Method for synchronously extracting and separating dihydromyricetin, raspberry tea flavone and raspberry tea polysaccharide

Abstract

The invention relates to a method for synchronously extracting and separating dihydromyricetin, raspberry tea flavone and raspberry tea polysaccharide, which comprises the following steps: by taking raspberry tea old leaves and / or branches as raw materials, extracting by adopting an organic solvent with the mass concentration of organic alcohol being 10-60wt%, filtering, extracting filter residues by adopting hot water, merging filtrates, treating, and separating to obtain filter residues A and filtrate B; dissolving dihydromyricetin in the filter residue A by adopting an organic solvent, filtering, recovering the organic solvent in the filtrate to obtain dihydromyricetin, dissolving the filter residue in hot water, mixing with the filtrate B, concentrating, extracting by adopting an organic extracting agent, recovering the organic extracting agent in the extracting solution to obtain raspberry tea flavone, and preparing raspberry tea polysaccharide from the raffinate by adopting an alcohol precipitation method. According to the method, the old leaves and branches of the raspberry tea can be used as raw materials, multiple active ingredients are synchronously extracted and separated, the content of the active ingredients in the extract is high, the extraction cost is low, and the comprehensive utilization degree of the raspberry tea is improved.

Description

Technical Field

[0001] The invention relates to the technical field of biological extraction, and more specifically to a method for synchronously extracting and separating dihydromyricetin, berry tea flavonoids and berry tea polysaccharides. Background Art

[0002] Berry tea is a tea product made from the tender stems and leaves of Ampelopsis grossedentata, a plant of the genus Ampelopsis, in the family Vitaceae, after withering, rolling and drying. It is also called Qingshuang Ancient Vine Tea. The chemical composition of berry tea mainly contains a variety of natural active ingredients such as flavonoids, polysaccharides and steroids, which have significant biological activities such as anti-inflammatory, antibacterial, antioxidant and antiviral. The total flavonoid content in its ingredients accounts for about 44%, which is the plant with the highest content of flavonoids discovered so far, and is known as the "king of flavonoids" in nature. Among them, dihydromyricetin is the main active ingredient of berry tea flavonoids, which has multiple effects such as antioxidant, antibacterial and anti-tumor. As a natural polysaccharide, berry tea polysaccharide has been proven to play a good antioxidant activity in scavenging free radicals, resisting reactive oxygen, protecting cells from oxidative stress damage, anti-tumor, and reducing lipid peroxidation. However, the young leaves of vines only account for 1-3% of the total biomass. After picking, they are mainly used to make berry (vine) tea and sold in the form of tea. The existing extraction technology usually uses vine tea or young leaves of vines as raw materials to extract active ingredients, which has high extraction costs and a single type of active ingredients.

[0003] Existing extraction technology generally uses a set of systems to simply extract dihydromyricetin, berry tea flavonoids or berry tea polysaccharides from berry tea. For example, in patent CN117447431A, rattan tea is used as raw material, and hot water ultrasound is used to extract dihydromyricetin; in CN109329524A, sterilized Maotai berry tea is used as raw material, cellulase is used for enzymolysis, and flavonoid crude extract is obtained by alkali dissolution and acid precipitation; in CN116217745B, rattan tea is used as raw material, and water extraction and alcohol precipitation are adopted to obtain crude polysaccharides, and then the crude polysaccharides are deproteinized and desalted, and finally ion exchange column chromatography and gel filtration column chromatography are used to separate and purify the crude polysaccharides to obtain two pure rattan tea polysaccharides, rattan tea polysaccharide AGP1 and rattan tea polysaccharide AGP2. However, these extraction methods only extract one active ingredient in berry (rattan) tea, which is easy to cause the loss of other active ingredients.

