CN116138382A - Method for preparing plant beverage by vacuum radio frequency assisted biological enzymolysis - Google Patents

Method for preparing plant beverage by vacuum radio frequency assisted biological enzymolysis Download PDF

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CN116138382A
CN116138382A CN202211710561.1A CN202211710561A CN116138382A CN 116138382 A CN116138382 A CN 116138382A CN 202211710561 A CN202211710561 A CN 202211710561A CN 116138382 A CN116138382 A CN 116138382A
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chinese medicine
traditional chinese
radio frequency
fermentation
steaming
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温青玉
孙东弦
乔曙光
张立涛
李斐
王洁
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Henan Ankang Future Food Technology Co ltd
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Henan Ankang Future Food Technology Co ltd
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
    • A23L2/38—Other non-alcoholic beverages
    • A23L2/382—Other non-alcoholic beverages fermented
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
    • A23L2/70—Clarifying or fining of non-alcoholic beverages; Removing unwanted matter
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
    • A23L2/70—Clarifying or fining of non-alcoholic beverages; Removing unwanted matter
    • A23L2/84—Clarifying or fining of non-alcoholic beverages; Removing unwanted matter using microorganisms or biological material, e.g. enzymes
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
    • A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
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    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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Abstract

The invention belongs to the technical field of food biology, and discloses a method for preparing a plant drink by vacuum radio frequency assisted biological enzymolysis, which comprises the following steps: mixing and crushing the dried traditional Chinese medicine raw materials to obtain traditional Chinese medicine mixed raw materials; uniformly mixing the traditional Chinese medicine mixed raw materials with pure water according to a proportion, heating by three sections of steam, cooling to room temperature, adding cellulose amylase for enzymolysis, and then carrying out auxiliary extraction by a vacuum radio frequency method to obtain a traditional Chinese medicine mixture; adding Eurotium cristatum and monascus into the traditional Chinese medicine mixture for fermentation to obtain fermentation liquor; centrifuging the fermentation liquor, clarifying the supernatant, and concentrating by reduced pressure evaporation to obtain extract; adding pure water into the extract, then adding honey, and fully stirring and uniformly mixing; and filling the obtained mixture, steaming, sterilizing and packaging to obtain the plant beverage. The vacuum radio frequency assisted biological enzymolysis technology is adopted, and meanwhile, active functional substances can be effectively prevented from being oxidized under the vacuum condition, so that the extraction rate is improved, and the obtained plant drink has strong functionality and high use value.

Description

Method for preparing plant beverage by vacuum radio frequency assisted biological enzymolysis
Technical Field
The invention belongs to the technical field of food biology, and relates to a method for preparing a plant drink by vacuum radio frequency assisted biological enzymolysis.
Background
Alcoholism and alcoholism can seriously affect physical and mental health and home and social stability. Acute consumption of alcohol can cause nausea, vomiting, hypomnesis, inattention, impaired fine motor ability and emotional instability, and in severe cases can be fatal due to paralysis of respiratory muscles. Long-term drinking can cause mental disorder, gastric ulcer, fatty liver, alcoholic hepatitis, liver cirrhosis and other diseases. About 80% of the ethanol in the body is metabolized by the liver, the ethanol is converted into acetaldehyde by alcohol dehydrogenase, the acetaldehyde is converted into acetic acid by acetaldehyde dehydrogenase, the acetic acid enters the citric acid cycle, and finally, the acetic acid is converted into water and carbon dioxide to be discharged out of the body. In the process, the alcohol dehydrogenase and the acetaldehyde dehydrogenase are used as two key enzyme systems in the anti-alcohol factors and are regulated and controlled by isoflavone compounds. When a large amount of alcohol is taken into the organism, the contents of alcohol dehydrogenase, glutathione peroxidase and the like in the liver are obviously reduced, so that the liver is directly damaged, and therefore alcohol dehydrogenase, glutathione, superoxide dismutase and the like are often used as detection indexes, and the alcohol dispelling and liver protecting functions of the product are verified. And secondly, malondialdehyde (MDA) is reactive aldehyde released in the lipid peroxidation reaction process, is a final product of the lipid peroxidation reaction, can cause cell necrosis, and the content of the reactive aldehyde can reflect the degree of liver injury.
