A blackcurrant and orange mixed soda

Abstract

This invention relates to a blackcurrant and orange juice mixed soft drink, belonging to the field of beverage technology. The mixed soft drink contains blackcurrant extract, orange juice, white hyacinth bean seed extract, mogrosides, steviol glycosides, etc. The blackcurrant extract is a product with a lower than 5kDa obtained by enzymatic hydrolysis of blackcurrants with hemicellulase and pectinase, followed by sequential enzymatic hydrolysis with nattokinase, fig protease, papain, and glucokinase. The white hyacinth bean seed extract is a product with a lower than 5kDa obtained by trypsin hydrolysis of white hyacinth bean seeds, followed by co-fermentation with Propionibacterium finigense, Lactobacillus brevis, and Bifidobacterium longum. The blackcurrant extract and white hyacinth bean seed extract have excellent anti-fatigue, antioxidant, and immune-enhancing effects, making the mixed soft drink both delicious and healthy.

Description

Technical Field

[0001] This invention belongs to the field of beverage technology, specifically relating to a blackcurrant and orange juice mixed soda. Background Technology

[0002] Mixed drinks typically refer to beverages made by blending various fruits, vegetables, spices, tea, or other ingredients. Common mixed fruit drinks include juices or fruit pieces from a variety of fruits such as apples, oranges, strawberries, and grapes, satisfying consumers' needs for rich flavors and diverse nutrition.

[0003] This type of beverage is characterized by its rich flavor and diverse combination of nutrients, which also offer certain health benefits. If the mixed beverage contains fruits and vegetables rich in antioxidants such as vitamin C, vitamin E, and flavonoids, like oranges, strawberries, blueberries, and carrots, it will have a certain antioxidant effect, helping to eliminate free radicals in the body, reduce oxidative stress damage to cells, and thus lower the risk of chronic diseases to some extent. Some mixed beverages also include ingredients with prebiotic properties, such as bananas and apples, which help regulate the balance of gut microbiota and promote intestinal health.

[0004] Blackcurrant, scientifically known as *Ribes nigrum* L., is also called blackcurrant, black bean fruit, or purple plum. It belongs to the genus *Ribes* in the family Ribsaceae, order Saxifragales. It is a small shrub whose ripe fruit is a small black berry, rich in vitamin C and anthocyanins, and is edible. It can also be processed into juice, jam, and other food products. Blackcurrants are very rich in vitamin C, phosphorus, magnesium, potassium, calcium, anthocyanins, and phenolic compounds.

[0005] Biotechnology has also played a crucial role in the beverage industry. By screening and cultivating specific probiotic strains, such as Bifidobacteria and Lactobacillus, and adding them to beverages, the balance of intestinal flora can be regulated, immunity enhanced, and nutrient absorption promoted. Furthermore, the use of biotechnology to extract effective components from plants, such as tea polyphenols, anthocyanins, and flavonoids, provides a rich source of raw materials for developing health beverages with antioxidant, anti-inflammatory, and lipid-lowering functions.

[0006] However, due to differences in raw materials and extraction methods, the resulting components vary, leading to different nutritional supplementation and health benefits for the human body. Therefore, improvements are needed in terms of health and efficacy. As people place increasing emphasis on the health of food and beverages, the beverage industry is continuously developing and improving technologies to provide higher quality and healthier, safer drinks, offering consumers more delicious and healthy beverage choices. Summary of the Invention

[0007] Existing beverages, due to differences in raw materials and extraction methods, produce different components, resulting in varying nutritional supplementation and health benefits for the human body, thus requiring improvement in terms of health and efficacy. This invention provides a blackcurrant and orange juice mixed soda, containing blackcurrant extract, orange juice, white hyacinth bean seed extract, mogrosides, steviol glycosides, etc. The blackcurrant extract is a product with a Da content below 5kDa obtained by enzymatic hydrolysis of blackcurrants using hemicellulase and pectinase, followed by sequential enzymatic hydrolysis using nattokinase, fig protease, papain, and glucokinase. The white hyacinth bean seed extract is a product with a Da content below 5kDa obtained by trypsin hydrolysis of white hyacinth bean seeds, followed by co-fermentation with Propionibacterium finigense, Lactobacillus brevis, and Bifidobacterium longum. Both the blackcurrant extract and white hyacinth bean seed extract have excellent anti-fatigue, antioxidant, and immune-enhancing effects, making the mixed soda both delicious and healthy. The specific technical solution is as follows:

[0008] A blackcurrant and orange juice mixed soft drink, comprising the following ingredients in parts by weight: 5 to 8 parts blackcurrant extract, 10 to 15 parts orange juice, 0.5 to 1.5 parts white hyacinth bean seed extract, 0.005 to 0.01 parts mogroside, 0.002 to 0.008 parts steviol glycoside, and 50 to 60 parts water.

[0009] The blackcurrant extract is a product with a value of less than 5 kDa obtained by enzymatic hydrolysis of blackcurrants with hemicellulase and pectinase, followed by sequential enzymatic hydrolysis with nattokinase, figase, papain and glucokinase; the white hyacinth bean seed extract is a product with a value of less than 5 kDa obtained by enzymatic hydrolysis of white hyacinth bean seeds with trypsin, followed by co-fermentation with Propionibacterium finigense, Lactobacillus brevis and Bifidobacterium longum.

