A forest frog skin and red ginseng composite functional beverage and a preparation method thereof

Abstract

The application discloses a kind of forest frog skin red ginseng composite functional drinks and preparation method thereof, belong to food technical field, wherein, the drink includes: including forest frog polypeptide liquid, red ginseng saponin liquid, deodorant, sweetening agent and whey protein;Forest frog polypeptide liquid is prepared by forest frog skin through defatting, enzymolysis processing;Red ginseng saponin liquid is prepared by red ginseng through alcohol extraction;Deodorant is Chinese wolfberry original pulp.The application uses forest frog processing by-product as raw material, meets green food and recycling economic concept.The application can significantly reduce fishy smell by the triple action of ethanol defatting, enzymolysis fishy smell protein, plant extract masking;By enzymolysis, the protein component in forest frog skin is improved, and the saponin component in red ginseng is extracted, with good antioxidant capacity;In addition, the application can optimize the ratio of deodorant, sweetening agent and protein through orthogonal experiment, and the prepared drink has smooth taste, moderate sweetness and natural aroma.

Description

Technical Field

[0001] This invention relates to the field of food technology, specifically to a functional beverage made from a combination of forest frog skin and red ginseng, and its preparation method. Background Technology

[0002] Forest frogs have a long history of consumption in Northeast my country, with their skin being a traditional ingredient widely used in local dishes such as braised forest frog and dried forest frog. In modern food processing, the oviducts of female forest frogs are often used to prepare frog oil. Forest frog skin, often a byproduct of frog oil processing, is rich in collagen, polypeptides, and various amino acids, possessing potential antioxidant and other bioactivities. However, the strong fishy smell and residual off-flavor oil components of forest frog skin limit its application in the food industry. Currently, there are no functional beverages on the market using forest frog skin as the main ingredient.

[0003] In existing technologies, methods for removing fishy odors often involve simple masking or high-temperature treatment, which can easily damage nutritional components. Although red ginseng-based beverages have a certain market presence, they are mostly extracted alone and lack combination with animal-derived polypeptide liquids.

[0004] Therefore, developing a compound functional beverage of frog skin and red ginseng that can effectively remove the fishy smell of frog skin, has a harmonious taste, good stability, and excellent antioxidant activity is of great practical significance and market value. Summary of the Invention

[0005] The purpose of this invention is to provide a functional beverage made from forest frog skin and red ginseng, and its preparation method, in order to solve the problems mentioned in the background art.

[0006] To achieve the above objectives, the present invention provides the following technical solution: A functional beverage made from frog skin and red ginseng includes frog polypeptide liquid, red ginseng saponin liquid, deodorizing agent, sweetener, and whey protein; the frog polypeptide liquid is obtained from frog skin through defatting and enzymatic hydrolysis; the red ginseng saponin liquid is obtained from red ginseng through alcohol extraction; and the deodorizing agent is wolfberry puree.

[0007] Another object of the present invention is to provide a method for preparing the above-mentioned functional beverage made from forest frog skin and red ginseng, comprising the following steps: Forest frog skin pretreatment: Forest frog skin powder is degreased with edible ethanol to obtain degreased forest frog skin material; Enzymatic hydrolysis of forest frog skin: Add defatted forest frog skin to water and adjust the pH to 6.5-7.5. Add compound flavor protease for enzymatic hydrolysis, filter after enzyme inactivation to obtain forest frog polypeptide liquid; Red ginsenoside extraction: Red ginseng powder was extracted with edible ethanol, filtered to remove the edible ethanol, and then redissolved in water to obtain red ginsenoside solution; Preparation: Mix frog polypeptide liquid with red ginseng saponin liquid, add deodorizing agent, sweetener and whey protein and homogenize to obtain frog skin and red ginseng compound functional beverage.

[0008] Furthermore, the pretreatment steps for the frog skin specifically include: The dried frog skin is crushed and sieved to obtain frog skin powder; Weigh out the frog skin powder, add edible ethanol, stir in a water bath at 50-70℃ for 20-40 minutes, sieve and discard the filtrate, rinse the filter residue with water and drain thoroughly to remove edible ethanol residue, and obtain defatted frog skin material; the mass-volume ratio of the frog skin powder to edible ethanol is (80-100) g:(400-500) mL.

[0009] Furthermore, the enzymatic hydrolysis step of the forest frog skin specifically includes: Add defatted frog skin to water, adjust the pH to 6.5-7.5 with baking soda and / or citric acid, add compound flavor protease, and enzymatically hydrolyze at 50-60℃ for 3-5 hours. Then, inactivate the enzyme by water bath at 85-95℃ for 5-15 minutes. Sieve to obtain frog polypeptide solution; the mass ratio of the compound flavor protease to frog skin powder is (1-2):(80-100).

[0010] Furthermore, the steps for extracting red ginsenosides specifically include: Weigh red ginseng powder, add edible ethanol, extract in a water bath at 50-70℃ for 30-50 minutes, filter, and then evaporate the filtrate in an open water bath at 50-70℃ until there is no alcohol odor, remove the edible ethanol, and then redissolve in water to obtain red ginseng saponin solution; the mass-volume ratio of red ginseng powder to edible ethanol is (80-100) g:(800-1000) mL.