There are various plant beverages with the functions of dispelling the effects of alcohol and protecting liver developed by taking kudzuvine root, hovenia dulcis thunb and the like as main raw materials on the market, and the plant beverages contain a large amount of carbohydrates, so that sugar can be rapidly supplemented in a short time, and symptoms of weakness of hands and feet, nausea and vomiting after excessive drinking can be relieved. Most of the anti-alcoholic products contain certain functional components (isoflavone compounds such as puerarin and daidzin), but the main active substances of the anti-alcoholic products have the problems of low extraction rate and easy oxidation. Therefore, there is a need to improve the preparation method of traditional Chinese medicine anti-alcohol products, increase the extraction rate of active substances and increase the functionality of the anti-alcohol products.
Disclosure of Invention
Aiming at the problems of low extraction rate and easy oxidization of active substances in the existing anti-alcohol products, the invention provides a method for preparing a plant beverage by vacuum radio frequency assisted biological enzymolysis.
In order to achieve the above purpose, the present invention adopts the following technical scheme:
the invention provides a method for preparing a plant drink by vacuum radio frequency assisted biological enzymolysis, which comprises the following steps:
1) Cleaning the fresh medicinal materials with pure water, and then naturally air-drying or airing;
2) Weighing the dried traditional Chinese medicine raw materials according to the following weight portions: 60-70 parts of kudzuvine root, 4-10 parts of hovenia dulcis thunb, 6-8 parts of cassia seed, 4-6 parts of chrysanthemum and 3-5 parts of liquorice, and mixing and crushing the materials to obtain a traditional Chinese medicine mixed raw material with the size of 0.5-1 cm;
3) Mixing the traditional Chinese medicine raw materials with pure water according to the following ratio of 1:25, heating for 60min by three sections of steam, cooling to room temperature, adding cellulose amylase, performing enzymolysis in water bath at 50deg.C for 2 hr, wherein the addition amount of the enzyme is 1% of the total amount of the Chinese medicinal materials, and performing vacuum radio frequency assisted extraction to obtain Chinese medicinal mixture;
4) Adding Eurotium cristatum and monascus into the traditional Chinese medicine mixture in the step 3) for fermentation, wherein the fermentation temperature is 28-37 ℃ and the fermentation time is 48 hours, so as to obtain fermentation liquor; the addition amount of the Eurotium cristatum and the monascus is 0.75% of the total amount of the traditional Chinese medicine mixture;
5) Centrifuging the fermentation liquor, clarifying the supernatant, and concentrating by reduced pressure evaporation to obtain extract;
6) Adding pure water to the extract until the concentration is 10%, then adding Mel, and stirring thoroughly; the weight ratio of the honey to the concentrated solution is 1:10;
7) Filling the mixture obtained in the step 6) into a 50ml brown bottle, steaming and sterilizing for 60min, and packaging to obtain the plant beverage.
In one technical scheme, the specific process of heating three sections of steam in the step 3) for 60min is as follows: heating at 80deg.C for 20min, then at 120deg.C for 30min, and finally at 100deg.C for 10min.
In one technical scheme, the distance between the radio frequency plates of the vacuum radio frequency method in the step 3) is 114mm, and the extraction time is 320s.
In one embodiment, the clarification in step 5) is performed with chitosan or gelatin, or a mixture of chitosan and gelatin.
In one embodiment, the concentration temperature in the step 5) is 55 ℃ and the density of the concentrated extract is 1.32.
In one technical scheme, the specific process of steaming and sterilizing for 60min in the step 7) is as follows: steaming at 90deg.C for 20min, steaming at 100deg.C for 30min, and steaming at 80deg.C for 10min.
