A sparkling water beverage and its production method

Abstract

This invention provides a sparkling water beverage and its production method. The beverage comprises the following raw materials in parts by weight: 2-5 parts of a compound fermented product, 0.5-1 part of cassia seed extract, 1-2 parts of monk fruit extract, 3-6 parts of a compound fruit enzyme freeze-dried powder, 0.01-0.02 parts of vitamin E, 100 parts of purified water, and carbon dioxide added at a rate of 1.5-2.5 liters per liter of beverage. This sparkling water beverage, through a scientifically designed formula and advanced preparation process, exhibits significant blood sugar-lowering and antioxidant functions, while also possessing a pleasant taste and color, providing consumers with a healthy functional beverage option.

Description

Technical Field

[0001] This invention belongs to the field of food production, and in particular relates to a sparkling water beverage and its production method. Background Technology

[0002] With changes in modern lifestyles, the incidence of chronic diseases such as diabetes and cardiovascular disease is rising year by year. These health problems are closely related to poor dietary habits, lack of exercise, and environmental factors. Type 2 diabetes, in particular, has become one of the major public health challenges worldwide. Meanwhile, oxidative stress plays a significant role in the development of many chronic diseases, including diabetic complications such as retinopathy, nephropathy, and neuropathy. Therefore, developing functional beverages that can help control blood sugar levels and provide antioxidant protection has become an important research direction.

[0003] Traditional carbonated beverages typically contain high levels of sugar and other additives, which not only increase calorie intake but may also negatively impact blood sugar levels and lack antioxidants. In contrast, sugar-free or low-sugar sparkling water is favored by consumers because it contains no extra calories. However, plain sugar-free sparkling water does not possess the active blood sugar-regulating and antioxidant effects. To meet the growing consumer demand for healthy drinks, researchers have begun exploring how to incorporate natural ingredients with blood sugar-lowering and antioxidant properties into sparkling water, thereby creating a new type of functional beverage that is both refreshing and healthy.

[0004] Many herbs, fruits, and their extracts have been shown to have potential hypoglycemic effects. For example, bitter melon (Momordica charantia) contains a variety of bioactive compounds, including peptide-P and chalcone, which are believed to mimic the action of insulin and help improve glucose metabolism. In addition, certain components in cinnamon (Cinnamomum verum) have been found to increase cellular sensitivity to insulin and may slow the rate of postprandial blood glucose rise.

[0005] In addition, studies have shown that certain types of tea, such as green tea (Camellia sinensis), are rich in catechin antioxidants. These substances not only have strong antioxidant capabilities, but have also been shown to promote blood sugar stability through different mechanisms.

[0006] Currently, some functional beverages on the market use bitter melon extract as one of their main ingredients, aiming to help consumers control blood sugar. Many other brands have launched sugar-free or low-sugar beverages based on green tea, emphasizing their antioxidant and potential blood sugar management benefits. However, the levels of active ingredients in many functional beverages are insufficient to produce significant physiological effects. Some natural ingredients with blood sugar-lowering and antioxidant properties may have a bitter or other unpleasant taste, affecting the consumer's drinking experience. To address this issue, the amount of active ingredient added may be reduced, thereby weakening its efficacy.

[0007] Based on the above issues, developing a new type of beverage that can both maintain the original flavor of sparkling water and exert blood sugar-lowering and antioxidant effects has become an innovative direction. Summary of the Invention

[0008] The purpose of this invention is to provide a sparkling water beverage and its production method. The sparkling water beverage has a blood sugar lowering effect and antioxidant capacity, while maintaining a good taste and flavor, providing consumers with a healthy functional beverage option.

[0009] To achieve the above objectives, the present invention provides the following technical solution:

[0010] A sparkling water beverage comprises the following ingredients in parts by weight: 2-5 parts of compound fermented product, 0.5-1 part of cassia seed extract, 1-2 parts of monk fruit extract, 3-6 parts of compound fruit enzyme freeze-dried powder, 0.01-0.02 parts of vitamin E, 100 parts of purified water, and carbon dioxide added at a rate of 1.5-2.5 liters / liter of beverage.

[0011] Furthermore, by weight, the raw materials for preparing the compound fermentation product include: 2-4 parts bitter melon, 4-5 parts mulberry leaves, and 1-2 parts turmeric.

[0012] Furthermore, the preparation method of the compound fermentation product includes the following steps:

[0013] (1) Wash bitter melon, mulberry leaves and turmeric, break them into small pieces, add deionized water at a material-to-liquid ratio of 1g:6-10mL, heat to boiling for 1-2 hours, adding water every 10-30 minutes until the original volume is reached, filter after heating to obtain filtrate and residue.