[0010] The preparation method of the blackcurrant extract in the above technical solution includes the following steps:

[0011] Take washed blackcurrants, crush them, add 6 to 8 times their weight of water, adjust the pH to 5.0 to 5.5, heat to 45℃ to 55℃, add 0.8% to 2.5% of the blackcurrant's weight of hemicellulase and pectinase respectively, and hydrolyze at a constant temperature for 2 to 4 hours. Cool to 40℃ to 50℃, adjust the pH to 8.0 to 9.0, add 0.8% to 1.5% of the blackcurrant's weight of nattokinase, and hydrolyze at a constant temperature for 1.5 to 2 hours. Heat to inactivate the enzymes, cool to 60℃ to 65℃, adjust the pH to 5.5 to 6.5, and add 0.8% to 1.5% of the blackcurrant's weight of fig protease. The enzyme was hydrolyzed at a constant temperature for 50-80 minutes, then heated to inactivate the enzyme, cooled to 55-65°C, and the pH was adjusted to 6.0-7.0. Papain (0.8%-1.5% by weight of blackcurrant) was added, and the enzyme was hydrolyzed at a constant temperature for 50-80 minutes. The enzyme was then heated to inactivate the enzyme, cooled to 35-38°C, and the pH was adjusted to 7.5-8.0. Glucokinase (0.05%-0.1% by weight of blackcurrant) was added, and the enzyme was hydrolyzed at a constant temperature for 1-1.5 hours. The enzyme was then heated to inactivate the enzyme, cooled to room temperature, and filtered through a 5kDa ultrafiltration membrane to obtain a filtrate with a concentration below 5kDa. This filtrate was concentrated and dried to obtain the blackcurrant extract.

[0012] In the above-mentioned method for preparing blackcurrant extract, the enzyme activities of hemicellulase, pectinase, fig protease, and papain are 80,000 U / g to 100,000 U / g, the enzyme activity of nattokinase is 5,000 U / g to 10,000 U / g, and the enzyme activity of glucokinase is 500 U / g.

[0013] In the above method for preparing blackcurrant extract, the temperature for inactivating the enzyme is 85℃~90℃ and the time is 15min~20min.

[0014] The preparation method of the white hyacinth bean seed extract in the above technical solution includes the following steps:

[0015] Take washed white hyacinth bean seeds, crush them, add 8 to 10 times the weight of the white hyacinth bean seeds in water, adjust the pH to 7.5 to 8.0, add 1% to 2% of the weight of the white hyacinth bean seeds in trypsin, and enzymatically hydrolyze at 36℃ to 39℃ for 3 to 5 hours. After heating to inactivate the enzyme, cool to room temperature, add 0.03% to 0.05% of the weight of the white hyacinth bean seeds in galactooligosaccharides, adjust the pH to 6.5, and add 0.2% to 0.5% of the weight of the white hyacinth bean seeds in Propionibacterium fibrillosa, Lactobacillus brevis, and Bifidobacterium longum, respectively. Perform anaerobic fermentation at 37℃ for 30 to 40 hours, sterilize by heating, cool to room temperature, filter using a 5kDa ultrafiltration membrane to obtain a filtrate with a strength below 5kDa, concentrate, and dry to obtain the white hyacinth bean seed extract.

[0016] In the above method for preparing white hyacinth bean seed extract, the trypsin activity is 100,000 U / g to 500,000 U / g; the bacterial activity of Propionibacterium fischeri, Lactobacillus brevis, and Bifidobacterium longum is 1 billion CFU / g to 3 billion CFU / g.

[0017] In the above-mentioned method for preparing white hyacinth bean seed extract, the temperature for inactivating the enzyme is 80℃~85℃ and the time is 15min~20min; the temperature for sterilization is 121℃~130℃ and the time is 10min~15min.

[0018] In the above technical solution, the mixed soft drink also contains non-sugar additives, including one or more combinations of sodium citrate, vitamins, baking soda, sodium chloride, ethyl maltol, flavorings and colorings.

[0019] The preparation method of the above-mentioned blackcurrant and orange juice mixed soda includes the following steps:

[0020] Add water to 35℃~45℃ according to the mass ratio, then add monk fruit glycosides and steviol glycosides and mix evenly. Then add orange juice, blackcurrant extract and white hyacinth bean seed extract and mix evenly. Perform negative pressure deoxygenation. Then perform instantaneous sterilization, cool, introduce sterile carbon dioxide gas, and package to obtain mixed soft drink.

[0021] In the above preparation method, the temperature of the negative pressure deoxygenation is 35℃~45℃, and the pressure is 0.04MPa~0.06MPa; the temperature of the instantaneous sterilization is 130℃~150℃, and the time is 10s~15s; the temperature of the cooling is 80℃~85℃.

[0022] The blackcurrant and orange juice mixed soda provided by this invention has the following advantages compared with the prior art:

[0023] I. This invention's assorted soft drink contains blackcurrant extract, orange juice, white hyacinth bean seed extract, mogrosides, steviol glycosides, etc., possessing excellent nutritional value. Blackcurrants are rich in vitamin C, B vitamins, anthocyanins, flavonoids, and other nutrients, offering numerous health benefits such as antioxidant and anti-inflammatory properties. Orange juice is rich in vitamin C, flavonoids, minerals, etc., helping to boost immunity and possessing antioxidant and anti-inflammatory properties. Minerals help maintain normal heart function and blood pressure levels. The acidity of orange juice stimulates the secretion of saliva and gastric juices, aiding digestion. The aroma of orange juice stimulates the olfactory nerves, creating a refreshing and comfortable feeling, relieving fatigue. Mogrosides and steviol glycosides enhance sweetness without raising blood sugar.