[0004] Patent CN102106931B discloses a production method of multiple extracts of Ampelopsis grossedentata. Using the young tender vine stems and leaves with a length of 30 - 50 cm after germination as raw materials, it adopts pure water microwave extraction, separates by macroporous resin adsorption, and after the column effluent is concentrated, it is precipitated with 95% ethanol, and dried to obtain the Ampelopsis grossedentata polysaccharide extract; then it is eluted with hot pure water, the eluate is collected, concentrated and spray-dried to obtain the total flavonoids extract of Ampelopsis grossedentata, or after collecting the eluate, it passes through a 5000Da spiral ultrafiltration membrane, reverse osmosis membrane, dialysis solution reflux, activated carbon decolorization, pressure filtration, cooling crystallization, and vacuum drying to obtain dihydromyricetin. Although this method can extract Ampelopsis grossedentata polysaccharide and total flavonoids of Ampelopsis grossedentata or dihydromyricetin, actually this method cannot extract dihydromyricetin and Ampelopsis grossedentata flavonoids simultaneously. And total flavonoids include various types such as flavones, flavonols, dihydroflavones, dihydroflavonols, and isoflavones. Among them, dihydromyricetin in dihydroflavonols is easily soluble in hot water, while flavonoids such as quercetin and kaempferol have poor solubility in water, and flavonols and isoflavones are insoluble in water. It can be seen that the total flavonoids of Ampelopsis grossedentata extracted by this method using pure water are water-soluble flavonoid compounds.

[0005] Although active ingredients such as flavonoids and polysaccharides can be extracted using Ampelopsis grossedentata or young leaves of vine plants as raw materials, the extraction cost is high and it is difficult to promote and apply. Compared with the young stems and leaves of Ampelopsis grossedentata, the content of active ingredients in its old leaves and branches is low and the extraction difficulty is great. Therefore, researching a method for extracting multiple Ampelopsis grossedentata extracts using the old leaves and branches of Ampelopsis grossedentata as raw materials is of great significance for improving the economic value and comprehensive utilization degree of Ampelopsis grossedentata. Summary of the Invention

[0006] In view of the above defects or improvement requirements of the prior art, the present invention provides a method for synchronously extracting and separating dihydromyricetin, Ampelopsis grossedentata flavonoids, and Ampelopsis grossedentata polysaccharides. The purpose is to discover that using the old leaves and / or branches of Ampelopsis grossedentata as raw materials, first extract with an organic solvent with an organic alcohol mass concentration of 10 - 60wt%, after filtration, the filter residue is then extracted with hot water, the filtrates are combined, and after treatment, the filter residue A and filtrate B are separated; the filter residue A is dissolved in acetone and filtered, the acetone in the filtrate is recovered to obtain the crude product of dihydromyricetin, the filter residue is dissolved in hot water and mixed with the filtrate B, and an organic extractant is used for extraction, the organic extractant in the extract is recovered to obtain Ampelopsis grossedentata flavonoids, and the raffinate phase is obtained by alcohol precipitation to obtain Ampelopsis grossedentata polysaccharides, thereby solving the technical problems of high extraction cost and single type of active ingredients extracted in the existing Ampelopsis grossedentata extracts.

[0007] To achieve the above purpose, according to one aspect of the present invention, a method for synchronously extracting and separating dihydromyricetin, Ampelopsis grossedentata flavonoids, and Ampelopsis grossedentata polysaccharides is provided, which specifically includes the following steps:

[0008] (1) Synchronous extraction: Using old leaves and / or branches of Ampelopsis grossedentata as raw materials, first extract at room temperature or slightly boiling with an organic solvent with an organic alcohol mass concentration of 10-60 wt%, filter to collect the filtrate and residue, then extract the residue with hot water and filter, and combine the filtrates to obtain a crude extract containing dihydromyricetin, Ampelopsis grossedentata flavonoids and Ampelopsis grossedentata polysaccharides;

[0009] (2) Separation and preparation of dihydromyricetin, Ampelopsis grossedentata flavonoids and Ampelopsis grossedentata polysaccharides: Recover the solvent in the crude extract of step (1), concentrate, and separate to obtain residue A and filtrate B; Dissolve dihydromyricetin in the residue A with an organic solvent, filter to obtain the filtrate and residue C, recover the organic solvent in the filtrate, and dry the solid to obtain dihydromyricetin;

[0010] Dissolve residue C in hot water and mix it with filtrate B, concentrate to a specific gravity of 1.05-1.15, add an organic extractant to the concentrated solution for extraction, separate to obtain an extract containing Ampelopsis grossedentata flavonoids and a raffinate containing Ampelopsis grossedentata polysaccharides, recover the organic extractant in the extract, and dry the solid to obtain Ampelopsis grossedentata flavonoids; The raffinate is subjected to alcohol precipitation to obtain Ampelopsis grossedentata polysaccharides.

