CN111493149A - Compound liquid containing protein polyphenol, preparation method thereof and double-protein milk - Google Patents
Compound liquid containing protein polyphenol, preparation method thereof and double-protein milk Download PDFInfo
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Abstract
The invention belongs to the technical field of double-protein milk, and particularly relates to compound liquid containing protein polyphenol, a preparation method of the compound liquid and double-protein milk. The compound liquid containing the protein polyphenol comprises the following components in parts by weight: 4-30 parts of coarse cereal protein, 2-30 parts of whey protein concentrate, 1 part of fruit and vegetable polyphenol, 8-80 parts of whole milk powder and 100-1000 parts of water. According to the invention, a double-protein concept and a protein-polyphenol interaction concept are combined, common and nutritious coarse cereals are selected, protein of the coarse cereals is extracted, and the coarse cereals, whey protein concentrate, fruit and vegetable polyphenol and whole milk powder are mixed according to a mass ratio of 4-30: 2-30: 1: 8-80; by adding the fruit and vegetable polyphenols in a specific proportion, the proteins and the polyphenols are combined through covalent or non-covalent, so that the functional properties such as the solubility, the foamability and the emulsibility of the plant proteins and the animal proteins can be improved, the oxidation resistance of the plant polyphenols can be improved, and the fruit and vegetable polyphenol beverage is very beneficial to the health of a human body.
Description
Technical Field
The invention belongs to the technical field of double-protein milk, and particularly relates to compound liquid containing protein polyphenol, a preparation method of the compound liquid and double-protein milk.
Background
The Chinese characteristic double-protein engineering aims to utilize global plant protein, animal protein and new protein resources, and applies modern high and new technology to promote the nutrition dose effect and the accurate interaction preparation of different source proteins, meets the key nutrition requirements of different groups, and has important scientific significance and practical significance for improving the health level of residents in China and solving the structural problem of modern agriculture-food industry-nutrition and health integrated collaborative development in China. The double-protein food is a functional nutritional food which is fused with modern science and technology, has accurate interaction nutrition, can accelerate the repair of human cell damage, regulate digestion and absorption metabolism and enhance the natural immunity of cells, and has an important supporting function for ensuring the health of human beings. Therefore, the food prospect of the double-protein is very wide.
In addition, the interaction mechanism of proteins and polyphenols has become a research hotspot since the seventies of the twentieth century. Protein and polyphenol are widely present in food systems, and the interaction of the protein and the polyphenol influences the structure and mouthfeel of food. With the increase of public health consciousness, the antioxidant property and health care function of polyphenol compounds are receiving increasing attention. The addition of polyphenolic substances to food systems is often seen as one of the effective ways to confer and improve antioxidant properties and biofunctional properties to foods, while proteins, by having properties that bind polyphenols, are seen as good carriers for their incorporation into food systems. At present, more and more products rich in protein and polyphenol are on the market.
Research shows that the double-protein milk has poor solubility, the solubility of protein is very important for the quality of milk, and how to improve the solubility of plant high-quality protein is a problem which is always researched in the prior stage.
Disclosure of Invention
The invention aims to combine the concept of double protein and the concept of protein-polyphenol interaction, overcome the problems in the prior art and provide the compound liquid containing protein polyphenol, the preparation method thereof and the double protein milk.
In order to achieve the above purpose, the invention provides the following technical scheme:
the invention provides a compound liquid containing protein polyphenol, which comprises the following components in parts by weight:
4-30 parts of coarse cereal protein, 2-30 parts of whey protein concentrate, 1 part of fruit and vegetable polyphenol, 8-80 parts of whole milk powder and 100-1000 parts of water;
the raw materials for preparing the coarse cereal protein comprise: the black bean, the mung bean, the soybean, the small red bean and the kidney bean are in a mass ratio of 15-25: 25-30: 20-25: 20-30: 10-20;
the raw materials for preparing the fruit and vegetable polyphenol comprise: the grape, the strawberry, the watermelon, the blueberry, the orange, the purple cabbage, the carrot and the tomato are mixed according to a mass ratio of 30-35: 25-35: 20-30: 25-35: 15-20: 10-20;
the whey protein concentrate includes β -lactoglobulin, α -lactalbumin, immunoglobulin and lactoferrin.
The invention provides a preparation method of the composite liquid, which comprises the following steps:
(1) drying, crushing and degreasing raw materials for preparing the coarse cereal protein to obtain degreased materials, mixing the degreased materials with water, and then performing alkali extraction and acid precipitation to obtain the coarse cereal protein;
(2) squeezing the raw materials for preparing the fruit and vegetable polyphenol, sequentially digesting the obtained juice with saliva α -amylase, pepsin, pancreatin and bile to obtain a treatment solution, centrifuging the treatment solution to obtain a supernatant, and removing sugar water compounds in the supernatant to obtain the fruit and vegetable polyphenol;
(3) mixing the coarse cereal protein obtained in the step (1) with whey protein concentrate, whole milk powder and water to obtain a mixture, mixing the mixture with glucose oxidase, carrying out enzymolysis to obtain an zymolyte, mixing the zymolyte with the fruit and vegetable polyphenol obtained in the step (2), and carrying out reaction to obtain a compound liquid containing protein polyphenol;
the steps (1) and (2) are not limited in time sequence.
Preferably, the step (1) is performed to obtain a crushed material after crushing, and the crushed material is degreased, wherein the particle size of the crushed material is 48-150 μm.
Preferably, the reagent used for degreasing in the step (1) comprises petroleum ether and/or n-hexane, and the mass ratio of the crushed material to the reagent is 1g: 3-6 ml.
Preferably, the mass ratio of the degreased substance to the water in the step (1) is 1g: 5-15 ml.
Preferably, in the step (2), the sugar water compound in the supernatant is removed by using an acetone solution with the mass percentage of 80%.