[0024] II. The blackcurrant extract of this invention is a product with a value of less than 5kDa obtained by enzymatic hydrolysis of blackcurrants by hemicellulase and pectinase, followed by sequential enzymatic hydrolysis by nattokinase, fig protease, papain and glucokinase. It can obtain small molecule peptide products with anti-fatigue, antioxidant and immune-enhancing effects. Small molecule peptides can effectively remove free radicals in the body, reduce oxidative stress damage to cells, help delay aging, stimulate the activity of immune system cells, improve the body's immune function, enhance resistance to disease, and effectively improve the anti-fatigue and immune-enhancing effects of soft drinks.

[0025] III. The white hyacinth bean seed extract of this invention is a product with a Da content below 5 kDa obtained by trypsin hydrolysis of white hyacinth bean seeds, followed by co-fermentation with Propionibacterium finigense, Lactobacillus brevis, and Bifidobacterium longum. Specific enzymatic hydrolysis and fermentation processes can yield components with good anti-fatigue, antioxidant, and immune-enhancing effects. The enzymatic fermentation product contains various antioxidants, which can effectively scavenge free radicals in the body, reduce oxidative damage, and delay aging; it helps protect cells from external environmental damage and reduces the risk of chronic diseases. It also has a regulatory effect on the immune system, enhances the body's immunity, improves resistance, and combats fatigue. Detailed Implementation

[0026] The present invention will be further described below with reference to specific implementation examples, but the present invention is not limited to these embodiments.

[0027] Example 1

[0028] A blackcurrant and orange juice mixed soft drink, comprising the following ingredients in parts by weight: 5 parts blackcurrant extract, 10 parts orange juice, 0.5 parts white hyacinth bean seed extract, 0.005 parts monk fruit glycoside, 0.002 parts steviol glycoside, and 50 parts water.

[0029] The preparation method of blackcurrant extract includes the following steps:

[0030] Take washed blackcurrants, crush them, add 6 times their weight of water, adjust the pH to 5.0, heat to 45℃, add 0.8% (by weight of the blackcurrants) of hemicellulase and pectinase respectively, and hydrolyze at a constant temperature for 2 hours. Cool to 40℃, adjust the pH to 8.0, add 0.8% (by weight of the blackcurrants) of nattokinase, and hydrolyze at a constant temperature for 1.5 hours. Heat to 85℃ to inactivate the enzymes for 15 minutes, cool to 60℃, adjust the pH to 5.5, add 0.8% (by weight of the blackcurrants) of fig protease, and hydrolyze at a constant temperature for 50 minutes. The enzyme was inactivated at 85℃ for 15 minutes, then cooled to 55℃ and the pH was adjusted to 6.0. Papain (0.8% by weight of blackcurrant) was added, and the mixture was hydrolyzed at a constant temperature for 50 minutes. The enzyme was then inactivated at 85℃ for 15 minutes, cooled to 35℃, and the pH was adjusted to 7.5. Glucokinase (0.05% by weight of blackcurrant) was added, and the mixture was hydrolyzed at a constant temperature for 1 hour. The enzyme was then inactivated at 85℃ for 15 minutes, cooled to room temperature, and filtered through a 5kDa ultrafiltration membrane to obtain a filtrate with a concentration below 5kDa. The filtrate was then concentrated and dried to obtain the blackcurrant extract.

[0031] The preparation method of white hyacinth bean seed extract includes the following steps:

[0032] Take the washed white hyacinth bean seeds, crush them, add 8 times the weight of water to adjust the pH to 7.5, add 1% of trypsin by weight of white hyacinth bean seeds, and enzymatically hydrolyze at 36℃ for 3 hours. Then, heat to 80℃ to inactivate the enzyme for 15 minutes. After cooling to room temperature, add 0.03% of galactooligosaccharides by weight of white hyacinth bean seeds, and adjust the pH to 6.5. Then, add 0.2% of Propionibacterium fischeri, Lactobacillus brevis, and Bifidobacterium longum by weight of white hyacinth bean seeds respectively. Perform anaerobic fermentation at 37℃ for 30 hours, then heat to 121℃ for sterilization for 10 minutes. After cooling to room temperature, filter using a 5kDa ultrafiltration membrane to obtain a filtrate with a strength below 5kDa. Concentrate and dry to obtain the white hyacinth bean seed extract.

[0033] The preparation method of the above-mentioned blackcurrant and orange juice mixed soda includes the following steps:

[0034] Add water to 35°C according to the specified mass ratio, then add mogrosides and steviol glycosides and mix well. Next, add orange juice, blackcurrant extract and white hyacinth bean seed extract and mix well. Perform negative pressure deoxygenation at 35°C and 0.04 MPa. Then, perform instantaneous sterilization at 130°C for 10 seconds, cool to 70°C, introduce sterile carbon dioxide gas, and package to obtain mixed soft drink.

[0035] The enzyme activities of the hemicellulase, pectinase, fig protease, papain, nattokinase, glucokinase, and trypsin used in this embodiment are 100,000 U / g, 100,000 U / g, 500 U / g, and 500,000 U / g; the bacterial activity of Propionibacterium fischeri is 3 billion CFU / g, Lactobacillus brevis is 3 billion CFU / g, and Bifidobacterium longum is 3 billion CFU / g.

[0036] Example 2

[0037] A blackcurrant and orange juice mixed soft drink, comprising the following ingredients in parts by weight: 6.5 parts blackcurrant extract, 12 parts orange juice, 1 part white hyacinth bean seed extract, 0.008 parts mogroside, 0.005 parts steviol glycoside, and 55 parts water.

