40° C. to 1 / 5 of the original volume to obtain brandy distillation residue; at 4° C., precipitate the brandy distillation residue with 96% by volume glacial acetic acid for 20 hours, and then filter the brandy distillation residue with a centrifuge to obtain a precipitate of the brandy distillation residue, wherein the volume ratio of the brandy distillation residue to the 96% by volume glacial acetic acid is 1:4.

[0044] S2. First, use a 96% volume fraction ethanol solution to clean the brandy distillation residual liquid precipitate for 2 hours, and after centrifugation, the obtained solid is an ethanol wash, wherein the mass volume ratio of the 96% volume fraction ethanol solution to the brandy distillation residual liquid precipitate is 3mL:1g; then use methanol to wash the ethanol wash for 2 hours, and after centrifugation, the obtained solid is a methanol wash; wherein the mass volume ratio of methanol to ethanol wash is 3mL:1g; then use a methanol / chloroform mixed solution to wash the methanol wash for 2 hours, and the obtained solid is a methanol / chloroform mixed solution wash, wherein the volume ratio of methanol to chloroform in the methanol / chloroform mixed solution is 4:6, and the mass volume ratio of the methanol / chloroform mixed solution to the methanol wash is 3mL:1g; then use chloroform to wash the methanol / chloroform mixed solution wash for 2 hours, and the obtained solid is a chloroform wash; wherein the mass volume ratio of chloroform to the methanol / chloroform mixed solution wash is 3mL:1g; finally, use acetone to wash the chloroform wash for 2 hours, and the obtained solid is an acetone wash, such as Figure 1 As shown; wherein, the mass volume ratio of acetone to chloroform washings is 2mL:1g.

[0045] S3. Put the acetone washings into a fume hood, open the tube cover, and air dry. Then add the same volume of ultrapure water, keep in a -80°C refrigerator for 16 hours, and then freeze-dry to obtain the brandy distillation residue crude polysaccharide. The brandy distillation residue crude polysaccharide is as follows: Figure 2 shown.

[0046] S4, such as Figure 3 As shown, 2 g of crude polysaccharide from brandy distillation residue is dissolved in 2 ml of ultrapure water to obtain an aqueous solution of crude polysaccharide from brandy distillation residue. Sevage reagent is added to the aqueous solution of crude polysaccharide from brandy distillation residue, and then the solution is placed in a shaker for full shaking. The solution is centrifuged at 5000 r / min for 10 min, and the supernatant is taken. The operation is repeated for more than 5 times until no protein layer appears in the supernatant. The supernatant is then collected and concentrated under reduced pressure at 40° C. to remove the organic reagent in the supernatant, and a deproteinized crude polysaccharide solution is obtained. Figure 5 As shown; wherein, the mass volume of Sevage reagent and crude polysaccharide from brandy distillation residue is 1mL:5g.

[0047] S5, such as Figure 4 As shown, the deproteinized crude polysaccharide solution was dialyzed in ultrapure water for 72 hours using a dialysis bag with a molecular weight cutoff of 3500Da, and then concentrated under reduced pressure and freeze-dried for 72 hours to obtain brandy distillation residue polysaccharide. The brandy distillation residue polysaccharide sample is as shown in Figure 5 shown.

[0048] Example 2

[0049] A method for extracting polysaccharides from brandy distillation residue is the same as the preparation method in Example 1, except that the volume ratio of brandy distillation residue to 96% glacial acetic acid in S1 is replaced from 1:4 to 1:5, and the temperature of precipitation treatment is replaced from 4°C to 10°C, comprising the following steps:

[0050] S1. Take 500 mL of brandy distillation residue and concentrate it under reduced pressure at 40° C. to 1 / 5 of the original volume using a rotary evaporator to obtain brandy distillation residue; at 10° C., precipitate the brandy distillation residue with 96% by volume glacial acetic acid for 20 hours, and then filter the brandy distillation residue using a centrifuge to obtain a precipitate of the brandy distillation residue, wherein the volume ratio of the brandy distillation residue to the 96% by volume glacial acetic acid is 1:5.