[0014] (2) Add deionized water to the filter residue at a mass ratio of 1:1 to 1.5, add pectinase, cellulase, hemicellulase and acidic protease, adjust the pH to 4.5 to 5, the temperature to 40 to 50°C, and enzymatically hydrolyze for 5 to 10 hours; the amount of pectinase, cellulase, hemicellulase and acidic protease added is 0.1% to 1% of the mass of the filter residue.

[0015] (3) After the enzymatic hydrolysis is completed, raise the temperature to 80-100℃ and keep it for 10-20 minutes to inactivate the enzyme and obtain the enzymatic hydrolysate.

[0016] (4) Add 1%-5% glucose and 0.05%-0.2% yeast extract to the filtrate of step (1) and the enzymatic hydrolysate of step (3) respectively, and then inoculate each with 2-5 wt% of compound bacteria, stir evenly, and anaerobic ferment at 37-40℃ for 2-3 days to obtain fermentation product A and fermentation product B respectively. The compound bacteria are Lactobacillus acidophilus, Bifidobacterium breve and Lactobacillus rhamnosus in a mass ratio of 1:2-4:1-2.

[0017] (5) Filter fermentation product B, mix the filtrate with fermentation product A, add 1%-5% glucose and 0.05%-0.2% yeast extract, then inoculate with 2-5 wt% of Lactobacillus plantarum, stir evenly, and anaerobic ferment at 35-37℃ for 3-4 days to obtain the compound fermentation product.

[0018] Bitter melon contains various active substances, such as momordicin, which has been proven to lower blood sugar. Mulberry leaves contain 1-deoxynojirimycin (DNJ), which can inhibit the activity of α-glucosidase in the intestine, slowing down the breakdown of carbohydrates into monosaccharides, thereby helping to control postprandial blood sugar rise. Turmeric has anti-inflammatory and antioxidant effects, and can also improve insulin sensitivity and indirectly help control blood sugar. This invention selects these three ingredients to work together, affecting blood sugar levels through different pathways. Bitter melon directly promotes blood sugar reduction, mulberry leaves work by slowing down the digestive process, and turmeric provides antioxidant support to help protect β cells from oxidative stress damage and maintain normal insulin secretion function. The blood sugar-lowering effect of the product is significantly enhanced after the three ingredients are combined.

[0019] This invention selects a specific ratio of Lactobacillus acidophilus, Bifidobacterium breve, and Lactobacillus rhamnosus as the compound bacteria for the first fermentation, followed by the addition of Lactobacillus plantarum for a second fermentation. The first fermentation pre-treats raw materials such as bitter melon, mulberry leaves, and turmeric, making their active ingredients easier to extract. The second fermentation further transforms the products from the first fermentation, especially by introducing Lactobacillus plantarum, which enriches the biodiversity of the final product and increases the variety of beneficial metabolites. The compound fermentation product also increases the bioavailability of active substances in cassia seed and monk fruit extracts, making them easier for the human body to absorb and exert their effects.

[0020] This invention utilizes enzymatic hydrolysis to treat the raw material filter residue, which allows the raw material to release more antioxidant components. The fermentation process further improves the bioavailability of these components and enhances the antioxidant capacity of the product.

[0021] Furthermore, by weight, the raw materials for preparing the compound fruit enzyme freeze-dried powder include: 4-5 parts blueberry, 4-5 parts raspberry, and 2-3 parts cherry.

[0022] Furthermore, the preparation method of the compound fruit enzyme freeze-dried powder includes the following steps:

[0023] (1) Wash blueberries, raspberries and cherries, break them into small pieces, add a mixture of citric acid and 60-80 wt% ethanol aqueous solution with a mass ratio of 1:20-50, and a material-to-liquid ratio of 1g:3-7mL, heat to 30-50℃, extract for 1-2 hours, filter after extraction, concentrate under vacuum to remove citric acid and ethanol, and obtain the extract.

[0024] (2) Adjust the pH of the extract in step (1) to 5-6, add 1.5-2.5% of β-glucosidase by mass of the extract, heat to 40-50℃, and hydrolyze for 3-5 hours. After the reaction is complete, filter, concentrate the filtrate, and vacuum dry to obtain the compound fruit enzyme freeze-dried powder.

[0025] Blueberries, raspberries, and cherries are all rich in anthocyanins and other antioxidants. In the preparation of the compound fruit enzyme freeze-dried powder, citric acid and ethanol are used for extraction, which fully extracts the anthocyanins from the raw materials and helps maintain the activity of these natural compounds. Further hydrolysis is performed using β-glucosidase, which hydrolyzes the glycosidic bonds on anthocyanin molecules, removing glycosidic or acylated groups and exposing more phenolic hydroxyl groups, thereby enhancing their ability to react with free radicals and further improving the antioxidant properties of anthocyanins. The hydrolyzed anthocyanin molecules are smaller in size, making it easier for them to cross cell membranes, thus improving their absorption rate and bioavailability in the body. The enzymatic hydrolysis process can also optimize the color and taste of the beverage.