[0038] The preparation method of blackcurrant extract includes the following steps:

[0039] Take washed blackcurrants, crush them, add 7 times their weight of water, adjust the pH to 5.2, heat to 50℃, add 1.5% hemicellulase and pectinase (by weight of the blackcurrants) respectively, and hydrolyze at this temperature for 3 hours. Cool to 45℃, adjust the pH to 8.5, add 1.2% nattokinase (by weight of the blackcurrants), and hydrolyze at this temperature for 1.5 hours. Heat to 88℃ to inactivate the enzymes for 18 minutes, cool to 63℃, adjust the pH to 6.0, add 1.2% fig protease (by weight of the blackcurrants), and hydrolyze at this temperature for 65 minutes. The enzyme was inactivated at 88℃ for 18 minutes, then cooled to 60℃, and the pH was adjusted to 6.5. Papain (1.2% by weight of blackcurrant) was added, and the mixture was hydrolyzed at a constant temperature for 65 minutes. The enzyme was then inactivated at 88℃ for 18 minutes, cooled to 37℃, and the pH was adjusted to 7.8. Glucokinase (0.08% by weight of blackcurrant) was added, and the mixture was hydrolyzed at a constant temperature for 1 hour. The enzyme was then inactivated at 88℃ for 18 minutes, cooled to room temperature, and filtered through a 5kDa ultrafiltration membrane to obtain a filtrate with a concentration below 5kDa. The filtrate was then concentrated and dried to obtain the blackcurrant extract.

[0040] The preparation method of white hyacinth bean seed extract includes the following steps:

[0041] Take the washed white hyacinth bean seeds, crush them, add 9 times the weight of water to adjust the pH to 7.8, add 1.5% trypsin by weight of white hyacinth bean seeds, and enzymatically hydrolyze at 37℃ for 4 hours. Then, heat to 82℃ to inactivate the enzyme for 18 minutes. After cooling to room temperature, add 0.04% galactooligosaccharides by weight of white hyacinth bean seeds, and adjust the pH to 6.5. Add 0.3% each of Propionibacterium fischeri, Lactobacillus brevis, and Bifidobacterium longum by weight of white hyacinth bean seeds, and anaerobic ferment at 37℃ for 35 hours. Then, heat to 125℃ to sterilize for 12 minutes. After cooling to room temperature, filter using a 5kDa ultrafiltration membrane to obtain a filtrate with a strength below 5kDa. Concentrate and dry to obtain the white hyacinth bean seed extract.

[0042] The preparation method of the above-mentioned blackcurrant and orange juice mixed soda includes the following steps:

[0043] Add water to 40°C according to the specified mass ratio, then add mogrosides and steviol glycosides and mix well. Next, add orange juice, blackcurrant extract and white hyacinth bean seed extract and mix well. Perform negative pressure deoxygenation at 40°C and 0.05 MPa. Then, perform instantaneous sterilization at 140°C for 12 seconds, cool to 75°C, introduce sterile carbon dioxide gas, and package to obtain assorted soft drink.

[0044] The enzyme activities of the hemicellulase, pectinase, fig protease, papain, nattokinase, glucokinase, and trypsin used in this embodiment are 100,000 U / g, 100,000 U / g, 100,000 U / g, 5,000 U / g, 500 U / g, and 300,000 U / g; the bacterial activity of Propionibacterium fischeri is 2 billion CFU / g, Lactobacillus brevis is 2 billion CFU / g, and Bifidobacterium longum is 2 billion CFU / g.

[0045] Example 3

[0046] A blackcurrant and orange juice mixed soft drink, comprising the following ingredients in parts by weight: 8 parts blackcurrant extract, 15 parts orange juice, 1.5 parts white hyacinth bean seed extract, 0.01 parts mogroside, 0.008 parts steviol glycoside, and 60 parts water.

[0047] The preparation method of blackcurrant extract includes the following steps:

[0048] Take washed blackcurrants, crush them, add 8 times their weight of water, adjust the pH to 5.5, heat to 55℃, add 2.5% (by weight of the blackcurrants) of hemicellulase and pectinase respectively, and hydrolyze at a constant temperature for 4 hours. Cool to 50℃, adjust the pH to 9.0, add 1.5% (by weight of the blackcurrants) of nattokinase, and hydrolyze at a constant temperature for 2 hours. Heat to 90℃ to inactivate the enzymes for 20 minutes, cool to 65℃, adjust the pH to 6.5, add 1.5% (by weight of the blackcurrants) of fig protease, and hydrolyze at a constant temperature for 80 minutes. The enzyme was inactivated at 90℃ for 20 min, then cooled to 65℃, and the pH was adjusted to 7.0. Papain (1.5% by weight of blackcurrant) was added, and the mixture was hydrolyzed at a constant temperature for 80 min. The temperature was then raised to 90℃ to inactivate the enzyme for 20 min, cooled to 38℃, and the pH was adjusted to 8.0. Glucokinase (0.1% by weight of blackcurrant) was added, and the mixture was hydrolyzed at a constant temperature for 1.5 h. The temperature was then raised to 90℃ to inactivate the enzyme for 20 min, cooled to room temperature, and filtered through a 5 kDa ultrafiltration membrane to obtain a filtrate with a concentration below 5 kDa. The filtrate was then concentrated and dried to obtain the blackcurrant extract.

