CN112457421A - Extraction and separation method of dihydromyricetin and polysaccharide - Google Patents
Extraction and separation method of dihydromyricetin and polysaccharide Download PDFInfo
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Abstract
The invention discloses an extraction and separation method of dihydromyricetin and polysaccharide. The extraction and separation method comprises the following steps: crushing the ampelopsis grossedentata raw material; carrying out at least one first leaching treatment on the crushed Ampelopsis grossedentata raw material in a hot first extracting solution to obtain a first leaching solution; performing first filtration treatment on the first leaching solution by adopting a ceramic microfiltration membrane to obtain a first filtrate; carrying out first concentration treatment on the first filtrate to obtain a first concentrated solution; carrying out first cooling precipitation treatment on the first concentrated solution, and then carrying out first solid-liquid separation treatment to obtain first filter residue and first filtrate; and purifying the first filter residue to obtain purified dihydromyricetin, and purifying the first filtrate to obtain purified polysaccharide. The extraction and separation method can ensure the extraction rate and purity of dihydromyricetin and polysaccharide to the maximum extent, and the extraction rate can ensure good stability.
Description
Technical Field
The invention belongs to the technical field of extraction and separation, and particularly relates to an extraction and separation method of dihydromyricetin and polysaccharide.
Background
The dihydromyricetin is extract of Ampelopsis grossedentata (Ampelopsis grossedentata) of Vitis, is flavonoid as main active ingredient in Ampelopsis grossedentata (Ampelopsis grossedentata), and has effects of scavenging free radicals, resisting oxidation, resisting thrombi, resisting tumor, and relieving inflammation; the dihydromyricetin is a special flavonoid compound, and has special effects in relieving alcoholism, preventing alcoholic liver and fatty liver, inhibiting hepatocyte deterioration, reducing incidence rate of liver cancer, resisting hypertension, inhibiting in vitro platelet aggregation and in vivo thrombosis, reducing blood lipid and blood glucose levels, improving SOD activity, protecting liver, etc.
At present, it is reported that extraction of dihydromyricetin is generally performed by taking ampelopsis grossedentata leaves and stems as raw materials, performing hot water extraction treatment by utilizing the heat capacity characteristic of dihydromyricetin, performing cooling precipitation treatment on an extraction solution after extraction treatment, and then performing purification, such as molecular sieve column purification.
However, in the actual extraction process, the raw material of the Ampelopsis grossedentata is adopted as the extraction raw material, and the raw material of the Ampelopsis grossedentata contains various components, such as protein, fiber and pectin, and also contains macromolecular impurities such as polysaccharide functional components. In addition, the extraction method generally comprises the step of crushing the extract, so that a plurality of macromolecular impurities are dissolved or dispersed in the extracting solution together in the process of carrying out heat extraction treatment, and the final target extract dihydromyricetin has low extraction rate, low purity, high pressure for subsequent purification and high cost.
Meanwhile, the ampelopsis grossedentata raw material contains abundant polysaccharide functional components, so that the raw material is generally removed as waste filtrate at present, which causes waste and high cost for extracting dihydromyricetin.
Disclosure of Invention
The invention aims to provide an extraction and separation method of dihydromyricetin and polysaccharide, aiming at solving the technical problems of low extraction purity, large purification pressure and single extraction target component of the existing dihydromyricetin, which causes high extraction cost.
In order to achieve the purpose, the invention provides a method for extracting and separating dihydromyricetin and polysaccharide. The extraction and separation method of dihydromyricetin and polysaccharide comprises the following steps:
crushing the ampelopsis grossedentata raw material;
carrying out at least one first leaching treatment on the crushed Ampelopsis grossedentata raw material in a hot first extracting solution to obtain a first leaching solution; wherein the first extract is acidic;
performing first filtration treatment on the first leaching solution by adopting a ceramic microfiltration membrane to obtain a first filtrate;
carrying out first concentration treatment on the first filtrate to obtain a first concentrated solution;
carrying out first cooling precipitation treatment on the first concentrated solution, and then carrying out first solid-liquid separation treatment to obtain first filter residue and first filtrate; and purifying the first filter residue to obtain purified dihydromyricetin, and purifying the first filtrate to obtain purified polysaccharide.