Compared with the prior art, the invention has the beneficial effects that:
the invention has the advantages that: (1) The raw materials are reasonable in design, convenient to manufacture and use, made of pure Chinese herbal medicines, free of other chemical additives, free of preservative in cooking sterilization and side effects, capable of manufacturing the functional plant beverage with good sense of export and rich nutrition, and good in popularization prospect and commercial medical value. (2) The vacuum radio frequency assisted biological enzymolysis technology is adopted, an electromagnetic field penetrates through the raw materials to damage plant cell walls, polar molecules in the raw materials are caused to move in a polarized mode and migrate through ion oscillation, active functional substances are easier to separate out, and meanwhile active functional substances such as flavonoid compounds can be effectively prevented from being oxidized under the vacuum condition, so that the extraction rate of the active functional substances is improved. (3) The microbial fermentation method is adopted to ferment the extracting stock solution, so that the activity of alcohol dehydrogenase is improved. (4) the invention is safe and has no toxic or side effect. And carrying out HepG2 cytotoxicity experimental study, and calculating according to a formula y= -0.188x+0.993 to prove that the half lethal dose is 2.62g/g, and 17L of the product is required to be taken for killing 65kg of people. The safety of the product is proved. And (5) the anti-alcohol effect of the invention is obvious. Oxidative stress is known to be an important mechanism for the development of alcoholic liver injury, and glutathione, superoxide dismutase and catalase all have antioxidant effects. By establishing a damage model group, the activity change rates of glutathione (> 35%), superoxide dismutase (> 40%) and catalase (> 12.5%) of the experimental group are proved to be far higher than that of the model group (< 0%); compared with a model group (80%), the content of malondialdehyde (< 5%) in the experimental group is obviously reduced, and the results show that the invention has good antioxidant capacity and can effectively relieve the body injury caused by alcohol. (6) The method of adding gelatin and chitosan clarifier for clarification is adopted, so that the activity of various active ingredients of the traditional Chinese medicine is greatly maintained, the loss of functional ingredients is reduced, and the nutritional value of the traditional Chinese medicine is maintained.
Drawings
FIG. 1 shows the effect of plant drink on intracellular MDA content and activity of SOD, GSH-Px and CAT in example 9.
Detailed Description
The following examples are illustrative of the present invention and are not intended to limit the scope of the invention. The technical means used in the examples are conventional means well known to those skilled in the art unless otherwise indicated. The test methods in the following examples are conventional methods unless otherwise specified.
Example 1
1) Cleaning the fresh medicinal materials with pure water, and then naturally air-drying or airing;
2) Weighing the dried traditional Chinese medicine raw materials according to the following weight: 60g of kudzuvine root, 4g of hovenia dulcis thunb, 6g of cassia seed, 4g of chrysanthemum and 3g of liquorice, and mixing and crushing the materials to obtain a traditional Chinese medicine mixed raw material with the size of 0.5-1 cm;
3) Mixing the traditional Chinese medicine raw materials with pure water according to the following ratio of 1:25, heating with steam at 80deg.C for 20min, heating with steam at 120deg.C for 30min, heating with steam at 100deg.C for 10min, cooling to room temperature, adding 0.77g cellulose amylase, performing enzymolysis in water bath at 50deg.C for 2 hr, setting the distance between radio frequency polar plates to 114mm, and extracting for 320s to obtain the Chinese medicinal mixture;
4) Adding 0.58g of Eurotium cristatum and Monascus purpureus into 77g of the traditional Chinese medicine mixture in the step 3) for fermentation, wherein the fermentation temperature is 37 ℃ and the fermentation time is 48 hours, so as to obtain fermentation liquor;
5) Centrifuging the fermentation liquor, adding 1% chitosan solution into the supernatant for clarification, and concentrating at 55deg.C under reduced pressure until the density of the extract is 1.32 to obtain extract;
6) Adding pure water to the extract until the concentration is 10%, then adding Mel, and stirring thoroughly; the weight ratio of the honey to the concentrated solution is 1:10;
7) Filling the mixture obtained in the step 6) into a 50ml brown bottle, steaming and sterilizing for 20min at 90 ℃, steaming and sterilizing for 30min at 100 ℃, steaming and sterilizing for 10min at 80 ℃, and packaging to obtain the plant beverage.