[0011] Furthermore, the volume percentage concentration of the edible ethanol is 65%-75%.

[0012] Furthermore, the volume ratio of the frog polypeptide liquid, red ginseng saponin liquid and wolfberry pulp is (20-30):(20-30):(45-55); the volume-to-mass ratio of the frog polypeptide liquid to sweetener and whey protein is (20-30) mL:(3-5):(0.5-1.5).

[0013] Furthermore, after the preparation step, a sterilization step is also included.

[0014] Furthermore, the sterilization step specifically includes sterilizing the prepared beverage by treating it at 80-90℃ for 10-20 minutes.

[0015] The functional beverage made from frog skin and red ginseng provided by this invention utilizes frog processing byproducts as raw materials, aligning with the concepts of green food and circular economy. Specifically, this invention significantly reduces the fishy odor through a triple process of ethanol defatting, enzymatic hydrolysis of fishy-smelling proteins, and masking with plant extracts. Enzymatic hydrolysis increases the protein content in frog skin, and the extraction of saponins from red ginseng yields components with excellent antioxidant capabilities. Furthermore, this invention optimizes the ratio of deodorizing agents, sweeteners, and proteins through orthogonal experiments, resulting in a beverage with a smooth texture, moderate sweetness, and natural aroma. Attached Figure Description

[0016] Figure 1 This is a schematic flowchart illustrating the preparation method of the forest frog skin and red ginseng compound functional beverage provided by the present invention.

[0017] Figure 2 The appearance and properties of the compound functional beverage sample made with forest frog skin and red ginseng with the optimal ratio.

[0018] Figure 3 The image shows the appearance and properties of the red ginsenoside solution.

[0019] Figure 4 The image shows the appearance and properties of the frog polypeptide solution. Detailed Implementation

[0020] The technical solutions of this invention will now be clearly and completely described in conjunction with the present invention. 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.

[0021] In one embodiment of the present invention, a method for preparing a safe, odorless, well-tasting, nutritious, and antioxidant-rich compound functional beverage made from frog skin and red ginseng is provided. This method addresses the problems of difficulty in removing the fishy smell of frog skin, poor product stability, and unpleasant taste in existing technologies. The beverage comprises frog polypeptide liquid, red ginseng saponin liquid, a deodorizing agent, a sweetener, and whey protein. The frog polypeptide liquid is obtained from frog skin through defatting and enzymatic hydrolysis. The red ginseng saponin liquid is obtained from red ginseng through alcohol extraction. The deodorizing agent includes one or more of wolfberry pulp, ginger juice, and licorice powder, preferably wolfberry pulp.

[0022] Specifically, such as Figure 1 As shown, the preparation method of the above-mentioned forest frog skin and red ginseng compound functional beverage includes the following steps: S1. Pretreatment of frog skin: Crush dried frog skin into powder and sieve to obtain frog skin powder; weigh 80-100g of frog skin powder, add 400-500mL of edible ethanol (volume percentage concentration of 65%-75%), stir in a water bath at 50-70℃ for 20-40min, sieve and discard the filtrate, rinse the filter residue with purified water and drain thoroughly to remove edible ethanol residue, and obtain defatted frog skin material; S2. Enzymatic hydrolysis of frog skin: Add the above defatted frog skin to 0.8-1.2L of purified water, adjust the pH to 6.5-7.5 with baking soda and / or citric acid, add 1-2g of compound flavor protease, hydrolyze at 50-60℃ for 3-5h, then inactivate the enzyme in a water bath at 85-95℃ for 5-15min, sieve, and obtain frog polypeptide solution. S3. Extraction of red ginsenosides: Weigh 80-100g of red ginseng powder, add 800-1000mL of edible ethanol (volume percentage concentration of 65%-75%), extract in a water bath at 50-70℃ for 30-50min, filter, and then evaporate the filtrate in an open water bath at 50-70℃ until there is no alcohol odor, remove the edible ethanol, and then redissolve in purified water to obtain red ginsenoside solution. S4. Preparation: Weigh 20-30 mL of the above-mentioned frog polypeptide solution and 20-30 mL of red ginseng saponin solution and mix them together. Add an appropriate amount of deodorizing agent (such as 45-55 mL of wolfberry puree), 3-5 g of sweetener (such as sucrose) and 0.5-1.5 g of whey protein and homogenize for 5-8 minutes until the system is uniform and smooth.

[0023] S5. Sterilization: Sterilize the above-prepared beverage at 80-90℃ for 10-20 minutes to obtain the forest frog skin red ginseng compound functional beverage.

[0024] Unless otherwise specified, all raw materials used in the following embodiments are commercially available products and can be purchased through commercial channels. The invention will be described in detail below through specific embodiments in practical applications.