Compared with the prior art, the invention has the following technical effects:
1. the extraction and separation method of dihydromyricetin and polysaccharide firstly utilizes the ceramic microfiltration membrane to filter the first extract, effectively ensures that the filtering treatment can be carried out at a relatively high temperature, thereby effectively ensuring the good solubility of the dihydromyricetin of the target extract, and simultaneously effectively separating and removing other impurities such as protein, fiber and the like, thereby effectively reducing the purification pressure of the subsequent purification of the dihydromyricetin and polysaccharide target extract, reducing the purification cost and improving the purification efficiency.
2. Meanwhile, the filtrate obtained by solid-liquid separation is purified, so that the extraction rate of dihydromyricetin is effectively improved, and the polysaccharide functional components can be simultaneously extracted and purified, thereby improving the utilization rate of the ampelopsis grossedentata raw material and effectively reducing the economic cost of the extraction and separation method.
3. The extraction and separation method of dihydromyricetin and polysaccharide has easily controlled process conditions, can effectively ensure stable extraction efficiency of dihydromyricetin and polysaccharide, and has high efficiency.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and other drawings can be obtained by those skilled in the art without creative efforts.
FIG. 1 is a schematic process flow diagram of the extraction and separation method of dihydromyricetin and polysaccharide in the embodiment of the invention.
Detailed Description
In order to make the technical problems, technical solutions and advantageous effects to be solved by the present invention more clearly apparent, the present invention is further described in detail below with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
The embodiment of the invention provides an extraction and separation method of dihydromyricetin and polysaccharide. The technological process of the extraction and separation method of dihydromyricetin and polysaccharide is shown in figure 1, and comprises the following steps:
s01, crushing the ampelopsis grossedentata raw material;
s02, performing at least one first leaching treatment on the crushed Ampelopsis grossedentata raw material in a hot first extracting solution to obtain a first leaching solution; wherein the first extract is acidic;
s03, performing first filtration treatment on the first leaching solution by adopting a ceramic microfiltration membrane to obtain first filtrate;
s04, carrying out first concentration treatment on the first filtrate to obtain a first concentrated solution;
s05, carrying out first cooling and precipitation treatment on the first concentrated solution, and then carrying out first solid-liquid separation treatment to obtain first filter residue and first filtrate; and purifying the first filter residue to obtain purified dihydromyricetin, and purifying the first filtrate to obtain purified polysaccharide.
Therefore, the extraction and separation method of dihydromyricetin and polysaccharide firstly utilizes the ceramic microfiltration membrane to filter the first extracting solution, effectively ensures that the filtering treatment can be carried out at a relatively high temperature, thereby effectively ensuring the good solubility of the dihydromyricetin of the target extract, and simultaneously effectively separating and removing other impurities such as protein, fiber and the like, thereby effectively reducing the purification pressure of subsequent purification on the target extract, reducing the purification cost and improving the purification efficiency. According to the extraction and separation method provided by the embodiment of the invention, the filter residue is purified to obtain the purified dihydromyricetin, and the filtrate is purified to obtain the polysaccharide functional component, so that the utilization rate of the ampelopsis grossedentata raw material is improved, and the economic cost of the extraction and separation method provided by the invention is effectively reduced. In addition, the extraction and separation method provided by the embodiment of the invention has the advantages that the process conditions in the steps from the step S01 to the step S013 are easy to control, the stable extraction efficiency of the dihydromyricetin and the polysaccharide can be effectively ensured, and the efficiency is high.