Example 2
1) Cleaning the fresh medicinal materials with pure water, and then naturally air-drying or airing;
2) Weighing the dried traditional Chinese medicine raw materials according to the following weight: 60g of kudzuvine root, 4g of hovenia dulcis thunb, 6g of cassia seed, 4g of chrysanthemum and 3g of liquorice, and mixing and crushing the materials to obtain a traditional Chinese medicine mixed raw material with the size of 0.5-1 cm;
3) Mixing the traditional Chinese medicine raw materials with pure water according to the following ratio of 1:25, heating with steam at 80deg.C for 20min, heating with steam at 120deg.C for 30min, heating with steam at 100deg.C for 10min, cooling to room temperature, adding 0.77g cellulose amylase, performing enzymolysis in water bath at 50deg.C for 2 hr, setting the distance between radio frequency polar plates to 114mm, and extracting for 320s to obtain the Chinese medicinal mixture;
4) Adding 0.58g of Eurotium cristatum and Monascus purpureus into 77g of the traditional Chinese medicine mixture in the step 3) for fermentation, wherein the fermentation temperature is 28 ℃, and the fermentation time is 48 hours, so as to obtain fermentation liquor;
5) Centrifuging the fermentation liquor, adding 1% chitosan solution into the supernatant for clarification, and concentrating at 55deg.C under reduced pressure until the density of the extract is 1.32 to obtain extract;
6) Adding pure water to the extract until the concentration is 10%, then adding Mel, and stirring thoroughly; the weight ratio of the honey to the concentrated solution is 1:10;
7) Filling the mixture obtained in the step 6) into a 50ml brown bottle, steaming and sterilizing for 20min at 90 ℃, steaming and sterilizing for 30min at 100 ℃, steaming and sterilizing for 10min at 80 ℃, and packaging to obtain the plant beverage.
Example 3
1) Cleaning the fresh medicinal materials with pure water, and then naturally air-drying or airing;
2) Weighing the dried traditional Chinese medicine raw materials according to the following weight: 60g of kudzuvine root, 4g of hovenia dulcis thunb, 6g of cassia seed, 4g of chrysanthemum and 3g of liquorice, and mixing and crushing the materials to obtain a traditional Chinese medicine mixed raw material with the size of 0.5-1 cm;
3) Mixing the traditional Chinese medicine raw materials with pure water according to the following ratio of 1:25, heating with steam at 80deg.C for 20min, heating with steam at 120deg.C for 30min, heating with steam at 100deg.C for 10min, cooling to room temperature, adding 0.77g cellulose amylase, performing enzymolysis in water bath at 50deg.C for 2 hr, setting the distance between radio frequency polar plates to 114mm, and extracting for 320s to obtain the Chinese medicinal mixture;
4) Adding 0.58g of Eurotium cristatum and Monascus purpureus into 77g of the traditional Chinese medicine mixture in the step 3) for fermentation, wherein the fermentation temperature is 37 ℃ and the fermentation time is 48 hours, so as to obtain fermentation liquor;
5) Centrifuging the fermentation liquid, adding 1% gelatin solution into the supernatant for clarification, and concentrating at 55deg.C under reduced pressure to obtain extract with density of 1.32;
6) Adding pure water to the extract until the concentration is 10%, then adding Mel, and stirring thoroughly; the weight ratio of the honey to the concentrated solution is 1:10;
7) Filling the mixture obtained in the step 6) into a 50ml brown bottle, steaming and sterilizing for 20min at 90 ℃, steaming and sterilizing for 30min at 100 ℃, steaming and sterilizing for 10min at 80 ℃, and packaging to obtain the plant beverage.
Example 4
1) Cleaning the fresh medicinal materials with pure water, and then naturally air-drying or airing;
2) Weighing the dried traditional Chinese medicine raw materials according to the following weight: 60g of kudzuvine root, 4g of hovenia dulcis thunb, 6g of cassia seed, 4g of chrysanthemum and 3g of liquorice, and mixing and crushing the materials to obtain a traditional Chinese medicine mixed raw material with the size of 0.5-1 cm;
3) Mixing the traditional Chinese medicine raw materials with pure water according to the following ratio of 1:25, heating with steam at 80deg.C for 20min, heating with steam at 120deg.C for 30min, heating with steam at 100deg.C for 10min, cooling to room temperature, adding 0.77g cellulose amylase, performing enzymolysis in water bath at 50deg.C for 2 hr, setting the distance between radio frequency polar plates to 114mm, and extracting for 320s to obtain the Chinese medicinal mixture;
4) Adding 0.58g of Eurotium cristatum and Monascus purpureus into 77g of the traditional Chinese medicine mixture in the step 3) for fermentation, wherein the fermentation temperature is 37 ℃ and the fermentation time is 48 hours, so as to obtain fermentation liquor;
5) Centrifuging the fermentation liquid, adding 1% gelatin solution and 1% chitosan solution into the supernatant for clarification, and concentrating at 55deg.C under reduced pressure to obtain extract with density of 1.32;
6) Adding pure water to the extract until the concentration is 10%, then adding Mel, and stirring thoroughly; the weight ratio of the honey to the concentrated solution is 1:10;
7) Filling the mixture obtained in the step 6) into a 50ml brown bottle, steaming and sterilizing for 20min at 90 ℃, steaming and sterilizing for 30min at 100 ℃, steaming and sterilizing for 10min at 80 ℃, and packaging to obtain the plant beverage.
