A method for preparing a deer penis wine with anti-fatigue efficacy

Abstract

This application discloses a method for making deer antler wine with anti-fatigue effects, relating to the field of health wine technology, including the following steps: Step 1, deer antler pretreatment: Fresh deer antler is washed, blanched, removed, sliced, dried, and soaked in water, then pulverized to obtain deer antler powder; Step 2, further processing: The deer antler powder is subjected to enzymatic hydrolysis, alcohol soaking to obtain the supernatant, and then the alcohol is evaporated to obtain a deer antler polypeptide aqueous solution; Step 3, compound deer antler wine: The deer antler polypeptide aqueous solution is mixed with white wine, and then red ginseng, yam, wolfberry, angelica, ophiopogon japonicus, cinnamon, and cloves are added to obtain... A finished deer antler wine containing 7-8 g / L deer antler, 9-10.5 g / L red ginseng, 4.5-5.5 g / L yam, 4.5-5.5 g / L angelica, 1.8-2.1 g / L ophiopogon japonicus, 0.95-1.05 g / L cinnamon, and 0.24-0.26 g / L cloves; wherein, during enzymatic hydrolysis, the mass concentration of the deer antler powder aqueous solution is controlled at 4.5-5.5%, and 5500-7000 U / g alkaline protease and 5500-7000 U / g neutral protease are used. This application has the effects of reducing fishy smell and anti-fatigue.

Description

Technical Field

[0001] This application relates to the field of health wine technology, and in particular to a method for making deer antler wine with anti-fatigue effects. Background Technology

[0002] In traditional Chinese medicine, deer antler velvet is considered a kidney-tonifying and aphrodisiac herb. Modern research has also found that it contains various active ingredients, providing a basis for its pharmacological effects. According to traditional Chinese medicine theory, deer antler velvet is sweet and salty in taste, warm in nature, and enters the liver, kidney, and bladder meridians. Its core functions are to tonify kidney essence, strengthen kidney yang, and fortify the lower back and knees. Modern research has found that deer antler velvet contains active ingredients such as proteins, polysaccharides, amino acids, and various trace elements. It has effects such as anti-fatigue, immune regulation, and strengthening kidney yang.

[0003] Currently, deer antler velvet is mainly used for soaking in alcohol.

[0004] However, directly soaking deer antler in alcohol has the following disadvantages: (1) Low dissolution rate of effective ingredients, resulting in waste of raw materials. The main active ingredients of deer antler are proteins and polypeptides, which are wrapped in dense connective tissue and cell membranes. In 40-60 degree liquor, macromolecules are difficult to diffuse out of the cells. Studies have shown that the dissolution rate of effective ingredients such as amino acids and trace elements in deer antler by traditional soaking method is usually about 12%, and most of the nutrients are still left in the discarded dregs, resulting in a great waste of raw materials. (2) Strong fishy smell and poor taste. The spongy tissue around the urethra of deer antler, residual blood and fat contain a large amount of fishy substances such as trimethylamine. After these substances dissolve in alcohol, they will make the liquor produce an unacceptable fishy smell, which greatly limits the acceptance of the product. (3) Low absorption and utilization rate and slow onset of action. The substances that are incorporated into the liquor are still mainly macromolecular proteins. After being ingested by the human body, they need to be digested and broken down into small molecule peptides and amino acids by the gastrointestinal tract before they can be absorbed. For middle-aged and elderly people with weak digestive function, this process is very inefficient. (4) Quality is uncontrollable and safety is not guaranteed. The size, part, freshness and processing method of each deer penis are different, resulting in large differences in the composition and concentration of the wine produced in each batch. Standardized production cannot be achieved and needs to be improved. Summary of the Invention

[0005] In view of this, the purpose of this application is to provide a method for making deer antler wine with anti-fatigue effects, so as to achieve the purpose of reducing fishy smell and anti-fatigue. The specific solution is as follows: A method for making deer antler wine with anti-fatigue effects includes the following steps: Step 1, Deer penis pretreatment: Fresh deer penis is washed, blanched, removed, sliced, dried and soaked in water, and then crushed to obtain deer penis powder; Step 2, further processing: The deer antler powder is sequentially enzymatically hydrolyzed, soaked in alcohol, and the supernatant is collected and the alcohol is evaporated to obtain an aqueous solution of deer antler polypeptide; Step 3, Compound Deer Antler Wine: Mix deer antler polypeptide aqueous solution with white wine, then add red ginseng, yam, wolfberry, angelica, ophiopogon japonicus, cinnamon and cloves to obtain a wine containing 7-8 g / L deer antler, 9-10.5 g / L red ginseng, 4.5-5.5 g / L yam, 4.5-5.5 g / L angelica, 1.8-2.1 g / L ophiopogon japonicus, 0.95-1.05 g / L cinnamon and 0.24-0.26 g / L cloves. During the enzymatic hydrolysis, the mass concentration of the deer antler powder aqueous solution is controlled at 4.5-5.5%, and alkaline protease with a concentration of 5500-7000 U / g and neutral protease with a concentration of 5500-7000 U / g are used.