[0051] S2. First, use a 96% ethanol solution with a volume fraction to clean the brandy distillation residual liquid precipitate for 2 hours, and after centrifugation, the obtained solid is an ethanol washing, wherein the mass volume ratio of the 96% ethanol solution with a volume fraction to the brandy distillation residual liquid precipitate is 3mL:1g; then use methanol to wash the ethanol washing for 2 hours, and after centrifugation, the obtained solid is a methanol washing; wherein the mass volume ratio of methanol to ethanol washing is 3mL:1g; then use a methanol / chloroform mixture to wash the methanol washing for 2 hours, and the obtained solid is a methanol / chloroform mixture. Chloroform mixed liquid washings, wherein the volume ratio of methanol to chloroform in the methanol / chloroform mixed liquid is 4:6, and the mass volume ratio of the methanol / chloroform mixed liquid to the methanol washings is 3mL:1g; then using chloroform to wash the methanol / chloroform mixed liquid washings for 2h, and the obtained solid is chloroform washings; wherein the mass volume ratio of chloroform to the methanol / chloroform mixed liquid washings is 3mL:1g; finally, using acetone to wash the chloroform washings for 2h, and the obtained solid is acetone washings, wherein the mass volume ratio of acetone to chloroform washings is 2mL:1g.

[0052] S3. Put the acetone washings into a fume hood, open the tube cover, wait until it is air-dried, add the same volume of ultrapure water, keep it in a -80°C refrigerator for 16 hours, and then freeze-dry to obtain the crude polysaccharide of brandy distillation residue.

[0053] S4. Dissolve 1 g of crude polysaccharide from brandy distillation residue in 2 ml of ultrapure water to obtain an aqueous solution of crude polysaccharide from brandy distillation residue, add Sevage reagent to the aqueous solution of crude polysaccharide from brandy distillation residue, put it into a shaker for sufficient shaking, centrifuge it at 5000 r / min for 10 min, take the supernatant, repeat the operation for more than 5 times until no protein layer appears in the supernatant; then collect the supernatant, and concentrate it under reduced pressure at 40°C to remove the organic reagent in the supernatant to obtain a deproteinized crude polysaccharide solution; wherein, the mass volume of Sevage reagent and crude polysaccharide from brandy distillation residue is 1 mL:5 g.

[0054] S5. The deproteinized crude polysaccharide solution was dialyzed in ultrapure water for 72 hours using a dialysis bag with a molecular weight cutoff of 3500 Da, and then concentrated under reduced pressure and freeze-dried for 72 hours to obtain brandy distillation residue polysaccharide.

[0055] Example 3

[0056] A method for extracting polysaccharides from brandy distillation residue is the same as the preparation method in Example 2, except that the volume ratio of brandy distillation residue to 96% glacial acetic acid in S1 is replaced from 1:5 to 1:6, and the precipitation treatment time is replaced from 20h to 24h, comprising the following steps:

[0057] S1. Take 500 mL of brandy distillation residue and concentrate it under reduced pressure on a rotary evaporator at 40° C. to 1 / 5 of the original volume to obtain brandy distillation residue; at 10° C., precipitate the brandy distillation residue with glacial acetic acid with a volume fraction of 96% for 24 hours, and then filter the brandy distillation residue with a centrifuge to obtain a precipitate of the brandy distillation residue, wherein the volume ratio of the brandy distillation residue to the glacial acetic acid with a volume fraction of 96% is 1:4.

[0058] S2. First, use a 96% ethanol solution with a volume fraction to clean the brandy distillation residual liquid precipitate for 2 hours, and after centrifugation, the obtained solid is an ethanol washing, wherein the mass volume ratio of the 96% ethanol solution with a volume fraction to the brandy distillation residual liquid precipitate is 3mL:1g; then use methanol to wash the ethanol washing for 2 hours, and after centrifugation, the obtained solid is a methanol washing; wherein the mass volume ratio of methanol to ethanol washing is 3mL:1g; then use a methanol / chloroform mixture to wash the methanol washing for 2 hours, and the obtained solid is a methanol / chloroform mixture. Chloroform mixed liquid washings, wherein the volume ratio of methanol to chloroform in the methanol / chloroform mixed liquid is 4:6, and the mass volume ratio of the methanol / chloroform mixed liquid to the methanol washings is 3mL:1g; then using chloroform to wash the methanol / chloroform mixed liquid washings for 2h, and the obtained solid is chloroform washings; wherein the mass volume ratio of chloroform to the methanol / chloroform mixed liquid washings is 3mL:1g; finally, using acetone to wash the chloroform washings for 2h, and the obtained solid is acetone washings, wherein the mass volume ratio of acetone to chloroform washings is 2mL:1g.

