阅读说明：本技术 一种藤茶中二氢杨梅素的提取工艺及藤茶功能饮料的制备工艺 (Extraction process of dihydromyricetin in vine tea and preparation process of vine tea functional beverage ) 是由 谭乔 代传文 于 2021-08-25 设计创作，主要内容包括：本发明公开了一种藤茶中二氢杨梅素的提取工艺及藤茶功能饮料的制备工艺,其中浸提方法包括预处理、逆流浸提处理、滤渣的预处理、滤渣的提取及合并、黄酮类提取物的减压浓缩、结晶等步骤。所述浸提方法既能够使得对原材料的浸提充分,还能够使得对滤渣中残留的黄酮类物质的浸提更加彻底,保证了黄酮类物质的高效提取,浸提率高、黄酮含量高。同时,提取方法采用水浸提、超声浸提以及高压浸提相结合的方法,避免了乙醇等有机溶剂的使用带来的潜在污染。所述方法二氢杨梅素提取率高,含量高。(The invention discloses an extraction process of dihydromyricetin in vine tea and a preparation process of vine tea functional beverage, wherein the extraction method comprises the steps of pretreatment, countercurrent extraction treatment, pretreatment of filter residue, extraction and combination of the filter residue, reduced pressure concentration of flavonoid extract, crystallization and the like. The leaching method can fully leach the raw materials, can more thoroughly leach residual flavonoid substances in filter residues, ensures high-efficiency extraction of the flavonoid substances, and has high leaching rate and high flavonoid content. Meanwhile, the extraction method adopts a method combining water extraction, ultrasonic extraction and high-pressure extraction, so that potential pollution caused by the use of organic solvents such as ethanol and the like is avoided. The method has high extraction rate and high content of dihydromyricetin.)

1. A method for extracting dihydromyricetin from vine tea is characterized by comprising the following steps:

(1) pretreatment: drying and crushing the vine tea leaves for later use;

(2) low-temperature countercurrent leaching treatment: performing low-temperature countercurrent leaching on the tea leaves crushed in the step (1) to obtain filtrate and filter residues; wherein the countercurrent leaching solvent is water solution containing surfactant;

(3) pretreatment of filter residue: adding deionized water into the filter residue obtained in the step (2), and then carrying out ultrasonic treatment for 8-15 min at the temperature of 60 ℃ and the power of 80-200W;

(4) extracting filter residues: adding the filter residue pretreated in the step (3) and deionized water into a high-pressure container, and keeping the mixture for 8-15 min under 0.4-1.0 MPa to obtain secondary filtrate;

(5) combining the filtrate obtained in the step (2) with the secondary filtrate obtained in the step (4) to obtain the vine tea flavonoid extract;

(6) concentrating the flavonoid extract of the vine tea obtained in the step (5) under reduced pressure, standing and centrifuging to obtain a precipitate;

(7) adding water to the precipitate obtained in the step (6) for multiple times for recrystallization to obtain a crystal; and finally, drying the obtained crystal in vacuum to obtain the dihydromyricetin product.

2. The method for extracting dihydromyricetin from vine tea as claimed in claim 1, wherein the low-temperature countercurrent leaching temperature in the step (2) is room temperature, and the countercurrent leaching time is 15-30 min.

3. The method for extracting dihydromyricetin from Ampelopsis grossedentata as claimed in claim 1, wherein the surfactant in the step (2) is selected from a mixture of Tween 80 and sodium dodecyl sulfate at a mass ratio of 3: 1.

4. The method for extracting dihydromyricetin from Ampelopsis grossedentata as claimed in claim 2 or 3, wherein the concentration of the surfactant is 1.4 g/L.

5. The method for extracting dihydromyricetin from vine tea as claimed in claim 4, wherein the liquid-solid ratio of the low-temperature countercurrent leaching in the step (2) is 25 mL/g.

6. The method for extracting dihydromyricetin from vine tea as claimed in claim 5, wherein the mass ratio of the filter residue to the deionized water in the step (3) is 1:1 to 2.