[0006] Preferably: In step 1, the cleaning involves rinsing with clean water to remove blood and impurities and removing surface hair, muscle, fat, and connective tissue; the blanching involves cutting the slices and boiling them with ginger and rice wine for 2-3 minutes, then removing and cooling; the removal involves removing spongy tissue; the slicing involves obtaining thin slices with a thickness of 2.8-3.2 mm; the drying involves drying at a controlled temperature of 38-42℃ until the moisture content is 45-55%; the soaking involves soaking the slices in rice wine for 1-1.2 hours, then draining off the rice wine and drying until the moisture content is 13-17%, and controlling the amount of rice wine soaked to be 50% of the mass of the dried slices.

[0007] Preferably, in step 2, 0.8-1.2% of the total mass of the deer antler powder aqueous solution is added to the aqueous solution of the deer antler powder. After stirring at a constant temperature for 35-40 minutes, the complex precipitate is removed by centrifugation to obtain a purified and activated deer antler powder aqueous solution. The sulfopropyl grafted modified composite weak organic acid is obtained by sulfopropyl grafting modification after mixing citric acid and L-malic acid in a mass ratio of 2:1.

[0008] Preferably, the preparation method of the sulfopropyl graft-modified composite weak organic acid includes first mixing citric acid and L-malic acid at a mass ratio of 2:1 to obtain the grafting system; then controlling the grafting system under normal pressure, inert gas and temperature of 55-60℃, adding 5% by mass of sodium 3-chloropropanesulfonate monomer, controlling the pH value to 5.0-5.6, and stirring at a constant temperature for 60-80 min to obtain the sulfopropyl graft-modified composite weak organic acid.

[0009] Preferably, in step 2, the enzymatic hydrolysis temperature is 50-55℃, the enzymatic hydrolysis time is 4.5-5.5h, and then the enzyme is inactivated by a water bath at 90-95℃ for 14-16min.

[0010] Preferably, in step 2, the alcohol soaking and supernatant collection involves controlling the alcohol content to 60° and the temperature to 4°C, allowing it to stand for 24 hours, and then centrifuging it at 8000 rpm / min for 20-22 minutes before collecting the supernatant; the evaporation to remove alcohol involves controlling the water bath temperature to 38-42°C, the vacuum degree to -0.09 MPa, and the rotation speed to 50-70 rpm / min until the alcohol content is below 3%.

[0011] Preferably, in step 3, the liquor is a high-concentration grain liquor with an alcohol content greater than 52°.

[0012] Preferably, in step 3, the red ginseng, yam, wolfberry, angelica, ophiopogon japonicus, cinnamon, and clove are all modified by multi-component site-linked chemical activation. The modification process includes sequentially removing impurities, air-drying at low temperature, and passing through a 40-mesh sieve to obtain composite medicinal powder. The composite medicinal powder is then placed in an alcohol-water system with a volume fraction of 15% ethanol and pre-swelled at 24-26°C for 28-32 minutes, heated to 52-55°C and kept at a constant temperature for 40-45 minutes, then quenched to 20°C. This process is repeated at least twice, followed by sealed static activation for 30-32 minutes to obtain the modified composite medicinal raw material.

[0013] Preferably, the modified composite medicinal material is added to a uniformly mixed deer antler polypeptide aqueous solution and white wine, and after being sealed and soaked at room temperature, it is filtered to obtain deer antler wine.

[0014] Preferably, the period of room temperature sealed soaking is 20-22 days, and the mixture is stirred once a day.

[0015] As can be seen from the above scheme, this application provides a method for making deer antler wine with anti-fatigue effects, which has the following beneficial effects: 1. By pre-treating deer antler, using a double-enzyme gradient hydrolysis process and alcohol-soluble enrichment and purification, as well as adjusting the proportion of medicinal materials, the traditional deer antler wine is made with a strong fishy smell, low utilization rate of effective ingredients, single anti-fatigue effect, and unstable batch quality. At the same time, the process controllability is improved, thereby enhancing the safety and taste of food and significantly improving the anti-fatigue effect. It is suitable for large-scale production. 2. By using sulfopropyl grafted modified composite weak organic acid to purify and activate deer antler velvet, the strong electrostatic complexing properties of the sulfopropyl functional group can target and capture alkaline odor-causing small molecules such as trimethylamine at multiple sites, thereby achieving deep deodorization at the molecular level. At the same time, the spatial conformation of deer antler velvet protein is reconstructed, which promotes the exposure of a large number of enzymatic active sites, thereby significantly improving the efficiency of dual-enzyme hydrolysis and achieving the goal of improving the yield and bioactivity of alcohol-soluble anti-fatigue peptides. 3. By adopting a multi-component site-linking chemical activation modification treatment, the active groups such as saponins, polysaccharides, and flavonoids inside are fully released under temperature change and alcohol-water system control, and hydrogen bond coordination cross-linking is formed between each active component to form a structurally stable composite active molecular group. This effectively solves the problems of low active dissolution, easy degradation of components, and dispersion of efficacy when traditional medicinal materials are directly soaked, and significantly improves the dissolution of active ingredients and storage stability. 4. By synergistically combining the highly active deer antler peptides in the deer antler peptide aqueous solution with the complex active molecular groups in the modified compound medicinal materials to form a multidimensional anti-fatigue system that provides rapid fatigue relief and long-term repair, the goal of highly efficient anti-fatigue can be achieved. Detailed Implementation

[0016] The technical solutions described below in conjunction with the embodiments of this application will be clearly and completely described. Obviously, the described embodiments are only a part of the embodiments of this application, and not all of the embodiments. Based on the embodiments of this application, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of this application.