[0059] S3. Put the acetone washings into a fume hood, open the tube cover, wait until it is air-dried, add the same volume of ultrapure water, keep it in a -80°C refrigerator for 16 hours, and then freeze-dry to obtain the crude polysaccharide of brandy distillation residue.

[0060] S4. Dissolve 1 g of crude polysaccharide from brandy distillation residue in 2 ml of ultrapure water to obtain an aqueous solution of crude polysaccharide from brandy distillation residue, add Sevage reagent to the aqueous solution of crude polysaccharide from brandy distillation residue, put it into a shaker for sufficient shaking, centrifuge it at 5000 r / min for 10 min, take the supernatant, repeat the operation for more than 5 times until no protein layer appears in the supernatant; then collect the supernatant, and concentrate it under reduced pressure at 40°C to remove the organic reagent in the supernatant to obtain a deproteinized crude polysaccharide solution; wherein, the mass volume of Sevage reagent and crude polysaccharide from brandy distillation residue is 1 mL:5 g.

[0061] S5. The deproteinized crude polysaccharide solution was dialyzed in ultrapure water for 72 hours using a dialysis bag with a molecular weight cutoff of 3500 Da, and then concentrated under reduced pressure and freeze-dried for 72 hours to obtain brandy distillation residue polysaccharide.

[0062] Example 4

[0063] A method for extracting polysaccharides from brandy distillation residue is the same as the preparation method of Example 1, except that the throttling molecular weight of the dialysis bag in S5 is replaced from 3500Da to 3000Da, comprising the following steps:

[0064] S1. Take 500 mL of brandy distillation residue and concentrate it under reduced pressure on a rotary evaporator at 40° C. to 1 / 5 of the original volume to obtain brandy distillation residue; at 4° C., precipitate the brandy distillation residue with 96% by volume glacial acetic acid for 20 hours, and then filter the brandy distillation residue with a centrifuge to obtain a precipitate of the brandy distillation residue, wherein the volume ratio of the brandy distillation residue to the 96% by volume glacial acetic acid is 1:4.

[0065] S2. First, use a 96% ethanol solution with a volume fraction to clean the brandy distillation residual liquid precipitate for 2 hours, and after centrifugation, the obtained solid is an ethanol washing, wherein the mass volume ratio of the 96% ethanol solution with a volume fraction to the brandy distillation residual liquid precipitate is 3mL:1g; then use methanol to wash the ethanol washing for 2 hours, and after centrifugation, the obtained solid is a methanol washing; wherein the mass volume ratio of methanol to ethanol washing is 3mL:1g; then use a methanol / chloroform mixture to wash the methanol washing for 2 hours, and the obtained solid is a methanol / chloroform mixture. Chloroform mixed liquid washings, wherein the volume ratio of methanol to chloroform in the methanol / chloroform mixed liquid is 4:6, and the mass volume ratio of the methanol / chloroform mixed liquid to the methanol washings is 3mL:1g; then using chloroform to wash the methanol / chloroform mixed liquid washings for 2h, and the obtained solid is chloroform washings; wherein the mass volume ratio of chloroform to the methanol / chloroform mixed liquid washings is 3mL:1g; finally, using acetone to wash the chloroform washings for 2h, and the obtained solid is acetone washings, wherein the mass volume ratio of acetone to chloroform washings is 2mL:1g.