[0025] Example 1: This example provides a method for preparing a compound functional beverage of forest frog skin and red ginseng, specifically including the following steps: S1. Pretreatment of frog skin: The dried frog skin is crushed using a pulverizer and passed through an 80-mesh sieve to obtain frog skin powder; 90g of frog skin powder is weighed and 450mL of edible ethanol (volume percentage concentration of 70%) is added. The mixture is stirred in a 60℃ water bath for 30min, passed through a 100-mesh sieve and the filtrate is discarded. The filter residue is rinsed with purified water and thoroughly drained to remove edible ethanol residue, thus obtaining defatted frog skin material. S2. Enzymatic hydrolysis of frog skin: Add the above defatted frog skin to 1L of purified water, adjust the pH to 7.0 with baking soda and citric acid, add 1.5g of compound flavor protease, hydrolyze at 55℃ for 4h, then inactivate the enzyme in a 90℃ water bath for 12min, and pass through a 100-mesh sieve to obtain frog polypeptide solution. S3. Extraction of red ginsenosides: Weigh 90g of red ginseng powder, add 900mL of edible ethanol (volume percentage concentration of 70%), extract in a 60℃ water bath for 40min, filter, and then evaporate the filtrate in a 60℃ water bath with the opening open until there is no alcohol odor. Remove the edible ethanol, and then add purified water to redissolve to obtain red ginsenoside solution. S4. Preparation: Weigh 25 mL of the above-mentioned frog polypeptide solution and 25 mL of red ginseng saponin solution and mix them together. Add 50 mL of wolfberry puree, 4 g of sucrose and 1 g of whey protein. Homogenize the mixture for 7 min using a homogenizer until the system is uniform and smooth.

[0026] S5. Sterilization: The above-prepared beverage is sterilized by treating it at 85℃ for 15 minutes to obtain the forest frog skin red ginseng compound functional beverage.

[0027] Example 2: This example provides a method for preparing a compound functional beverage of forest frog skin and red ginseng, specifically including the following steps: S1. Pretreatment of frog skin: The dried frog skin is crushed using a pulverizer and passed through an 80-mesh sieve to obtain frog skin powder; 80g of frog skin powder is weighed and 400mL of edible ethanol (volume percentage concentration of 65%) is added. The mixture is stirred in a 50℃ water bath for 20min, passed through a 100-mesh sieve and the filtrate is discarded. The filter residue is rinsed with purified water and drained thoroughly to remove edible ethanol residue, thus obtaining defatted frog skin material. S2. Enzymatic hydrolysis of frog skin: Add the above defatted frog skin to 0.8L of purified water, adjust the pH to 6.5 with baking soda and citric acid, add 1g of compound flavor protease, hydrolyze at 50℃ for 3h, then inactivate the enzyme in a water bath at 85℃ for 5min, and pass through a 100-mesh sieve to obtain frog polypeptide solution. S3. Extraction of red ginsenosides: Weigh 80g of red ginseng powder, add 800mL of edible ethanol (volume percentage concentration of 65%), extract in a 50℃ water bath for 30min, filter, and then evaporate the filtrate in a 50℃ water bath with the opening open until there is no alcohol odor. Remove the edible ethanol, and then add purified water to redissolve to obtain red ginsenoside solution. S4. Preparation: Weigh 20 mL of the above-mentioned frog polypeptide solution and 20 mL of red ginseng saponin solution and mix them together. Add 45 mL of wolfberry puree, 3 g of sucrose and 0.5 g of whey protein. Homogenize the mixture for 5 min using a homogenizer until the system is uniform and smooth.

[0028] S5. Sterilization: The above-prepared beverage is sterilized by treating it at 80℃ for 10 minutes to obtain the forest frog skin red ginseng compound functional beverage.

[0029] Example 3: This example provides a method for preparing a compound functional beverage of forest frog skin and red ginseng, specifically including the following steps: S1. Pretreatment of frog skin: The dried frog skin is crushed using a pulverizer and passed through an 80-mesh sieve to obtain frog skin powder; 100g of frog skin powder is weighed and 500mL of edible ethanol (volume percentage concentration of 75%) is added. The mixture is stirred in a 70℃ water bath for 40min, passed through a 100-mesh sieve and the filtrate is discarded. The filter residue is rinsed with purified water and thoroughly drained to remove edible ethanol residue, resulting in defatted frog skin material. S2. Enzymatic hydrolysis of frog skin: Add the above defatted frog skin to 1.2L of purified water, adjust the pH to 7.5 with baking soda and citric acid, add 2g of compound flavor protease, hydrolyze at 60℃ for 5h, then inactivate the enzyme in a 95℃ water bath for 15min, and pass through a 100-mesh sieve to obtain frog polypeptide solution. S3. Extraction of red ginsenosides: Weigh 100g of red ginseng powder, add 1000mL of edible ethanol (volume percentage concentration of 75%), extract in a 70℃ water bath for 50min, filter, and then evaporate the filtrate in a 70℃ water bath with the opening open until there is no alcohol odor. Remove the edible ethanol, and then add purified water to redissolve to obtain red ginsenoside solution. S4. Preparation: Weigh 30 mL of the above-mentioned frog polypeptide solution and 30 mL of red ginseng saponin solution and mix them together. Add 55 mL of wolfberry puree, 5 g of sucrose and 1.5 g of whey protein. Homogenize the mixture for 8 minutes until the system is uniform and smooth.

