CN111057161A - Extraction method of tremella polysaccharide - Google Patents
Extraction method of tremella polysaccharide Download PDFInfo
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- CN111057161A CN111057161A CN201811274269.3A CN201811274269A CN111057161A CN 111057161 A CN111057161 A CN 111057161A CN 201811274269 A CN201811274269 A CN 201811274269A CN 111057161 A CN111057161 A CN 111057161A
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- tremella
- tremella polysaccharide
- polysaccharide
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- coarse powder
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0003—General processes for their isolation or fractionation, e.g. purification or extraction from biomass
Abstract
The invention discloses a method for extracting tremella polysaccharide, which comprises the following steps: freeze drying fresh tremella, and pulverizing to obtain coarse powder; carrying out superfine grinding on the tremella coarse powder and a physical auxiliary agent together to prepare micro-cutting auxiliary powder; then the tremella polysaccharide is obtained by ultrasonic-assisted extraction, decolorization, ultrafiltration, alcohol precipitation and vacuum drying. The invention can better retain the biological activity of the tremella polysaccharide by freeze drying, and can effectively improve the extraction rate of the tremella polysaccharide by adopting micro-cutting assisted interaction and ultrasonic assisted extraction of the tremella polysaccharide. The Tremella polysaccharide has wide application in health food and cosmetic field.
Description
Technical Field
The invention belongs to the field of plant polysaccharide extraction, and particularly relates to a method for extracting tremella polysaccharide.
Background
The tremella polysaccharide is the most important component in tremella, mainly comes from tremella sporophore and submerged fermentation liquid of tremella cell, and has complex structure, main chain is composed of mannose connected by α - (1-3) glycosidic bond, 2, 4, 6 positions are connected with residues of glucose, xylose, fucose, general uronic acid and the like to form side chains, wherein α - (1-3) mannan is the active center of the tremella polysaccharide.
At present, the tremella polysaccharide can be extracted by methods such as an ultrasonic-assisted extraction method, a microwave-assisted extraction method, an enzymatic reverse extraction method, acid-base extraction and the like, the extraction rate is obviously improved compared with the traditional extraction method, but the defects of polysaccharide active structure damage, strict extraction condition requirements, complex operation and the like still exist. Therefore, the extraction rate of the tremella polysaccharide needs to be further improved, the higher biological activity of the tremella polysaccharide is reserved, and the industrial production of the tremella polysaccharide is realized. The micro-cutting assisted interaction mode is a new method for efficiently extracting active ingredients of animals and plants in recent years, and the basic principle is as follows: animal and plant raw material particle group is broken through micro shear cell, fresh section is generated on the cell surface, group or intermolecular interaction occurs between the physical auxiliary agent and the effective component, the microscopic performance of the effective component is changed, the surface energy is increased, the specific surface area is increased, the adsorbability and the polarity are increased, the dissolubility of the effective component in the raw material is increased, and then the extraction rate of the effective component is improved. In addition, the freeze-dried product has a complete tissue structure and better nutrient retention due to the direct sublimation of the ice crystals among the cells, and the extract has higher biological activity.
Disclosure of Invention
The invention aims to provide a method for extracting tremella polysaccharide with high extraction rate and high biological activity. The fresh tremella fuciformis is freeze-dried, so that the biological activity of tremella fuciformis polysaccharide can be well kept, and the tremella fuciformis polysaccharide can be extracted by micro-cutting assisted interaction and ultrasonic assistance, so that the extraction rate of the tremella fuciformis polysaccharide can be effectively improved.
The technical scheme of the invention is as follows: a method for extracting tremella polysaccharide comprises the following steps:
(1) cleaning fresh tremella, carrying out vacuum freeze drying at the freezing temperature of-20 to-80 ℃, preferably-45 ℃, crushing the dried tremella, and sieving with a sieve of 80 to 200 meshes to obtain tremella coarse powder;
(2) adding a physical assistant into the tremella coarse powder, and putting the tremella coarse powder into a vibration mill for ultramicro grinding for 10-30 minutes to obtain micro-cutting assistant powder; the addition amount of the physical auxiliary agent is 1-5% of the mass of the tremella coarse powder, and preferably 2%; the milling time is preferably 20 minutes;
(3) the micro-cutting auxiliary powder and water are subjected to ultrasonic-assisted extraction for 1-3 times, preferably 2 times, with the extraction time of 0.5 hour each time under the conditions of 100W of power, 40KHz of frequency and 80-90 ℃ according to the material-liquid ratio of 1: 40-1: 70(W/W), preferably 1: 60(W/W), and then the extract is obtained by filtering.
(4) Decolorizing the extractive solution with active carbon, and ultrafiltering to remove impurities to obtain ultrafiltrate.