Preferably, the mass ratio of the juice to the α -amylase in the step (2) is 100: 0.1-0.3;
the mass ratio of the juice to the pepsin is 100: 0.05-2.4;
the mass ratio of the juice to the pancreatin is 100: 0.1-2.2;
the mass ratio of the juice to the bile is 100: 5-7.
Preferably, the conditions for enzymolysis in step (3) include: the temperature of the enzymolysis is 80-90 ℃, and the time of the enzymolysis is 25-35 min.
Preferably, the reaction conditions of the step (3) include: the reaction temperature is 20-30 ℃, and the reaction time is 1.5-2.5 h.
The invention also provides double-protein milk which comprises the compound liquid, lactose, sucrose and vegetable oil in the technical scheme, wherein the mass ratio of the compound liquid to the lactose to the sucrose to the vegetable oil is 100-120: 1-2: 8-10: 1-2.
Has the advantages that:
according to the compound liquid containing protein polyphenol and the double-protein milk, the double-protein concept and the protein-polyphenol interaction concept are combined, common and nutritious coarse cereals are selected, the protein of the coarse cereals is extracted, and the coarse cereals, the whey protein concentrate, the fruit and vegetable polyphenol and the whole milk powder are mixed according to the mass ratio of 4-30: 2-30: 1: 8-80; by adding the fruit and vegetable polyphenols in a specific proportion, the proteins and the polyphenols are combined through covalent or non-covalent, so that the functional properties such as the solubility, the foamability and the emulsibility of the plant proteins and the animal proteins can be improved, the oxidation resistance of the plant polyphenols can be improved, and the fruit and vegetable polyphenol beverage is very beneficial to the health of a human body.
Furthermore, the preparation method of the protein polyphenol-containing compound liquid further limits the method for obtaining the polyphenol of the fruit and vegetable juice by simulating in vitro digestion, so that the structure of macromolecular substances such as protein and polysaccharide in the fruit and vegetable juice can be damaged, and micromolecular phenolic substances combined with the macromolecular substances can be better released; and the fruit and vegetable polyphenol obtained by an in vitro digestion method can reduce the damage to the phenolic substances and the inactivation of the phenol.
Detailed Description
The invention provides a compound liquid containing protein polyphenol, which comprises the following components in parts by weight:
4-30 parts of coarse cereal protein, 2-30 parts of whey protein concentrate, 1 part of fruit and vegetable polyphenol, 8-80 parts of whole milk powder and 100-1000 parts of water;
the raw materials for preparing the coarse cereal protein comprise: the black bean, the mung bean, the soybean, the small red bean and the kidney bean are in a mass ratio of 15-25: 25-30: 20-25: 20-30: 10-20;
the raw materials for preparing the fruit and vegetable polyphenol comprise: the grape, the strawberry, the watermelon, the blueberry, the orange, the purple cabbage, the carrot and the tomato are mixed according to a mass ratio of 30-35: 25-35: 20-30: 25-35: 15-20: 10-20;
the whey protein concentrate includes β -lactoglobulin, α -lactalbumin, immunoglobulin and lactoferrin.
If not specifically stated, the present invention has no special requirements for the sources of the raw materials of various coarse cereals and the raw materials of fruits and vegetables in the protein and polyphenol containing composite liquid, and the present invention only needs to adopt commercial products which are well known to those skilled in the art.
The fruit and vegetable polyphenol-containing composite liquid provided by the invention comprises 4-30 parts of coarse cereal protein, preferably 4-10 parts, and more preferably 6 parts by weight. The raw materials for preparing the coarse cereal protein comprise: the black bean, the mung bean, the soybean, the small red bean and the kidney bean are in a mass ratio of 15-25: 25-30: 20-25: 20-30: 10-20, and preferably 20:28:23:25: 15. The coarse cereal raw materials are common coarse cereals with rich nutritional ingredients, the coarse cereals have high protein content and rich amino acid types, good plant protein can be provided for the product, the human body requirements are met, the solubility and the functional characteristics of the coarse cereal protein can be improved, and the oxidation resistance of fruit and vegetable polyphenol can be improved through the interaction of the coarse cereal protein and fruit and vegetable polyphenol with different proportions.
The composite liquid containing fruit and vegetable polyphenols provided by the invention comprises 2-30 parts of whey protein, preferably 4-10 parts of whey protein, more preferably 6 parts of whey protein, wherein the source of the whey protein is not particularly limited, and the source of the whey protein known by the skilled person is adopted.
Based on the mass parts of the coarse cereal protein, the fruit and vegetable polyphenol-containing compound liquid provided by the invention comprises 1 part of fruit and vegetable polyphenol. The raw materials for preparing the fruit and vegetable polyphenol comprise: the grape, the strawberry, the watermelon, the blueberry, the orange, the purple cabbage, the carrot and the tomato are in a mass ratio of 30-35: 25-35: 20-30: 25-35: 15-20: 10-20, and preferably 33:33:30:25:30:18:15: 15. The fruit and vegetable polyphenol raw materials are common fruits and vegetables with rich polyphenol content, and through the interaction of fruit and vegetable polyphenol and coarse cereal protein in different proportions, the oxidation resistance of the fruit and vegetable polyphenol can be improved, and the solubility and functional characteristics of the coarse cereal protein can be improved.
Based on the weight parts of the coarse cereal protein, the fruit and vegetable polyphenol-containing composite liquid provided by the invention comprises 8-80 parts of whole milk powder, preferably 10-40 parts, and more preferably 16 parts. The source of the whole milk powder is not particularly limited, and a source of whole milk powder known to those skilled in the art may be used. The whole milk powder is a supply source of animal protein.
Based on the weight parts of the coarse cereal protein, the fruit and vegetable polyphenol-containing compound liquid provided by the invention comprises 100-1000 parts of water, preferably 100-300 parts, and more preferably 195 parts.