[0017] It should be mentioned that the liquor in this embodiment is a high-concentration grain liquor with an alcohol content greater than 52°, which is commercially available.

[0018] The following will describe in detail the method for making a deer antler wine with anti-fatigue effects according to this application.

[0019] A method for making deer antler wine with anti-fatigue effects includes the following steps: Step 1, Deer penis pretreatment: Fresh deer penis is washed, blanched, removed, sliced, dried and soaked in water, and then crushed to obtain deer penis powder; Step 2, further processing: The deer antler powder is sequentially enzymatically hydrolyzed, soaked in alcohol, and the supernatant is collected and the alcohol is evaporated to obtain an aqueous solution of deer antler polypeptide; Step 3, Compound Deer Antler Wine: Mix deer antler polypeptide aqueous solution with white wine, then add red ginseng, yam, wolfberry, angelica, ophiopogon japonicus, cinnamon and cloves to obtain a finished deer antler wine containing 7-8g / L deer antler, 9-10.5g / L red ginseng, 4.5-5.5g / L yam, 4.5-5.5g / L angelica, 1.8-2.1g / L ophiopogon japonicus, 0.95-1.05g / L cinnamon and 0.24-0.26g / L cloves. in: In step 1, washing involves rinsing with clean water to remove blood and impurities, and removing surface hair, muscle, fat, and connective tissue. Blanching involves cutting the slices into sections and boiling them with ginger and rice wine for 2-3 minutes, then removing and cooling. Removal refers to excising the spongy tissue. Slicing yields thin slices with a thickness of 2.8-3.2 mm. Drying involves drying at a controlled temperature of 38-42℃ until the moisture content reaches 45-55%. Soaking involves soaking the slices in rice wine for 1-1.2 hours, then draining the rice wine and drying until the moisture content reaches 13-17%, with the amount of rice wine used for soaking controlled to be 50% of the mass of the dried slices.

[0020] In step 2, the concentration of the deer antler powder aqueous solution is controlled at 4.5-5.5% during enzymatic hydrolysis. 0.8-1.2% (by weight of the total aqueous solution) of a sulfopropyl-grafted modified weak organic acid is added to the deer antler powder aqueous solution. After stirring at a constant temperature for 35-40 minutes, the complex precipitate is removed by centrifugation to obtain a purified and activated deer antler powder aqueous solution. Finally, enzymatic hydrolysis is performed using 5500-7000 U / g of alkaline protease and 5500-7000 U / g of neutral protease. It should be noted that the enzymatic hydrolysis temperature is 50-55℃, the hydrolysis time is 4.5-5.5 hours, and the enzyme is then inactivated by a 90-95℃ water bath for 14-16 minutes.

[0021] The sulfonyl-grafted modified composite weak organic acid is obtained by sulfonyl grafting modification of a mixture of citric acid and L-malic acid at a mass ratio of 2:1. The preparation method of the sulfonyl-grafted modified composite weak organic acid includes: firstly, mixing citric acid and L-malic acid at a mass ratio of 2:1 to obtain the grafting system; then, controlling the grafting system under normal pressure, inert gas, and a temperature of 55-60℃, adding 5% (w / w) of sodium 3-chloropropanesulfonate monomer, controlling the pH value to 5.0-5.6, and stirring at a constant temperature for 60-80 min to obtain the sulfonyl-grafted modified composite weak organic acid.

[0022] Meanwhile, the supernatant was collected after alcohol immersion, with the alcohol content controlled at 60° and the temperature at 4°C, and allowed to stand for 24 hours. It was then centrifuged at 8000 rpm / min for 20-22 minutes before collecting the supernatant. Alcohol was removed by evaporation, with the water bath temperature controlled at 38-42°C, the vacuum degree at -0.09 MPa, and the centrifugation speed at 50-70 rpm / min until the alcohol content was below 3%.

[0023] In step 3, red ginseng, yam, wolfberry, angelica, ophiopogon japonicus, cinnamon, and cloves all undergo multi-component site-linked chemical activation modification treatment. The modification process includes sequentially removing impurities, low-temperature air drying, and passing through a 40-mesh sieve to obtain a composite medicinal powder. The powder is then placed in a 15% (v / v) ethanol-water system and pre-swelled at 24-26°C for 28-32 minutes. The temperature is then raised to 52-55°C and held constant for 40-45 minutes. The mixture is then quenched to 20°C, and this cycle is repeated at least twice. Afterward, the mixture is sealed and allowed to stand for 30-32 minutes to activate, yielding the modified composite medicinal raw material. This modified raw material is added to a uniformly mixed deer antler peptide aqueous solution and baijiu (Chinese liquor). After a 20-22 day period of sealed soaking at room temperature with daily stirring, the mixture is filtered to obtain deer antler wine.