The plant drink of example 4 was subjected to assay test results, and each index is shown in table 1.
TABLE 1 organoleptic, physicochemical, and microbial indicators of example 4 plant beverage
Figure BDA0004027246350000051
Figure BDA0004027246350000061
Example 5
1) Cleaning the fresh medicinal materials with pure water, and then naturally air-drying or airing;
weighing the dried traditional Chinese medicine raw materials according to the following weight: 60g of kudzuvine root, 4g of hovenia dulcis thunb, 6g of cassia seed, 4g of chrysanthemum and 3g of liquorice, and mixing and crushing the materials to obtain a traditional Chinese medicine mixed raw material with the size of 0.5-1 cm;
3) Mixing the traditional Chinese medicine raw materials with pure water according to the following ratio of 1:25, heating with steam at 80deg.C for 20min, heating with steam at 120deg.C for 30min, heating with steam at 100deg.C for 10min, cooling to room temperature, adding 0.77g cellulose amylase, performing enzymolysis in water bath at 50deg.C for 2 hr, setting the distance between radio frequency polar plates to 114mm, and extracting for 320s to obtain the Chinese medicinal mixture;
4) Adding 0.58g of Eurotium cristatum into 77g of the traditional Chinese medicine mixture in the step 3) for fermentation, wherein the fermentation temperature is 37 ℃ and the fermentation time is 48 hours, so as to obtain fermentation liquor;
5) Centrifuging the fermentation liquid, adding 1% gelatin solution and 1% chitosan solution into the supernatant for clarification, and concentrating at 55deg.C under reduced pressure to obtain extract with density of 1.32;
6) Adding pure water to the extract until the concentration is 10%, then adding Mel, and stirring thoroughly; the weight ratio of the honey to the concentrated solution is 1:10;
7) Filling the mixture obtained in the step 6) into a 50ml brown bottle, steaming and sterilizing for 20min at 90 ℃, steaming and sterilizing for 30min at 100 ℃, steaming and sterilizing for 10min at 80 ℃, and packaging to obtain the plant beverage.
Example 6
1) Cleaning the fresh medicinal materials with pure water, and then naturally air-drying or airing;
weighing the dried traditional Chinese medicine raw materials according to the following weight: 60g of kudzuvine root, 4g of hovenia dulcis thunb, 6g of cassia seed, 4g of chrysanthemum and 3g of liquorice, and mixing and crushing the materials to obtain a traditional Chinese medicine mixed raw material with the size of 0.5-1 cm;
3) Mixing the traditional Chinese medicine raw materials with pure water according to the following ratio of 1:25, heating with steam at 80deg.C for 20min, heating with steam at 120deg.C for 30min, heating with steam at 100deg.C for 10min, cooling to room temperature, adding 0.77g cellulose amylase, performing enzymolysis in water bath at 50deg.C for 2 hr, setting the distance between radio frequency polar plates to 114mm, and extracting for 320s to obtain the Chinese medicinal mixture;
4) Adding 0.58g of monascus into 77g of the traditional Chinese medicine mixture in the step 3) for fermentation, wherein the fermentation temperature is 37 ℃ and the fermentation time is 48 hours, so as to obtain fermentation liquor;
5) Centrifuging the fermentation liquid, adding 1% gelatin solution and 1% chitosan solution into the supernatant for clarification, and concentrating at 55deg.C under reduced pressure to obtain extract with density of 1.32;
6) Adding pure water to the extract until the concentration is 10%, then adding Mel, and stirring thoroughly; the weight ratio of the honey to the concentrated solution is 1:10;
7) Filling the mixture obtained in the step 6) into a 50ml brown bottle, steaming and sterilizing for 20min at 90 ℃, steaming and sterilizing for 30min at 100 ℃, steaming and sterilizing for 10min at 80 ℃, and packaging to obtain the plant beverage.