According to the invention, by adding the fruit and vegetable polyphenols in a specific proportion, the proteins and the polyphenols are combined through covalent or non-covalent, so that the functional properties such as the solubility, the foamability and the emulsibility of the plant proteins and the animal proteins can be improved, the oxidation resistance of the plant polyphenols can be improved, and the fruit and vegetable polyphenol compound is very beneficial to the health of a human body.
The invention also provides a preparation method of the composite liquid, which comprises the following steps:
(1) drying, crushing and degreasing raw materials for preparing the coarse cereal protein to obtain degreased materials, mixing the degreased materials with water, and then performing alkali extraction and acid precipitation to obtain the coarse cereal protein;
(2) squeezing the raw materials for preparing the fruit and vegetable polyphenol, sequentially digesting the obtained juice with saliva α -amylase, pepsin, pancreatin and bile to obtain a treatment solution, centrifuging the treatment solution to obtain a supernatant, and removing sugar water compounds in the supernatant to obtain the fruit and vegetable polyphenol;
(3) mixing the coarse cereal protein obtained in the step (1) with whey protein concentrate, whole milk powder and water to obtain a mixture, mixing the mixture with glucose oxidase, carrying out enzymolysis to obtain an zymolyte, mixing the zymolyte with the fruit and vegetable polyphenol obtained in the step (2), and carrying out reaction to obtain a compound liquid containing protein polyphenol;
the steps (1) and (2) are not limited in time sequence.
In the invention, the drying is preferably carried out in a forced air drying oven, and the drying temperature is preferably 40-50 ℃, more preferably 45 ℃; the drying time is preferably 24 h. In the invention, the drying can remove part of water and volatile oil and fat in the coarse cereals.
The coarse cereal protein is prepared by crushing raw materials for preparing the coarse cereal protein to obtain a crushed material, and degreasing the crushed material, wherein the particle size of the crushed material is preferably 48-150 μm, more preferably 50-90 μm, and most preferably 74 μm. The invention crushes the coarse cereals, can increase the contact area of the coarse cereals and the degreasing agent, and further promotes the degreasing of the coarse cereals.
In the present invention, the reagent used for degreasing preferably includes petroleum ether and/or n-hexane, and the ratio of the mass of the pulverized material to the volume of the reagent is preferably 1g:3 to 6ml, more preferably 1g:5 ml. The source of the petroleum ether or n-hexane in the present invention is not particularly limited, and those known to those skilled in the art can be used.
In the present invention, the mass to volume ratio of the degreased matter to water is preferably 1g: 5-15 ml, and more preferably 1g:10 ml. The alkali extraction and acid precipitation method is not particularly limited in the present invention, and an alkali extraction and acid precipitation method known to those skilled in the art may be used.
In the present invention, the rotation speed of the centrifugation is preferably 10000rpm, and the time of the centrifugation is preferably 20 min.
The sugar water compound in the supernatant is removed by using the acetone solution with the mass percentage of 80%.
In the invention, the mass ratio of the juice to α -amylase is preferably 100: 0.1-0.3, more preferably 100:0.1, the mass ratio of the juice to pepsin is preferably 100: 0.05-2.4, more preferably 100:2.4, the mass ratio of the juice to pancreatin is preferably 100: 0.1-2.2, more preferably 100:2.2, and the mass ratio of the juice to bile is preferably 100: 5-7, more preferably 100: 7. the invention has no special limitation on the sources of α -amylase, pepsin, pancreatin and bile, and can be sold on the market.
In the invention, the mass ratio of the mixture to the glucose oxidase is preferably 100: 0.2-0.6, and more preferably 100: 0.4. The source of the glucose oxidase is not particularly limited, and the glucose oxidase can be obtained by adopting conventional commercial products. In the present invention, the glucose oxidase functions as: deoxidizing and preventing browning of the composite liquid.
In the present invention, the conditions for the enzymatic hydrolysis preferably include: the enzymolysis temperature is preferably 80-90 ℃, and more preferably 85 ℃; the enzymolysis time is preferably 25-35 min, and more preferably 30 min.
In the present invention, the conditions of the reaction preferably include: the reaction temperature is preferably 20-30 ℃, and more preferably 25 ℃; the reaction time is preferably 1.5-2.5 h, and more preferably 2 h. In the invention, after the reaction, the protein and the polyphenol are combined through covalent or non-covalent, so that the functional properties such as the solubility, the foamability and the emulsibility of the plant protein and the animal protein can be improved, and the oxidation resistance of the plant polyphenol can be improved.
According to the preparation method of the protein polyphenol-containing compound liquid, provided by the invention, the fruit and vegetable juice polyphenol is obtained by a method of simulating in-vitro digestion, so that not only can the structures of macromolecular substances such as protein and polysaccharide in the fruit and vegetable juice be damaged, but also micromolecular phenolic substances combined with the macromolecular substances can be better released; and the fruit and vegetable polyphenol obtained by an in vitro digestion method can reduce the damage to the phenolic substances and the inactivation of the phenol.
The invention also provides double-protein milk which comprises the compound liquid, lactose, sucrose and vegetable oil in the technical scheme, wherein the mass ratio of the compound liquid to the lactose to the sucrose to the vegetable oil is 100-120: 1-2: 8-10: 1-2, and preferably 1115:15:90: 16. The sources of the lactose, sucrose and vegetable oil are not particularly limited and may be those conventionally commercially available. In the invention, the lactose and the sucrose increase the mouthfeel of the double-protein milk; the addition of the vegetable oil can not only ensure the constancy of fatty acid in the double-protein milk, but also ensure that the double-protein milk is easier to digest and absorb.
The preparation method of the double-protein milk is not particularly limited, and the double-protein milk is prepared by adopting the conditions of homogenization, sterilization and the like of conventional prepared protein milk.