(5) Concentrating the ultrafiltrate to 1/3-1/6 of the original volume, preferably 1/5, repeatedly washing and precipitating with ethanol for 3 times, and drying in vacuum to obtain the tremella polysaccharide product.
The physical auxiliary agent in the step (2) is sodium carbonate, sodium bicarbonate, potassium carbonate or potassium bicarbonate.
The aperture of the micropores of the ultrafiltration filter element in the step (4) is 0.1-0.22 μm, preferably 0.22 μm.
The concentration of the ethanol in the step (5) is 75-95%; the volume ratio of the ethanol to the ultrafiltrate is 2: 1-5: 1, and the preferable ratio is 3: 1.
compared with the prior art, the invention has the beneficial effects that: (1) the fresh tremella is processed by freeze drying, so that high-quality and high-efficiency tremella polysaccharide can be obtained; (2) the extraction rate of the tremella polysaccharide can be effectively improved by micro-cutting interaction and ultrasonic-assisted extraction of the tremella polysaccharide; (3) solid alkaline substances such as sodium carbonate or sodium bicarbonate are adopted in the micro-shearing process, the leaching liquor is alkalescent, and the extraction rate of the tremella polysaccharide is further improved. (4) The production process is simple, the selected solvent is water and ethanol, the green and pollution-free effects are achieved, and the industrial production is facilitated.
Detailed Description
Example 1
Freeze drying fresh Tremella at-45 deg.C under vacuum degree of less than 40Pa for 48 hr, pulverizing, and sieving with 80 mesh sieve to obtain Tremella coarse powder;
taking 500g of tremella coarse powder and 10g of sodium bicarbonate, uniformly mixing, placing into a vibration mill, and carrying out superfine grinding for 20 minutes to obtain micro-cutting auxiliary powder;
weighing 50g of micro-cutting auxiliary powder, adding the micro-cutting auxiliary powder into 3L of water for mixing, performing ultrasonic-assisted extraction for 2 times under the conditions of 100W power, 40KHz frequency and 80 ℃ temperature, wherein the extraction time is 0.5 hour each time, and filtering to obtain an extracting solution;
decolorizing the extractive solution with active carbon, and ultrafiltering with 0.22 μm filter core to obtain ultrafiltrate;
concentrating the ultrafiltrate in vacuum to 1/5 of the original volume, adding 3 times of 75% ethanol into the concentrated solution for precipitation, and repeatedly washing the precipitate with 75% ethanol for 3 times; and (3) carrying out vacuum drying on the washed precipitate at the temperature of 40 ℃ to finally obtain the tremella polysaccharide product.
Example 2
Freeze drying fresh Tremella at-45 deg.C under vacuum degree of less than 40Pa for 48 hr, pulverizing, and sieving with 80 mesh sieve to obtain Tremella coarse powder;
taking 500g of tremella coarse powder and 5g of sodium bicarbonate, uniformly mixing, placing into a vibration mill, and carrying out superfine grinding for 20 minutes to obtain micro-cutting auxiliary powder;
weighing 50g of micro-cutting auxiliary powder, adding the micro-cutting auxiliary powder into 3L of water for mixing, performing ultrasonic-assisted extraction for 2 times under the conditions of 100W power, 40KHz frequency and 80 ℃ temperature, wherein the extraction time is 0.5 hour each time, and filtering to obtain an extracting solution;
decolorizing the extractive solution with active carbon, and ultrafiltering with 0.22 μm filter core to obtain ultrafiltrate;
concentrating the ultrafiltrate in vacuum to 1/5 of the original volume, adding 3 times of 75% ethanol into the concentrated solution for precipitation, and repeatedly washing the precipitate with 75% ethanol for 3 times; and (3) carrying out vacuum drying on the washed precipitate at the temperature of 40 ℃ to finally obtain the tremella polysaccharide product.
Example 3
Freeze drying fresh Tremella at-45 deg.C under vacuum degree of less than 40Pa for 48 hr, pulverizing, and sieving with 80 mesh sieve to obtain Tremella coarse powder;
taking 500g of tremella coarse powder and 25g of sodium bicarbonate, uniformly mixing, placing into a vibration mill, and carrying out superfine grinding for 20 minutes to obtain micro-cutting auxiliary powder;
weighing 50g of micro-cutting auxiliary powder, adding the micro-cutting auxiliary powder into 3L of water for mixing, performing ultrasonic-assisted extraction for 2 times under the conditions of 100W power, 40KHz frequency and 80 ℃ temperature, wherein the extraction time is 0.5 hour each time, and filtering to obtain an extracting solution;
decolorizing the extractive solution with active carbon, and ultrafiltering with 0.22 μm filter core to obtain ultrafiltrate;
concentrating the ultrafiltrate in vacuum to 1/5 of the original volume, adding 3 times of 75% ethanol into the concentrated solution for precipitation, and repeatedly washing the precipitate with 75% ethanol for 3 times; and (3) carrying out vacuum drying on the washed precipitate at the temperature of 40 ℃ to finally obtain the tremella polysaccharide product.