7. The method for extracting dihydromyricetin from vine tea as claimed in claim 1, wherein the standing temperature in the step (6) is 4 ℃, and the standing time is 8-12 h.

8. The method for extracting dihydromyricetin from vine tea as claimed in claim 1, wherein the vacuum drying temperature in step (7) is 38 ℃.

9. Use of a vine tea flavonoid extract obtained by the extraction method of any one of claims 1-8 in the preparation of a functional beverage.

10. The use as claimed in claim 9, wherein the functional beverage comprises 40-100 parts of vine tea flavonoid extract, 10-60 parts of water, 0.08 part of beta-cyclodextrin and 0.1 part of sodium carboxymethylcellulose.

Technical Field

The invention belongs to the technical field of vine tea processing, and particularly relates to an extraction process of dihydromyricetin in vine tea and a preparation process of vine tea functional beverage.

Background

The Ampelopsis grossedentata is called Maoyan Qingshuang tea, Qingshuang ancient Ampelopsis grossedentata, Shenxian tea, dragon whiskers tea, etc. The vine tea is not only a pure natural green drink, but also a wonderful flower in tea with strong drug effect and health care performance, and contains abundant flavonoids. The flavonoids have the effects of protecting liver, treating chronic hepatitis and resisting oxidation, and also have the effects of preventing and treating hypertension and coronary heart disease, and reducing blood lipid and cholesterol.

At present, the main methods for extracting the flavonoid substances in the vine tea include a hot water extraction method, an ultrafiltration method, a dilute ethanol extraction method, an acetone extraction method, an ultrasonic extraction method, a microwave extraction method, an enzymolysis method and the like. For example, patent document CN101810735A adopts microwave radiation and alcohol extraction method to extract total flavonoids from ampelopsis grossedentata; patent document CN109692248A adopts a semi-bionic-biological enzyme extraction method to extract total flavonoids in ampelopsis grossedentata; patent document CN107307255A adopts cellulase and microwave assisted method to extract ampelopsis grossedentata to obtain selenium-rich granular beverage. As can be seen from the prior art, the conventional extraction method is mostly adopted for extracting the flavonoid substances in the vine tea, and the problems of low extraction rate and low content of total flavonoids exist; and the extraction method such as ethanol extraction method and acetone extraction method has the defect of solvent residue. The dihydromyricetin is the main component in the vine tea flavone extract, and the defects of the existing vine tea flavone extraction method also greatly restrict the subsequent extraction of the dihydromyricetin in the vine tea.

Disclosure of Invention

Aiming at the defects in the prior art, the invention provides an extraction process of dihydromyricetin in vine tea and a preparation process of vine tea functional beverage, wherein the extraction method adopts low-temperature countercurrent extraction, and a solvent adopts a surfactant, so that the extraction amount is greatly improved; the residual flavone substances in the filter residue are fully extracted by the combined use of ultrasonic and high-pressure extraction of the filter residue, so that the extraction efficiency of the dihydromyricetin is improved. The method is simple and has wide application prospect.

In order to achieve the purpose, the invention adopts the following technical scheme:

a method for extracting dihydromyricetin from Ampelopsis grossedentata comprises the following steps:

(1) pretreatment: drying and crushing the vine tea leaves for later use;

(2) and (3) countercurrent leaching treatment: performing low-temperature countercurrent leaching on the tea leaves crushed in the step (1) to obtain filtrate and filter residues; wherein the countercurrent leaching solvent is water solution containing surfactant;

(3) pretreatment of filter residue: adding deionized water into the filter residue obtained in the step (2), and then carrying out ultrasonic treatment for 8-15 min at the temperature of 60 ℃ and the power of 80-200W;

(4) extracting filter residues: adding the filter residue pretreated in the step (3) and deionized water into a high-pressure container, and keeping the mixture for 8-15 min under 0.4-1.0 MPa to obtain secondary filtrate;

(5) combining the filtrate obtained in the step (2) with the secondary filtrate obtained in the step (4) to obtain the vine tea flavonoid extract;

(6) concentrating the flavonoid extract of the vine tea obtained in the step (5) under reduced pressure, standing and centrifuging to obtain a precipitate;

(7) adding water to the precipitate obtained in the step (6) for multiple times for recrystallization to obtain a crystal; and finally, drying the obtained crystal in vacuum to obtain the dihydromyricetin product.