[0049] The preparation method of white hyacinth bean seed extract includes the following steps:

[0050] Take the washed white hyacinth bean seeds, crush them, add 10 times the weight of water to adjust the pH to 8.0, add 2% trypsin by weight of white hyacinth bean seeds, and enzymatically hydrolyze at 39℃ for 5 hours. Then, heat to 85℃ to inactivate the enzyme for 20 minutes. After cooling to room temperature, add 0.05% galactooligosaccharides by weight of white hyacinth bean seeds and adjust the pH to 6.5. Then, add 0.5% Propionibacterium fibrillosa, Lactobacillus brevis, and Bifidobacterium longum by weight of white hyacinth bean seeds respectively. Perform anaerobic fermentation at 37℃ for 40 hours, then heat to 130℃ for sterilization for 15 minutes. After cooling to room temperature, filter using a 5kDa ultrafiltration membrane to obtain a filtrate with a strength below 5kDa. Concentrate and dry to obtain the white hyacinth bean seed extract.

[0051] The preparation method of the above-mentioned blackcurrant and orange juice mixed soda includes the following steps:

[0052] Add water to 45°C according to the specified mass ratio, then add mogrosides and steviol glycosides and mix well. Next, add orange juice, blackcurrant extract and white hyacinth bean seed extract and mix well. Perform negative pressure deoxygenation at 45°C and 0.06 MPa. Then, perform instantaneous sterilization at 150°C for 15 seconds, cool to 80°C, introduce sterile carbon dioxide gas, and package to obtain assorted soft drink.

[0053] The enzyme activities of the hemicellulase, pectinase, fig protease, papain, nattokinase, glucokinase, and trypsin used in this embodiment are 100,000 U / g, 100,000 U / g, 500 U / g, and 500,000 U / g; the bacterial activity of Propionibacterium fischeri is 3 billion CFU / g, Lactobacillus brevis is 3 billion CFU / g, and Bifidobacterium longum is 3 billion CFU / g.

[0054] Example 4

[0055] A blackcurrant and orange juice mixed soft drink, comprising the following ingredients in parts by weight: 5 parts blackcurrant extract, 15 parts orange juice, 0.5 parts white hyacinth bean seed extract, 0.01 parts mogroside, 0.002 parts steviol glycoside, and 60 parts water; and also containing 0.0002 parts sodium citrate, 0.0001 parts vitamin E, 0.0005 parts baking soda, and 0.0001 parts flavoring.

[0056] The preparation method of blackcurrant extract includes the following steps:

[0057] Take washed blackcurrants, crush them, add 6 times their weight of water, adjust the pH to 5.5, heat to 45℃, add 2.5% (by weight of the blackcurrants) of hemicellulase and pectinase respectively, and hydrolyze at a constant temperature for 2 hours. Cool to 50℃, adjust the pH to 8.0, add 1.5% (by weight of the blackcurrants) of nattokinase, and hydrolyze at a constant temperature for 1.5 hours. Heat to 90℃ to inactivate the enzymes for 15 minutes, cool to 65℃, adjust the pH to 5.5, add 1.5% (by weight of the blackcurrants) of fig protease, and hydrolyze at a constant temperature for 50 minutes. The enzyme was inactivated at 90℃ for 15 minutes, then cooled to 65℃ and the pH was adjusted to 6.0. Papain (1.5% by weight of blackcurrant) was added, and the mixture was hydrolyzed at a constant temperature for 50 minutes. The enzyme was then inactivated at 90℃ for 15 minutes, cooled to 38℃, and the pH was adjusted to 7.5. Glucokinase (0.1% by weight of blackcurrant) was added, and the mixture was hydrolyzed at a constant temperature for 1 hour. The enzyme was then inactivated at 90℃ for 15 minutes, cooled to room temperature, and filtered through a 5kDa ultrafiltration membrane to obtain a filtrate with a concentration below 5kDa. The filtrate was then concentrated and dried to obtain the blackcurrant extract.

[0058] The preparation method of white hyacinth bean seed extract includes the following steps:

[0059] Take the washed white hyacinth bean seeds, crush them, add 10 times the weight of water to adjust the pH to 7.5, add 2% trypsin by weight of white hyacinth bean seeds, and enzymatically hydrolyze at 36℃ for 5 hours. Then, heat to 80℃ to inactivate the enzyme for 20 minutes. After cooling to room temperature, add 0.03% galactooligosaccharides by weight of white hyacinth bean seeds and adjust the pH to 6.5. Then, add 0.5% Propionibacterium fischeri, Lactobacillus brevis, and Bifidobacterium longum by weight of white hyacinth bean seeds respectively. Perform anaerobic fermentation at 37℃ for 30 hours, then heat to 130℃ for sterilization for 10 minutes. After cooling to room temperature, filter using a 5kDa ultrafiltration membrane to obtain a filtrate with a strength below 5kDa. Concentrate and dry to obtain the white hyacinth bean seed extract.

[0060] The preparation method of the above-mentioned blackcurrant and orange juice mixed soda includes the following steps:

[0061] Add water to 45°C according to the specified mass ratio, then add mogrosides and steviol glycosides and mix well. Next, add orange juice, blackcurrant extract and white hyacinth bean seed extract and mix well. Perform negative pressure deoxygenation at 35°C and 0.06 MPa. Then, perform instantaneous sterilization at 130°C for 15 seconds, cool to 70°C, introduce sterile carbon dioxide gas, and package to obtain mixed soft drink.

[0062] The enzyme activities of the hemicellulase, pectinase, fig protease, papain, nattokinase, glucokinase, and trypsin used in this embodiment are 100,000 U / g, 80,000 U / g, 100,000 U / g, 80,000 U / g, 10,000 U / g, 500 U / g, and 100,000 U / g; the bacterial activity of Propionibacterium fischeri is 3 billion CFU / g, Lactobacillus brevis is 1 billion CFU / g, and Bifidobacterium longum is 3 billion CFU / g.