[0026] Furthermore, the preparation method of the cassia seed extract includes the following steps:

[0027] (1) Wash the cassia seeds, dry them, and crush them into small particles;

[0028] (2) Add the cassia seed granules to a 60-80 wt% ethanol aqueous solution, with a material-to-liquid ratio of 1 g: 10-20 mL, and heat under reflux for 1-2 hours;

[0029] (3) After extraction, filter and concentrate the filtrate under vacuum to obtain the cassia seed extract.

[0030] Furthermore, the preparation method of the monk fruit extract includes the following steps:

[0031] (1) Wash the monk fruit, dry it, and crush it into small particles;

[0032] (2) Add the monk fruit granules to deionized water at a material-to-liquid ratio of 1g:10-20mL, heat to boiling, maintain for 1-2 hours, and filter to obtain the first extract.

[0033] (3) Add the residue back into deionized water at a ratio of 1g:10-20mL, heat to boiling, maintain for 1-2 hours, and filter to obtain the second extract.

[0034] (4) The two extracts were combined and concentrated under vacuum to obtain the monk fruit extract.

[0035] Cassia seeds contain various active ingredients, such as emodin and cassiaside, which help improve intestinal function and have a certain auxiliary effect in lowering blood sugar levels. Monk fruit is rich in mogroside V, a non-sugar sweetener that is much sweeter than sucrose but contains almost no calories, making it suitable for diabetics or those who need to control their blood sugar. In addition, other polyphenols in monk fruit also exhibit strong antioxidant properties, helping to eliminate free radicals in the body and reduce the damage caused by oxidative stress.

[0036] In terms of taste and color, the sparkling water of this invention, through the addition of compound fruit enzyme freeze-dried powder, gives the beverage a vibrant color and a rich berry aroma, enhancing the enjoyable drinking experience. Anthocyanins treated with β-glucosidase are more easily absorbed by the body, and the color and taste of the beverage are optimized, making it smoother and more delicate. Bitter melon, mulberry leaves, and turmeric, while inherently possessing a certain bitterness or distinctive odor, undergo significant flavor enhancement through fermentation, making them more mellow and palatable. Cassia seeds have a slight herbal aroma, adding a refreshing botanical touch to the beverage. Monk fruit naturally contains the sweetener mogroside V, which enhances sweetness while retaining a refreshing feel, avoiding the heavy, cloying taste of traditional sugary drinks. Furthermore, monk fruit also carries a subtle fruity aroma, enhancing the overall flavor profile of the beverage. The appropriate addition of carbon dioxide adds a lively effervescence, providing a refreshing and pleasant sensation. Overall, the sparkling water of this invention maintains its refreshing and non-greasy characteristics while also possessing a sense of layering and complexity, making it very suitable for daily consumption by people who pursue a healthy lifestyle.

[0037] The present invention also provides a method for preparing the above-mentioned sparkling water beverage, comprising the following steps:

[0038] (1) Mix the compound fruit enzyme freeze-dried powder with pure water, heat to 40-50℃, stir for 30-60 minutes to fully dissolve the compound fruit enzyme freeze-dried powder, and obtain material 1;

[0039] (2) Add compound fermentation product, cassia seed extract, monk fruit extract and vitamin E to material 1, stir well to obtain material 2;

[0040] (3) The material 2 is subjected to high-temperature instantaneous sterilization at a temperature of 110-120°C for 5-10 seconds to obtain material 3.

[0041] (4) Reduce the temperature of material 3 to 4-8°C and introduce carbon dioxide gas into it to obtain the sparkling water beverage. The amount of carbon dioxide added is 1.5-2.5 liters / liter of beverage.

[0042] Compared with the prior art, the advantages and beneficial effects of the present invention are as follows:

[0043] 1. Highly Effective Blood Sugar Lowering Effect: This invention uses bitter melon, mulberry leaves, and turmeric as the main raw materials for the compound fermentation product. Momordicin in bitter melon can directly lower blood sugar levels; 1-deoxynojirimycin (DNJ) in mulberry leaves can inhibit the activity of α-glucosidase in the intestine, slowing down the rate of carbohydrate breakdown into monosaccharides, thereby controlling postprandial blood sugar rise; curcumin in turmeric not only has anti-inflammatory and antioxidant effects but also improves insulin sensitivity, indirectly helping to control blood sugar. Through two fermentation processes, using specific proportions of Lactobacillus acidophilus, Bifidobacterium breve, Lactobacillus rhamnosus, and Lactobacillus plantarum, the extraction rate and bioavailability of the effective components of these raw materials are further improved, enhancing the blood sugar lowering effect of the product.