[0024] Example 1 A method for making deer antler wine with anti-fatigue effects includes the following steps: Step 1, Deer penis pretreatment: Fresh deer penis is washed, blanched, removed, sliced, dried and soaked in water, and then crushed to obtain deer penis powder; Step 2, further processing: The deer antler powder is sequentially enzymatically hydrolyzed, soaked in alcohol, and the supernatant is collected and the alcohol is evaporated to obtain an aqueous solution of deer antler polypeptide; Step 3, Compound Deer Antler Wine: Mix deer antler polypeptide aqueous solution with white wine, then add red ginseng, yam, wolfberry, angelica, ophiopogon japonicus, cinnamon and cloves to obtain a finished deer antler wine containing 7g / L deer antler, 9g / L red ginseng, 4.5g / L yam, 4.5g / L angelica, 1.8g / L ophiopogon japonicus, 0.95g / L cinnamon and 0.24g / L cloves. in: In step 1, washing involves rinsing with clean water to remove blood and impurities, and removing surface hair, muscle, fat, and connective tissue. Blanching involves cutting the slices into sections and boiling them with ginger and rice wine for 2 minutes, then removing and cooling. Removal involves excising the spongy tissue. Slicing yields thin slices with a thickness of 2.8 mm. Drying involves drying at a controlled temperature of 38°C until the moisture content reaches 45%. Soaking involves soaking the slices in rice wine for 1 hour, then draining the rice wine and drying until the moisture content reaches 13%, with the amount of rice wine used for soaking controlled to be 50% of the mass of the dried slices.

[0025] In step 2, the concentration of the deer antler powder aqueous solution was controlled at 4.5% during enzymatic hydrolysis, and enzymatic hydrolysis was performed using 5500 U / g alkaline protease and 5500 U / g neutral protease. It should be noted that the enzymatic hydrolysis temperature was 50℃, the hydrolysis time was 4.5 h, and then the enzyme was inactivated by a 90℃ water bath for 14 min.

[0026] Meanwhile, the supernatant was collected after alcohol immersion, with the alcohol content controlled at 60° and the temperature at 4°C, and allowed to stand for 24 hours. It was then centrifuged at 8000 rpm / min for 20 minutes before the supernatant was collected. Alcohol was removed by evaporation, with the water bath temperature controlled at 38°C, the vacuum degree at -0.09 MPa, and the centrifugation speed at 50 rpm / min until the alcohol content was below 3%.

[0027] Example 2 The difference between Example 2 and Example 1 is that in step 2 of Example 2, the mass concentration of the deer antler powder aqueous solution is controlled at 4.5% during enzymatic hydrolysis, and 0.8% of the total mass of the deer antler powder aqueous solution is added to the solution. After stirring at a constant temperature for 35 minutes, the complex precipitate is removed by centrifugation to obtain a purified and activated deer antler powder aqueous solution. Finally, enzymatic hydrolysis is performed using 5500 U / g alkaline protease and 5500 U / g neutral protease. The sulfopropyl-grafted modified composite weak organic acid is obtained by sulfopropyl grafting modification after mixing citric acid and L-malic acid in a mass ratio of 2:1. The preparation method of sulfopropyl graft-modified composite weak organic acid includes first mixing citric acid and L-malic acid at a mass ratio of 2:1 to obtain the grafting system; then controlling the grafting system at normal pressure, inert gas and temperature of 55℃, adding 5% sodium 3-chloropropanesulfonate monomer, controlling the pH value range of 5.0-5.6, and stirring at a constant temperature for 60 min to obtain sulfopropyl graft-modified composite weak organic acid.

[0028] Example 3 The difference between Example 3 and Example 1 is that in step 3 of Example 3, red ginseng, yam, wolfberry, angelica, ophiopogon japonicus, cinnamon, and cloves are all modified through multi-component site-linked chemical activation. The modification process includes sequentially removing impurities, low-temperature air drying, and passing through a 40-mesh sieve to obtain a composite medicinal powder. The powder is then placed in a 15% (v / v) ethanol-water system and pre-swelled at 24°C for 28 minutes. The temperature is then gradually increased to 52°C and held constant for 40 minutes, followed by rapid quenching to 20°C. This cycle is repeated twice, followed by sealed static activation for 30 minutes to obtain the modified composite medicinal raw material. This modified composite medicinal raw material is added to a uniformly mixed deer antler peptide aqueous solution and baijiu (Chinese liquor). After a 20-day period of sealed soaking at room temperature with daily stirring, the mixture is filtered to obtain deer antler wine.

[0029] Furthermore, in this embodiment of the application, the gradient heating is controlled to increase the temperature by 10°C per minute, and the rapid quenching is carried out within 20 seconds.