[0063] Example 5

[0064] A blackcurrant and orange juice mixed soft drink, comprising the following ingredients in parts by weight: 8 parts blackcurrant extract, 10 parts orange juice, 1.5 parts white hyacinth bean seed extract, 0.005 parts mogroside, 0.008 parts steviol glycoside, and 60 parts water; and also containing 0.005 parts sodium chloride, 0.0001 parts ethyl maltol, 0.0001 parts flavoring and 0.0003 parts coloring.

[0065] The preparation method of blackcurrant extract includes the following steps:

[0066] Take washed blackcurrants, crush them, add 8 times their weight of water, adjust the pH to 5.0, heat to 55℃, add 0.8% hemicellulase and pectinase (by weight of the blackcurrants) respectively, and hydrolyze at a constant temperature for 4 hours. Cool to 40℃, adjust the pH to 9.0, add 0.8% nattokinase (by weight of the blackcurrants), and hydrolyze at a constant temperature for 2 hours. Heat to 85℃ to inactivate the enzymes for 20 minutes, cool to 60℃, adjust the pH to 6.5, add 0.8% fig protease (by weight of the blackcurrants), and hydrolyze at a constant temperature for 80 minutes. The enzyme was inactivated at 85℃ for 20 min, then cooled to 55℃, and the pH was adjusted to 7.0. Papain (0.8% by weight of blackcurrant) was added, and the mixture was hydrolyzed at a constant temperature for 80 min. The temperature was then raised to 85℃ to inactivate the enzyme for 20 min, cooled to 35℃, and the pH was adjusted to 8.0. Glucokinase (0.05% by weight of blackcurrant) was added, and the mixture was hydrolyzed at a constant temperature for 1.5 h. The temperature was then raised to 85℃ to inactivate the enzyme for 20 min, cooled to room temperature, and filtered using a 5 kDa ultrafiltration membrane to obtain a filtrate with a concentration below 5 kDa. The filtrate was then concentrated and dried to obtain the blackcurrant extract.

[0067] The preparation method of white hyacinth bean seed extract includes the following steps:

[0068] Take the washed white hyacinth bean seeds, crush them, add 8 times the weight of water to adjust the pH to 8.0, add 1% of trypsin by weight of white hyacinth bean seeds, and enzymatically hydrolyze at 39℃ for 3 hours. Then, heat to 85℃ to inactivate the enzyme for 15 minutes. After cooling to room temperature, add 0.05% of galactooligosaccharides by weight of white hyacinth bean seeds and adjust the pH to 6.5. Then, add 0.2% of Propionibacterium fischeri, Lactobacillus brevis, and Bifidobacterium longum by weight of white hyacinth bean seeds respectively. Perform anaerobic fermentation at 37℃ for 40 hours, then heat to 121℃ to sterilize for 15 minutes. After cooling to room temperature, filter using a 5kDa ultrafiltration membrane to obtain a filtrate with a strength below 5kDa. Concentrate and dry to obtain the white hyacinth bean seed extract.

[0069] The preparation method of the above-mentioned blackcurrant and orange juice mixed soda includes the following steps:

[0070] Add water to 35°C according to the specified mass ratio, then add mogrosides and steviol glycosides and mix well. Next, add orange juice, blackcurrant extract and white hyacinth bean seed extract and mix well. Perform negative pressure deoxygenation at 45°C and 0.04 MPa. Then, perform instantaneous sterilization at 150°C for 10 seconds, cool to 80°C, introduce sterile carbon dioxide gas, and package to obtain assorted soft drink.

[0071] The enzyme activities of the hemicellulase, pectinase, fig protease, papain, nattokinase, glucokinase, and trypsin used in this embodiment are 80,000 U / g, 100,000 U / g, 80,000 U / g, 100,000 U / g, 5,000 U / g, 500 U / g, and 500,000 U / g; the bacterial activity of Propionibacterium fischeri is 1 billion CFU / g, Lactobacillus brevis is 3 billion CFU / g, and Bifidobacterium longum is 1 billion CFU / g.

[0072] Comparative Example 1

[0073] In the preparation method of blackcurrant extract, nattokinase enzymatic hydrolysis is not performed; other parameters and methods are the same as in Example 1.

[0074] Comparative Example 2

[0075] In the preparation method of blackcurrant extract, enzymatic hydrolysis with fig protease is not performed; other parameters and methods are the same as in Example 1.

[0076] Comparative Example 3

[0077] In the preparation method of blackcurrant extract, papain enzymatic hydrolysis is not performed; other parameters and methods are the same as in Example 1.

[0078] Comparative Example 4

[0079] No white hyacinth bean seed extract was added; the components of the white hyacinth bean seed extract were replaced with water. Other parameters and methods were the same as in Example 1.

[0080] Comparative Example 5

[0081] No white hyacinth bean seed extract was added, and the white hyacinth bean seeds were not subjected to trypsin hydrolysis; they were directly fermented. Other parameters and methods were the same as in Example 1.

[0082] Comparative Example 6

[0083] In the preparation method of white hyacinth bean seed extract, fermentation is not carried out, and the trypsin hydrolysate is directly filtered after adjusting the pH to 6.5; other parameters and methods are the same as in Example 1.

[0084] Comparative Example 7

[0085] In the preparation method of white hyacinth bean seed extract, no Propionibacterium fischeri or Lactobacillus brevis are added during fermentation; other parameters and methods are the same as in Example 1.

[0086] The anti-fatigue, antioxidant, and immune-enhancing effects of the semi-finished carbonated beverage products (which were samples of non-carbonated water without added carbon dioxide) in the above embodiments and comparative examples were tested.

[0087] I. Fatigue resistance

[0088] Each semi-finished soft drink product was concentrated to 50% of its original volume to obtain a sample.