In order to further illustrate the present invention, the following examples are provided to describe the protein polyphenol-containing complex liquid, the preparation method thereof and the double-protein milk in detail, but they should not be construed as limiting the scope of the present invention.
Example 1
Step 1: 200g of soybean, 150g of black bean, 250g of mung bean, 100g of kidney bean and 200g of small red bean are subjected to impurity removal and cleaning, then are put into a forced air drying oven to be dried for 24h at 40 ℃, are respectively put into a pulverizer to be primarily pulverized after being dried, then are put into a superfine pulverizer to be superfine pulverized, and are sieved by a 100-mesh sieve, so that various coarse cereal powders are obtained for later use.
Step 2: putting the various coarse cereal powders obtained in the step 1 into a beaker, mixing the coarse cereal powders with petroleum ether in a ratio of 1:5(g: ml), degreasing for 3 times with stirring of a stirrer for 5 hours each time, and airing and bagging the various coarse cereal powders for later use after degreasing.
Extracting protein by using an alkali extraction and acid precipitation method, mixing coarse cereal bean powder with distilled water according to a ratio of 1:10(g: ml), adjusting pH to 9 by using 1 mol/L NaOH solution, stirring and extracting for 60min in a water bath at 50 ℃, centrifuging for 10min by using a centrifuge at 3500r/min, taking supernatant after centrifuging, adding an antifoaming agent (the mass ratio of the supernatant to the antifoaming agent is 100:0.06) for degassing, adjusting the pH value of the supernatant to the isoelectric point of each bean protein by using 1 mol/L HCl solution for acid precipitation, standing for 1h, centrifuging for 10min at 3500r/min in the centrifuge, separating the protein solution after acid precipitation by using a separator, performing neutral adjustment by using 0.1 mol/L sodium hydroxide solution, performing deacidification and desalination treatment by using a dialysis method, washing with water, taking precipitate, performing spray drying, and storing at-20 ℃ for later use to obtain coarse cereal protein powder.
And step 3: 300g of grapes, 300g of strawberries, 200g of blueberries, 150g of purple cabbage and 100g of tomatoes are adopted: cleaning, blanching, cooling, removing core, pulping, squeezing, filtering with 80 mesh filter cloth, subpackaging, and preserving to obtain fruit and vegetable juice; 250g of watermelon, 250g of orange and 100g of carrot are adopted: cleaning, peeling, cutting into pieces, pulping, squeezing, filtering with 80 mesh filter cloth, subpackaging, and preserving to obtain fruit and vegetable juice.
Step 4, selecting a method for simulating in vitro digestion to obtain fruit and vegetable juice polyphenol filtrate, mixing the fruit and vegetable juice obtained in the step 3, putting the mixture into a conical flask, and then adding saliva α -amylase (the mass ratio of the fruit and vegetable juice to the saliva α -amylase is 100:0.1) to dissolve in CaCl2(1 mmol/L, pH7.0), reacting in a shaking table of constant temperature water bath at 37 ℃, after digesting for 10min, adjusting the pH of the enzymolysis liquid to 2 by using 6 mol/L HCl, adding pepsin (the mass ratio of the fruit and vegetable juice to the pepsin is 100:2.4) and dissolving in 5m L HCl (0.1 mol/L), reacting in a shaking table of constant temperature water bath at 37 ℃, digesting for 1h again, adjusting the pH to 6.80 by using 6 mol/L NaOH, and then adding Na2HCO3Pancreatin (the mass ratio of fruit and vegetable juice to pancreatin is 100:2.2) and bile (the mass ratio of fruit and vegetable juice to bile is 100:7) in a solution (0.5 mol/L, 10m L), reacting in a constant-temperature water bath shaker at 37 ℃, finally digesting for 2 hours, transferring the solution into a 1kDa dialysis bag for dialysis overnight, centrifuging the obtained digestive juice at 4 ℃ (10000rpm,20min), taking supernatant, precipitating with 80% acetone to remove sugar water compounds in the supernatant, centrifuging (10000rpm,10min), taking the supernatant as polyphenol filtrate of the fruit and vegetable juice, performing rotary evaporation at 45 ℃, freeze drying, and storing at-20 ℃ for later use to obtain the polyphenol fruit and vegetable extract.
And 5, taking 30g of the plant coarse cereal protein in the step 2, adding 30g of whey protein concentrate into 80g of whole milk powder. Adding 975g of purified water, shearing and mixing uniformly by using a high-speed shearing machine, and storing at 4 ℃ overnight to fully hydrate protein molecules. Adding glucose oxidase (the mass ratio of protein to glucose oxidase is 100:0.2), reacting at 85 deg.C for 30min, adding fruit and vegetable polyphenol extract 1g, and reacting at 25 deg.C for 2 hr to obtain double protein milk-fruit and vegetable polyphenol composite liquid.
And 6, adding 15g of lactose, 90g of cane sugar and 16g of vegetable oil into the double-protein milk-fruit and vegetable polyphenol compound liquid, and carrying out high-pressure homogenization for 2 times under the conditions of 25MPa of pressure and 50 ℃ of homogenization temperature. Then sterilizing at high temperature, heating at 121 deg.C for 10min, and cooling to 45 deg.C in cold water bath to obtain double protein milk.
Example 2
Step 1: 230g of soybean, 200g of black bean, 280g of mung bean, 150g of kidney bean and 250g of small red bean are subjected to impurity removal and cleaning, then are put into a forced air drying oven to be dried for 24h at 45 ℃, are respectively put into a pulverizer to be primarily pulverized after being dried, then are put into an ultrafine pulverizer to be ultrafine pulverized, and are sieved by a 200-mesh sieve, so that various coarse cereal powders are obtained for later use.