Preferably, the low-temperature countercurrent leaching temperature in the step (2) is room temperature, and the countercurrent leaching time is 15-30 min.

Preferably, in the step (2), the surfactant is selected from a mixture of tween 80 and sodium dodecyl sulfate in a mass ratio of 3: 1.

Preferably, the concentration of the surfactant is 1.4 g/L.

Preferably, the liquid-solid ratio of the low-temperature countercurrent leaching in the step (2) is 25 mL/g.

Preferably, the mass ratio of the filter residue to the deionized water in the step (3) is 1:1 to 2.

Preferably, the standing temperature in the step (6) is 4 ℃, and the standing time is 8-12 h.

Preferably, the vacuum drying temperature in step (7) is 38 ℃.

The invention also claims application of the vine tea flavonoid extract prepared by the method in preparing functional beverages.

More preferably, the raw materials of the functional beverage comprise 40-100 parts of vine tea flavonoid extract, 10-60 parts of water, 0.08 part of beta-cyclodextrin and 0.1 part of sodium carboxymethylcellulose.

Compared with the prior art, the invention has the following beneficial effects:

(1) the extraction method of dihydromyricetin in vine tea provided by the invention adopts a method combining water extraction, ultrasonic extraction and high-pressure extraction, and avoids potential pollution caused by the use of organic solvents such as ethanol and the like.

(2) The vine tea is subjected to low-temperature countercurrent leaching treatment, and the thought of countercurrent heat exchange in the field of heat exchangers is adopted, so that flavonoids can be fully leached, and the leaching efficiency is greatly improved; meanwhile, the applicant unexpectedly finds that the countercurrent leaching solvent adopts the aqueous solution containing the surfactant, so that the high-efficiency extraction at room temperature can be realized, and the defects of heating leaching in the existing leaching process can be avoided. In the aspect of selecting specific surfactant substances, the applicant conducts a plurality of screens, and finally finds that the better extraction efficiency can be realized by the mixed use of tween 80 and sodium dodecyl sulfate, the tween 80 is a nonionic surfactant, and the sodium dodecyl sulfate is an anionic surfactant, and the tween 80 and the sodium dodecyl sulfate are matched with each other to achieve a good extraction effect cooperatively. Meanwhile, the two substances are safe and edible, and the use requirement of directly preparing the functional beverage from the intermediate product flavone extract is met.

(3) According to the invention, for the treatment of the filter residue, the extraction difficulty of flavonoid substances in the filter residue is increased due to the efficient extraction of the flavonoid substances in the countercurrent extraction stage, so that the combined use method of firstly performing ultrasonic treatment and then performing high-pressure extraction is provided, the components in the filter residue can be preliminarily extracted by firstly performing simple ultrasonic treatment, and then the flavonoid substances can be fully dissolved out by adopting high-pressure treatment, so that the extraction rate is higher and the efficiency is better by adopting the combined extraction method.

In a word, the extraction method can fully extract the raw materials, can more thoroughly extract the residual flavonoid substances in the filter residue, ensures the high-efficiency extraction of the flavonoid substances, and has high extraction rate (more than 60 percent) and high flavonoid content (more than 27 percent); meanwhile, the yield of the dihydromyricetin product is up to 18 percent, and the content of the dihydromyricetin in the product is more than or equal to 86 percent.

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail 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. In addition, the technical features involved in the embodiments of the present invention described below may be combined with each other as long as they do not conflict with each other.

It should be noted that tween 80 and sodium lauryl sulfate used in the present invention are commercially available.