[0089] Female ICR mice (weighing 20g-22g) were randomly divided into three groups of six: a blank control group, an example group, and a comparative group. After 3 days of acclimatization, the experiment was conducted: the blank control group was administered 0.5mL of distilled water by gavage daily; the example group was administered 0.5mL of the sample corresponding to each example by gavage daily; and the comparative group was administered 0.5mL of the sample corresponding to each comparative example by gavage daily. This gavage continued for 20 days. 30 minutes after the last gavage, a weighted swimming test was conducted. Lead sheets of 3% of the mouse's body weight were attached to the mouse's tail, and the mouse was placed in 25℃ water at a depth of 25cm. Timing was started. The first sinking was defined as the mouse's nose tip first submerging below the water surface; the exhaustion was defined as the mouse remaining submerged for 10 seconds. The time of the first sinking and the time of exhaustion were recorded. The longer the time, the better the endurance, i.e., the better the fatigue resistance. The test results are shown in Table 1 below.

[0090] Table 1 Fatigue resistance (average value)

[0091]

[0092] The results above indicate that the semi-finished carbonated beverage samples from Examples 1 to 5 exhibit excellent anti-fatigue effects. Specifically, the results from Comparative Examples 1 to 3 show that changing the preparation method of the blackcurrant extract—specifically, omitting enzymatic hydrolysis with nattokinase, figase, or papain—results in different product compositions and a decrease in the anti-fatigue effect. The results from Comparative Example 4 show that the white hyacinth bean seed extract possesses good anti-fatigue effects. The results from Comparative Examples 5 to 7 show that changing the preparation method of the white hyacinth bean seed extract—specifically, omitting trypsin hydrolysis, omitting fermentation, or omitting the addition of Propionibacterium fischeri and Lactobacillus brevis during fermentation—all affect the product composition and decrease the anti-fatigue effect.

[0093] II. Antioxidant properties

[0094] After decolorizing each semi-finished soft drink product using activated carbon adsorption, it was concentrated to 30% of its volume to obtain a sample.

[0095] Add 2 mL of the sample to a cuvette, then add 2 mL of 1.0 × 10⁴ mol / L DPPH solution, mix well, and let stand for 30 min. Use a 95% (v / v) ethanol aqueous solution as a reference solution to measure the absorbance value A at 517 nm. Add 2.0 mL of the sample to a cuvette, then add 2 mL of 95% (v / v) ethanol aqueous solution, mix well, and let stand for 30 min. Use a 95% (v / v) ethanol aqueous solution as a reference solution to measure the absorbance value A0 at 517 nm. Add 2 mL of 1.0 × 10⁴ mol / L DPPH solution to a cuvette, then add 2.0 mL of 95% (v / v) ethanol aqueous solution, mix well, and let stand for 30 min. Use a 95% (v / v) ethanol aqueous solution as a reference solution to measure the absorbance value A1 at 517 nm. Calculate the DPPH scavenging rate: Scavenging rate (%) = [1 - (A - A0) / A1] × 100. The detection results are shown in Table 2 below.

[0096] Table 2. DPPH free radical scavenging rate (average)

[0097]

[0098] The results above indicate that the semi-finished carbonated beverage samples from Examples 1 to 5 exhibit excellent antioxidant activity in scavenging free radicals. Specifically, the results from Comparative Examples 1 to 3 show that changing the preparation method of the blackcurrant extract—specifically, omitting enzymatic hydrolysis with nattokinase, figase, or papain—decreases the free radical scavenging rate. The results from Comparative Example 4 demonstrate that the white hyacinth bean seed extract possesses good free radical scavenging activity. The results from Comparative Examples 5 to 7 show that changing the preparation method of the white hyacinth bean seed extract—specifically, omitting trypsin hydrolysis, omitting fermentation, or omitting the addition of Propionibacterium fischeri and Lactobacillus brevis during fermentation—affects the product composition and decreases the free radical scavenging rate.

[0099] III. Effects on Enhancing Immunity

[0100] Each semi-finished soft drink product was concentrated to 40% of its original volume to obtain a sample.

[0101] Female ICR mice (weighing 20g-22g) were randomly divided into groups of 6 mice each. After acclimatization for 3 days, the experiment was conducted: the normal control group was administered 0.5mL of distilled water by gavage daily; the model control group was administered 0.5mL of distilled water by gavage daily; the spleen peptide group was administered 1.2g / kg of spleen peptide by gavage daily; the example groups were administered 0.5mL of the sample corresponding to each example by gavage daily; and the comparative groups were administered 0.5mL of the sample corresponding to each comparative example by gavage daily.

[0102] Except for the normal control group, the other groups were administered cyclophosphamide 40 mg / kg intraperitoneally for two consecutive days after 3 weeks of continuous gavage. After 4 weeks of continuous gavage, 1 ml of whole blood was collected from each mouse, and the total white blood cell count and lymphocyte count were detected using a complete blood cell analyzer; the results are shown in Table 3 below.

[0103] Table 3. Boosting Immunity (Average)

[0104]

[0105] The results above indicate that the components of the semi-finished carbonated beverage samples from Examples 1 to 5 have a certain effect on enhancing immunity. Specifically, the results from Comparative Examples 1 to 3 show that changing the preparation method of the blackcurrant extract—that is, omitting the enzymatic hydrolysis with nattokinase, figase, or papain—significantly reduces the effect on enhancing immunity. The results from Comparative Example 4 show that the white hyacinth bean seed extract also has a good effect on enhancing immunity. The results from Comparative Examples 5 to 7 show that changing the preparation method of the white hyacinth bean seed extract—that is, omitting trypsin hydrolysis, omitting fermentation, and omitting the addition of Propionibacterium fischeri and Lactobacillus brevis during fermentation—affects the degree of enhancement of immunity.