Example 4
Freeze drying fresh Tremella at-20 deg.C under vacuum degree of less than 40Pa for 48 hr, pulverizing, and sieving with 80 mesh sieve to obtain Tremella coarse powder;
taking 500g of tremella coarse powder and 10g of sodium bicarbonate, uniformly mixing, placing into a vibration mill, and carrying out superfine grinding for 10 minutes to obtain micro-cutting auxiliary powder;
weighing 50g of micro-cutting auxiliary powder, adding the micro-cutting auxiliary powder into 2L of water for mixing, performing ultrasonic-assisted extraction for 1 time under the conditions of 100W power, 40KHz frequency and 80 ℃ temperature, wherein the extraction time is 0.5 hour each time, and filtering to obtain an extracting solution;
decolorizing the extractive solution with active carbon, and ultrafiltering with 0.1 μm filter core to obtain ultrafiltrate;
concentrating the ultrafiltrate in vacuum to 1/6 of the original volume, adding 2 times of 75% ethanol into the concentrated solution for precipitation, and repeatedly washing the precipitate with 75% ethanol for 3 times; and (3) carrying out vacuum drying on the washed precipitate at the temperature of 40 ℃ to finally obtain the tremella polysaccharide product.
Example 5
Freeze drying fresh Tremella at-80 deg.C under vacuum degree of less than 40Pa for 48 hr, pulverizing, and sieving with 80 mesh sieve to obtain Tremella coarse powder;
taking 500g of tremella coarse powder and 10g of sodium bicarbonate, uniformly mixing, placing into a vibration mill, and carrying out superfine grinding for 30 minutes to obtain micro-cutting auxiliary powder;
weighing 50g of micro-cutting auxiliary powder, adding the micro-cutting auxiliary powder into 3.5L of water for mixing, then carrying out ultrasonic auxiliary extraction for 3 times under the conditions of 100W power, 40KHz frequency and 80 ℃ temperature, wherein the extraction time is 0.5 hour each time, and filtering to obtain an extracting solution;
decolorizing the extractive solution with active carbon, and ultrafiltering with 0.22 μm filter core to obtain ultrafiltrate;
concentrating the ultrafiltrate in vacuum to 1/3 of the original volume, adding 75% ethanol with 5 times volume of the concentrated solution for precipitation, and repeatedly washing the precipitate with 75% ethanol for 3 times; and (3) carrying out vacuum drying on the washed precipitate at the temperature of 40 ℃ to finally obtain the tremella polysaccharide product.
Comparative example 1
Drying fresh Tremella fuciformis at 50 deg.C for 48 hr with traditional hot air, pulverizing, and sieving with 80 mesh sieve to obtain Tremella fuciformis coarse powder;
taking 500g of tremella coarse powder and 10g of sodium bicarbonate, uniformly mixing, placing into a vibration mill, and carrying out superfine grinding for 20 minutes to obtain micro-cutting auxiliary powder;
weighing 50g of micro-cutting auxiliary powder, adding the micro-cutting auxiliary powder into 3L of water for mixing, performing ultrasonic-assisted extraction for 2 times under the conditions of 100W power, 40KHz frequency and 80 ℃ temperature, wherein the extraction time is 0.5 hour each time, and filtering to obtain an extracting solution;
decolorizing the extractive solution with active carbon, and ultrafiltering with 0.22 μm filter core to obtain ultrafiltrate;
concentrating the ultrafiltrate in vacuum to 1/5 of the original volume, adding 3 times of 75% ethanol into the concentrated solution for precipitation, and repeatedly washing the precipitate with 75% ethanol for 3 times; and (3) carrying out vacuum drying on the washed precipitate at the temperature of 40 ℃ to finally obtain the tremella polysaccharide product.
Comparative example 2
Drying fresh Tremella fuciformis at 50 deg.C for 48 hr with traditional hot air, pulverizing, and sieving with 80 mesh sieve to obtain Tremella fuciformis coarse powder;
weighing 50g of tremella coarse powder, adding the tremella coarse powder into 3L of water for mixing, performing ultrasonic-assisted extraction for 2 times under the conditions of 100W power, 40KHz frequency and 80 ℃, wherein the extraction time is 0.5 hour each time, and filtering to obtain an extracting solution;
decolorizing the extractive solution with active carbon, and ultrafiltering with 0.22 μm filter core to obtain ultrafiltrate;
concentrating the ultrafiltrate in vacuum to 1/5 of the original volume, adding 3 times of 75% ethanol into the concentrated solution for precipitation, and repeatedly washing the precipitate with 75% ethanol for 3 times; and (3) carrying out vacuum drying on the washed precipitate at the temperature of 40 ℃ to finally obtain the tremella polysaccharide product.