Example 1

A method for extracting dihydromyricetin from Ampelopsis grossedentata comprises the following steps:

(1) pretreatment: drying and crushing 1kg of vine tea leaves for later use;

(2) and (3) countercurrent leaching treatment: carrying out low-temperature countercurrent extraction on the tea leaves crushed in the step (1) in low-temperature countercurrent equipment, wherein the extraction temperature is room temperature, the extraction time is 15min, the extraction solvent is an aqueous solution containing 1.4g/L of surfactant (a mixture of Tween 80 and sodium dodecyl sulfate in a mass ratio of 3: 1), the solid ratio of the extraction solution is 25mL/g, and after the extraction is finished, filtrate and filter residues are obtained;

(3) pretreatment of filter residue: adding deionized water into the filter residue obtained in the step (2) according to the mass ratio of 1:1, and then carrying out ultrasonic treatment for 10min at 60 ℃ under the conditions of 40khz and 150W;

(4) extracting filter residues: transferring the filter residue pretreated in the step (3) and water into a high-pressure container, and keeping the high-pressure container for 8min under 0.5MPa to obtain secondary filtrate;

(5) mixing the filtrate obtained in the step (2) with the secondary filtrate obtained in the step (4) to obtain the vine tea flavonoid extract;

(6) concentrating the flavonoid extract of Ampelopsis grossedentata obtained in the step (5) under reduced pressure to 1/3 volume, standing at 4 ℃ for 8h, and centrifuging at 3800rpm for 18min to obtain precipitate;

(7) adding water to the precipitate obtained in the step (6) for 3 times for recrystallization to obtain a crystal; and finally, carrying out vacuum drying on the obtained crystal at 38 ℃ to obtain the dihydromyricetin product.

Example 2

A method for extracting dihydromyricetin from Ampelopsis grossedentata comprises the following steps:

(1) pretreatment: drying and crushing 1kg of vine tea leaves for later use;

(2) and (3) countercurrent leaching treatment: carrying out low-temperature countercurrent extraction on the tea leaves crushed in the step (1) in low-temperature countercurrent equipment, wherein the extraction temperature is room temperature, the extraction time is 18min, the extraction solvent is a mixture containing 1.4g/L of surfactant (Tween 80 and sodium dodecyl sulfate in a mass ratio of 3: 1), the solid ratio of the extraction liquid is 25mL/g, and after the extraction is finished, filtrate and filter residues are obtained;

(3) pretreatment of filter residue: adding deionized water into the filter residue obtained in the step (2) according to the mass ratio of 1:1.5, and then carrying out ultrasonic treatment for 8min at the temperature of 60 ℃, 40khz and 180W;

(4) extracting filter residues: transferring the filter residue pretreated in the step (3) and water into a high-pressure container, and keeping the high-pressure container for 8min under 0.6MPa to obtain secondary filtrate;

(5) mixing the filtrate obtained in the step (2) with the secondary filtrate obtained in the step (4) to obtain the vine tea flavonoid extract;

(6) concentrating the flavonoid extract of Ampelopsis grossedentata obtained in the step (5) under reduced pressure to 1/3 volume, standing at 4 ℃ for 10h, and centrifuging at 3800rpm for 20min to obtain precipitate;

(7) adding water to the precipitate obtained in the step (6) for 3 times for recrystallization to obtain a crystal; and finally, carrying out vacuum drying on the obtained crystal at 38 ℃ to obtain the dihydromyricetin product.