Claims

1. A blackcurrant and orange juice mixed soda, characterized in that, The soft drink comprises the following ingredients in parts by weight: 5 to 8 parts blackcurrant extract, 10 to 15 parts orange juice, 0.5 to 1.5 parts white hyacinth bean seed extract, 0.005 to 0.01 parts mogroside, 0.002 to 0.008 parts steviol glycoside, and 50 to 60 parts water; the mixed soft drink also contains non-sugar additives, including one or more combinations of sodium citrate, vitamins, baking soda, sodium chloride, ethyl maltol, flavorings, and colorings. The preparation method of the blackcurrant extract includes the following steps: Take washed blackcurrants, crush them, add 6 to 8 times the mass of the blackcurrants in water, adjust the pH to 5.0 to 5.5, raise the temperature to 45℃ to 55℃, add 0.8% to 2.5% of the blackcurrants' mass of hemicellulase and pectinase respectively, and enzymatically hydrolyze at a constant temperature for 2 to 4 hours. Cool down to 40℃ to 50℃, adjust the pH to 8.0 to 9.0, add 0.8% to 1.5% of the blackcurrants' mass of nattokinase, and enzymatically hydrolyze at a constant temperature for 1.5 to 2 hours. Raise the temperature to inactivate the enzyme, lower the temperature to 60℃ to 65℃, adjust the pH to 5.5 to 6.5, and add 0.8% to 1% of the blackcurrants' mass of nattokinase. 5% fig protease was added and hydrolyzed at a constant temperature for 50-80 minutes. The enzyme was then inactivated by heating and cooled to 55-65°C. The pH was adjusted to 6.0-7.

0. Papain (0.8%-1.5% by weight of blackcurrant) was added and hydrolyzed at a constant temperature for 50-80 minutes. The enzyme was then inactivated by heating and cooled to 35-38°C. The pH was adjusted to 7.5-8.

0. Glucokinase (0.05%-0.1% by weight of blackcurrant) was added and hydrolyzed at a constant temperature for 1-1.5 hours. The enzyme was then inactivated by heating and cooled to room temperature. The mixture was filtered through a 5kDa ultrafiltration membrane to obtain a filtrate with a concentration below 5kDa. The filtrate was concentrated and dried to obtain the blackcurrant extract. The preparation method of the white hyacinth bean seed extract includes the following steps: Take washed white hyacinth bean seeds, crush them, add 8 to 10 times the weight of the white hyacinth bean seeds in water, adjust the pH to 7.5 to 8.0, add 1% to 2% of the weight of the white hyacinth bean seeds in trypsin, and enzymatically hydrolyze at 36℃ to 39℃ for 3 to 5 hours. After heating to inactivate the enzyme, and cooling to room temperature, add 0.03% to 0.05% of the weight of the white hyacinth bean seeds in galactooligosaccharides, adjust the pH to 6.5, and add 0.2% to 0.5% of the weight of the white hyacinth bean seeds in Propionibacterium fibrillosa, Lactobacillus brevis, and Bifidobacterium longum, respectively. Perform anaerobic fermentation at 37℃ for 30 to 40 hours, sterilize by heating, cool to room temperature, filter using a 5kDa ultrafiltration membrane to obtain a filtrate with a strength below 5kDa, concentrate, and dry to obtain the white hyacinth bean seed extract.

2. The blackcurrant and orange juice mixed soda according to claim 1, characterized in that, In the preparation method of blackcurrant extract, the enzyme activities of hemicellulase, pectinase, fig protease, and papain are 80,000 U / g to 100,000 U / g, the enzyme activity of nattokinase is 5,000 U / g to 10,000 U / g, and the enzyme activity of glucokinase is 500 U / g.

3. The blackcurrant and orange juice mixed soda according to claim 1, characterized in that, In the preparation method of blackcurrant extract, the temperature for inactivating the enzyme is 85℃~90℃ and the time is 15min~20min.

4. The blackcurrant and orange juice mixed soda according to claim 1, characterized in that, In the preparation method of white hyacinth bean seed extract, the trypsin enzyme activity is 100,000 U / g to 500,000 U / g; the bacterial activity of Propionibacterium fischeri, Lactobacillus brevis and Bifidobacterium longum is 1 billion CFU / g to 3 billion CFU / g.

5. A blackcurrant and orange juice mixed soda according to claim 1, characterized in that, In the preparation method of white hyacinth bean seed extract, the temperature for inactivating enzymes is 80℃~85℃ and the time is 15min~20min; the temperature for sterilization is 121℃~130℃ and the time is 10min~15min.

6. A blackcurrant and orange juice mixed soda according to claim 1, characterized in that, The preparation method of mixed soft drinks includes the following steps: Add water to 35℃~45℃ according to the mass ratio, then add monk fruit glycosides and steviol glycosides and mix evenly. Then add orange juice, blackcurrant extract and white hyacinth bean seed extract and mix evenly. Perform negative pressure deoxygenation. Then perform instantaneous sterilization, cool, introduce sterile carbon dioxide gas, and package to obtain mixed soft drink.

7. A blackcurrant and orange juice mixed soda according to claim 6, characterized in that, The negative pressure deoxygenation temperature is 35℃~45℃, and the pressure is 0.04MPa~0.06MPa; the instantaneous sterilization temperature is 130℃~150℃, and the time is 10s~15s; the cooling temperature is 80℃~85℃.

Images