[0030] Example 4 A method for making deer antler wine with anti-fatigue effects includes the following steps: Step 1, Deer penis pretreatment: Fresh deer penis is washed, blanched, removed, sliced, dried and soaked in water, and then crushed to obtain deer penis powder; Step 2, further processing: The deer antler powder is sequentially enzymatically hydrolyzed, soaked in alcohol, and the supernatant is collected and the alcohol is evaporated to obtain an aqueous solution of deer antler polypeptide; Step 3, Compound Deer Antler Wine: Mix deer antler polypeptide aqueous solution with white wine, then add red ginseng, yam, wolfberry, angelica, ophiopogon japonicus, cinnamon and cloves to obtain a finished deer antler wine containing 7.5g / L deer antler, 10g / L red ginseng, 5g / L yam, 5g / L angelica, 2g / L ophiopogon japonicus, 1g / L cinnamon and 0.25g / L cloves. in: In step 1, washing involves rinsing with clean water to remove blood and impurities, and removing surface hair, muscle, fat, and connective tissue. Blanching involves cutting the slices into sections and boiling them with ginger and rice wine for 2.5 minutes, then removing and cooling. Removal involves excising the spongy tissue. Slicing yields thin slices with a thickness of 3 mm. Drying involves drying at a controlled temperature of 40℃ until the moisture content reaches 50%. Soaking involves soaking the slices in rice wine for 1.1 hours, then draining the rice wine and drying until the moisture content reaches 15%, with the amount of rice wine used for soaking controlled to be 50% of the mass of the dried slices.

[0031] In step 2, the concentration of the deer antler powder aqueous solution was controlled at 5% during enzymatic hydrolysis. 1% (by weight) of a sulfopropyl-grafted modified weak organic acid was added to the deer antler powder aqueous solution. After stirring at a constant temperature for 38 minutes, the complex precipitate was removed by centrifugation to obtain a purified and activated deer antler powder aqueous solution. Finally, enzymatic hydrolysis was performed using 6000 U / g alkaline protease and 6000 U / g neutral protease. It should be noted that the enzymatic hydrolysis temperature was 52℃, the hydrolysis time was 5 hours, and the enzyme was then inactivated by a 92℃ water bath for 15 minutes.

[0032] The sulfonyl-grafted modified composite weak organic acid is obtained by sulfonyl grafting modification of a mixture of citric acid and L-malic acid at a mass ratio of 2:1. The preparation method of the sulfonyl-grafted modified composite weak organic acid includes: firstly, mixing citric acid and L-malic acid at a mass ratio of 2:1 to obtain the grafting system; then, controlling the grafting system under normal pressure, inert gas, and a temperature of 52℃, adding 5% (w / w) of sodium 3-chloropropanesulfonate monomer, controlling the pH value within the range of 5.0-5.6, and stirring at a constant temperature for 70 min to obtain the sulfonyl-grafted modified composite weak organic acid.

[0033] Meanwhile, the supernatant was collected after alcohol immersion, with the alcohol content controlled at 60° and the temperature at 4°C, and allowed to stand for 24 hours. It was then centrifuged at 8000 rpm / min for 21 minutes before the supernatant was collected. Alcohol was removed by evaporation, with the water bath temperature controlled at 40°C, the vacuum degree at -0.09 MPa, and the centrifugation speed at 60 rpm / min until the alcohol content was below 3%.

[0034] In step 3, red ginseng, yam, wolfberry, angelica, ophiopogon japonicus, cinnamon, and cloves all underwent multi-component site-linked chemical activation modification treatment. The modification process included sequentially removing impurities, low-temperature air drying, and passing through a 40-mesh sieve to obtain a composite medicinal powder. The powder was then placed in a 15% (v / v) ethanol-water system and pre-swelled at 25°C for 30 minutes, followed by heating to 54°C and holding the temperature for 42 minutes, then quenching to 20°C. This cycle was repeated twice, followed by sealed static activation for 31 minutes to obtain the modified composite medicinal raw material. This modified composite medicinal raw material was added to a uniformly mixed deer antler peptide aqueous solution and baijiu (Chinese liquor), and after a 21-day period of sealed soaking at room temperature with daily stirring, the mixture was filtered to obtain deer antler wine.

[0035] Example 5 A method for making deer antler wine with anti-fatigue effects includes the following steps: Step 1, Deer penis pretreatment: Fresh deer penis is washed, blanched, removed, sliced, dried and soaked in water, and then crushed to obtain deer penis powder; Step 2, further processing: The deer antler powder is sequentially enzymatically hydrolyzed, soaked in alcohol, and the supernatant is collected and the alcohol is evaporated to obtain an aqueous solution of deer antler polypeptide; Step 3, Compound Deer Antler Wine: Mix deer antler polypeptide aqueous solution with white wine, then add red ginseng, yam, wolfberry, angelica, ophiopogon japonicus, cinnamon and cloves to obtain the finished deer antler wine containing 8g / L deer antler, 10.5g / L red ginseng, 5.5g / L yam, 5.5g / L angelica, 2.1g / L ophiopogon japonicus, 1.05g / L cinnamon and 0.26g / L cloves. in: In step 1, washing involves rinsing with clean water to remove blood and impurities, and removing surface hair, muscle, fat, and connective tissue. Blanching involves cutting the slices into sections and boiling them with ginger and rice wine for 3 minutes, then removing and cooling. Removal involves excising the spongy tissue. Slicing yields thin slices with a thickness of 3.2 mm. Drying is performed at a controlled temperature of 42℃ until the moisture content reaches 55%. Soaking involves soaking the slices in rice wine for 1.2 hours, then draining off the rice wine and drying until the moisture content reaches 17%, with the amount of rice wine used for soaking controlled to be 50% of the mass of the dried slices.