[0011] Preferably, in the method, in step (1), the raw materials are first crushed to 10-20 mesh, the organic solvent is added according to the mass ratio of raw materials to organic solvent of 1:20, and extracted at slightly boiling for 1-2 times, 1-2 h each time.

[0012] Preferably, in the method, the mass concentration of organic alcohol in the organic solvent is 30 wt%-50 wt%, and it is extracted at slightly boiling for 1-2 times, 1 h each time.

[0013] Preferably, in the method, the organic alcohol is methanol or ethanol.

[0014] Preferably, in the method, the hot water extraction is to add water to the residue according to the mass ratio of raw materials to pure water of 1:10-15, heat to slightly boiling and extract for 1-2 times, 1-2 h each time.

[0015] Preferably, in the method, in step (2), residue A is repeatedly dissolved with acetone and the filtrate is collected by filtration.

[0016] Preferably, in the method, the concentration is vacuum decompression concentration, the vacuum degree is 0.01-0.1 Mpa, the temperature is 40-80 °C, and the concentration is concentrated to a specific gravity of 1.05-1.10 at 20 °C of the concentrated solution.

[0017] Preferably, in the method, for the extraction, the organic extractant is added according to the volume ratio of the concentrated solution to the organic extractant of 1:1-3, and extracted at room temperature or at 4-10 °C for 1-3 times.

[0018] Preferably, in the method, the organic extractant includes any one of n-butanol, ethyl acetate, n-pentanol or chloroform.

[0019] Preferably, in the method, in the alcohol precipitation method, 95% ethanol is added to the raffinate phase to make the volume concentration of ethanol in the mixed solution 50-80%, and it is left standing for 8-24 h, filtered, and the filter residue is dried to obtain ampelopsis grossedentata polysaccharide.

[0020] Generally speaking, compared with the prior art by the above technical solution conceived by the present invention, the following beneficial effects can be achieved:

[0021] The extraction method provided by the present invention uses old leaves and / or branches of ampelopsis grossedentata as raw materials. First, it is extracted with an organic solvent with an organic alcohol mass concentration of 10-60 wt%, filtered, and the filter residue is then extracted with hot water. The filtrates are combined to obtain a crude extract. The crude extract is treated and separated to obtain filter residue A and filtrate B. The filter residue A is dissolved with an organic solvent to dissolve dihydromyricetin, and filtered to obtain dihydromyricetin. The filter residue is dissolved in hot water and mixed with filtrate B, and extracted with an organic extractant. The solvent in the extract is recovered to obtain ampelopsis grossedentata flavonoids. The raffinate phase is subjected to alcohol precipitation to obtain ampelopsis grossedentata polysaccharide, which can simultaneously extract and separate dihydromyricetin, ampelopsis grossedentata flavonoids and ampelopsis grossedentata polysaccharide. Using old leaves and branches of ampelopsis grossedentata as raw materials, the extraction cost of active ingredients is low, and it is also beneficial to improve the comprehensive utilization degree of ampelopsis grossedentata. Detailed embodiments

[0022] To further elaborate on the technical means and effects adopted by the present invention to achieve the predetermined invention purpose, the following combines the drawings and preferred embodiments to describe in detail the specific embodiments, structures, features and their effects according to the present invention as follows.

[0023] Ampelopsis grossedentata, also known as Qing Shuang Guteng tea or small-leaf species rattan tea, is a perennial vine plant growing in the Wuling Mountains with Zhangjiajie as the core area, belonging to the Ampelopsis grossedentata plant of the Ampelopsis genus in the Vitaceae family. To promote the germination of new branches in the coming year and obtain more tender leaves to prepare ampelopsis grossedentata tea, tea farmers generally carry out appropriate mowing of old leaves and branches in autumn. However, these old leaves and branches are discarded because they cannot be used for making tea for sale and are not well utilized.