Step 2: and (3) putting the various coarse cereal powders obtained in the step (1) into a beaker, mixing the various coarse cereal powders with petroleum ether in a ratio of 1:5, degreasing the mixture for 3 times under the stirring of a stirrer, and drying the various coarse cereal powders after degreasing, and bagging the various coarse cereal powders for later use.
The extraction of protein is carried out by adopting an alkali extraction and acid precipitation method, mixing the coarse cereal bean powder with distilled water according to a ratio of 1:10, adjusting the pH to 9.5 by using 1 mol/L NaOH solution, stirring and extracting for 60min in a water bath at 50 ℃, centrifuging for 15min by a centrifuge at 3500r/min, taking supernatant after centrifugation, adding an antifoaming agent (the mass ratio of the supernatant to the antifoaming agent is 100:0.09) for degassing, adjusting the pH value of the supernatant to the isoelectric point of each bean protein by using 1 mol/L HCl solution for acid precipitation, standing for 1h, centrifuging for 15min at 3500r/min in the centrifuge, separating the protein liquid after acid precipitation by a separator, carrying out neutral adjustment by using 0.1 mol/L sodium hydroxide solution, carrying out deacidification and desalination treatment by using a dialysis method, washing with water, taking precipitate, carrying out spray drying, and storing at-20 ℃ for later use to obtain the coarse cereal protein powder.
And step 3: 330g of grapes, 330g of strawberries, 250g of blueberries, 180g of purple cabbages and 150g of tomatoes are adopted: cleaning, blanching, cooling, removing core, pulping, squeezing, filtering with 90 mesh filter cloth, subpackaging, and preserving to obtain fruit and vegetable juice; 300g of watermelon, 300g of orange and 150g of carrot are adopted: cleaning, peeling, cutting into pieces, pulping, squeezing, filtering with 90 mesh filter cloth, subpackaging, and preserving to obtain fruit and vegetable juice.
Step 4, selecting a method for simulating in vitro digestion to obtain fruit and vegetable juice polyphenol filtrate, mixing the fruit and vegetable juice obtained in the step 3, putting the mixture into a conical flask, and then adding saliva α -amylase (the mass ratio of the fruit and vegetable juice to the saliva α -amylase is 100:0.1) to dissolve in CaCl2(1 mmol/L, pH7.0) in a solution of 3Reacting in a constant-temperature water bath shaker at 7 ℃, after digesting for 20min, adjusting the pH of the enzymolysis liquid to 2 by using 6 mol/L HCl, adding pepsin (the mass ratio of the fruit and vegetable juice to the pepsin is 100:2.4) to be dissolved in 5m L HCl (0.1 mol/L), reacting in a constant-temperature water bath shaker at 37 ℃, digesting for 1.5h again, adjusting the pH to 6.80 by using 6 mol/L NaOH, and then adding Na2HCO3Pancreatin (the mass ratio of fruit and vegetable juice to pancreatin is 100:2.2) and bile (the mass ratio of fruit and vegetable juice to bile is 100:7) in a solution (0.5 mol/L, 10m L), reacting in a constant-temperature water bath shaker at 37 ℃, finally transferring the solution into a 1kDa dialysis bag for dialysis overnight after digesting for 3 hours, centrifuging the obtained digestive juice at 4 ℃ (10000rpm,20 minutes) to obtain a supernatant, precipitating with 80% acetone to remove sugar water compounds in the supernatant, centrifuging (10000rpm,10 minutes) to obtain a supernatant which is polyphenol filtrate of the fruit and vegetable juice, performing rotary evaporation at 45 ℃, freeze drying, and storing at-20 ℃ for later use to obtain the polyphenol fruit and vegetable extract.
And 5, taking 30g of the plant coarse cereal protein in the step 2, adding 30g of whey protein concentrate into 80g of whole milk powder. Adding 975g of purified water, shearing and mixing uniformly by using a high-speed shearing machine, and storing at 4 ℃ overnight to fully hydrate protein molecules. Adding glucose oxidase (the mass ratio of protein to glucose oxidase is 100:0.4), reacting at 85 deg.C for 30min, adding fruit and vegetable polyphenol extract 5g, and reacting at 25 deg.C for 2 hr to obtain double protein milk-fruit and vegetable polyphenol composite liquid.
And 6, adding 15g of lactose, 90g of cane sugar and 16g of vegetable oil into the double-protein milk-fruit and vegetable polyphenol compound liquid, and carrying out high-pressure homogenization for 2 times under the conditions of 25MPa of pressure and 50 ℃ of homogenization temperature. Then sterilizing at high temperature, heating at 121 deg.C for 10min, and cooling to 45 deg.C in cold water bath to obtain double protein milk.
Example 3
Step 1: removing impurities from 250g of soybean, 250g of black bean, 300g of mung bean, 200g of kidney bean and 300g of small red bean, cleaning, then placing into a forced air drying oven for drying at 50 ℃ for 24h, respectively placing into a pulverizer for primary grinding after drying, then placing into an ultrafine grinder for ultrafine grinding, and sieving with a 300-mesh sieve to obtain various coarse cereal powders for later use.
Step 2: and (3) putting the various coarse cereal powders obtained in the step (1) into a beaker, mixing the various coarse cereal powders with petroleum ether in a ratio of 1:5, degreasing the mixture for 3 times under the stirring of a stirrer, and drying the various coarse cereal powders after degreasing, and bagging the various coarse cereal powders for later use.
The extraction of protein is carried out by adopting an alkali extraction and acid precipitation method, mixing the coarse cereal bean powder with distilled water according to a ratio of 1:10, adjusting pH to 10 by using 1 mol/L NaOH solution, stirring and extracting for 60min in a water bath at 50 ℃, centrifuging for 20min by a centrifuge at 3500r/min, taking supernatant after centrifugation, adding an antifoaming agent (the mass ratio of the supernatant to the antifoaming agent is 100:0.12) for degassing treatment, adjusting the pH value of the supernatant to the isoelectric point of various bean proteins by using 1 mol/L HCl solution for acid precipitation, standing for 1h, centrifuging for 20min at 3500r/min in the centrifuge, separating the protein liquid after acid precipitation by a separator, carrying out neutral adjustment by using 0.1 mol/L sodium hydroxide solution, deacidifying and desalting by using a dialysis method, washing with water, taking precipitate, carrying out spray drying, and storing at-20 ℃ for later use to obtain the coarse cereal protein powder.