[0044] 2. Powerful Antioxidant Properties: The compound fruit enzyme freeze-dried powder is made from blueberries, raspberries, and cherries, fruits rich in natural antioxidants such as anthocyanins. Extracted using a mixed solvent of citric acid and ethanol, and enzymatically hydrolyzed using β-glucosidase, it removes glycosides or acylated groups that hinder the reaction of phenolic hydroxyl groups with free radicals, thus significantly enhancing the antioxidant capacity of anthocyanins. The enzymatically hydrolyzed anthocyanin molecules are smaller in size, making them easier for the body to absorb, improving their bioavailability and further enhancing the overall antioxidant properties of the beverage.

[0045] 3. Optimized taste and color: The sparkling water beverage of the present invention presents a natural and bright color, while having a refreshing and pleasant flavor, enhancing the consumer's drinking experience. Detailed Implementation

[0046] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention. In addition, it is worth noting that the raw materials involved in the present invention are all commercially available products unless otherwise specified.

[0047] The following sources of raw materials are provided as examples:

[0048] Pectinase was purchased from Angel Yeast Co., Ltd., with an enzyme activity of 50 U / mg;

[0049] Cellulase was purchased from Angel Yeast Co., Ltd., with an enzyme activity of 20 U / mg;

[0050] Hemicellulase was purchased from Angel Yeast Co., Ltd., with an enzyme activity of 100 U / mg;

[0051] Acidic protease was purchased from Angel Yeast Co., Ltd., with an enzyme activity of 1000 U / mg;

[0052] β-glucosidase was purchased from Angel Yeast Co., Ltd., with an enzyme activity of 50 U / mg;

[0053] Lactobacillus acidophilus was purchased from Beijing Zhongke Quality Inspection Biotechnology Co., Ltd., product number CICC 6074;

[0054] Bifidobacterium breve was purchased from Beijing Zhongke Quality Inspection Biotechnology Co., Ltd., product number CICC 6079;

[0055] Lactobacillus rhamnosus was purchased from Beijing Zhongke Quality Inspection Biotechnology Co., Ltd., product number ZKCC 15463.

[0056] Example 1

[0057] This embodiment provides a method for preparing a compound ferment, including the following steps:

[0058] (1) Wash 2 parts by weight of bitter melon, 5 parts by weight of mulberry leaves and 1 part by weight of turmeric, break them into small pieces, add deionized water at a material-liquid ratio of 1g:8mL to obtain a mixture; heat and boil for 2 hours, adding water every 10 minutes to the original volume (volume of the mixture), filter after heating to obtain filtrate and residue.

[0059] (2) Add deionized water to the filter residue at a mass ratio of 1:1, add pectinase, cellulase, hemicellulase and acidic protease, adjust the pH to 4.8, set the temperature to 45℃, and enzymatically hydrolyze for 8 hours; the amount of pectinase, cellulase, hemicellulase and acidic protease added is 0.5% of the mass of the filter residue.

[0060] (3) After the enzymatic hydrolysis is completed, the temperature is raised to 100℃ and kept for 10 minutes to inactivate the enzyme, and the enzymatic hydrolysate is obtained.

[0061] (4) Add 2% glucose and 0.1% yeast extract by mass to the filtrate of step (1) and the enzymatic hydrolysate of step (3) respectively, then inoculate each with 3wt% of compound bacteria, stir evenly, and anaerobic ferment at 37°C for 3 days to obtain fermentation product A and fermentation product B respectively. The compound bacteria are Lactobacillus acidophilus, Bifidobacterium breve and Lactobacillus rhamnosus in a mass ratio of 1:3:2.

[0062] (5) Filter fermentation product B, mix the filtrate with fermentation product A, add 1% glucose and 0.05% yeast extract relative to its mass, then inoculate with 4 wt% of Lactobacillus plantarum, stir evenly, and anaerobic ferment at 36°C for 4 days to obtain the compound fermentation product.

[0063] Example 2

[0064] This embodiment provides a method for preparing compound fruit enzyme freeze-dried powder, including the following steps:

[0065] (1) Wash 5 parts by weight of blueberries, 4 parts by weight of raspberries and 2 parts by weight of cherries, break them into small pieces, add a mixture of citric acid and 60wt% ethanol aqueous solution with a mass ratio of 1:30, the material-liquid ratio is 1g:5mL, heat to 40℃, extract for 2 hours, filter after extraction, vacuum concentrate to remove citric acid and ethanol, and obtain the extract.