[0024] The present invention attempts to utilize the difference between dihydromyricetin and other flavonoids in ampelopsis grossedentata to achieve the extraction and separation of dihydromyricetin and ampelopsis grossedentata flavonoids; and then utilize the chemical property differences between ampelopsis grossedentata flavonoids and ampelopsis grossedentata polysaccharide to achieve the extraction and separation of ampelopsis grossedentata flavonoids and ampelopsis grossedentata polysaccharide. Through experimental exploration, it is found that first extracting with an organic solvent containing 10-60 wt% of organic alcohol, filtering, and then extracting the filter residue with hot water, and combining the filtrates as a crude extract can efficiently extract multiple active ingredients from old leaves and / or branches of ampelopsis grossedentata at the same time, and separate and obtain dihydromyricetin, ampelopsis grossedentata flavonoids and ampelopsis grossedentata polysaccharide. Specifically:

[0025] The old leaves and / or branches of Ampelopsis grossedentata are crushed and used as raw materials. An organic solvent with a methanol or ethanol mass concentration of 10 - 60 wt% is used as the organic extraction medium, and extraction is carried out 1 - 2 times. The filtrate and filter residue are collected by filtration. The filter residue is then extracted with water as the inorganic extraction medium by heating 1 - 2 times, and the filtrate is collected by filtration. The filtrates are combined to obtain a crude extract; the obtained crude extract is concentrated under reduced pressure to recover the solvent and concentrated to 1 / 2 - 2 / 3 of the original volume, and then centrifuged while standing to obtain filter residue A and filtrate B.

[0026] Among them, for filter residue A, a strongly polar organic solvent such as acetone is first used to dissolve dihydromyricetin in the filter residue, and then filtered to obtain a filtrate and filter residue C. The filtrate is concentrated to recover the organic solvent, and the solid matter is dried to obtain dihydromyricetin; filter residue C is dissolved in hot water and then mixed with filtrate B, and concentrated to a specific gravity of 1.05 - 1.15 (20 °C), and then an organic extractant is added for extraction. The extract containing Ampelopsis grossedentata flavonoids and the raffinate containing Ampelopsis grossedentata polysaccharides are separated; the extraction solvent in the extract is recovered, and the solid matter is dried to obtain Ampelopsis grossedentata flavonoids; ethanol is added to the raffinate to precipitate the polysaccharides, and then filtered, and the filter residue is dried to obtain Ampelopsis grossedentata polysaccharides; the organic extractant includes any one of n-butanol, ethyl acetate, n-pentanol or chloroform.

[0027] By using this method, dihydromyricetin, Ampelopsis grossedentata flavonoids and Ampelopsis grossedentata polysaccharides can be synchronously and efficiently extracted and separated from the old leaves and branches of Ampelopsis grossedentata. Among them, the content of dihydromyricetin is stably maintained at 45 - 70 wt%, the content of Ampelopsis grossedentata flavonoids is stably maintained at 40 - 65 wt%, and the polysaccharide content is stably maintained at 40 - 55 wt%. This can reduce the extraction cost of the active ingredients of Ampelopsis grossedentata, and at the same time improve the comprehensive utilization value of Ampelopsis grossedentata.

[0028] Based on this, the present invention provides a method for synchronously extracting and separating dihydromyricetin, Ampelopsis grossedentata flavonoids and Ampelopsis grossedentata polysaccharides, which comprises the following steps:

[0029] (1) Synchronous extraction: The old leaves and / or branches of Ampelopsis grossedentata are crushed. First, an organic solvent with an organic alcohol mass concentration of 10 - 60 wt% is used as the extraction medium, and extraction is carried out at room temperature or slightly boiling for 1 - 2 times, 1 - 2 h each time, mainly for extracting dihydromyricetin and hydrophobic Ampelopsis grossedentata flavonoids. The filtrate and filter residue are collected by filtration; the filter residue is then extracted with hot water 1 - 2 times, 1 - 2 h each time, mainly for extracting water-soluble Ampelopsis grossedentata flavonoids and Ampelopsis grossedentata polysaccharides. The filtrate is collected by filtration, and the filtrates are combined to obtain a crude extract containing dihydromyricetin, Ampelopsis grossedentata flavonoids and Ampelopsis grossedentata polysaccharides.