[0030] S5. Sterilization: The above-prepared beverage is sterilized by treating it at 90℃ for 20 minutes to obtain the forest frog skin red ginseng compound functional beverage.

[0031] Example 4: This example is a single-factor analysis experiment, as detailed below: 1. The sensory evaluation criteria used in the experiment are shown in Table 1: Table 1

[0032] The formula for calculating sensory scores is: ; In the formula: X i For the evaluator's score on a certain indicator i, W i The corresponding weights.

[0033] The sensory evaluation involved five healthy volunteers aged 18-25 who provided sensory evaluation scores for each formula, and then the average score for each score was taken.

[0034] 2. Single-factor experiment on the amount of wolfberry puree added: According to the preparation method provided in Example 1, 25 mL of frog polypeptide solution and red ginseng saponin solution were mixed, and then the corresponding wolfberry puree, 3g sucrose and 1g whey protein were added and homogenized and sterilized to obtain the final product.

[0035] As shown in Table 2, a single-factor experiment was conducted on the amount of wolfberry puree added, and the sensory evaluation score was calculated based on the collected sensory evaluation data and weighted weights.

[0036] Table 2

[0037] Based on the results of the single-factor experiment, the addition amounts of wolfberry juice were selected as 40mL, 45mL, and 50mL as the range for the orthogonal experiment.

[0038] 3. Single-factor experiment on the amount of licorice powder added: According to the preparation method provided in Example 1, 25 mL of frog polypeptide solution and red ginseng saponin solution were mixed, and then the corresponding licorice powder (replacing wolfberry puree), 3g sucrose and 1g whey protein were added. After homogenization and sterilization, the solution was obtained, as shown in Table 3.

[0039] Table 3

[0040] Based on the results of the single-factor experiment, the amounts of licorice powder added were selected as 0.2g, 0.4g, and 0.6g as the range for the orthogonal experiment.

[0041] 4. Single-factor experiment on the amount of ginger juice added: According to the preparation method provided in Example 1, 25 mL of frog polypeptide liquid and red ginseng saponin liquid were mixed together, and then the corresponding ginger juice (replacing wolfberry puree), 3g sucrose and 1g whey protein were added. After homogenization and sterilization, the product was obtained, as shown in Table 4.

[0042] Table 4

[0043] Based on the results of the single-factor experiment, the amounts of ginger juice added were selected as 0.5 mL, 0.75 mL, and 1 mL as the range for the orthogonal experiment.

[0044] Example 5: This example is an orthogonal experimental design and formulation optimization experiment, as detailed below: 1. Orthogonal experimental design: L9(3) 4 An orthogonal experimental design was used to investigate the effects of different types and amounts of deodorizing agents, sucrose, and whey protein on sensory quality, using pre-treated frog polypeptide solutions and red ginseng saponin solutions as substrates. Levels: A. Type of deodorizing agent; B. Amount of deodorizing agent; C. Sucrose; D. Whey protein. The orthogonal experimental design is shown in Table 5. Table 5

[0045] Based on the above design, a compound functional beverage of forest frog skin and red ginseng was prepared and its sensory evaluation was conducted. The specific results are shown in Table 6 below: Table 6

[0046] 2. The order of influence of factors: Sort by range R value from largest to smallest: R A (1.223)>R C (0.244)>R D (0.147)>R B (0.067); Therefore, the factors are: type of deodorant (A) > amount of sucrose added (C) > amount of whey protein added (D) > amount of deodorant added (B); indicating that the type of deodorant has the greatest impact on sensory quality, followed by the amount of sucrose added, while the amount of deodorant added has the least impact.

[0047] 3. Determine the optimal level combination: Based on the principle that the higher the sensory score, the better, select the level with the largest k-value for each factor: A: Since k1 is the largest, the type of deodorant to choose is wolfberry puree.

[0048] B: k3 is the largest, so a high amount of deodorizing agent should be added.

[0049] C: k2 is the largest, so the amount of sucrose added is 4g.

[0050] D: k1 is the largest, so the amount of whey protein added is 1g.

[0051] Therefore, the optimal ratio is 25 mL of frog polypeptide solution, 25 mL of red ginseng saponin solution, 50 mL of wolfberry puree, 4 g of sucrose, and 1 g of whey protein.

[0052] Example 6: This example is a quality evaluation experiment, as detailed below: The forest frog skin and red ginseng compound functional beverage was prepared according to the optimal ratio in Example 5 (i.e., Example 1), and samples were taken for the following tests.

[0053] 1. Appearance and properties: such as Figure 2 As shown, this beverage is a homogeneous and stable reddish-brown (brown) liquid that does not separate, flocculate, or precipitate after standing.

[0054] This beverage has the unique, delicate medicinal aroma of red ginseng and the natural fragrance of goji berry juice. It is free of the fishy, ​​pungent, or other unpleasant odors of frog skin, indicating that the fishy smell of frog skin has been successfully removed. The taste is dominated by the sweetness of goji berries and the unique aroma of red ginseng, accompanied by a slightly acceptable astringent sensation, and the overall taste is smooth.