In step S01, the raw material of ampelopsis grossedentata may be pulverized into powder according to a conventional pulverization process to increase the contact surface between the powder and the first extract in step S01, thereby increasing the extraction efficiency. Such as but not limited to, size reduction to 30 mesh particle size.
In step S02, the powdered material of ampelopsis grossedentata is subjected to a first extraction treatment in a hot first extraction solution, and the target extract dihydromyricetin is extracted from the powdered material of ampelopsis grossedentata by utilizing the hot water solubility of the target extract, and simultaneously, other water-soluble substances such as polysaccharides are also extracted and filtered. In one embodiment, the temperature of the first extraction liquid is set at 60-80 deg.C, such as 70 deg.C. Namely, the powder raw material of the ampelopsis grossedentata is subjected to first leaching treatment in a first extracting solution at 60-80 ℃, so that the target extract can be fully dissolved in the first extracting solution. In addition, the first leach treatment should be sufficient, e.g., at the first leach treatment in the temperature range, the time of the first leach treatment may be, but is not limited to, 2 hours. In addition, the first leaching treatment may be multiple times of extraction, that is, after the first leaching solution treatment is performed on the powdered raw material of the ampelopsis grossedentata by using the first extracting solution, the first leaching treatment is performed on the filter residue respectively and repeatedly by using a plurality of other parts of the first extracting solution.
In addition, in order to increase the leaching amount of the target extract in the first extract, the first extract in step S02 is acidic. As one example, the pH of the first extract is 6.0-7.0. The pH value of the first extracting solution is regulated to be acidic, so that the stability of the dihydromyricetin of the target extract and the dissolving amount of polysaccharide can be effectively improved.
In a further embodiment, the pH of the first extract and/or the second extract is adjusted to 6.0-7.0 with at least one of hydrochloric acid, acetic acid, sorbic acid. The acidic components are added into the first extracting solution, so that the first extracting solution has stable acidity, an acidic leaching environment is constructed for the first leaching treatment, the dissolving amount of dihydromyricetin and polysaccharide of a target extract is increased, the stability of the dihydromyricetin can be effectively improved, and the biological activity of the dihydromyricetin is ensured.
In addition, the first extract in step S02 may be a conventional extract, such as water. In an embodiment of the invention, the first extraction liquid is an aqueous solution of ethanol. Wherein, in the first extracting solution, the volume concentration of the ethanol is controlled to be more than 0 and less than or equal to 30 percent. Through the optimization of the components of the first extracting solution, ethanol is added into water, and the synergistic effect is generated between the ethanol and the contained acid components, so that the dissolving amount of the dihydromyricetin and the polysaccharide of the target extract is further improved.
In step S03, the first leaching solution in step S02 inevitably contains other impurities, such as macromolecular impurities like pectin, protein, cellulose, etc., which are leached together, in addition to the dihydromyricetin and polysaccharide components of the target extract. Therefore, the first filtering treatment in step S03 can effectively filter impurities in the first leachate. Since the leaching temperature of the first leaching process is generally maintained in the first leaching solution in step S02, the first filtering process is performed by using a ceramic microfiltration membrane, so that on one hand, the impurities in the first leaching solution can be effectively removed by filtration; on the other hand, the high temperature resistant characteristic of ceramic microfiltration membrane can guarantee the stability of first filtration, promptly can effectively realize first filtration under the prerequisite that does not reduce first leaching liquor to avoided if traditional adoption resin film layer filter lead to need treat the filtrate cooling treatment or lead to the resin film layer lead to the not enough of filtration efficiency variation under high temperature. Therefore, in one embodiment, the temperature of the first filtering process, i.e. the temperature of the first leaching solution, is preferably 60-80 ℃, particularly 70 ℃ of the temperature of the first leaching process in step S02. Therefore, the ceramic microfiltration membrane is adopted to realize that the first filtration treatment can be directly carried out on the first leaching solution, and the first filtration treatment is not required to be carried out firstly, so that the stable solubility of the dihydromyricetin and the polysaccharide of the target extract in the filtration process is effectively ensured, the filtration treatment steps are simplified, and the filtration efficiency is improved. In a preferred embodiment, the pore size of the ceramic microfiltration membrane is 100-500nm, in particular a 100nm pore size ceramic microfiltration membrane
The filter residue after the first filtration treatment contains rich protein, fiber and pectin, etc., and can be used as other raw materials according to requirements, and the obtained first filtrate is rich in target extract such as dihydromyricetin.