[0036] In step 2, the concentration of the deer antler powder aqueous solution was controlled at 5.5% during enzymatic hydrolysis. 1.2% (by weight of the total aqueous solution) of a sulfopropyl-grafted modified weak organic acid was added to the deer antler powder aqueous solution. After stirring at a constant temperature for 40 minutes, the complex precipitate was removed by centrifugation to obtain a purified and activated deer antler powder aqueous solution. Finally, enzymatic hydrolysis was performed using 7000 U / g alkaline protease and 7000 U / g neutral protease. It should be noted that the enzymatic hydrolysis temperature was 55℃, the hydrolysis time was 5.5 hours, and the enzyme was then inactivated by a 95℃ water bath for 16 minutes.

[0037] The sulfonyl-grafted modified composite weak organic acid is obtained by sulfonyl grafting modification of a mixture of citric acid and L-malic acid at a mass ratio of 2:1. The preparation method of the sulfonyl-grafted modified composite weak organic acid includes: firstly, mixing citric acid and L-malic acid at a mass ratio of 2:1 to obtain the grafting system; then, controlling the grafting system under normal pressure, inert gas, and a temperature of 60℃, adding 5% (w / w) of sodium 3-chloropropanesulfonate monomer, controlling the pH value within the range of 5.0-5.6, and stirring at a constant temperature for 80 min to obtain the sulfonyl-grafted modified composite weak organic acid.

[0038] Meanwhile, the supernatant was collected after alcohol soaking, with the alcohol content controlled at 60° and the temperature at 4°C, and allowed to stand for 24 hours. It was then centrifuged at 8000 rpm / min for 22 minutes before the supernatant was collected. Alcohol was removed by evaporation, with the water bath temperature controlled at 42°C, the vacuum degree at -0.09 MPa, and the centrifugation speed at 70 rpm / min until the alcohol content was below 3%.

[0039] In step 3, red ginseng, yam, wolfberry, angelica, ophiopogon japonicus, cinnamon, and cloves all underwent multi-component site-specific cross-linking chemical activation modification treatment. The modification process included sequentially removing impurities, low-temperature air drying, and passing through a 40-mesh sieve to obtain composite medicinal powder. The composite medicinal powder was then placed in an alcohol-water system with a 15% (v / v) ethanol concentration, pre-swelled at 26°C for 32 minutes, heated to 55°C and held at that temperature for 45 minutes, then quenched to 20°C. This cycle was repeated twice, followed by sealed static activation for 32 minutes to obtain the modified composite medicinal raw material. This modified composite medicinal raw material was added to a uniformly mixed deer antler peptide aqueous solution and baijiu (Chinese liquor), and after a 22-day period of sealed soaking at room temperature with daily stirring, the mixture was filtered to obtain deer antler wine.

[0040] Comparative Example 1 The difference between Comparative Example 1 and Example 4 is that the enzymatic hydrolysis in Comparative Example 1 uses an alkaline protease with a concentration of 6000 U / g.

[0041] Comparative Example 2 The difference between Comparative Example 2 and Example 4 is that in step 1 of Comparative Example 2, fresh deer antlers are washed, sliced ​​and dried in sequence, and then crushed to obtain deer antler powder.

[0042] Performance testing: 1. Sensory test for fishy and gamey flavor: Referring to GB / T 10345-2022 "Sensory Evaluation Method of Baijiu", the scoring standard is set from 1 to 10 points. The lower the score, the stronger the fishy and gamey flavor and the lower the acceptance. The higher the score, the lower the fishy and gamey flavor and the higher the acceptance. Ten random people will give scores without prompting. 2. Active ingredient dissolution rate test: Weigh an equal amount of deer antler raw material, soak it for the corresponding time, filter it, dry the filter residue and weigh it, and calculate the dissolution rate (%) of the active ingredient.

[0043] The performance test results are shown in Table 1 below.

[0044] Table 1 Performance Test Results

[0045] In addition, the deer antler wine from Example 1 and Comparative Example 2 was rotary distilled and then freeze-dried to obtain the active ingredients of the deer antler wine. The recommended dosage for normal adults is 50-150 ml / day. The following test set up a normal dose group of 100 ml / day and a high dose group of 200 ml / day, with the recommended dosage of 100 ml / day of Jinjiu (a type of Chinese liquor) as the control group. According to K... m The coefficient conversion method was used to adjust the daily dose for mice. The experimental procedure was conducted according to the "Test Method for Relieving Physical Fatigue" in the "Methods for Functional Testing and Evaluation of Health Foods (2023 Edition)". The experimental results are shown in Tables 2 to 5 below: Table 2. Swimming time under load for mice in each group (mean ± standard deviation)

[0046] Table 3. BUN concentration in mice of each group (mean ± standard deviation, mmol / L)

[0047] Table 4. Liver glycogen content in mice of each group (mean ± standard deviation, mg / g)

[0048] Table 5. Area under the lactate curve for each group of mice

[0049] In Tables 2 to 5, A indicates a highly significant difference compared to group BK (P < 0.01); a indicates a significant difference compared to group BK (P < 0.05).