[0055] 2. pH value measurement: The pH value of the forest frog skin and red ginseng compound functional beverage was measured three times using a calibrated pH meter and was 4.71±0.046.

[0056] 3. Moisture content determination: A certain amount of the frog skin and red ginseng compound functional beverage sample was weighed and placed in a drying oven at 101-105℃. The sample was weighed every hour until the difference between two weighings did not exceed 2mg, at which point the weight was considered constant. The moisture content of the sample was then calculated. The moisture content of the frog skin and red ginseng compound functional beverage sample was determined to be 75.2g / 100g.

[0057] 4. Protein Content Determination: The protein content in various samples (frog skin soaking solution, frog skin enzymatic hydrolysate, and the frog skin and red ginseng compound functional beverage prepared according to the optimal ratio of this invention) was quantitatively determined using the diquinoline formic acid method. Bovine serum albumin (BSA) was used as the standard, and therefore the determined protein content was calculated based on BSA. 20 μL of sample was mixed with 200 μL of BCA working solution and incubated at 37°C for 15-30 min. The A562 nm concentration was measured using an ELISA reader, and the protein concentration was calculated based on the standard curve. Each group was repeated three times, as shown in Table 7.

[0058] The standard curve is: y = 0.7547x + 0.1358, R0 2 =0.987.

[0059] Table 7

[0060] As can be seen from the table above, the compound flavor protease hydrolyzes insoluble proteins such as collagen, elastin, and reticular protein in the skin of the forest frog into soluble polypeptides, thereby increasing the content of soluble proteins in the skin of the forest frog.

[0061] 5. Antioxidant capacity determination: The in vitro antioxidant capacity of the sample (optimally formulated forest frog skin and red ginseng compound functional beverage) was evaluated using the DPPH free radical scavenging method. Because the color of the sample could affect the final results, the sample was diluted 10-fold and 20-fold with distilled water, respectively. 50 μL of the sample was mixed with 150 μL of anhydrous ethanol to prepare 0.1 mM DPPH working solution. After mixing, the solution was incubated at room temperature in the dark for 30 min, and the absorbance was measured at 517 nm in triplicate. The DPPH free radical scavenging activity of the sample was calculated using the following formula: ; In the formula: As is the absorbance of the sample and DPPH at 517 nm; Aso is the absorbance of the sample and ethanol at 517 nm; Aec is the absorbance of the sample solution and DPPH at 517 nm; Aeb is the absorbance of the sample solution and ethanol at 517 nm.

[0062] The free radical scavenging rate of the samples was determined using the DPPH method. The results are shown in Table 8 below. The DPPH free radical scavenging rates of the 10-fold and 20-fold dilutions were 47.76±1.82% and 26.29±0.89%, respectively.

[0063] Table 8

[0064] Example 7: This example is a verification experiment on the antioxidant synergistic effect and pharmacological synergistic effect of the three components: frog polypeptide liquid, red ginseng saponin liquid, and wolfberry pulp. The details are as follows: I. Mechanism of Action: 1. Synergistic Antioxidant Pathways, Breaking Through the Limits of Single Component Efficacy: Small-molecule collagen peptides obtained from the enzymatic hydrolysis of frog skin are rich in thiol groups and hydrophobic amino acids. They can directly capture and scavenge reactive oxygen free radicals through proton donor action, which is a rapid and direct antioxidant pathway; Saponins extracted from red ginseng can upregulate the activity of endogenous antioxidant enzymes (such as SOD and GSH-Px) in the body, continuously alleviating oxidative damage, which is a long-term regulatory pathway; The polyphenols and flavonoids contained in wolfberry have strong electron donor capabilities, which can efficiently scavenge free radicals. Its polysaccharide components can chelate transition metal ions such as iron and copper, blocking the source of free radicals generated by the Fenton reaction, and can reduce and regenerate oxidized antioxidant components, forming a free radical regeneration cycle, which is a source blocking and cycle enhancement pathway. The three components act on different stages and targets of oxidative stress, forming a multidimensional antioxidant network of "direct scavenging - source blocking and regeneration - endogenous regulation." Their pathways are complementary and their effects are synergistic. The combined free radical scavenging capacity is significantly better than the simple sum of the individual components, and even better than the sum of the tested binary combinations, achieving excellent antioxidant effects at low doses. Particularly noteworthy is that while red ginseng has a relatively weak direct free radical scavenging capacity, it significantly amplifies the overall antioxidant activity in the combined system, indicating that each component plays an irreplaceable role in this synergistic network.

[0065] 2. Synergistic effects in formulation enhance the dispersion stability and sensory quality of beverages: Red ginsenosides are highly hydrophobic, and their pure aqueous solutions tend to aggregate and precipitate over long-term storage. Forest frog small-molecule peptides possess an amphiphilic structure, which can encapsulate saponin molecules through hydrogen bonds and hydrophobic interactions, forming a stable colloidal dispersion system. Lycium barbarum polysaccharides are hydrophilic macromolecules that can adsorb onto the surface of the peptide-saponin complex, forming a hydration protective layer and increasing the viscosity of the continuous phase. This steric hindrance inhibits particle aggregation and sedimentation, while its antioxidant activity slows down the oxidative degradation of active ingredients. The synergistic effect of these three factors significantly improves the water solubility of red ginsenosides and the uniformity of dispersion during storage, solving the problem of easy stratification and precipitation in single-origin red ginseng extracts.