Example 3

A method for extracting dihydromyricetin from Ampelopsis grossedentata comprises the following steps:

(1) pretreatment: drying and crushing 1kg of vine tea leaves for later use;

(2) and (3) countercurrent leaching treatment: carrying out low-temperature countercurrent extraction on the tea leaves crushed in the step (1) in low-temperature countercurrent equipment, wherein the extraction temperature is room temperature, the extraction time is 20min, the extraction solvent is a mixture containing 1.4g/L of surfactant (Tween 80 and sodium dodecyl sulfate in a mass ratio of 3: 1), the solid ratio of the extraction liquid is 25mL/g, and after the extraction is finished, filtrate and filter residues are obtained;

(3) pretreatment of filter residue: adding deionized water into the filter residue obtained in the step (2) according to the mass ratio of 1:1, and then carrying out ultrasonic treatment for 8min at 60 ℃, 40khz and 200W;

(4) extracting filter residues: transferring the filter residue pretreated in the step (3) and water into a high-pressure container, and keeping the high-pressure container for 10min under 0.5MPa to obtain secondary filtrate;

(5) mixing the filtrate obtained in the step (2) with the secondary filtrate obtained in the step (4) to obtain the vine tea flavonoid extract;

(6) concentrating the flavonoid extract of Ampelopsis grossedentata obtained in the step (5) under reduced pressure to 1/3 volume, standing at 4 ℃ for 10h, and centrifuging at 3800rpm for 18min to obtain precipitate;

(7) adding water to the precipitate obtained in the step (6) for 2 times for recrystallization to obtain a crystal; and finally, carrying out vacuum drying on the obtained crystal at 38 ℃ to obtain the dihydromyricetin product.

Example 4

A vine tea functional beverage comprises the following raw materials: 80g of the flavonoid extract obtained in the example 1, 50g of water, 0.08g of beta-cyclodextrin and 0.1g of sodium carboxymethyl cellulose, and the raw materials are uniformly mixed under sterile conditions at normal temperature, sterilized and packaged to obtain a beverage product.

Comparative example 1

A method for extracting dihydromyricetin from Ampelopsis grossedentata comprises the following steps:

(1) pretreatment: drying and crushing 1kg of vine tea leaves for later use;

(2) and (3) countercurrent leaching treatment: carrying out low-temperature countercurrent extraction on the tea leaves crushed in the step (1) in low-temperature countercurrent equipment, wherein the extraction temperature is room temperature, the extraction time is 15min, an extraction solvent is an aqueous solution, the solid ratio of an extraction solution is 25mL/g, and after the extraction is finished, filtrate and filter residues are obtained;

(3) pretreatment of filter residue: adding deionized water into the filter residue obtained in the step (2) according to the mass ratio of 1:1, and then carrying out ultrasonic treatment for 10min at 60 ℃ under the conditions of 40khz and 150W;

(4) extracting filter residues: transferring the filter residue pretreated in the step (3) and water into a high-pressure container, and keeping the high-pressure container for 8min under 0.5MPa to obtain secondary filtrate;

(5) mixing the filtrate obtained in the step (2) with the secondary filtrate obtained in the step (4) to obtain the vine tea flavonoid extract;

(6) concentrating the flavonoid extract of Ampelopsis grossedentata obtained in the step (5) under reduced pressure to 1/3 volume, standing at 4 ℃ for 8h, and centrifuging at 3800rpm for 18min to obtain precipitate;

(7) adding water to the precipitate obtained in the step (6) for 3 times for recrystallization to obtain a crystal; and finally, carrying out vacuum drying on the obtained crystal at 38 ℃ to obtain the dihydromyricetin product.

Comparative example 2

A method for extracting dihydromyricetin from Ampelopsis grossedentata comprises the following steps:

(1) pretreatment: drying and crushing 1kg of vine tea leaves for later use;

(2) and (3) countercurrent leaching treatment: carrying out low-temperature countercurrent leaching on the tea leaves crushed in the step (1) in low-temperature countercurrent equipment, wherein the leaching temperature is room temperature, the leaching time is 15min, the leaching solvent is an aqueous solution containing 1.4g/L of surfactant Tween 80, the solid ratio of the leaching solution is 25mL/g, and after leaching is finished, filtrate and filter residues are obtained;

(3) pretreatment of filter residue: adding deionized water into the filter residue obtained in the step (2) according to the mass ratio of 1:1, and then carrying out ultrasonic treatment for 10min at 60 ℃ under the conditions of 40khz and 150W;