[0066] S3. Put the acetone washings into a fume hood, open the tube cover, wait until it is air-dried, add the same volume of ultrapure water, keep it in a -80°C refrigerator for 16 hours, and then freeze-dry to obtain the crude polysaccharide of brandy distillation residue.

[0067] S4. Dissolve the crude polysaccharide from the distillation residue of brandy in water to obtain an aqueous solution of the crude polysaccharide from the distillation residue of brandy, add Sevage reagent to the aqueous solution of the crude polysaccharide from the distillation residue of brandy, put it into a shaker for sufficient shaking, centrifuge it at 5000r / min for 10min, take the supernatant, repeat the operation for more than 5 times until no protein layer appears in the supernatant; then collect the supernatant, and concentrate it under reduced pressure at 40°C to remove the organic reagent in the supernatant to obtain a deproteinized crude polysaccharide solution; wherein the mass volume of Sevage reagent and crude polysaccharide from the distillation residue of brandy is 2mL:5g.

[0068] S5. The deproteinized crude polysaccharide solution was dialyzed in ultrapure water for 72 hours using a dialysis bag with a molecular weight cutoff of 3000 Da, and then concentrated under reduced pressure and freeze-dried for 72 hours to obtain brandy distillation residue polysaccharide.

[0069] Example 5

[0070] A method for extracting polysaccharides from brandy distillation residue comprises the following steps:

[0071] S1. Take 500 mL of brandy distillation residue and concentrate it under reduced pressure on a rotary evaporator at 40° C. to 1 / 5 of the original volume to obtain brandy distillation residue; at 4° C., precipitate the brandy distillation residue with 96% by volume glacial acetic acid for 20 hours, and then filter the brandy distillation residue with a centrifuge to obtain a precipitate of the brandy distillation residue, wherein the volume ratio of the brandy distillation residue to the 96% by volume glacial acetic acid is 1:4.

[0072] S2. First, use a 96% ethanol solution by volume to clean the brandy distillation residue precipitate for 2 hours, and after centrifugation, the obtained solid is an ethanol wash, wherein the mass volume ratio of the 96% ethanol solution by volume to the brandy distillation residue precipitate is 2mL:1g; then use methanol to wash the ethanol wash for 2 hours, and after centrifugation, the obtained solid is a methanol wash; wherein the mass volume ratio of methanol to ethanol wash is 2mL:1g; then use a methanol / chloroform mixture to wash the methanol wash for 2 hours, and the obtained solid is a methanol / chloroform mixture. Chloroform mixed liquid washings, wherein the volume ratio of methanol to chloroform in the methanol / chloroform mixed liquid is 4:6, and the mass volume ratio of the methanol / chloroform mixed liquid to the methanol washings is 2mL:1g; then using chloroform to wash the methanol / chloroform mixed liquid washings for 2h, and the obtained solid is chloroform washings; wherein the mass volume ratio of chloroform to the methanol / chloroform mixed liquid washings is 2mL:1g; finally, using acetone to wash the chloroform washings for 2h, and the obtained solid is acetone washings, wherein the mass volume ratio of acetone to chloroform washings is 3mL:1g.

[0073] S3. Put the acetone washings into a fume hood, open the tube cover, wait until it is air-dried, add the same volume of ultrapure water, keep it in a -80°C refrigerator for 16 hours, and then freeze-dry to obtain the crude polysaccharide of brandy distillation residue.

[0074] S4. Dissolve 2 g of crude polysaccharide from brandy distillation residue in 1 ml of ultrapure water to obtain an aqueous solution of crude polysaccharide from brandy distillation residue, add Sevage reagent to the aqueous solution of crude polysaccharide from brandy distillation residue, put it into a shaker for sufficient shaking, centrifuge it at 5000 r / min for 10 min, take the supernatant, repeat the operation for more than 5 times until no protein layer appears in the supernatant; then collect the supernatant, and concentrate it under reduced pressure at 40°C to remove the organic reagent in the supernatant to obtain a deproteinized crude polysaccharide solution; wherein, the mass volume of Sevage reagent and crude polysaccharide from brandy distillation residue is 1 mL:5 g.