[0066] II. Experimental Basis: The experimental groups are as follows: (1) Forest frog polypeptide solution group (a medium volume of pure forest frog polypeptide solution compared with the compound group). (2) Red ginsenoside solution group (a medium volume of pure red ginsenoside solution compared with the compound group); (3) Goji berry puree group (a medium volume of pure goji berry puree compared with the compound group); (4) Theoretical addition group 1 (mixed group of frog polypeptide solution and red ginseng saponin solution, binary interaction); (5) Theoretical addition group 2 (mixed group of frog polypeptide liquid, red ginseng saponin liquid and wolfberry pulp, ternary interaction); (6) Actual summation group 1 (mixed group of frog polypeptide solution and red ginseng saponin solution, binary interaction effect). (7) The compound group of the present invention (i.e., the forest frog skin and red ginseng compound functional beverage prepared in Example 1, with the effect of three-element interaction). The DPPH radical scavenging rate of each group was measured (the method for measuring DPPH was the same as in Example 6), and the structures are shown in Table 9. The effect was quantified by calculating the Synergy Ratio (SR). ; Table 9

[0067] As can be seen from the table above, the single-component active bases are as follows: the wolfberry pulp group showed the strongest DPPH free radical scavenging ability (0.2052), followed by the frog polypeptide liquid group (0.0813), while the red ginseng saponin liquid showed a weak direct scavenging ability at this concentration (0.0303).

[0068] Binary synergistic verification: After combining the frog polypeptide liquid group and the red ginseng saponin liquid group, the measured scavenging rate (0.1806) was higher than the theoretical summation value (0.1116), and the synergistic rate reached SR=0.1806 / 0.1116≈1.62. This indicates that although red ginseng saponins have limited antioxidant activity on their own, they can enhance the antioxidant efficacy of frog polypeptides.

[0069] Ternary Synergy: After introducing wolfberry puree, the scavenging rate of the ternary system increased to 0.4776, which is higher than the theoretical sum value (0.3167, SR=0.4776 / 0.3167≈1.51).

[0070] Nonlinear synergistic effect: Data shows that the antioxidant activity of this composition is not a simple linear additive of the components. The addition of red ginseng saponin liquid enhances the antioxidant capacity of frog polypeptide liquid, while the addition of wolfberry puree further constructs a stable antioxidant network, producing an unexpected technical effect of "1+1+1>3".

[0071] In summary, the compounding solution provided by this invention has the following beneficial effects: 1. Significant synergistic antioxidant effect: The ternary composition of forest frog-goji berry-red ginseng provided by this invention has a measured DPPH free radical scavenging rate that is 1.51 times the theoretical sum. This synergistic effect overcomes the shortcomings of limited activity or poor stability of single components, and achieves improved antioxidant efficacy.

[0072] 2. Multi-target antioxidant mechanism: Based on the above data, it is inferred that frog peptides (providing hydrogen protons), wolfberry polysaccharides / polyphenols (electron donors / metal chelators) and red ginsenosides (endogenous regulation) constitute a multi-dimensional antioxidant defense network, thereby achieving efficient and long-lasting scavenging of free radicals.

[0073] Furthermore, red ginsenosides are highly hydrophobic, making it difficult to form a stable dispersion system in pure aqueous solution; the inherent color of the frog polypeptide hydrolysate is grayish-black, lacking sensory appeal in beverages. This invention overcomes both of these formulation defects through a three-component compound: Regarding dispersion stability, such as Figure 3 As shown, after seven days of standing, the red ginseng saponin solution exhibited a significant sediment layer at the bottom of the bottle, accompanied by floating aggregates on the surface. This indicates that the hydrophobic saponin components aggregated and settled, the system was unstable, and the active ingredients were unevenly distributed. In contrast, the beverage prepared by this invention, after standing under the same conditions, was uniform throughout, without sedimentation, stratification, or floating aggregates, forming a stable colloidal dispersion system. The mechanism is as follows: the small molecule polypeptides of the forest frog possess an amphiphilic structure, which can encapsulate saponin molecules through hydrogen bonds and hydrophobic interactions, forming a micellar complex; the wolfberry polysaccharide is a hydrophilic macromolecule, which adsorbs onto the surface of the complex to form a hydration protective layer and increases the viscosity of the continuous phase. This steric hindrance inhibits particle aggregation and sedimentation, allowing the hydrophobic components to be uniformly and stably dispersed in the aqueous phase in colloidal form.