[0066] (2) Adjust the pH of the extract in step (1) to 5, add 2% of β-glucosidase by mass of the extract, heat to 45°C, hydrolyze for 4 hours, filter after the reaction, concentrate the filtrate, and vacuum dry to obtain the compound fruit enzyme freeze-dried powder.

[0067] Comparative Example 1

[0068] This comparative example provides a method for preparing a compound ferment, which differs from Example 1 in that step (5) is omitted. That is, after fermentation in step (4), ferment B is filtered, and the filtrate is mixed with ferment A to obtain the compound ferment.

[0069] Comparative Example 2

[0070] This comparative example provides a method for preparing a compound ferment, which differs from Example 1 in that step (4) uses 3wt% inoculum of Bifidobacterium breve for fermentation.

[0071] Comparative Example 3

[0072] This comparative example provides a method for preparing a compound fruit enzyme freeze-dried powder. The difference from Example 2 is that step (2) is omitted, that is, the extract in step (1) is vacuum dried to obtain the compound fruit enzyme freeze-dried powder.

[0073] Comparative Example 4

[0074] This comparative example provides a method for preparing a compound fruit enzyme freeze-dried powder, which differs from Example 2 in that step (2) replaces β-glucosidase with cellulase.

[0075] Example 3

[0076] This embodiment provides a method for preparing cassia seed extract, including the following steps:

[0077] (1) Wash the cassia seeds, dry them, and crush them into small particles;

[0078] (2) Add the cassia seed granules to a 60wt% ethanol aqueous solution with a material-to-liquid ratio of 1g:15mL and heat under reflux for 2 hours.

[0079] (3) After extraction, filter and concentrate the filtrate under vacuum to obtain the cassia seed extract.

[0080] Example 4

[0081] This embodiment provides a method for preparing monk fruit extract, including the following steps:

[0082] (1) Wash the monk fruit, dry it, and crush it into small particles;

[0083] (2) Add the monk fruit granules to deionized water at a material-to-liquid ratio of 1g:20mL, heat to boiling, maintain for 2 hours, and filter to obtain the first extract.

[0084] (3) Add the residue back into deionized water at a ratio of 1g:10mL, heat to boiling, maintain for 2 hours, and filter to obtain the second extract.

[0085] (4) The two extracts were combined and concentrated under vacuum to obtain the monk fruit extract.

[0086] Example 5

[0087] This embodiment provides a method for preparing sparkling water beverage, the raw materials of which include, by weight: 4 parts of compound fermentation product, 0.5 parts of cassia seed extract, 2 parts of monk fruit extract, 6 parts of compound fruit enzyme freeze-dried powder, 0.02 parts of vitamin E, 100 parts of purified water, and carbon dioxide added at a rate of 2 liters / liter of beverage.

[0088] The preparation method includes the following steps:

[0089] (1) Mix the compound fruit enzyme freeze-dried powder with pure water, heat to 48°C, stir for 40 minutes to fully dissolve the compound fruit enzyme freeze-dried powder, and obtain material 1;

[0090] (2) Add compound fermentation product, cassia seed extract, monk fruit extract and vitamin E to material 1, stir well to obtain material 2;

[0091] (3) Material 2 is subjected to high-temperature instantaneous sterilization at a temperature of 110°C for 8 seconds to obtain material 3.

[0092] (4) Reduce the temperature of material 3 to 4°C and introduce carbon dioxide gas into it to obtain the sparkling water beverage. The amount of carbon dioxide added is 2 liters / liter of beverage, that is, 2 liters of carbon dioxide are dissolved in every 1 liter of beverage.

[0093] In this embodiment, the compound ferment, compound fruit enzyme freeze-dried powder, cassia seed extract, and monk fruit extract were prepared using the methods described in Examples 1-4, respectively.

[0094] Example 6

[0095] This embodiment provides a method for preparing sparkling water beverage, the raw materials of which include, by weight: 5 parts of compound fermentation product, 1 part of cassia seed extract, 1 part of monk fruit extract, 3 parts of compound fruit enzyme freeze-dried powder, 0.02 parts of vitamin E, 100 parts of purified water, and the amount of carbon dioxide added is 2 liters / liter of beverage.

[0096] The preparation method includes the following steps:

[0097] (1) Mix the compound fruit enzyme freeze-dried powder with pure water, heat to 40°C, stir for 50 minutes to fully dissolve the compound fruit enzyme freeze-dried powder, and obtain material 1;

[0098] (2) Add compound fermentation product, cassia seed extract, monk fruit extract and vitamin E to material 1, stir well to obtain material 2;

[0099] (3) Material 2 is subjected to high-temperature instantaneous sterilization at a temperature of 110°C for 8 seconds to obtain material 3.