Effect example 1
1mL of DPPH (10)-4mol/L) and a tremella polysaccharide sample with the mass concentration of 1mg/mL and VC positive control solution are mixed evenly and then the absorbance is measured at the wavelength of 517 nm. The extraction rate and DPPH free radical clearance of tremella polysaccharide by different extraction methods are shown in Table 1.
TABLE 1
| Extraction ratio (%) | DPPH radical scavenging ratio (%) | |
| Example 1 | 49.3 | 82.6 |
| Example 2 | 38.9 | 71.9 |
| Example 3 | 40.4 | 78.5 |
| Example 4 | 41.6 | 72.8 |
| Example 5 | 49.5 | 81.9 |
| Comparative example 1 | 32.5 | 53.2 |
| Comparative example 2 | 28.3 | 46.8 |
Claims (10)
1. The extraction method of the tremella polysaccharide is characterized by comprising the following steps:
(1) cleaning fresh tremella, carrying out vacuum freeze drying at the freezing temperature of-20 to-80 ℃, crushing the dried tremella, and sieving with a sieve of 80 to 200 meshes to obtain tremella coarse powder;
(2) adding a physical additive into the tremella fuciformis coarse powder, and putting the tremella fuciformis coarse powder into a vibration mill for ultramicro grinding for 10-30 minutes to obtain micro-cutting auxiliary powder, wherein the addition amount of the physical additive is 1-5% of the mass of the tremella fuciformis coarse powder;
(3) carrying out ultrasonic-assisted extraction on the micro-cutting auxiliary powder and water according to the material-liquid ratio of 1: 40-1: 70(W/W) for 1-3 times under the conditions of 100W of power, 40KHz of frequency and 80-90 ℃ of temperature, wherein the extraction time is 0.5 hour each time, and then filtering to obtain an extracting solution;
(4) decolorizing the extractive solution with active carbon, ultrafiltering to remove impurities to obtain ultrafiltrate;
(5) concentrating the ultrafiltrate to 1/3-1/6 of the original volume, repeatedly washing and precipitating with ethanol for 3 times, and performing vacuum drying to obtain the tremella polysaccharide product.
2. The method for extracting tremella polysaccharide as claimed in claim 1, wherein the freezing temperature in step (1) is-45 ℃.
3. The method for extracting tremella polysaccharide as claimed in claim 1, wherein the step (2) is: adding a physical additive into the tremella fuciformis coarse powder, and putting the tremella fuciformis coarse powder into a vibration mill for ultramicro grinding for 20 minutes to obtain micro-cutting auxiliary powder, wherein the addition amount of the physical additive is 2% of the mass of the tremella fuciformis coarse powder.
4. The method for extracting tremella polysaccharide as claimed in claim 1, wherein the step (3) comprises performing ultrasonic assisted extraction on the fine cutting powder and water at a ratio of 1: 60(W/W) at a power of 100W and a frequency of 40KHz at a temperature of 80-90 ℃ for 2 times, wherein the extraction time is 0.5 hour each time, and filtering to obtain the extract.
5. The method for extracting tremella polysaccharide as claimed in claim 1, wherein the step (5) is: concentrating the ultrafiltrate to 1/5 of the original volume, repeatedly washing the precipitate with ethanol for 3 times, and vacuum drying to obtain the tremella polysaccharide product.
6. The method for extracting tremella polysaccharide as claimed in claim 1, wherein the physical auxiliary agent in step (2) is at least one of sodium carbonate, sodium bicarbonate, potassium carbonate, and potassium bicarbonate.
7. The method for extracting the tremella polysaccharide as claimed in claim 1, wherein the pore size of the filter element used in the step (4) of ultrafiltration is 0.1-0.22 μm.
8. The method for extracting tremella polysaccharide as claimed in claim 7, wherein the pore size of the pores of the filter element is 0.22 μm.
9. The method for extracting tremella polysaccharide as claimed in claim 5, wherein the concentration of ethanol is 75-95%, and the volume ratio of ethanol to the ultrafiltration concentrate is 2: 1-5: 1.
10. The method for extracting tremella polysaccharide as claimed in claim 9, wherein the volume ratio of ethanol to the ultrafiltration concentrate is 3: 1.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN111803526A (en) * | 2020-07-22 | 2020-10-23 | 威海凯菲尔生物科技有限公司. | Probiotic formula for preventing diarrhea |
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Application publication date: 20200424 |