[0030] (2) Separation and preparation of dihydromyricetin, Ampelopsis grossedentata flavonoids and Ampelopsis grossedentata polysaccharides

[0031] The crude extract obtained in step (1) is concentrated under reduced pressure to recover the solvent, and separated to obtain residue A and filtrate B. Among them, residue A is repeatedly dissolved in acetone, filtered to collect the filtrate and residue C, so that dihydromyricetin in the residue is fully dissolved in acetone. The filtrates are combined, acetone is recovered, and the solid is dried to obtain dihydromyricetin;

[0032] Then, residue C that is insoluble or hardly soluble in the above organic solvent acetone is dissolved in hot water and mixed with filtrate B. The mixed solution is concentrated to a specific gravity of 1.05 - 1.15, and an organic extractant is added for extraction. The extract containing flavonoids from Ampelopsis grossedentata and the raffinate containing polysaccharides from Ampelopsis grossedentata are separated. The organic extractant in the extract is recovered, and the solid is dried to obtain flavonoids from Ampelopsis grossedentata; ethanol is added to the obtained raffinate to precipitate the polysaccharides, filtered, and the residue is dried to obtain polysaccharides from Ampelopsis grossedentata.

[0033] In step (1) of some embodiments, the old leaves and / or branches of Ampelopsis grossedentata are crushed into 10 - 20 meshes. The organic alcohol with a mass concentration of 10 - 60 wt% is a mixed solvent of methanol or ethanol and water, where the mass concentration of methanol or ethanol is 10 - 60 wt%, preferably the mass concentration of ethanol in the organic solvent is 30% - 50%, and it is heated to slightly boiling for extraction twice; the hot water extraction is heated to slightly boiling for 1 - 2 h.

[0034] In step (1), the crude extract is concentrated under reduced pressure to recover the solvent, and the vacuum concentration is carried out under a vacuum of 0.01 - 0.1 Mpa and a temperature of 40 - 80 °C, and concentrated to a specific gravity of 1.05 - 1.10 at 20 °C of the concentrated solution.

[0035] In the extraction in step (2), the organic extractant is added according to the volume ratio of the concentrated solution to the organic extractant of 1:1 - 3, and extracted at room temperature or at a low temperature of 4 - 10 °C for 1 - 3 times, or extracted by an organic solvent continuous flow extraction separator for 1 - 3 times. The organic extractant includes any one of n-butanol, ethyl acetate, n-pentanol, or chloroform.

[0036] In some embodiments, 95% ethanol is added to the raffinate to make the volume concentration of ethanol in the mixed solution 50 - 80%, and left standing until no precipitation occurs, filtered to collect the residue, and the residue is dried to obtain polysaccharides from Ampelopsis grossedentata. In the present invention, the drying includes natural drying, vacuum concentration drying, freeze drying, or spray drying.

[0037] The yield of the pure dihydromyricetin obtained by extraction and separation according to this method can be stably above 10.0% (the yield can increase with the improvement of the quality of old leaves and branches of Ampelopsis grossedentata), and the content of dihydromyricetin in the extract is 45 - 70 wt%; the yield of the pure total flavonoids of Ampelopsis grossedentata is above 15.0% (the yield can increase with the improvement of the quality of old leaves and branches of Ampelopsis grossedentata), and the content of Ampelopsis grossedentata flavonoids in the extract is 40 - 65 wt%; the yield of the pure Ampelopsis grossedentata polysaccharide can be stably above 1.5% (the yield can increase with the improvement of the quality of old leaves and branches of Ampelopsis grossedentata), and the content of Ampelopsis grossedentata polysaccharide in the extract is 40 - 55 wt%.

[0038] The inventor of the present invention realized that there are losses in the flavonoids of Ampelopsis grossedentata extracted by the existing methods. By making full use of the chemical structural characteristics of dihydromyricetin, Ampelopsis grossedentata flavonoids and Ampelopsis grossedentata polysaccharides, an organic alcohol solvent that can dissolve dihydromyricetin and hydrophobic Ampelopsis grossedentata flavonoids is first used as the first extraction medium for extraction, and then pure water that can dissolve water-soluble Ampelopsis grossedentata flavonoids and Ampelopsis grossedentata polysaccharides is used as the second extraction medium for extraction. The extraction solutions are combined to obtain a crude extraction solution containing dihydromyricetin, Ampelopsis grossedentata flavonoids and Ampelopsis grossedentata polysaccharides, and then different treatment methods are used to separate dihydromyricetin, Ampelopsis grossedentata flavonoids and Ampelopsis grossedentata polysaccharides, so as to achieve the purpose of simultaneously extracting and separating dihydromyricetin, Ampelopsis grossedentata flavonoids and Ampelopsis grossedentata polysaccharides from old leaves and branches of Ampelopsis grossedentata.