In step S04, the first filtrate in step S03 is subjected to a first concentration treatment so that a first extract liquid portion is removed, thereby making the solubility of the objective extract such as dihydromyricetin and polysaccharide contained in the first filtrate saturated, so as to facilitate the efficiency and effect of the first cooling precipitation treatment in step S05. For example, the first filtrate may be concentrated to 20% of the original volume by the first concentration treatment so that the amount of dihydromyricetin solution is saturated. In one embodiment, the first concentration treatment is a concentration treatment using a nanofiltration membrane. The nanofiltration membrane is adopted to enter the nanofiltration membrane, so that the concentration efficiency can be effectively improved, small molecular impurities can be removed, and the subsequent purification pressure is reduced.
In step S05, during the cooling and precipitating process of the first concentrated solution, the solubility of dihydromyricetin is reduced due to the reduction of temperature, so that precipitation occurs in the first concentrated solution. In order to sufficiently precipitate the dihydromyricetin, in one embodiment, the temperature of the first cooling precipitation treatment is 5-20 ℃, specifically 15 ℃. By controlling the temperature within this range, precipitation of dihydromyricetin can be caused, and energy consumption can be reduced.
The first solid-liquid separation treatment in step S05 can effectively separate dihydromyricetin precipitated in the first filtrate subjected to the first cooling precipitation treatment from the solution. The first filter residue obtained by the first solid-liquid separation treatment is a dihydromyricetin crude product; the first filtrate obtained is rich in polysaccharides. Wherein, the first solid-liquid separation treatment can be realized by separating precipitates in a conventional manner.
In a further embodiment, the method for purifying the first filter residue in step S05 includes the following steps:
s051: and dissolving the first filter residue in a hot second extracting solution, performing second cooling precipitation treatment, and performing second solid-liquid separation treatment to obtain a second filter residue and a second filtrate, wherein the second filter residue is purified dihydromyricetin, and the second filtrate is mixed with the first filtrate.
The polysaccharide component remained in the crude dihydromyricetin product can be further separated by purifying the second filter residue according to the purification step in the step S051, and if the second filter residue obtained by separation is purified dihydromyricetin, the second filtrate contains the residual polysaccharide component, so that the second filter residue and the first filtrate in the step S05 are mixed, the polysaccharide can be extracted to the maximum extent, and the extraction rate of the polysaccharide is improved.
Wherein the composition, temperature and pH of the second extract in step S051 are the same as those of the first extract in the above step S02, and the second cooling precipitation treatment and second solid-liquid separation treatment conditions and effects are the first cooling precipitation treatment and first solid-liquid separation treatment conditions in step S05, respectively. Therefore, both the second cooling precipitation treatment and the second solid-liquid separation treatment are preferably carried out in accordance with the first cooling precipitation treatment and the first solid-liquid separation treatment.
In addition, the purification process of step S051 may be performed once or more than once to increase the purity of dihydromyricetin.
In another further embodiment, the method of subjecting the first filtrate to purification treatment in step S05 includes the steps of:
s052: neutralizing the first filtrate, and then performing second concentration treatment to obtain a second concentrated solution;
s053: performing third cooling precipitation treatment on the second concentrated solution, and performing third solid-liquid separation treatment to obtain third filter residue and third filtrate; and mixing the third filter residue with the first filter residue, and drying the third filtrate to obtain the purified polysaccharide.