[0050] As shown in Tables 1 to 5 above, Comparative Example 2, made from fresh deer antler without blanching, trimming, or processing with rice wine, contained a large amount of trimethylamine, a strong odorous substance, which was not removed, resulting in a sensory score as low as 2.0 ± 0.82. Simultaneously, the dense connective tissue of the deer antler was not softened, and the loose structure was not pretreated, leading to a low solubility rate of only 12.1% due to the natural extraction of active ingredients by the rice wine. Example 1, on the other hand, employed deer antler pretreatment and dual-enzyme hydrolysis. Although it lacked the modification treatment of sulfopropyl grafted modified composite weak organic acid and multi-component site-linked chemical activation, it achieved a significant reduction in odorous substances and a significantly higher dissolution rate than Comparative Example 2.

[0051] Example 2 utilizes sulfopropyl grafting to modify a weak organic acid, thereby effectively deodorizing small amine-based odor molecules through electrostatic complexation of the sulfopropyl groups. Simultaneously, it modifies the spatial conformation of deer antler protein, extending the peptide chains and exposing enzymatic sites, thus promoting peptide dissolution. Therefore, the solubility is significantly improved compared to Example 1. Example 3 employs a multi-component, site-linked, chemically activated modification treatment. The cell walls of the raw material are pore-created through temperature-induced quenching, allowing the formed polysaccharide and saponin active groups to become free and coordinate, synergistically enhancing the sustained anti-fatigue properties of deer antler. Examples 4 and 5, while significantly reducing odor, also synergistically enhance peptide dissolution and anti-fatigue function.

[0052] Meanwhile, compared with Example 4, Comparative Example 1 did not use a neutral protease, which prevented it from performing segmented enzymatic hydrolysis and cleavage of long protein chains. As a result, the amount of small molecule peptides generated decreased, and the dissolution rate and anti-fatigue index were significantly lower than those of Example 4.

[0053] In summary, this application provides a method for producing deer antler wine with anti-fatigue effects. This method, through deer antler pretreatment, a dual-enzyme gradient hydrolysis process, and alcohol-soluble enrichment and purification reprocessing, along with the appropriate ratio of medicinal materials, overcomes the problems of strong fishy smell, low utilization rate of effective components, single anti-fatigue effect, and unstable batch quality in traditional deer antler wine. It also improves process controllability, thereby enhancing food safety and taste, and significantly improving anti-fatigue efficacy, making it suitable for large-scale production. To further improve the quality stability of this deer antler wine, sulfopropyl grafted modified composite weak organic acid is used to purify and activate the deer antler. This allows the strong electrostatic complexing properties of the sulfopropyl functional groups to target and capture alkaline fishy small molecules such as trimethylamine at multiple sites, achieving deep deodorization at the molecular level. Simultaneously, the spatial conformation of the deer antler protein is reconstructed, exposing a large number of enzymatic active sites, thus significantly improving the efficiency of dual-enzyme hydrolysis and increasing the yield and bioactivity of alcohol-soluble anti-fatigue peptides. Meanwhile, through multi-component cross-linking chemical activation modification, the active groups such as saponins, polysaccharides, and flavonoids are fully released under controlled temperature changes and alcohol-water systems. This allows for hydrogen bond coordination and cross-linking between the active components, forming structurally stable composite active molecular clusters. This effectively solves the problems of low active ingredient dissolution, easy degradation, and dispersed efficacy associated with traditional direct soaking of medicinal materials, significantly improving the dissolution rate and storage stability of active ingredients. Therefore, this method for producing deer antler wine with anti-fatigue effects achieves its goal of highly efficient anti-fatigue by synergistically combining highly active deer antler peptides in an aqueous solution with composite active molecular clusters in modified medicinal materials to form a multi-dimensional anti-fatigue system that provides rapid fatigue relief and long-term repair.

[0054] The terms “first,” “second,” “third,” “fourth,” etc., used in this application (if applicable) are used to distinguish similar objects and are not necessarily used to describe a specific order or sequence. It should be understood that such data can be interchanged where appropriate so that the embodiments described herein can be implemented in orders other than those described herein. Furthermore, the terms “comprising” and “having,” and any variations thereof, are intended to cover non-exclusive inclusion; for example, a process, method, or apparatus that includes a series of steps or units is not necessarily limited to those explicitly listed, but may include other steps or units not explicitly listed or inherent to such processes, methods, or apparatus.

[0055] It should be noted that the use of terms such as "first" and "second" in this application is for descriptive purposes only and should not be construed as indicating or implying their relative importance or implicitly specifying the number of technical features indicated. Therefore, a feature defined as "first" or "second" may explicitly or implicitly include at least one of those features. Furthermore, the technical solutions of the various embodiments can be combined with each other, but this must be based on the ability of those skilled in the art to implement them. If the combination of technical solutions is contradictory or impossible to implement, such a combination of technical solutions should be considered non-existent and not within the scope of protection claimed in this application.

[0056] This document uses specific examples to illustrate the principles and implementation methods of this application. The descriptions of the above embodiments are only for the purpose of helping to understand the methods and core ideas of this application. At the same time, for those skilled in the art, there will be changes in the specific implementation methods and application scope based on the ideas of this application. Therefore, the content of this specification should not be construed as a limitation of this application.