[0039] The following are examples

[0040] Example 1 Simultaneous extraction and separation of dihydromyricetin, Ampelopsis grossedentata flavonoids and Ampelopsis grossedentata polysaccharides

[0041] In this example, old leaves and branches of Ampelopsis grossedentata are used as the extraction object, and the following extraction method is adopted to achieve simultaneous extraction and separation, and dihydromyricetin, Ampelopsis grossedentata flavonoids and Ampelopsis grossedentata polysaccharides are obtained in sequence. The specific extraction method is as follows:

[0042] (1) Simultaneous extraction

[0043] The old leaves and branches of Ampelopsis grossedentata are crushed and passed through a 10-mesh sieve. 1 kg of the powder of old leaves and branches of Ampelopsis grossedentata is taken, 20 kg of 30 wt% methanol (30 g of methanol and 70 g of water are contained in 100 g of the solution) is added, and it is heated to extract at a gentle boil for 1 h. It is filtered while it is hot to obtain the first filtrate and the first filter residue; 15 times of tap water is added to the obtained first filter residue, and it is extracted at a gentle boil for 2 h. It is filtered while it is hot to obtain the second filtrate and the second filter residue; the first filtrate and the second filtrate are mixed, and methanol is recovered by reduced pressure concentration. The concentration is stopped when the mixed solution is concentrated to half of the original volume, and it is left standing for 8 h, and then filtered to obtain filter residue A and filtrate B.

[0044] (2) Separation and preparation of dihydromyricetin, Ampelopsis grossedentata flavonoids and Ampelopsis grossedentata polysaccharides

[0045] The filter residue A was refluxed and dissolved with 5 times of acetone, and filtered to obtain the third filtrate and filter residue C. The third filtrate was concentrated under reduced pressure to recover acetone, and dried to obtain 180 g of dihydromyricetin. The content of dihydromyricetin was determined to be 65%. The pure product of dihydromyricetin can be obtained through purification treatment, and the yield was 11.7%.

[0046] The filter residue C was dissolved with hot water and mixed with the filtrate B, and the mixture was concentrated to a specific gravity of 1.08 (20 °C). N-butanol saturated with water was added in a volume ratio of 1:1, and stirred and extracted 3 times to obtain an organic solvent extract and a raffinate phase. The extraction liquid was combined and the n-butanol was recovered under reduced pressure. The solid obtained by vacuum drying was 133.5 g of crude myricetin of vine tea, and the content of myricetin of vine tea was measured to be 60.0%. The drying temperature of the vacuum drying oven was 60 °C, and it was dried until the mass of the solid was constant. The pure product of myricetin of vine tea can be obtained through purification treatment, and the yield was 8%.

[0047] 95% ethanol was added to the raffinate phase until the ethanol concentration was 60%, and left to stand at room temperature for 24 h to completely precipitate the polysaccharide. It was vacuum filtered, and the solid obtained by vacuum drying was 52.1 g of crude polysaccharide of vine tea. The content of polysaccharide of vine tea was measured to be 53.0%. The pure product of polysaccharide of vine tea can be obtained through purification treatment, and the yield was 2.76%. The drying temperature of the vacuum drying was 60 °C, and it was dried until the mass of the solid was constant.

[0048] Example 2

[0049] (1) Synchronous extraction

[0050] 1 kg of old leaves and branches of vine tea were crushed to 20 mesh, 20 times of 50 wt% ethanol was added, heated to a gentle boil and extracted for 1 h, and filtered while it was hot to obtain the first filtrate and the first filter residue. The first filter residue was added with 15 times of 50 wt% ethanol, heated to a gentle boil and extracted for 1 h, and filtered while it was hot to obtain the second filtrate and the second filter residue. The second filter residue was added with 15 times of tap water, gently boiled and extracted for 2 h, and filtered while it was hot to obtain the third filtrate. The first filtrate, the second filtrate and the third filtrate were combined, concentrated under reduced pressure to recover ethanol, and the concentration was stopped when the mixture was concentrated to half of the original volume. It was left to stand for 8 h, and filtered to obtain filter residue A and filtrate B.