And step 3: 350g of grapes, 350g of strawberries, 300g of blueberries, 200g of purple cabbages and 200g of tomatoes are adopted: cleaning, blanching, cooling, removing core, pulping, squeezing, filtering with 100 mesh filter cloth, subpackaging, and preserving to obtain fruit and vegetable juice; 350g of watermelon, 350g of orange and 200g of carrot are adopted: cleaning, peeling, cutting into pieces, pulping, squeezing, filtering with 100 mesh filter cloth, subpackaging, and preserving to obtain fruit and vegetable juice.
Step 4, selecting a method for simulating in vitro digestion to obtain fruit and vegetable juice polyphenol filtrate, mixing the fruit and vegetable juice obtained in the step 3, putting the mixture into a conical flask, and then adding saliva α -amylase (the mass ratio of the fruit and vegetable juice to the saliva α -amylase is 100:0.1) to dissolve in CaCl2(1 mmol/L, pH7.0), reacting in a shaking table of constant temperature water bath at 37 ℃, after digesting for 30min, adjusting the pH of the enzymolysis liquid to 2 by using 6 mol/L HCl, adding pepsin (the mass ratio of the fruit and vegetable juice to the pepsin is 100:2.4) and dissolving in 5m L HCl (0.1 mol/L), reacting in a shaking table of constant temperature water bath at 37 ℃, digesting for 2h again, adjusting the pH to 6.80 by using 6 mol/L NaOH, and then adding Na2HCO3Pancreatin (the mass ratio of the fruit and vegetable juice to the pancreatin is 100:2.2) and bile (the mass ratio of the fruit and vegetable juice to the bile is 100:2.2) in the solution (0.5 mol/L, 10m L)100:7), reacting in a shaking table with constant temperature water bath at 37 ℃; after a final digestion for 4h the solution was transferred to a 1kDa dialysis bag for dialysis overnight. Centrifuging the digestive juice at 4 deg.C (10000rpm,20min), collecting supernatant, precipitating with 80% acetone to remove sugar water compounds in the supernatant, centrifuging (10000rpm,10min), collecting supernatant as fruit and vegetable juice polyphenol filtrate, rotary evaporating at 45 deg.C, freeze drying, and storing at-20 deg.C to obtain fruit and vegetable polyphenol extract.
And 5, taking 30g of the plant coarse cereal protein in the step 2, adding 30g of whey protein concentrate into 80g of whole milk powder. Adding 975g of purified water, shearing and mixing uniformly by using a high-speed shearing machine, and storing at 4 ℃ overnight to fully hydrate protein molecules. Adding glucose oxidase (the mass ratio of protein to glucose oxidase is 100:0.6), reacting at 85 deg.C for 30min, adding fruit and vegetable polyphenol extract 10g, and reacting at 25 deg.C for 2 hr to obtain double protein milk-fruit and vegetable polyphenol composite liquid.
And 6, adding 15g of lactose, 90g of cane sugar and 16g of vegetable oil into the double-protein milk-fruit and vegetable polyphenol compound liquid, and carrying out high-pressure homogenization for 2 times under the conditions of 25MPa of pressure and 50 ℃ of homogenization temperature. Then sterilizing at high temperature, heating at 121 deg.C for 10min, and cooling to 45 deg.C in cold water bath to obtain double protein milk.
Comparative example 1
Step 1: 230g of soybean, 200g of black bean, 280g of mung bean, 150g of kidney bean and 250g of small red bean are subjected to impurity removal and cleaning, then are put into a forced air drying oven to be dried for 24h at 40 ℃, are respectively put into a pulverizer to be primarily pulverized after being dried, then are put into an ultrafine pulverizer to be ultrafine pulverized, and are sieved by a 200-mesh sieve, so that various coarse cereal powders are obtained for later use.
Step 2: and (3) putting the various coarse cereal powders obtained in the step (1) into a beaker, mixing the various coarse cereal powders with petroleum ether in a ratio of 1:5, degreasing the mixture for 3 times under the stirring of a stirrer, and drying the various coarse cereal powders after degreasing, and bagging the various coarse cereal powders for later use.
The extraction of protein is carried out by adopting an alkali extraction and acid precipitation method, mixing the coarse cereal bean powder with distilled water according to a ratio of 1:10, adjusting the pH to 9.5 by using 1 mol/L NaOH solution, stirring and extracting for 60min in a water bath at 50 ℃, centrifuging for 15min by a centrifuge at 3500r/min, taking supernatant after centrifugation, adding an antifoaming agent (the mass ratio of the supernatant to the antifoaming agent is 100:0.09) for degassing, adjusting the pH value of the supernatant to the isoelectric point of each bean protein by using 1 mol/L HCl solution for acid precipitation, standing for 1h, centrifuging for 15min at 3500r/min in the centrifuge, separating the protein liquid after acid precipitation by a separator, carrying out neutral adjustment by using 0.1 mol/L sodium hydroxide solution, carrying out deacidification and desalination treatment by using a dialysis method, washing with water, taking precipitate, carrying out spray drying, and storing at-20 ℃ for later use to obtain the coarse cereal protein powder.
And 3, taking 30g of the plant coarse cereal protein in the step 2, adding 30g of whey protein concentrate into 80g of whole milk powder. Adding 975g of purified water, shearing and mixing uniformly by using a high-speed shearing machine, and storing at 4 ℃ overnight to fully hydrate protein molecules. Adding glucose oxidase (the mass ratio of the protein to the glucose oxidase is 100:0.4), and reacting at 85 deg.C for 30min to obtain double-protein milk.