Example 7
1) Cleaning the fresh medicinal materials with pure water, and then naturally air-drying or airing;
2) Weighing the dried traditional Chinese medicine raw materials according to the following weight: 65g of kudzuvine root, 7g of hovenia dulcis thunb, 7g of cassia seed, 5g of chrysanthemum and 4g of liquorice, and mixing and crushing the materials to obtain a traditional Chinese medicine mixed raw material with the size of 0.5-1 cm;
3) Mixing the traditional Chinese medicine raw materials with pure water according to the following ratio of 1:25, heating with steam at 80deg.C for 20min, heating with steam at 120deg.C for 30min, heating with steam at 100deg.C for 10min, cooling to room temperature, adding 0.88g cellulose amylase, performing enzymolysis in water bath at 50deg.C for 2 hr, setting the distance between radio frequency polar plates to 114mm, and extracting for 320s to obtain the Chinese medicinal mixture;
4) Adding 0.66g of Eurotium cristatum and Monascus into 88g of the traditional Chinese medicine mixture in the step 3) for fermentation, wherein the fermentation temperature is 37 ℃ and the fermentation time is 48 hours, so as to obtain fermentation liquor;
5) Centrifuging the fermentation liquid, adding 1% gelatin solution and 1% chitosan solution into the supernatant for clarification, and concentrating at 55deg.C under reduced pressure to obtain extract with density of 1.32;
6) Adding pure water to the extract until the concentration is 10%, then adding Mel, and stirring thoroughly; the weight ratio of the honey to the concentrated solution is 1:10;
7) Filling the mixture obtained in the step 6) into a 50ml brown bottle, steaming and sterilizing for 20min at 90 ℃, steaming and sterilizing for 30min at 100 ℃, steaming and sterilizing for 10min at 80 ℃, and packaging to obtain the plant beverage.
Example 8
1) Cleaning the fresh medicinal materials with pure water, and then naturally air-drying or airing;
2) Weighing the dried traditional Chinese medicine raw materials according to the following weight: 70g of kudzuvine root, 10g of hovenia dulcis thunb, 8g of cassia seed, 6g of chrysanthemum and 5g of liquorice, and mixing and crushing the materials to obtain a traditional Chinese medicine mixed raw material with the size of 0.5-1 cm;
3) Mixing the traditional Chinese medicine raw materials with pure water according to the following ratio of 1:25, heating with steam at 80deg.C for 20min, heating with steam at 120deg.C for 30min, heating with steam at 100deg.C for 10min, cooling to room temperature, adding 0.99g cellulose amylase, performing enzymolysis in water bath at 50deg.C for 2 hr, setting the distance between radio frequency polar plates to 114mm, and extracting for 320s to obtain the Chinese medicinal mixture;
4) Adding 0.74g of Eurotium cristatum and Monascus purpureus into the 99g of the traditional Chinese medicine mixture in the step 3) for fermentation, wherein the fermentation temperature is 37 ℃ and the fermentation time is 48 hours, so as to obtain fermentation liquor;
5) Centrifuging the fermentation liquid, adding 1% gelatin solution and 1% chitosan solution into the supernatant for clarification, and concentrating at 55deg.C under reduced pressure to obtain extract with density of 1.32;
6) Adding pure water to the extract until the concentration is 10%, then adding Mel, and stirring thoroughly; the weight ratio of the honey to the concentrated solution is 1:10;
7) Filling the mixture obtained in the step 6) into a 50ml brown bottle, steaming and sterilizing for 20min at 90 ℃, steaming and sterilizing for 30min at 100 ℃, steaming and sterilizing for 10min at 80 ℃, and packaging to obtain the plant beverage.