[0100] (4) Reduce the temperature of material 3 to 4°C and introduce carbon dioxide gas into it to obtain the sparkling water beverage. The amount of carbon dioxide added is 2 liters / liter of beverage, that is, 2 liters of carbon dioxide are dissolved in every 1 liter of beverage.

[0101] In this embodiment, the compound ferment, compound fruit enzyme freeze-dried powder, cassia seed extract, and monk fruit extract were prepared using the methods described in Examples 1-4, respectively.

[0102] Comparative Example 5

[0103] This comparative example provides a method for preparing sparkling water beverage, which is the same as that in Example 5, except that the compound fermentation product is prepared using the method described in Comparative Example 1.

[0104] Comparative Example 6

[0105] This comparative example provides a method for preparing sparkling water beverage, which is the same as that in Example 5, except that the compound fermentation product is prepared using the method described in Comparative Example 2.

[0106] Comparative Example 7

[0107] This comparative example provides a method for preparing sparkling water beverage, which is the same as that in Example 5, except that the compound fruit enzyme freeze-dried powder is prepared using the method described in Comparative Example 3.

[0108] Comparative Example 8

[0109] This comparative example provides a method for preparing sparkling water beverage, which is the same as that in Example 5, except that the compound fruit enzyme freeze-dried powder is prepared using the method described in Comparative Example 4.

[0110] Comparative Example 9

[0111] This comparative example provides a method for preparing a sparkling water beverage, which is the same as that in Example 5, except that the raw materials, by weight, include: 1 part of compound fermentation product, 1 part of cassia seed extract, 1 part of monk fruit extract, 9 parts of compound fruit enzyme freeze-dried powder, 0.02 parts of vitamin E, 100 parts of purified water, and the amount of carbon dioxide added is 2 liters / liter of beverage.

[0112] Performance Test 1: Evaluation of Blood Sugar Lowering Effect

[0113] Experimental animals: 90 Kunming mice, weighing 25±2g, in an environment with a temperature of 20-25℃ and a relative humidity of 60%.

[0114] Experimental grouping: Mice were acclimatized for 3 days and then randomly divided into 9 groups of 10 each: one normal control group, one diabetic control group, and seven experimental groups.

[0115] Diabetic mouse modeling: After fasting for 16 hours, mice in the diabetic control group and experimental group were injected intraperitoneally with sodium citrate buffer solution containing 0.1 mg / g·bw streptozotocin. The normal control group was injected with the same dose of physiological saline. 30 μL of tail vein blood was collected 72 hours later to measure blood glucose. Animals with fasting blood glucose values ​​greater than 10 mmol / L were considered successful models.

[0116] Experimental methods: The normal control group and the diabetic control group were administered physiological saline by gavage. The experimental groups were administered sparkling water beverages from Examples 5-6 and Comparative Examples 5-9 by gavage, respectively. All groups had free access to food. The gavage was continued for 3 weeks, and fasting blood glucose levels were measured. The results are shown in Table 1.

[0117] Table 1: Fasting blood glucose levels

[0118]

[0119] *** indicates p<0.05 compared to the diabetic control group, and ** indicates p<0.05 compared to Example 5 or Example 6.

[0120] The above results show that the sparkling water beverages of Examples 5 and 6 exhibit significant hypoglycemic effects, reducing fasting blood glucose levels in diabetic mice by approximately 40% and 38%, respectively. This indicates that these formulations can effectively lower blood glucose levels. While the sparkling water beverages of Comparative Examples 5-9 also showed some hypoglycemic effects, the effects were not as pronounced as those of Examples 5-6. This demonstrates that the formulation and preparation process of the present invention are crucial for improving the hypoglycemic effect. In particular, the two fermentation processes and the selection of specific bacterial strains significantly improved the bioavailability of the active ingredients and the hypoglycemic effect. The different preparation methods of the compound fermentation products in Comparative Examples 5-6, the different preparation methods of the compound fruit enzyme freeze-dried powder in Comparative Examples 7-8, and the different raw material ratios in Comparative Example 9 all affected the hypoglycemic effect of the final product.

[0121] Performance Test 2: Evaluation of Antioxidant Effect

[0122] Test samples: Sparkling water beverages from Examples 5-6 and Comparative Examples 5-9

[0123] Reagents: 1,1-Diphenyl-2-trinitrophenylhydrazine (DPPH), anhydrous ethanol

[0124] Test steps:

[0125] 1. Dissolve 0.04g of DPPH in 100mL of anhydrous ethanol to prepare a 0.1mM DPPH solution. Store the solution away from light for later use.