[0074] In terms of color and sensory quality, such as Figure 4 As shown, the frog polypeptide liquid is grayish-black, and its direct application to beverages can easily evoke negative sensory associations in consumers, constituting a sensory barrier to its beverage application. When combined with wolfberry puree, the beverage of this invention exhibits a uniform reddish-brown color. Figure 2 On the one hand, the natural pigments (carotenoids, polyphenols, etc.) contained in goji berries mask the undesirable grayish-black color of the frog polypeptide liquid, achieving natural coloring without the need for additional exogenous pigments. On the other hand, the reddish-brown color highly matches consumers' perception and psychological expectations of the color of nourishing ingredients such as red ginseng and goji berries, conveying a natural, mild, and healthy sensory impression, significantly improving the product's sensory quality and consumer acceptance. Furthermore, after standing for seven days, the beverage's color remained uniform and stable, with no pigment layering or fading, indicating that the pigment components were also stably contained within the aforementioned colloidal dispersion system, further confirming the formulation synergy of the three components.

[0075] In summary, the three components work synergistically in formulation, solving the dual problems of aggregation and sedimentation of single red ginseng extract and poor inherent color of single forest frog polypeptide extract, thereby improving the dispersion uniformity of active ingredients, storage retention rate, and sensory quality of the product.

[0076] Example 8: This example is a verification experiment of the synergistic effect of the three-stage deodorization system, as detailed below: I. Mechanism of Action: The three-stage deodorization system refers to a three-stage synergistic deodorization system consisting of defatting, enzymatic hydrolysis, and masking with goji berry juice, wherein: Level 1: Degreasing with edible ethanol water bath removes most of the fat tissue from the frog skin, cutting off the path from which fat oxidation produces hexanal, octanal and other fishy-smelling substances. The second stage involves controlled enzymatic hydrolysis of complex flavor proteases, which releases bound fishy odor precursors into free states, creating conditions for masking the original wolfberry pulp. The third stage involves masking the odor with goji berry puree. The polyphenols, flavonoids, and polysaccharides in goji berries can encapsulate trace amounts of residual hydrophobic fishy substances through hydrophobic interactions, reducing their volatility. Simultaneously, the natural sweetness of goji berries neutralizes any remaining off-flavors from a sensory perspective, balancing the beverage's taste. The whey protein added during the blending stage further fixes residual fishy substances through hydrophobic binding, while sucrose and the sweetness of goji berries together mask the sweetness. The three-stage process works together to cover the entire process of removing fishy odors, from their "source of formation to their form of existence to their sensory release," resulting in a more thorough removal of fishy odors and better taste than a single removal process.

[0077] II. Experimental Basis: The scoring method is the same as that in Example 4. The fishy smell sensory scores in Table 11 below are average scores. The experimental groups are as follows. Except for the components mentioned that were removed, all other components in the following groups are the same as in Example 1: (1) Blank control group: a mixture of enzymatic hydrolysate of frog skin without any deodorization treatment and red ginseng saponin solution; (2) Degreasing only group: only ethanol degreasing, without frog skin hydrolysate and wolfberry pulp; (3) Defatted + enzymatic hydrolysis group: After pretreatment, no wolfberry juice was added; (4) Group with only wolfberry puree: wolfberry puree was added directly without pretreatment; (5) The present invention group (Example 1): defatting + enzymatic hydrolysis + wolfberry juice whole process; Table 11

[0078] The experimental results are as follows: 1. Single deodorization measures have limited effectiveness and cannot independently impart sensory acceptability to the product. The mixture of untreated frog skin enzymatic hydrolysate and red ginseng saponin solution had a strong fishy smell (0.00 points); ethanol defatting to remove lipid-based fishy smell precursors only increased the score to 0.60 points; further enzymatic hydrolysis to degrade fishy smell-related substrates only increased the score to 1.00 points, still far below the acceptable level; adding wolfberry puree directly without pretreatment increased the score to 2.40 points by masking the fishy smell with the aroma of wolfberry, indicating that masking is the strongest single method, but its masking capacity is overwhelmed when the fishy smell load is too high, and it is still unacceptable.

[0079] 2. This invention achieves a qualitative leap in sensory acceptability throughout the entire process, with each step being indispensable. The group of this invention (defatting + enzymatic hydrolysis + goji berry puree) scored 3.80 points, reaching a level of virtually no fishy smell and sensory acceptability, while the scores of the other groups were no higher than 2.40 points. The above results indicate that source control (defatting, enzymatic hydrolysis) and end-stage masking (goji berry puree) are both indispensable; any missing process will result in a sensory-acceptable product.

[0080] 3. The combined effect of this invention surpasses the simple sum of the effects of individual measures, exhibiting multi-level synergy. The theoretical expectation is that the sum of "defatting + enzymatic hydrolysis" (1.00 point) and "adding only goji berry juice" (2.40 point) is 3.40 points. The actual measured value of this invention is 3.80 points, exceeding the theoretical expectation by 0.40 points, with a synergy rate of approximately 11.8%. Furthermore, the same goji berry juice increases the effect by 2.40 points without pretreatment and by 2.80 points with pretreatment; the same pretreatment increases the effect by 1.00 points without goji berry juice and by 1.40 points with goji berry juice, with the two mutually enhancing each other by 0.40 points, indicating that the combination is not a simple superposition of independent deodorizing effects. Degreasing and enzymatic hydrolysis reduce the generation and residue of fishy odor substances from the source, reducing the residual fishy odor load to below the masking and binding capacity threshold of wolfberry juice; the polyphenols and flavonoids contained in wolfberry juice bind and fix aldehyde fishy odor substances, and the organic acids contained in it protonate volatile amine fishy odor substances and inhibit their volatilization, while its inherent aroma and reddish-brown color further mask the aroma and sensory effects; the overall deodorizing effect of "source control" and "end masking" is non-linearly superimposed.