[0051] (2) Separation and preparation of dihydromyricetin, myricetin of vine tea and polysaccharide of vine tea

[0052] The filter residue A was refluxed and dissolved with 5 times of acetone, filtered, and repeated 1 time. The filtrate and filter residue C were collected. The combined filtrate was concentrated under reduced pressure to recover acetone, and dried to obtain 200 g of dihydromyricetin. The content of dihydromyricetin was measured to be 70%. The pure product of dihydromyricetin can be obtained through purification treatment, and the yield was 14%.

[0053] The filter residue C is dissolved in hot water and mixed with the filtrate B, and the mixture is concentrated to a specific gravity of 1.10 (20 °C). Then, water-saturated n-butanol is added in a volume ratio of 1:1, and the mixture is stirred and extracted 3 times to obtain an organic solvent extract and a raffinate. The extract is combined and the n-butanol is recovered under reduced pressure. The solid is vacuum-dried to obtain 140 g of crude ampelopsis grossedentata flavonoids. The content of ampelopsis grossedentata flavonoids is measured to be 62%. The pure ampelopsis grossedentata flavonoids can be obtained through purification, and the yield is 8.68%. The vacuum drying temperature is 60 °C, and the drying is continued until the mass of the solid is constant.

[0054] 95% ethanol is added to the raffinate until the ethanol concentration reaches 70%. It is left standing at room temperature (for 24 h) or at a low temperature of 4 - 10 °C until no precipitate is formed, and then vacuum filtered. The solid is vacuum-dried to obtain 50 g of crude ampelopsis grossedentata polysaccharides. The content of ampelopsis grossedentata polysaccharides is measured to be 55.0%. The pure ampelopsis grossedentata polysaccharides can be obtained through purification, and the yield is 2.75%. The vacuum drying temperature is 60 °C, and the drying is continued until the mass of the solid is constant.

[0055] Example 3

[0056] (1) Synchronous extraction

[0057] 1 kg of old ampelopsis grossedentata leaves (without branches) is crushed to 20 mesh, and extracted with 20 times 30 wt% ethanol by heating to a gentle boil for 1 h. It is filtered while hot to obtain the first filtrate and the first filter residue. The first filter residue is added with 15 times 30 wt% ethanol, heated to a gentle boil for 1 h, and filtered while hot to obtain the second filtrate and the second filter residue. The second filter residue is added with 10 times tap water, heated to a gentle boil for 2 h, and filtered while hot to obtain the third filtrate. The first filtrate, the second filtrate, and the third filtrate are mixed, and the ethanol is recovered by reduced pressure concentration. The concentration is stopped when the filtrate is concentrated to half of the original volume, and it is left standing for 8 h, and then filtered to obtain filter residue A and filtrate B.

[0058] (2) Separation and preparation of dihydromyricetin, ampelopsis grossedentata flavonoids, and ampelopsis grossedentata polysaccharides

[0059] Among them, filter residue A is refluxed and dissolved with 5 times acetone, filtered, and the operation is repeated once. The filtrate and filter residue C are collected. The filtrate is combined and the acetone is recovered by reduced pressure concentration. The solid is dried to obtain 210 g of dihydromyricetin. The content of dihydromyricetin is measured to be 66%. The yield of pure dihydromyricetin is 14%. The drying temperature is 68 °C, and the drying is continued until the mass of the solid is constant.

[0060] The filter residue C is dissolved in hot water and mixed with the filtrate B, and the mixed filtrate is concentrated to a specific gravity of 1.10 (20 °C). Then, ethyl acetate is added in a volume ratio of 1:1, and the mixture is stirred and extracted 3 times to obtain an organic solvent extract and a raffinate. The extract is combined and the ethyl acetate is recovered under reduced pressure. The solid is vacuum-dried to obtain 140 g of crude ampelopsis grossedentata flavonoids. The content of ampelopsis grossedentata flavonoids is measured to be 62%. The yield of pure ampelopsis grossedentata flavonoids is 9%. The vacuum drying temperature is 68 °C, and the drying is continued until the mass of the solid is constant.

[0061] Add 95% ethanol to the raffinate phase until the ethanol concentration reaches 60%. Let it stand at room temperature for 24 h to precipitate the polysaccharide. Filter it with a cloth bag and hang it to filter. Dry the filter residue to obtain 52.1 g of crude ampelopsis grossedentata polysaccharide. The content of ampelopsis grossedentata polysaccharide is measured to be 53.0%, and the yield of pure ampelopsis grossedentata polysaccharide is 3%. The drying temperature is 68 °C, and it is dried until the mass of the solid matter is constant.