[0126] 2. Degassing treatment: Take appropriate amounts of sparkling water beverage samples from Examples 5-6 and Comparative Examples 5-9 respectively, place them in a constant temperature water bath, and let them stand at room temperature for 30 minutes to remove carbon dioxide gas.

[0127] 3. Preparation of sample diluents: Dilute the deaerated sparkling water beverage with distilled water at a volume ratio of 1:10 to obtain a series of sample diluents of different concentrations.

[0128] 4. Reaction Procedure: Add 2 mL of DPPH solution to a test tube, then add 0.5 mL of sample diluent and mix gently. The control group consists of 2 mL of DPPH solution with 0.5 mL of distilled water; the blank group consists of 2 mL of sample diluent with 2 mL of anhydrous ethanol. Place all test tubes in the dark and let them stand at room temperature for 30 minutes.

[0129] 5. Absorbance measurement: The absorbance values ​​of the reaction mixture and the control group in each test tube were measured at a wavelength of 517 nm using a spectrophotometer.

[0130] 6. Calculate antioxidant activity: Calculate the DPPH scavenging rate for each sample using the following formula:

[0131] DPPH removal rate = [1 - (A1 - A2) / A0] × 100%

[0132] A0: Absorbance of 2ml distilled water with 2ml DPPH solution (control group);

[0133] A1: Absorbance of 2 mL of test sample with 2 mL of DPPH ethanol solution;

[0134] A2: Absorbance of 2 mL of test sample with 2 mL of anhydrous ethanol (blank group).

[0135] Each test sample was tested in triplicate, and the average value was calculated. The results are shown in Table 2 below:

[0136] Table 2: DPPH removal rate

[0137]

[0138] The results above indicate that the sparkling water beverages of Examples 5 and 6 exhibited very high DPPH scavenging rates, at 84.6% and 83.9%, respectively. This demonstrates that these formulations possess strong antioxidant capabilities, effectively scavenging free radicals and protecting cells from oxidative stress damage. Examples 5-6 used specific proportions of bitter melon, mulberry leaves, and turmeric, and improved the extraction rate and bioavailability of active ingredients through a two-stage fermentation process. These components themselves possess strong antioxidant effects, which were further enhanced after the optimized fermentation process. Blueberries, raspberries, and cherries are rich in natural antioxidants such as anthocyanins. Extraction with citric acid and ethanol, followed by enzymatic hydrolysis using β-glucosidase, removed glycosides or acylated groups that hinder the reaction of phenolic hydroxyl groups with free radicals, thereby significantly enhancing the antioxidant capacity of anthocyanins. The hydrolyzed anthocyanin molecules are smaller in size, making them easier for the body to absorb and improving their bioavailability in vivo. Cassia seed extract and monk fruit extract also contain a variety of antioxidants, which work synergistically with compound fermentation products and compound fruit enzyme freeze-dried powder to enhance the overall antioxidant effect.

[0139] Comparative Examples 5-9 showed poor antioxidant effects due to differences in fermentation processes, enzyme selection and application, and formulation ratios. This demonstrates that the two fermentation processes and the selection of specific microbial strains in the preparation method of this invention are crucial for improving the bioavailability and antioxidant performance of the active ingredients. Fermentation with a single microbial strain or the absence of a second fermentation step significantly reduces the antioxidant effect of the product. In the preparation of compound fruit enzyme freeze-dried powder, correct enzyme selection and application can effectively remove groups that hinder the function of antioxidant components, improving their bioavailability and antioxidant performance. Incorrect enzyme selection or omitting the enzymatic hydrolysis step will lead to a decrease in antioxidant effect. Furthermore, a reasonable formulation ratio is key to ensuring the synergistic effect of various components and achieving optimal results. An imbalance in the proportion of any component may affect the overall antioxidant performance.