[0075] S5. The deproteinized crude polysaccharide solution was dialyzed in ultrapure water for 72 hours using a dialysis bag with a molecular weight cutoff of 3500 Da, and then concentrated under reduced pressure and freeze-dried for 72 hours to obtain brandy distillation residue polysaccharide.

[0076] Example 6

[0077] A method for extracting polysaccharides from brandy distillation residue comprises the following steps:

[0078] S1. Take 500 mL of brandy distillation residue and concentrate it under reduced pressure on a rotary evaporator at 40° C. to 1 / 5 of the original volume to obtain brandy distillation residue; at 4° C., precipitate the brandy distillation residue with 96% by volume glacial acetic acid for 20 hours, and then filter the brandy distillation residue with a centrifuge to obtain a precipitate of the brandy distillation residue, wherein the volume ratio of the brandy distillation residue to the 96% by volume glacial acetic acid is 1:4.

[0079] S2. First, use a 96% ethanol solution by volume to clean the brandy distillation residue precipitate for 3 hours, and after centrifugation, the obtained solid is an ethanol wash, wherein the mass volume ratio of the 96% ethanol solution by volume to the brandy distillation residue precipitate is 4mL:1g; then use methanol to wash the ethanol wash for 3 hours, and after centrifugation, the obtained solid is a methanol wash; wherein the mass volume ratio of methanol to ethanol wash is 4mL:1g; then use a methanol / chloroform mixture to wash the methanol wash for 3 hours, and the obtained solid is a methanol / Chloroform mixed liquid washings, wherein the volume ratio of methanol to chloroform in the methanol / chloroform mixed liquid is 4:6, and the mass volume ratio of the methanol / chloroform mixed liquid to the methanol washings is 4mL:1g; then using chloroform to wash the methanol / chloroform mixed liquid washings for 3h, and the obtained solid is chloroform washings; wherein the mass volume ratio of chloroform to the methanol / chloroform mixed liquid washings is 4mL:1g; finally, using acetone to wash the chloroform washings for 3h, and the obtained solid is acetone washings, wherein the mass volume ratio of acetone to chloroform washings is 3mL:1g.

[0080] S3. Put the acetone washings into a fume hood, open the tube cover, wait until it is air-dried, add the same volume of ultrapure water, keep it in a -80°C refrigerator for 16 hours, and then freeze-dry to obtain the crude polysaccharide of brandy distillation residue.

[0081] S4. Dissolve 1 g of crude polysaccharide from brandy distillation residue in 2 ml of ultrapure water to obtain an aqueous solution of crude polysaccharide from brandy distillation residue, add Sevage reagent to the aqueous solution of crude polysaccharide from brandy distillation residue, put it into a shaker for sufficient shaking, centrifuge it at 5000 r / min for 10 min, take the supernatant, repeat the operation for more than 5 times until no protein layer appears in the supernatant; then collect the supernatant, and concentrate it under reduced pressure at 40°C to remove the organic reagent in the supernatant to obtain a deproteinized crude polysaccharide solution; wherein, the mass volume of Sevage reagent and crude polysaccharide from brandy distillation residue is 1 mL:5 g.

[0082] S5. The deproteinized crude polysaccharide solution was dialyzed in ultrapure water for 72 hours using a dialysis bag with a molecular weight cutoff of 3500 Da, and then concentrated under reduced pressure and freeze-dried for 72 hours to obtain brandy distillation residue polysaccharide.

[0083] Figure 6This is the infrared spectrum of polysaccharide from brandy distillation residue in Example 1 of the present invention. Figure 6 It was concluded that the polysaccharide in the brandy distillation residue showed a series of characteristic absorption peaks in the infrared spectrum. The main absorption peaks include: cellulose at 1025cm -1 The absorption of xyloglucan at 1041 cm -1 Absorption at 1017cm -1 The absorption of amide at 1650cm -1 and 1550cm -1 The absorption of esterified pectin and non-esterified pectin at 1740 cm -1 and 1610cm -1 The absorption at 400 nm indicated that the polysaccharide extracted from the brandy distillation residue in Example 1 of the present invention had a complex chemical structure and diverse biological activities, and these characteristics were clearly reflected in the infrared spectrum.