(4) extracting filter residues: transferring the filter residue pretreated in the step (3) and water into a high-pressure container, and keeping the high-pressure container for 8min under 0.5MPa to obtain secondary filtrate;

(5) mixing the filtrate obtained in the step (2) with the secondary filtrate obtained in the step (4) to obtain the vine tea flavonoid extract;

(6) concentrating the flavonoid extract of Ampelopsis grossedentata obtained in the step (5) under reduced pressure to 1/3 volume, standing at 4 ℃ for 8h, and centrifuging at 3800rpm for 18min to obtain precipitate;

(7) adding water to the precipitate obtained in the step (6) for 3 times for recrystallization to obtain a crystal; and finally, carrying out vacuum drying on the obtained crystal at 38 ℃ to obtain the dihydromyricetin product.

Comparative example 3

A method for extracting dihydromyricetin from Ampelopsis grossedentata comprises the following steps:

(1) pretreatment: drying and crushing 1kg of vine tea leaves for later use;

(2) and (3) countercurrent leaching treatment: carrying out low-temperature countercurrent extraction on the tea leaves crushed in the step (1) in low-temperature countercurrent equipment, wherein the extraction temperature is room temperature, the extraction time is 15min, the extraction solvent is an aqueous solution containing 1.4g/L of surfactant sodium dodecyl sulfate, the solid ratio of the extraction solution is 25mL/g, and after the extraction is finished, filtrate and filter residues are obtained;

(3) pretreatment of filter residue: adding deionized water into the filter residue obtained in the step (2) according to the mass ratio of 1:1, and then carrying out ultrasonic treatment for 10min at 60 ℃ under the conditions of 40khz and 150W;

(4) extracting filter residues: transferring the filter residue pretreated in the step (3) and water into a high-pressure container, and keeping the high-pressure container for 8min under 0.5MPa to obtain secondary filtrate;

(5) mixing the filtrate obtained in the step (2) with the secondary filtrate obtained in the step (4) to obtain the vine tea flavonoid extract;

(6) concentrating the flavonoid extract of Ampelopsis grossedentata obtained in the step (5) under reduced pressure to 1/3 volume, standing at 4 ℃ for 8h, and centrifuging at 3800rpm for 18min to obtain precipitate;

(7) adding water to the precipitate obtained in the step (6) for 3 times for recrystallization to obtain a crystal; and finally, carrying out vacuum drying on the obtained crystal at 38 ℃ to obtain the dihydromyricetin product.

The extraction rate and the total flavone extraction rate in examples 1 to 3 and comparative examples 1 to 3 were calculated respectively according to the following formula, and the calculation results are summarized in table 1.

The extraction rate (g/g) is the extract amount/crude drug amount multiplied by 100 percent;

the total flavone content (g/g) is the total flavone amount/crude drug amount x 100%.

Wherein, the total flavone content is measured by ultraviolet spectrophotometry, and the model of the spectrophotometer is UV-1800 produced by Shanghai precision instruments and meters.

TABLE 1

	Extraction ratio/%	Total flavone content/%)
			Example 1	61.35	28.37
Example 2	62.08	27.96
			Example 3	61.82	28.05
Comparative example 1	39.82	18.26
			Comparative example 2	52.63	24.55
Comparative example 3	49.05	23.01

And (3) respectively taking dihydromyricetin as a standard sample, determining the content of the dihydromyricetin by adopting an aluminum nitrate-sodium nitrite colorimetric method, and simultaneously calculating the yield of the dihydromyricetin. The results are shown in Table 2.

TABLE 2

	Yield/% of dihydromyricetin	Content of dihydromyricetin/%)
			Example 1	18.63	86.3
Example 2	18.25	86.5
			Example 3	18.46	86.0
Comparative example 1	8.35	83.8
			Comparative example 2	11.85	85.4
Comparative example 3	10.08	85.0

It will be understood by those skilled in the art that the foregoing is only a preferred embodiment of the present invention, and is not intended to limit the invention, and that any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the scope of the present invention.