By purifying the first filtrate according to the purification steps of step S052 and step S053, the dihydromyricetin remaining in the first filtrate can be recovered again, and the polysaccharide can be separated and purified, for example, the third filtrate obtained by separation contains polysaccharide components, and the third residue is the residual dihydromyricetin. The third filtrate may be directly dried, such as spray-dried, to obtain a polysaccharide extract, and the third residue is mixed with the first residue in step S05, so that dihydromyricetin can be extracted to the maximum extent, and the extraction rate of dihydromyricetin is increased.
Therefore, the extraction and separation method of dihydromyricetin and polysaccharide in each embodiment can ensure that dihydromyricetin and polysaccharide are leached from raw materials to the maximum extent and can ensure the extraction to the maximum extent in separation and purification, so that the extraction rate of the dihydromyricetin and polysaccharide in the target extract is high, and the extraction rate can ensure good stability. In addition, the extraction rate of the dihydromyricetin and the polysaccharide of the target extract can be further improved by optimizing the purification steps and the process conditions of the steps, and the extraction cost is effectively reduced.
The following examples are provided to illustrate the extraction and separation method of dihydromyricetin and polysaccharide according to the present invention. In the following examples, the purity of dihydromyricetin was confirmed by HPLC (high performance liquid chromatography) and the purity of polysaccharide was confirmed by anthrone-sulfuric acid spectrophotometry.
Example 1
This example provides a method for extracting and separating dihydromyricetin and polysaccharide. The extraction and separation method comprises the following steps:
s1: crushing one kilogram of dried Hunan xi Yongshun county blackberry tea into 30 meshes, adding 10 liters of an extracting solution containing 30% alcohol and having the pH value of 6.5, heating to 70 ℃, extracting for 2 hours at constant temperature, filtering, circularly extracting for three times, combining the extracting solutions, and discarding residues; wherein acetic acid is also added into the extractive solution, and the concentration of acetic acid is such that pH of the extractive solution is 6.5;
s2: and (3) circularly filtering the filtrate at a constant temperature of 70 ℃ by using a tubular ceramic membrane with the aperture of 100nm, stopping when no permeate flows out, discarding trapped fluid, and introducing the permeate into a tubular nanofiltration membrane for circularly concentrating until the volume of the permeate is 20% of that of the original solution.
S3: cooling the nanofiltration trapped fluid, keeping the temperature at 15 ℃, standing for 12 hours for precipitation, and performing first filtration treatment, wherein the light yellow precipitate is a dihydromyricetin crude product (containing a small amount of extracting solution), and the supernatant is a polysaccharide-containing solution. The crude dihydromyricetin product was dried in a freeze dryer to obtain 264 g of powder (purity 64%). Neutralizing the obtained polysaccharide solution with a neutralizing solution until the pH value is 7.0, and then pouring the neutralized polysaccharide solution into a spray dryer for drying to obtain 172 g of polysaccharide powder;
s4: adding three times of extracting solution containing 20% alcohol with pH of 6.5 into the crude dihydromyricetin product obtained in the step S3, heating to 70 ℃ for dissolving, cooling, keeping the temperature constant at 15 ℃ for precipitation for 12 hours, carrying out second filtration treatment, and freeze-drying the precipitate to obtain 210 g (yellowish) dihydromyricetin powder (purity 83%) which is refined once; the supernatant obtained by the second filtration treatment was combined with the polysaccharide solution in step S3, and then neutralized with a neutralizing solution to pH7.0, and then concentrated to 20% by volume of the stock solution with a nanofiltration membrane, and introduced into a spray dryer to be dried, whereby 226 g of polysaccharide powder was obtained.