Comparative example 1
The procedure of comparative example 1 was substantially the same as in example 4, except that: the supernatant in the step 5) is directly evaporated and concentrated under reduced pressure without clarification.
The turbidity values of each of the groups of example 1, example 3, example 4 and comparative example 1 were measured after clarification by adding 3mL of the clarifying agent to 250mL of the supernatant obtained in step 5) of example 1, example 3, example 4 and comparative example 1, respectively, and the results are shown in Table 2. The result shows that the compound beverage has better clarifying effect by adopting 1% gelatin and 1% chitosan, and can better improve the quality of the compound beverage.
TABLE 2 different clarification treatments effects
Clarification treatment mode Turbidity (NTU)
Comparative example 1 369
Example 1 253
Example 3 274
Example 4 190
Comparative example 2
Comparative example 2 is substantially the same as example 4 except that: and 4) adding no strain into the traditional Chinese medicine mixture.
Comparative example 3
Comparative example 3 is substantially the same as example 4 except that: and 3) adding no cellulase into the traditional Chinese medicine mixture.
The contents of total isoflavone and alcohol dehydrogenase in each of examples 2, 4 to 8 and comparative examples 2 and 3 were measured, and the results are shown in Table 3. The result shows that the fermentation product has better fermentation effect by adopting the Eurotium cristatum and the monascus to cooperate with each other; the enzymatic hydrolysis can be carried out by adding cellulase so as to effectively improve the activity of alcohol dehydrogenase.
TABLE 3 fermentation results for different fermentates
Figure BDA0004027246350000081
Figure BDA0004027246350000091
Example 9 cytotoxicology assay
HepG2 cells were counted and formulated as 1X 10 cells 5 cell suspensions of cells/mL were added to each well, after 24. 24 h of the cell suspension was subjected to wall-attached culture, the old medium (high sugar medium+10% bovine serum) was discarded, 100. Mu.L of the plant drink of the present invention was added to each well of the experimental group at mass concentrations of 50, 100, 200, 250, 500, 750, 1200, 1500. Mu.g/mL, 6 duplicate wells were set, the normal control group was added to the same volume of complete medium, after 24. 24 h of continuous culture, MTT solution was added to each well at final mass concentration of 0.5mg/mL, 4. 4 h of supernatant was discarded, 150. Mu.L of dimethyl sulfoxide was added to each well, and after shaking for 10min, the OD value was measured by an enzyme-labeled instrument at 490nm, and cell viability was calculated.
Establishing a damage model group: that is, 100. Mu.L of H at a concentration of 2, 5, 10, 20, 50, 100mmol/L was added to each well 2 O 2 The solution was prepared in 6 parallel wells, and after further culturing for 2h, the MTT method was used to measure the cell viability.
On the basis of the preliminary experiment established by the model, 20 mmol/L H and 50mmol/L H are screened out 2 O 2 After determining the optimal damage conditions for the cells respectively 2h, the PPH1 concentration without significant toxicity to the cells was selected as follows: blank group: untreated sample and H 2 O 2 A treated cell suspension;
experimental group: sequentially passing through the sample and H 2 O 2 A treated cell suspension;
model group: through only H 2 O 2 A treated cell suspension.
In 96-well plates, 100. Mu.L of 1X 10 density was added to each well 5 cell suspension of cells/mL was cultured in a constant temperature incubator for 24. 24 h, and then the medium in the wells was discarded, and 1%, 1.5% and 2% of the corresponding concentration was added to each well of the experimental groupThe plant drink prepared in example 4 of the present invention was 100. Mu.L per well. The blank and model groups were incubated with 100. Mu.L of serum-free medium for 24 h, respectively, and the medium was discarded, and 100. Mu. L H was added to each well of the experimental and model groups 2 O 2 Solution, blank were incubated with 100 μl of serum-free medium for 2 h. After the incubation, the MTT method determines the concentration of the plant beverage required when the cell viability is 99%, 90% and 50% respectively. The cell survival rate is 99% when the plant drink concentration is 0.016 g/g; cell viability was 90% at a plant drink concentration of 0.49 g/g; the cell viability was 50% at a plant drink concentration of 2.62 g/g.