[0140] The above description represents the preferred embodiments of the present invention. It should be noted that those skilled in the art can make various improvements and modifications without departing from the principles of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A sparkling water beverage, characterized in that, The ingredients are as follows by weight: 2-5 parts of compound fermented product, 0.5-1 part of cassia seed extract, 1-2 parts of monk fruit extract, 3-6 parts of compound fruit enzyme freeze-dried powder, 0.01-0.02 parts of vitamin E, 100 parts of purified water, and 1.5-2.5 liters of carbon dioxide added per liter of beverage. Raw materials for preparing compound fermentation product: 2-4 parts bitter melon, 4-5 parts mulberry leaves, and 1-2 parts turmeric; Raw materials for preparing compound fruit enzyme freeze-dried powder: 4-5 parts blueberry, 4-5 parts raspberry and 2-3 parts cherry; The preparation method of compound fermentation product includes: (1) Wash bitter melon, mulberry leaves and turmeric, break them into small pieces, add deionized water at a ratio of 1g:6-10mL, heat and boil for 1-2 hours, add water every 10-30 minutes to the original volume, filter after heating to obtain filtrate and filter residue; (2) Add deionized water to the filter residue at a mass ratio of 1:1-1.5, then add pectinase, cellulase, hemicellulase and acidic protease, adjust the pH to 4.5-5, the temperature to 40-50℃, and enzymatically hydrolyze for 5-10 hours. (3) After the enzymatic hydrolysis is complete, raise the temperature to 80-100℃ and keep it for 10-20 minutes to inactivate the enzyme and obtain the enzymatic hydrolysate. (4) Add 1%-5% glucose and 0.05%-0.2% yeast extract to the filtrate of step (1) and the enzymatic hydrolysate of step (3) respectively, then inoculate each with 2-5 wt% of the compound bacteria, stir evenly, and anaerobic ferment at 37-40℃ for 2-3 days to obtain fermentation product A and fermentation product B respectively. (5) Filter fermentation product B, mix the filtrate with fermentation product A, add 1%-5% glucose and 0.05%-0.2% yeast extract, then inoculate with 2-5 wt% of Lactobacillus plantarum, stir evenly, and anaerobic ferment at 35-37℃ for 3-4 days to obtain the compound fermentation product. The compound bacteria consist of Lactobacillus acidophilus, Bifidobacterium breve, and Lactobacillus rhamnosus in a mass ratio of 1:2-4:1-2. The preparation method of compound fruit enzyme freeze-dried powder includes: (1) washing blueberries, raspberries and cherries, breaking them into small pieces, adding extractant at a material-liquid ratio of 1g:3-7mL, heating to 30-50℃, extracting for 1-2 hours, filtering after extraction, vacuum concentrating to obtain extract; (2) Adjust the pH of the extract from step (1) to 5-6, add β-glucosidase, heat to 40-50℃, and hydrolyze for 3-5 hours. After the reaction is complete, filter, concentrate the filtrate, and vacuum dry to obtain the compound fruit enzyme freeze-dried powder.

2. The sparkling water beverage according to claim 1, characterized in that, The amount of each of the pectinase, cellulase, hemicellulase and acidic protease added is independently 0.1-1% of the mass of the filter residue.

3. The sparkling water beverage according to claim 1, characterized in that, The extractant is a mixture of citric acid and 60-80 wt% aqueous ethanol solution in a mass ratio of 1:20-50.

4. The sparkling water beverage according to claim 1, characterized in that, The amount of β-glucosidase added is 1.5-2.5% of the extract mass.

5. The sparkling water beverage according to claim 1, characterized in that, The preparation method of the cassia seed extract includes: (1) Wash the cassia seeds, dry them, and crush them into small particles; (2) Add the cassia seed granules to a 60-80wt% ethanol aqueous solution, with a material-to-liquid ratio of 1g:10-20mL, and heat and reflux for 1-2 hours. (3) After extraction, filter and concentrate the filtrate under vacuum to obtain the cassia seed extract.

6. The sparkling water beverage according to claim 1, characterized in that, The preparation method of the monk fruit extract includes: (1) Wash the monk fruit, dry it, and crush it into small particles; (2) Add the monk fruit granules to deionized water at a material-to-liquid ratio of 1g:10-20mL, heat to boiling, maintain for 1-2 hours, and filter to obtain the first extract. (3) Add the residue back into deionized water at a ratio of 1g:10-20mL, heat to boiling, maintain for 1-2 hours, and filter to obtain the second extract. (4) The two extracts were combined and concentrated under vacuum to obtain the monk fruit extract.

7. The method for preparing sparkling water beverage according to any one of claims 1-6, characterized in that, include: (1) Mix the compound fruit enzyme freeze-dried powder with pure water, heat to 40-50℃, stir for 30-60 minutes to fully dissolve the compound fruit enzyme freeze-dried powder, and obtain material 1; (2) Add compound fermentation product, cassia seed extract, monk fruit extract and vitamin E to material 1, stir well to obtain material 2; (3) Sterilize material 2 at high temperature for 5-10 seconds at a temperature of 110-120℃ to obtain material 3. (4) Reduce the temperature of material 3 to 4-8℃ and introduce carbon dioxide gas into it to obtain the sparkling water beverage. The amount of carbon dioxide added is 1.5-2.5 liters / liter of beverage.

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