Claims

1. A method for making deer antler wine with anti-fatigue effects, characterized in that, Includes the following steps: Step 1, Deer penis pretreatment: Fresh deer penis is washed, blanched, removed, sliced, dried and soaked in water, and then crushed to obtain deer penis powder; Step 2, further processing: The deer antler powder is sequentially enzymatically hydrolyzed, soaked in alcohol, and the supernatant is collected and the alcohol is evaporated to obtain an aqueous solution of deer antler polypeptide; Step 3, Compound Deer Antler Wine: Mix deer antler polypeptide aqueous solution with white wine, then add red ginseng, yam, wolfberry, angelica, ophiopogon japonicus, cinnamon and cloves to obtain a finished deer antler wine containing 7-8g / L deer antler, 9-10.5g / L red ginseng, 4.5-5.5g / L yam, 4.5-5.5g / L angelica, 1.8-2.1g / L ophiopogon japonicus, 0.95-1.05g / L cinnamon and 0.24-0.26g / L cloves. During the enzymatic hydrolysis, the mass concentration of the deer antler powder aqueous solution is controlled at 4.5-5.5%, and alkaline protease with a concentration of 5500-7000 U / g and neutral protease with a concentration of 5500-7000 U / g are used.

2. The method for preparing deer antler wine with anti-fatigue effects according to claim 1, characterized in that: In step 1, the cleaning involves rinsing with clean water to remove blood and dirt, and removing surface hair, muscle, fat, and connective tissue; the blanching involves cutting the meat into sections and boiling it with ginger and rice wine for 2-3 minutes, then removing it and cooling it. The removal refers to the excision of spongy tissue; the slicing refers to obtaining thin slices with a thickness of 2.8-3.2 mm; the drying refers to drying at a controlled temperature of 38-42℃ until the moisture content is 45-55%; the soaking refers to soaking in rice wine for 1-1.2 hours, then draining off the rice wine and drying until the moisture content is 13-17%, and controlling the amount of rice wine soaked to be 50% of the mass of the dried slices.

3. The method for preparing deer antler wine with anti-fatigue effects according to claim 1, characterized in that: In step 2, 0.8-1.2% of the total mass of the deer antler powder aqueous solution is added to the aqueous solution of the deer antler powder. After stirring at a constant temperature for 35-40 minutes, the complex precipitate is removed by centrifugation to obtain a purified and activated deer antler powder aqueous solution. The sulfopropyl grafted modified composite weak organic acid is obtained by sulfopropyl grafting modification after mixing citric acid and L-malic acid in a mass ratio of 2:

1.

4. The method for preparing deer antler wine with anti-fatigue effects according to claim 3, characterized in that: The preparation method of the sulfopropyl graft-modified composite weak organic acid includes first mixing citric acid and L-malic acid at a mass ratio of 2:1 to obtain the grafting system; then controlling the grafting system at normal pressure, inert gas and temperature of 55-60℃, adding 5% sodium 3-chloropropanesulfonate monomer, controlling the pH value to 5.0-5.6, and stirring at a constant temperature for 60-80 min to obtain the sulfopropyl graft-modified composite weak organic acid.

5. The method for preparing deer antler wine with anti-fatigue effects according to claim 1, characterized in that: In step 2, the enzymatic hydrolysis temperature is 50-55℃, the enzymatic hydrolysis time is 4.5-5.5h, and then the enzyme is inactivated by a water bath at 90-95℃ for 14-16min.

6. The method for preparing deer antler wine with anti-fatigue effects according to claim 1, characterized in that: In step 2, the alcohol soaking and supernatant collection involves controlling the alcohol content to 60° and the temperature to 4°C, allowing it to stand for 24 hours, and then centrifuging it at 8000 rpm / min for 20-22 minutes before collecting the supernatant. The alcohol evaporation involves controlling the water bath temperature to 38-42°C, the vacuum degree to -0.09 MPa, and the rotation speed to 50-70 rpm / min until the alcohol content is below 3%.

7. The method for making deer antler wine with anti-fatigue effects according to claim 1, characterized in that: In step 3, the liquor is a high-concentration grain liquor with an alcohol content greater than 52°.

8. The method for preparing deer antler wine with anti-fatigue effects according to claim 1, characterized in that: In step 3, the red ginseng, yam, wolfberry, angelica, ophiopogon japonicus, cinnamon, and cloves are all modified by multi-component site-linked chemical activation. The modification process includes sequentially removing impurities, air-drying at low temperature, and passing through a 40-mesh sieve to obtain composite medicinal powder. The composite medicinal powder is then placed in an alcohol-water system with a volume fraction of 15% ethanol and pre-swelled at 24-26℃ for 28-32 minutes, heated to 52-55℃ and kept at a constant temperature for 40-45 minutes, quenched to 20℃, and the process is repeated at least twice. After that, the mixture is sealed and allowed to stand for 30-32 minutes to activate, thus obtaining the modified composite medicinal raw material.

9. A method for preparing deer antler wine with anti-fatigue effects according to claim 8, characterized in that: The modified composite medicinal material is added to a well-mixed aqueous solution of deer antler peptides and baijiu (Chinese liquor), and after being sealed and soaked at room temperature, it is filtered to obtain deer antler wine.

10. A method for preparing deer antler wine with anti-fatigue effects according to claim 9, characterized in that: The period of room temperature sealed soaking is 20-22 days, and it is stirred once a day.

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