Example 2
This example provides a method for extracting and separating dihydromyricetin and polysaccharide. The extraction and separation method comprises the following steps:
s1: pulverizing 2kg of Ampelopsis grossedentata tea into 30 mesh powder, adding 6 times of extractive solution containing 20% alcohol with pH of 6.2, heating, extracting at 80 deg.C for 3 hr, filtering to obtain filtrate, repeatedly extracting for three times, and mixing filtrates; wherein the extractive solution is further added with hydrochloric acid, and the concentration of acetic acid is such that pH of the extractive solution is 6.2;
s2: circulating filtering the filtrate at constant temperature of 80 deg.C with tubular ceramic membrane with pore diameter of 200nm, stopping when no permeate flows out, discarding trapped fluid, and introducing the permeate into tubular nanofiltration membrane for cyclic concentration to 20% of the volume of the stock solution;
s3: cooling the nanofiltration trapped fluid, keeping the temperature at 15 ℃, standing for 12 hours for precipitation, and performing first filtration treatment, wherein the light yellow precipitate is a dihydromyricetin crude product (containing a small amount of extracting solution), and the supernatant is a solution containing polysaccharide; putting the crude dihydromyricetin product into a freeze dryer for drying to obtain 568 g of powder (with the purity of 68%); neutralizing the obtained polysaccharide solution with a neutralizing solution until the pH value is 7.0, and then introducing into a spray dryer for drying to obtain 357 g polysaccharide powder;
s4: adding four times of extracting solution containing 10% alcohol and having pH of 7.0 into the crude dihydromyricetin product obtained in the step S3, heating to 65 ℃ for dissolving, cooling, keeping the temperature constant at 15 ℃ for precipitation for 12 hours, carrying out second filtration treatment, and freeze-drying the precipitate to obtain 505 g (purity 83%) of once-refined dihydromyricetin powder; mixing the supernatant with the polysaccharide solution obtained in the step S3, neutralizing with a neutralizing solution until the pH value is 7.0, concentrating with a nanofiltration membrane to 20% of the volume of the stock solution, and introducing into a spray dryer for drying to obtain 412 g of polysaccharide powder;
s5: the twice-concentrated extract containing 5% alcohol and having a ph of 7.0 was added to the once-purified dihydromyricetin obtained in step S3, and the mixture was heated to 70 ℃ to dissolve the dihydromyricetin, and then cooled and precipitated at a constant temperature of 15 ℃ for 12 hours, followed by a third filtration treatment, and the precipitate was freeze-dried to obtain 441 g (white) of a twice-purified dihydromyricetin powder (94% purity).
Example 3
This example provides a method for extracting and separating dihydromyricetin and polysaccharide. The extraction and separation method comprises the following steps:
s1: refer to step S1 of example 2;
s2: refer to step S2 of example 2;
s3: refer to step S3 of example 2;
s4: adding four times of extracting solution containing 10% alcohol and having pH of 7.0 into the crude dihydromyricetin product obtained in the step S3, heating to 65 ℃ for dissolving, cooling, keeping the temperature constant at 15 ℃ for precipitation for 12 hours, carrying out second filtration treatment, and freeze-drying the precipitate to obtain 505 g (purity 83%) of once-refined dihydromyricetin powder; combining the second filtered supernatant with the first filtered supernatant;
s5: the twice-concentrated extract containing 5% alcohol and having a ph of 7.0 was added to the once-purified dihydromyricetin obtained in step S3, and the mixture was heated to 70 ℃ to dissolve the dihydromyricetin, and then cooled and precipitated at a constant temperature of 15 ℃ for 12 hours, followed by a third filtration treatment, and the precipitate was freeze-dried to obtain 441 g (white) of a twice-purified dihydromyricetin powder (94% purity). Combining the third filtered supernatant with the first filtered supernatant;
s6: neutralizing the combined solution of the supernatant in the step S5 with a neutralizing solution until the pH value is 7.0, concentrating the solution to 20% of the volume of the stock solution through a nanofiltration membrane, and introducing the solution into a spray dryer for drying to obtain 471 g of polysaccharide powder;
comparative example 1
The comparative example provides an extraction and separation method of dihydromyricetin and polysaccharide. The extraction and separation method comprises the following steps:
s1: crushing one kilogram of dried Hunan xi Yongshun county blackberry tea into 30 meshes, adding 10 liters of an extracting solution containing 30% alcohol and having the pH value of 6.5, heating to 70 ℃, extracting for 2 hours at constant temperature, filtering, circularly extracting for three times, combining the extracting solutions, and discarding residues; wherein acetic acid is also added into the extractive solution, and the concentration of acetic acid is such that pH of the extractive solution is 6.5;
s2: filtering the filtrate at constant temperature of 70 deg.C with 0.1 μm polyvinylidene fluoride ultrafiltration membrane, discarding the trapped fluid, and introducing the permeated fluid into tubular nanofiltration membrane for cyclic concentration to 20% of the volume of the stock solution.