[0062] The crude dihydromyricetin, ampelopsis grossedentata flavonoids and ampelopsis grossedentata polysaccharide obtained in the present invention can also be purified according to the existing corresponding purification methods to improve the purity of dihydromyricetin, ampelopsis grossedentata flavonoids and ampelopsis grossedentata polysaccharide.

[0063] The above are only the preferred embodiments of the present invention, and do not impose any form of limitation on the present invention. Although the present invention has been disclosed above with the preferred embodiments, it is not intended to limit the present invention. Any person skilled in the art can make some changes or modifications to equivalent embodiments by using the technical content disclosed above without departing from the technical solution of the present invention. However, as long as it does not depart from the technical solution content of the present invention, any brief modifications, equivalent changes and modifications made to the above embodiments based on the technical essence of the present invention still fall within the scope of the technical solution of the present invention.

Claims

1. A method for synchronously extracting and separating multiple extracts of berry tea, characterized in that: The specific steps include: (1) Synchronous extraction: using old leaves and / or branches of berry tea as raw materials, first extracting with an organic solvent having an organic alcohol mass concentration of 10 to 60 wt% at room temperature or slightly boiling, filtering and collecting the filtrate and filter residue, extracting the filter residue with hot water, filtering, and combining the filtrate to obtain a crude extract containing dihydromyricetin, berry tea flavonoids and berry tea polysaccharides; (2) Separating and preparing dihydromyricetin, berry tea flavonoids and berry tea polysaccharides: recovering the solvent in the crude extract of step (1), concentrating, and separating to obtain a filter residue A and a filtrate B; using an organic solvent to dissolve dihydromyricetin in the filter residue A, filtering to obtain a filtrate and a filter residue C, recovering the organic solvent in the filtrate, and drying the solid to obtain dihydromyricetin; The filter residue C is dissolved in hot water and mixed with the filtrate B, and concentrated to a specific gravity of 1.05-1.15, and an organic extractant is added to the concentrated solution for extraction, and an extract containing berry tea flavonoids and a raffinate phase containing berry tea polysaccharides are separated to obtain the extract, the organic extractant in the extract is recovered, and the solid is dried to obtain berry tea flavonoids; the raffinate phase is subjected to an alcohol precipitation method to obtain berry tea polysaccharides.

2. The method according to claim 1, characterized in that In step (1), the raw material is first crushed into 10-20 meshes, and the organic solvent is added according to the mass ratio of the raw material to the organic solvent of 1:20, and the extraction is performed with slight boiling for 1-2 times, 1-2 hours per time.

3. The method according to claim 2, characterized in that The mass concentration of the organic alcohol in the organic solvent is 30wt% to 50wt%, and the extraction is performed by slight boiling for 1 to 2 times, 1 hour per time.

4. The method according to claim 3, characterized in that The organic alcohol is methanol or ethanol.

5. The method according to any one of claims 1 to 4, characterized in that: The hot water extraction is performed by adding water to the filter residue according to a mass ratio of raw material to pure water of 1:10-15, heating to a slight boiling point, and extracting 1-2 times, 1-2 hours per time.

6. The method according to claim 5, characterized in that In step (2), the filter residue A is repeatedly dissolved in acetone, and the filtrate is collected by filtration.

7. The method according to claim 6, characterized in that The concentration is vacuum concentration with a vacuum degree of 0.01-0.1 MPa and a temperature of 40-80° C., and the specific gravity of the concentrated solution at 20° C. is 1.05-1.

10.

8. The method according to claim 7, characterized in that The extraction comprises adding an organic extractant in a volume ratio of the concentrate to the organic extractant of 1:1 to 3, and performing extraction at room temperature or at a low temperature of 4 to 10°C for 1 to 3 times.

9. The method according to claim 8, characterized in that The organic extractant includes any one of n-butanol, ethyl acetate, n-pentanol or chloroform.

10. The method according to claim 9, characterized in that In the alcohol precipitation method, 95% ethanol is added to the raffinate phase to make the volume concentration of ethanol in the mixed solution 50-80%, the mixture is allowed to stand for 8-24 hours, filtered, and the filter residue is dried to obtain berry tea polysaccharide.