And 4, adding 15g of lactose, 90g of cane sugar and 16g of vegetable oil into the double-protein milk, and carrying out high-pressure homogenization for 2 times under the conditions that the pressure is 25MPa and the homogenization temperature is 50 ℃. Then sterilizing at high temperature, heating at 121 deg.C for 10min, and cooling to 45 deg.C in cold water bath to obtain double protein milk.
Application example 1
The invention adopts an improved Miller et al in vitro simulated digestion method: the method included 3 hours of pepsin-hydrochloric acid digestion (simulated gastric digestion), and 2 hours of pepsin-hydrochloric acid digestion and 4 hours of pancreatin-bile digestion (simulated gastrointestinal digestion). Observing the antioxidant capacity of the double-protein milk after gastrointestinal tract digestion.
(1) Simulated gastric digestion, namely adding 200ml of normal saline into 20M L double-protein milk prepared in example 1, stirring uniformly by using a glass rod, boiling in a water bath for 15min, continuously stirring by using the glass rod, cooling, then using deionized water to constantly weigh the sample suspension to 220g, using 1M HCl to adjust the pH of the sample suspension to 2.0, then adding 2.5M L gastric juice (0.2g pepsin is dissolved in 5M L0.01M HCl), using equal volume of deionized water to replace hydrochloric acid and gastric juice in a blank control group, using equal volume of 0.01M HCl to replace gastric juice in a hydrochloric acid control group, covering a conical flask with tinfoil to avoid light, filling nitrogen, digesting for 3h in a constant-temperature water bath shaker at 37 ℃, taking out certain mass of suspension at reaction time of 0, 0.5, 1, 2 and 3h, respectively, 12000r/min, centrifuging for 15min at 4 ℃, taking supernatant, precipitating by 80% of acetone (concentration), removing protein and polysaccharide, and subpackaging for storage at-80 ℃ for later use.
(2) Simulated intestinal fluid digestion was performed by adding 20M L of the double protein milk prepared in example 1 to 200ml of physiological saline, stirring with a glass rod, boiling in a water bath for 15min while stirring with the glass rod, then adding deionized water to constantly weigh the sample suspension to 220g, adjusting the pH of the sample suspension to 2.0 with 1M HCl, adding 2.5M L of gastric juice (0.2g pepsin dissolved in 5M L0.01 MHCl), wrapping the flask with tinfoil to protect from light, introducing nitrogen, reacting in a 37 ℃ constant temperature water bath shaker for 2h, adding 1MNaHCO to the suspension3To pH7.0, 5m L simulated intestinal fluid (4g pancreatin, 25g bile extract in 1L 0.1.1 mol/L NaHCO) was added3/Na2CO3Buffer solution). Blank control with equal volume of NaHCO3/Na2CO3Buffer solution was substituted for simulated intestinal fluid. Wrapping with tinfoil, charging nitrogen gas, and digesting in a constant temperature water bath shaker at 37 deg.C for 4 hr. Taking out certain mass of suspension at 0 time of gastric digestion and 0, 0.5, 1, 2, 3 and 4h of intestinal digestion, centrifuging at 12000r/min at 4 deg.C for 15min, collecting supernatant, precipitating with 80% acetone (final concentration) to remove protein and polysaccharide, packaging, and storing at-80 deg.C.
The DPPH, ABTS, ORAC free radical scavenging ability, FRAP antioxidant ability of the different products after treatment with the in vitro simulated digestion method in application examples 1 and 2, and the degree of in vitro digestion of the protein of each product were determined using methods conventional in the art. The free radical scavenging ability and FRAP antioxidant ability of DPPH, ABTS and ORAC of different products are shown in table 1, and the in vitro protein digestion degree of each product is shown in table 2.
Application example 2
The antioxidant ability of the double-protein milk prepared in example 2, example 3 and comparative example 1 after gastrointestinal digestion was observed in the same manner as in application example 1.
DPPH, ABTS, ORAC of different products remove free radical, FRAP antioxidant capacity, and degree of protein digestion in vitro of each product. The free radical scavenging ability and FRAP antioxidant ability of DPPH, ABTS and ORAC of different products are shown in table 1, and the in vitro protein digestion degree of each product is shown in table 2.
TABLE 1 DPPH, ABTS, ORAC free radical scavenging, FRAP antioxidant of different products
Capability of
TABLE 2 in vitro protein digestion of the respective products
As can be seen from Table 1, the DPPH, ABTS and ORAC free radical scavenging ability and FRAP oxidation resistance of the double-protein milk group containing fruit and vegetable polyphenols in the examples 1-3 are higher than that of the double-protein milk group containing no fruit and vegetable polyphenols in the comparative example 1; in addition, DPPH, ORAC free radical scavenging capacity and FRAP antioxidant capacity of the double-protein milk group containing fruit and vegetable polyphenol in the embodiment 2 are higher than those of the fruit and vegetable polyphenol extract prepared in the embodiment 2, and the antioxidant capacity of the double-protein milk group containing fruit and vegetable polyphenol is close to that of the conventional Vc antioxidant capacity. Therefore, the double-protein milk containing fruit and vegetable polyphenols provided by the invention can obviously improve the oxidation resistance of plant polyphenols, and is very beneficial to the health of human bodies.
As can be seen from Table 2, the TCA precipitates of the double-protein milk groups containing fruit and vegetable polyphenols in examples 1-3 after digestion by pepsin are all lower than that of the double-protein milk group containing no fruit and vegetable polyphenols in comparative example 1. Therefore, the double-protein milk containing fruit and vegetable polyphenols provided by the invention can promote the digestion and absorption of pepsin on the protein of the double-protein milk, and is very beneficial to the health of human bodies.