Taking cells of each treatment group, washing for 2 times by PBS, adding 0.25% pancreatin to digest, preparing cell suspension, centrifuging for 5 min at 1000 r/min, collecting cells, washing for 1 time by PBS, adding cell lysate to lyse the cells, centrifuging for 10min at 12000 r/min, collecting supernatant, and measuring MDA (malondialdehyde) content and SOD (superoxide dismutase), GSH-Px (glutathione peroxidase) and CAT (catalase) activities in the cells according to the instruction of the kit. The results are shown in FIG. 1.
As can be seen from FIG. 1, the plant beverage prepared by the invention can improve the functional effects of intracellular superoxide dismutase, glutathione peroxidase and catalase activities, and the enzymes have antioxidant effects, can catalyze the decomposition of alcohol, avoid the damage of organism tissues, and greatly improve the anti-alcoholic effect, and meanwhile, the plant beverage prepared by the invention has the effect of removing the cancerogenic substance malondialdehyde in cells.
The above-mentioned embodiments are merely preferred embodiments of the present invention, which are not intended to limit the scope of the present invention, and other embodiments can be easily made by those skilled in the art through substitution or modification according to the technical disclosure in the present specification, so that all changes and modifications made in the principle of the present invention shall be included in the scope of the present invention.

Claims (6)

1. The method for preparing the plant beverage by using the vacuum radio frequency assisted biological enzymolysis is characterized by comprising the following steps of:
1) Cleaning the fresh medicinal materials with pure water, and then naturally air-drying or airing;
2) Weighing the dried traditional Chinese medicine raw materials according to the following weight portions: 60-70 parts of kudzuvine root, 4-10 parts of hovenia dulcis thunb, 6-8 parts of cassia seed, 4-6 parts of chrysanthemum and 3-5 parts of liquorice, and mixing and crushing the raw materials into a traditional Chinese medicine mixed raw material with the size of 0.5-1 cm;
3) Mixing the traditional Chinese medicine raw materials with pure water according to the following ratio of 1:25, heating for 60min by three sections of steam, cooling to room temperature, adding cellulose amylase, performing enzymolysis in water bath at 50deg.C for 2 hr, wherein the addition amount of the enzyme is 1% of the total amount of the Chinese medicinal materials, and performing vacuum radio frequency assisted extraction to obtain Chinese medicinal mixture;
4) Adding Eurotium cristatum and monascus into the traditional Chinese medicine mixture in the step 3) for fermentation, wherein the fermentation temperature is 28-37 ℃ and the fermentation time is 48 hours, so as to obtain fermentation liquor; the addition amount of the Eurotium cristatum and the monascus is 0.75% of the total amount of the traditional Chinese medicine mixture;
5) Centrifuging the fermentation liquor, clarifying the supernatant, and concentrating by reduced pressure evaporation to obtain extract;
6) Adding pure water to the extract until the concentration is 10%, then adding Mel, and stirring thoroughly; the weight ratio of the honey to the concentrated solution is 1:10;
7) Filling the mixture obtained in the step 6) into a 50ml brown bottle, steaming and sterilizing for 60min, and packaging to obtain the plant beverage.
2. The method according to claim 1, wherein the specific process of heating three steam sections in the step 3) for 60min is: heating at 80deg.C for 20min, then at 120deg.C for 30min, and finally at 100deg.C for 10min.
3. The method according to claim 1, wherein the distance between the radio frequency plates in the vacuum radio frequency method in the step 3) is 114mm, and the extraction time is 320s.
4. The method according to claim 1, wherein the clarification treatment in step 5) is chitosan or gelatin or a mixture of chitosan and gelatin.
5. The method according to claim 1, wherein the concentration temperature in the reduced pressure evaporation concentration in the step 5) is 55 ℃, and the concentration density of the concentrated extract is 1.32.
6. The method according to claim 1, wherein the specific process of steaming and sterilizing for 60min in the step 7) is: steaming at 90deg.C for 20min, steaming at 100deg.C for 30min, and steaming at 80deg.C for 10min.
CN202211710561.1A 2022-12-29 2022-12-29 Method for preparing plant beverage by vacuum radio frequency assisted biological enzymolysis Pending CN116138382A (en)

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