S3: cooling the nanofiltration trapped fluid, keeping the temperature at 15 ℃, standing for 12 hours for precipitation, and performing first filtration treatment, wherein the light yellow precipitate is a dihydromyricetin crude product (containing a small amount of extracting solution), and the supernatant is a polysaccharide-containing solution. The crude dihydromyricetin product was dried in a freeze dryer to obtain 241 g of powder (purity 46%). Neutralizing the obtained polysaccharide solution with a neutralizing solution until the pH value is 7.0, and then pouring the neutralized polysaccharide solution into a spray dryer for drying to obtain 166 g of polysaccharide powder;
s4: adding three times of extracting solution containing 20% alcohol with pH of 6.5 into the crude dihydromyricetin product obtained in the step S3, heating to 70 ℃ for dissolving, cooling, keeping the temperature constant at 15 ℃ for precipitation for 12 hours, carrying out second filtration treatment, and freeze-drying the precipitate to obtain 201 g (yellowish) dihydromyricetin powder (with purity of 76%) which is refined once; the supernatant obtained by the second filtration treatment was combined with the polysaccharide solution in step S3, and then neutralized with a neutralizing solution to pH7.0, and then concentrated to 20% by volume of the stock solution with a nanofiltration membrane, and introduced into a spray dryer to be dried, whereby 215 g of polysaccharide powder was obtained.
Comparative example 2
The comparative example provides an extraction and separation method of dihydromyricetin and polysaccharide. The extraction and separation method comprises the following steps:
s1: one kilogram of dried Hunan xi Yongshun county strawberry powder is ground to 30 meshes, 10 liters of extracting solution containing 30 percent alcohol with the pH value of 7 (compared with the extracting solution containing no acetic acid in example 1) is added, the extracting solution is heated to 70 ℃ and extracted for 2 hours at constant temperature, the filtering is carried out, the extraction solution is circularly extracted for three times, the extracting solution is combined, and the residue is discarded;
s2: filtering the filtrate at constant temperature of 70 deg.C with 0.1 μm polyvinylidene fluoride ultrafiltration membrane, discarding the trapped fluid, and introducing the permeated fluid into tubular nanofiltration membrane for cyclic concentration to 20% of the volume of the stock solution.
S3: cooling the nanofiltration trapped fluid, keeping the temperature at 15 ℃, standing for 12 hours for precipitation, and performing first filtration treatment, wherein the light yellow precipitate is a dihydromyricetin crude product (containing a small amount of extracting solution), and the supernatant is a polysaccharide-containing solution. The crude dihydromyricetin product was dried in a freeze dryer to obtain 246 g of powder (52% purity). Neutralizing the obtained polysaccharide solution with a neutralizing solution until the pH value is 7.0, and then pouring the solution into a spray dryer for drying to obtain 147 g of polysaccharide powder;
s4: adding three times of extracting solution containing 20% alcohol with pH of 6.5 into the crude dihydromyricetin product obtained in the step S3, heating to 70 ℃ for dissolving, cooling, keeping the temperature constant at 15 ℃ for precipitation for 12 hours, carrying out second filtration treatment, and freeze-drying the precipitate to obtain 186 g (purity of 78%) of once-refined dihydromyricetin powder; the supernatant obtained by the second filtration treatment was combined with the polysaccharide solution in step S3, and then neutralized with a neutralizing solution to pH7.0, and then concentrated to 20% by volume of the stock solution with a nanofiltration membrane, and introduced into a spray dryer to be dried, whereby 206 g of polysaccharide powder was obtained.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents and improvements made within the spirit and principle of the present invention are intended to be included within the scope of the present invention.