[0084] Although the preferred embodiments of the present invention have been described, those skilled in the art may make other changes and modifications to these embodiments once they have learned the basic creative concept. Therefore, the appended claims are intended to be interpreted as including the preferred embodiments and all changes and modifications that fall within the scope of the present invention.

[0085] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include these modifications and variations.

Claims

1. A method for extracting polysaccharides from brandy distillation residue, characterized in that: The following steps are involved: The brandy distillation residue is mixed with a precipitant and then subjected to precipitation treatment to obtain a brandy distillation residue precipitate; wherein the precipitant is selected from glacial acetic acid; washing the precipitate of the brandy distillation residue with ethanol, methanol, a methanol / chloroform mixed solution, chloroform and acetone in sequence to obtain a washed precipitate; adding Sevage reagent to the washed precipitate for deproteinization to obtain a deproteinized crude polysaccharide solution; The deproteinized crude polysaccharide solution is dialyzed to obtain brandy distillation residue polysaccharide.

2. The method for extracting polysaccharides from brandy distillation residue according to claim 1, characterized in that: The operation method of the washing treatment is: washing the brandy distillation residue precipitate with a 96% by volume ethanol solution for 2 h to 3 h to obtain an ethanol washing, wherein the mass volume ratio of the 96% by volume ethanol solution to the brandy distillation residue precipitate is 2 mL to 4 mL: 1 g; The ethanol washings are washed with methanol for 2 h to 3 h to obtain methanol washings; wherein the mass volume ratio of methanol to ethanol washings is 2 mL to 4 mL: 1 g; The methanol washings are washed with a methanol / chloroform mixed solution for 2 h to 3 h to obtain a methanol / chloroform mixed solution washing, wherein the volume ratio of methanol to chloroform in the methanol / chloroform mixed solution is 4:6; wherein the mass volume ratio of the methanol / chloroform mixed solution to the methanol washings is 2 mL to 4 mL: 1 g; Wash the methanol / chloroform mixed solution washings with chloroform for 2 h to 3 h to obtain chloroform washings; wherein the mass volume ratio of chloroform to the methanol / chloroform mixed solution washings is 2 mL to 4 mL: 1 g; The chloroform washings are washed with acetone for 2 h to 3 h to obtain acetone washings, wherein the mass volume ratio of acetone to chloroform washings is 2 mL to 3 mL: 1 g.

3. The method for extracting polysaccharides from brandy distillation residue according to claim 1, characterized in that: The mass volume ratio of the washed precipitate to the Sevage reagent is 5 g:1 mL to 2 mL.

4. The method for extracting polysaccharides from brandy distillation residue according to claim 1, characterized in that: The specific operation of the deproteinization treatment is: Sevage reagent is added to the washed precipitate several times, and the solution is shaken and centrifuged until no protein layer appears in the supernatant, and then concentrated to obtain a deproteinized crude polysaccharide solution.

5. The method for extracting polysaccharides from brandy distillation residue according to claim 1, characterized in that: The dialysis treatment conditions are: dialysis treatment at a molecular weight cutoff of 3000Da to 3500Da for 48h to 72h.

6. The method for extracting polysaccharides from brandy distillation residue according to claim 1, characterized in that: The conditions of the precipitation treatment are: standing at 4°C to 10°C for 20h to 24h.

7. The method for extracting polysaccharides from brandy distillation residue according to claim 1, characterized in that: The brandy distillation residue is also concentrated before use.

8. The method for extracting polysaccharides from brandy distillation residue according to claim 1, characterized in that: Before the deproteinization treatment, a rinsing treatment is also carried out. The operation of the rinsing treatment is: ultrapure water is added to the washed precipitate, and then freeze-dried to obtain crude polysaccharide from the brandy distillation residue.

9. The method for extracting polysaccharides from brandy distillation residue according to claim 8, characterized in that: During the rinsing process, the freeze-drying conditions are: first freezing at -20°C to -80°C for 16h to 18h, and then freezing at -6°C to 10°C for 48h to 72h.

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