[0081] 4. Deodorization and coloring work synergistically to jointly determine the sensory quality of the finished product. The wolfberry pulp in this deodorization system simultaneously masks the inherent gray-black color of the forest frog enzymatic hydrolysate, making the product present a uniform reddish-brown color that conforms to consumer perception. That is, the wolfberry pulp has multiple functions, including antioxidant synergy, formulation stability, natural coloring, and flavor masking, and works synergistically with the defatting and enzymatic hydrolysis processes to remove the fishy smell.

[0082] Based on the above-described preferred embodiments of the present invention, and through the foregoing description, those skilled in the art can make various changes and modifications without departing from the inventive concept. The technical scope of this invention is not limited to the contents of the specification.

Claims

1. A functional beverage made from a combination of forest frog skin and red ginseng, characterized in that, The product includes frog polypeptide liquid, red ginseng saponin liquid, deodorizing agent, sweetener, and whey protein; the frog polypeptide liquid is obtained by defatting and enzymatic hydrolysis of frog skin; the red ginseng saponin liquid is obtained by alcohol extraction of red ginseng; and the deodorizing agent is wolfberry puree.

2. A method for preparing a compound functional beverage of forest frog skin and red ginseng as described in claim 1, characterized in that, Includes the following steps: Forest frog Skin pretreatment: The frog skin powder is degreased with edible ethanol to obtain degreased frog skin material; Enzymatic hydrolysis of forest frog skin: Add defatted forest frog skin to water and adjust the pH to 6.5-7.

5. Add compound flavor protease for enzymatic hydrolysis, filter after enzyme inactivation to obtain forest frog polypeptide liquid; Red ginsenoside extraction: Red ginseng powder was extracted with edible ethanol, filtered to remove the edible ethanol, and then redissolved in water to obtain red ginsenoside solution; Preparation: Mix frog polypeptide liquid with red ginseng saponin liquid, add deodorizing agent, sweetener and whey protein and homogenize to obtain frog skin and red ginseng compound functional beverage.

3. The preparation method of the forest frog skin and red ginseng compound functional beverage according to claim 2, characterized in that, The pretreatment steps for the frog skin specifically include: The dried forest frog skin is crushed and sieved to obtain forest frog skin powder; Weigh out the frog skin powder, add edible ethanol, stir in a water bath at 50-70℃ for 20-40 minutes, sieve and discard the filtrate, rinse the filter residue with water and drain thoroughly to remove edible ethanol residue, and obtain defatted frog skin material; the mass-volume ratio of the frog skin powder to edible ethanol is (80-100) g:(400-500) mL.

4. The preparation method of the forest frog skin and red ginseng compound functional beverage according to claim 3, characterized in that, The enzymatic hydrolysis steps of the forest frog skin specifically include: Add defatted frog skin to water, adjust the pH to 6.5-7.5 with baking soda and / or citric acid, add compound flavor protease, and enzymatically hydrolyze at 50-60℃ for 3-5 hours. Then, inactivate the enzyme by water bath at 85-95℃ for 5-15 minutes. Sieve to obtain frog polypeptide solution; the mass ratio of the compound flavor protease to frog skin powder is (1-2):(80-100).

5. The preparation method of the forest frog skin and red ginseng compound functional beverage according to claim 2, characterized in that, The steps for extracting red ginsenosides specifically include: Weigh red ginseng powder, add edible ethanol, extract in a water bath at 50-70℃ for 30-50 minutes, filter, and then evaporate the filtrate in an open water bath at 50-70℃ until there is no alcohol odor, remove the edible ethanol, and then redissolve in water to obtain red ginseng saponin solution; the mass-volume ratio of red ginseng powder to edible ethanol is (80-100) g:(800-1000) mL.

6. The preparation method of the forest frog skin and red ginseng compound functional beverage according to claim 2, characterized in that, The volume percentage concentration of the edible ethanol is 65%-75%.

7. The preparation method of the forest frog skin and red ginseng compound functional beverage according to claim 2, characterized in that, The volume ratio of the frog polypeptide solution, red ginseng saponin solution and wolfberry puree is (20-30):(20-30):(45-55); the volume-to-mass ratio of the frog polypeptide solution to sweetener and whey protein is (20-30) mL:(3-5):(0.5-1.5).

8. The preparation method of the forest frog skin and red ginseng compound functional beverage according to claim 2, characterized in that, Following the preparation step, a sterilization step is also included.

9. The preparation method of the forest frog skin and red ginseng compound functional beverage according to claim 8, characterized in that, The sterilization step specifically includes sterilizing the prepared beverage at 80-90℃ for 10-20 minutes.

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