Claims (10)
1. A method for extracting and separating dihydromyricetin and polysaccharide comprises the following steps:
crushing the ampelopsis grossedentata raw material;
carrying out at least one first leaching treatment on the crushed Ampelopsis grossedentata raw material in a hot first extracting solution to obtain a first leaching solution; wherein the first extract is acidic;
performing first filtration treatment on the first leaching solution by adopting a ceramic microfiltration membrane to obtain a first filtrate;
carrying out first concentration treatment on the first filtrate to obtain a first concentrated solution;
carrying out first cooling precipitation treatment on the first concentrated solution, and then carrying out first solid-liquid separation treatment to obtain first filter residue and first filtrate; and purifying the first filter residue to obtain purified dihydromyricetin, and purifying the first filtrate to obtain purified polysaccharide.
2. The extraction and separation method according to claim 1, wherein the method for purifying the first filter residue comprises the following steps:
dissolving the first filter residue in a hot second extracting solution, performing second cooling precipitation treatment, and performing second solid-liquid separation treatment to obtain a second filter residue and a second filtrate, wherein the second filter residue is the purified dihydromyricetin, and the second filtrate is mixed with the first filtrate; wherein the second extract is acidic;
and/or
The method for purifying the first filtrate comprises the following steps:
neutralizing the first filtrate, and then performing second concentration treatment to obtain a second concentrated solution;
performing third cooling precipitation treatment on the second concentrated solution, and performing third solid-liquid separation treatment to obtain third filter residue and third filtrate; and mixing the third filter residue with the first filter residue, and drying the third filtrate to obtain the purified polysaccharide.
3. The extraction separation method according to any one of claims 1 to 2, characterized in that: the temperature of at least one of the first extract, the second extract and the first filtration treatment is 60-80 ℃.
4. The extraction separation method according to claim 3, characterized in that: the temperature of the first extracting solution, the second extracting solution and the first filtering treatment is 70 ℃.
5. The extraction separation method according to any one of claims 1, 2 and 4, wherein: the temperature of at least one of the first cooling precipitation treatment, the second cooling precipitation treatment and the third cooling precipitation treatment is 5-20 ℃.
6. The extraction separation method according to claim 5, wherein: the temperature of the first cooling precipitation treatment, the second cooling precipitation treatment and the third cooling precipitation treatment is 15 ℃.
7. The extraction and separation method according to any one of claims 1, 2, 4 and 6, wherein: and the first concentration treatment and/or the second concentration treatment is/are concentration treatment by adopting a nanofiltration membrane.
8. The extraction and separation method according to any one of claims 1, 2, 4 and 6, wherein: the pore diameter of the ceramic microfiltration membrane is 100-500 nm.
9. The extraction and separation method according to any one of claims 1, 2, 4 and 6, wherein: at least one of the first extractive solution and the second extractive solution has pH of 6.0-7.0; and/or
Adjusting pH of the first extractive solution and/or the second extractive solution to 6.0-7.0 with at least one of hydrochloric acid, acetic acid, and sorbic acid.
10. The extraction and separation method according to any one of claims 1, 2, 4 and 6, wherein: at least one of the first extract and the second extract contains ethanol, and the volume concentration of the ethanol is more than 0 and less than or equal to 30%.
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