Application example 3
The functional properties of the different double-protein milk of the examples 1-3 and the comparative example 1 are measured and sensory evaluation is carried out by adopting a conventional method in the field. The functional properties of the different products are shown in table 3, and the sensory evaluation of the respective products is shown in table 4.
TABLE 3 functional Properties of the product
| Group of | Foaming Property (%) | Emulsifiability (m)2/g) | Solubility (%) |
| Example 1 | 16.49±0.16a | 0.0031±0.00025c | 19.05±0.87c |
| Example 2 | 15.20±0.42 | 0.0092±0.00048 | 47.72±0.79 |
| Example 3 | 16.75±0.93a | 0.0086±0.00055b | 48.62±0.36a |
| Comparative example 1 | 14.35±0.76 | 0.0025±0.00037 | 17.06±0.54 |
TABLE 4 sensory evaluation of the products
As can be seen from table 3, the two-protein milk groups containing fruit and vegetable polyphenols in examples 1 to 3 are higher in foaming property, emulsifying property, solubility and the like than the two-protein milk group containing no fruit and vegetable polyphenols in comparative example 1. Therefore, the double-protein milk containing fruit and vegetable polyphenol provided by the invention can increase the solubility, foamability and emulsibility of vegetable protein and animal protein, and solves the problem of low solubility of vegetable protein.
As can be seen from Table 4, the comprehensive scores of the double-protein milk groups containing fruit and vegetable polyphenols in the examples 1-3 are higher than that of the double-protein milk group containing no fruit and vegetable polyphenols in the comparative example 1; and the color, smell, tissue form, taste, flavor and the like of the double-protein milk group containing fruit and vegetable polyphenol in the example 2 are higher than those of the double-protein milk group without fruit and vegetable polyphenol in the comparative example 1. Therefore, the double-protein milk containing fruit and vegetable polyphenol provided by the invention has better color, smell, tissue form, taste, flavor and the like.
Although the present invention has been described in detail with reference to the above embodiments, it is only a part of the embodiments of the present invention, not all of the embodiments, and other embodiments can be obtained without inventive step according to the embodiments, and the embodiments are within the scope of the present invention.
Claims (10)
1. The compound liquid containing protein polyphenol is characterized by comprising the following components in parts by weight:
4-30 parts of coarse cereal protein, 2-30 parts of whey protein concentrate, 1 part of fruit and vegetable polyphenol, 8-80 parts of whole milk powder and 100-1000 parts of water;
the raw materials for preparing the coarse cereal protein comprise: the black bean, the mung bean, the soybean, the small red bean and the kidney bean are in a mass ratio of 15-25: 25-30: 20-25: 20-30: 10-20;
the raw materials for preparing the fruit and vegetable polyphenol comprise: the grape, the strawberry, the watermelon, the blueberry, the orange, the purple cabbage, the carrot and the tomato are mixed according to a mass ratio of 30-35: 25-35: 20-30: 25-35: 15-20: 10-20;
the whey protein concentrate includes β -lactoglobulin, α -lactalbumin, immunoglobulin and lactoferrin.
2. The method for preparing the complex liquid according to claim 1, comprising the steps of:
(1) drying, crushing and degreasing raw materials for preparing the coarse cereal protein to obtain degreased materials, mixing the degreased materials with water, and then performing alkali extraction and acid precipitation to obtain the coarse cereal protein;
(2) squeezing the raw materials for preparing the fruit and vegetable polyphenol, sequentially digesting the obtained juice with saliva α -amylase, pepsin, pancreatin and bile to obtain a treatment solution, centrifuging the treatment solution to obtain a supernatant, and removing sugar water compounds in the supernatant to obtain the fruit and vegetable polyphenol;
(3) mixing the coarse cereal protein obtained in the step (1) with whey protein concentrate, whole milk powder and water to obtain a mixture, mixing the mixture with glucose oxidase, carrying out enzymolysis to obtain an zymolyte, mixing the zymolyte with the fruit and vegetable polyphenol obtained in the step (2), and carrying out reaction to obtain a compound liquid containing protein polyphenol;
the steps (1) and (2) are not limited in time sequence.
3. The production method according to claim 2, wherein the step (1) is carried out to obtain a pulverized material after pulverization, and the pulverized material is degreased and has a particle size of 48 to 150 μm.
4. The preparation method according to claim 2 or 3, wherein the reagent used for degreasing in the step (1) comprises petroleum ether and/or n-hexane, and the mass-to-volume ratio of the crushed material to the reagent is 1g: 3-6 ml.
5. The preparation method according to claim 2, wherein the mass-to-volume ratio of the degreased matter to water in the step (1) is 1g: 5-15 ml.
6. The method according to claim 2, wherein the sugar water compound in the supernatant is removed by using an acetone solution having a content of 80% by mass in the step (2).
7. The production method according to claim 2, wherein in the step (2):
the mass ratio of the juice to the α -amylase is 100: 0.1-0.3;
the mass ratio of the juice to the pepsin is 100: 0.05-2.4;
the mass ratio of the juice to the pancreatin is 100: 0.1-2.2;
the mass ratio of the juice to the bile is 100: 5-7.
8. The preparation method according to claim 2, wherein the conditions of the enzymolysis in the step (3) comprise: the temperature of the enzymolysis is 80-90 ℃, and the time of the enzymolysis is 25-35 min.
9. The method according to claim 2, wherein the reaction conditions of the step (3) include: the reaction temperature is 20-30 ℃, and the reaction time is 1.5-2.5 h.
10. A double-protein milk is characterized by comprising the compound liquid as described in claim 1 or 2, lactose, sucrose and vegetable oil, wherein the mass ratio of the compound liquid to the lactose to the sucrose to the vegetable oil is 100-120: 1-2: 8-10: 1-2.
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