Kidney-tonifying spleen-strengthening intelligence-developing brain peptide oral liquid and preparation method thereof
By utilizing a multi-protease system and the sequential action of microbial fermentation, sheep brain peptides, oyster peptides, and soybean-walnut complex peptides were prepared. Combined with traditional Chinese medicine extracts, the problems of slow absorption and poor stability of existing preparations were solved, achieving synergistic effects of tonifying the kidney and spleen and improving intelligence, and enhancing the bioavailability and stability of the product.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- 湖北研妆实业有限公司
- Filing Date
- 2026-04-13
- Publication Date
- 2026-05-12
AI Technical Summary
Existing kidney-tonifying and spleen-strengthening preparations and brain-boosting products suffer from slow absorption, low bioavailability, and poor stability. Furthermore, existing brain peptide products have low retention rates of active ingredients, making it difficult to achieve the synergistic effects of simultaneously regulating the spleen and kidneys and nourishing the brain and body.
By employing a multi-protein system and the sequential action of microbial fermentation, sheep brain peptides, oyster peptides, and soybean-walnut complex peptides were prepared. Combined with traditional Chinese medicine extracts, DHA algal oil, and other ingredients, the content and targeting of active peptides were enhanced through precise compounding enzymatic hydrolysis and fermentation processes, resulting in a synergistic effect.
It significantly improves the content and absorption efficiency of active peptides in peptide components, achieving the dual effects of tonifying the kidneys and spleen and improving intelligence, and enhances the stability and bioavailability of the formulation, making it suitable for large-scale production.
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Abstract
Description
Technical Field
[0001] This application relates to the field of health drink preparation technology, and more specifically, it relates to a brain peptide oral liquid that tonifies the kidneys, strengthens the spleen, and improves intelligence, and its preparation method. Background Technology
[0002] With the aging population and the expansion of the sub-healthy group, symptoms such as memory decline, cognitive decline, lower back and knee pain, and spleen and stomach weakness are becoming increasingly common. This has created an urgent market demand for health products that can both regulate the internal organs and improve brain function. Traditional Chinese medicine theory holds that "the kidneys store essence, produce marrow, and connect to the brain," and "the spleen is the source of qi and blood." Therefore, the strength of brain function is closely related to the function of the spleen and kidneys, and tonifying the kidneys and strengthening the spleen is the core approach to improving brain function.
[0003] In existing technologies, kidney-tonifying and spleen-strengthening preparations are mostly in the form of pills or powders, which have the problems of slow absorption and low bioavailability; while brain-boosting products mostly rely on chemically synthesized ingredients, which are prone to causing side effects with long-term use.
[0004] Brain peptides, as naturally occurring active substances in the brain, play a crucial role in nerve cell repair and cognitive function regulation. However, directly extracted brain peptides suffer from poor stability and are easily destroyed by digestive juices. Existing brain peptide products mostly employ single extraction processes, failing to incorporate traditional Chinese medicine's theory of organ-based regulation, and suffer from drawbacks such as low retention of active ingredients and insufficient formulation stability, making it difficult to achieve the synergistic effect of "simultaneous regulation of the spleen and kidneys, and simultaneous nourishment of the brain and body." Therefore, developing a scientifically formulated, advancedly prepared brain peptide oral liquid with dual effects of tonifying the kidneys and spleen while improving intelligence and brain function, and possessing good stability and high bioavailability, is of great significance for meeting clinical needs and enhancing product competitiveness. Summary of the Invention
[0005] To address the technical problems mentioned in the background section, this application provides a brain peptide oral liquid that tonifies the kidneys, strengthens the spleen, and improves intelligence, as well as a method for its preparation.
[0006] In the first aspect, this application provides a brain peptide oral liquid that tonifies the kidneys, strengthens the spleen, and improves intelligence, employing the following technical solution:
[0007] A brain peptide oral liquid for tonifying the kidneys, strengthening the spleen, and improving intelligence contains the following raw material components in parts by weight per 100 servings: 3-5 parts longan pulp, 3-6 parts wolfberry, 3-8 parts mulberry, 3-6 parts red dates, 2-6 parts poria cocos, 0.5-2 parts alpinia oxyphylla, 0.05-0.1 parts dried ginger, 8-12 parts peptide components, 0.01-0.05 parts DHA algal oil, 0.05-0.1 parts N-acetylneuraminic acid, 1-3 parts γ-aminobutyric acid, and 0.002-0.004 parts vitamin B. 12 0.05-0.1 parts steviol glycosides, 12-15 parts concentrated apple juice, and the remainder is purified water.
[0008] Preferably, the preparation method of the peptide component includes the following preparation steps:
[0009] Step 1: Homogenize sheep brain and add deionized water to adjust the pH to 6.5-7.0. Add complex protease and hydrolyze for 2.5-3 hours. After inactivating the enzyme, centrifuge and collect the supernatant to obtain sheep brain peptides.
[0010] Step 2: Crush the oyster meat and add deionized water to obtain a fermentation substrate. Inoculate the fermentation substrate with Bacillus natto and ferment for 18-24 hours to obtain a fermentation broth. Add neutral protease to the fermentation broth and hydrolyze for 2-5 hours. Centrifuge to remove the supernatant and obtain oyster peptides.
[0011] Step 3: Mix walnuts and soybeans, grind and sieve them, add deionized water and mix evenly, adjust the pH to 8.0-8.5 to obtain soybean-walnut liquid; first add alkaline protease to soybean-walnut liquid and hydrolyze for 2-3 hours, then add flavor protease and hydrolyze for 1-3 hours, inactivate the enzyme, centrifuge and take the supernatant to obtain soybean-walnut complex peptide.
[0012] Step 4: Mix sheep brain peptides, oyster peptides and soybean-walnut complex peptides, vacuum concentrate to a solid content of 20-25%, and then freeze-dry to obtain peptide components.
[0013] Preferably, in step 1, the complex protease is composed of subtilisin, elastase and aminopeptidase in a mass ratio of 10-12:5-8:2-5.
[0014] Preferably, the mass ratio of the complex protease to sheep brain in step 1 is 1-3:200-300.
[0015] Preferably, the amount of Bacillus natto inoculated in step 2 is 2-5% of the mass of the fermentation substrate.
[0016] Preferably, in step 2, the mass ratio of fermentation broth to neutral protease is 100-200:3-5.
[0017] Preferably, in step 3, the mass ratio of soybean-walnut liquid, alkaline protease, and flavor protease is 200-300:3-5:1-3.
[0018] Preferably, in step 4, the mass ratio of sheep brain peptide, oyster peptide, and soybean-walnut complex peptide is 3-6:2-5:1-2.
[0019] Secondly, this application provides a method for preparing a brain peptide oral liquid that tonifies the kidneys, strengthens the spleen, and improves intelligence, using the following technical solution:
[0020] A method for preparing a brain peptide oral liquid that tonifies the kidneys, strengthens the spleen, and improves intelligence, specifically including the following preparation steps:
[0021] Mix longan pulp, goji berries, mulberries, red dates, poria cocos, alpinia oxyphylla, dried ginger, and purified water, and decoct 2-3 times, 1-2 hours each time. Combine the decoctions, filter and concentrate to obtain a herbal extract. Add peptide components and DHA algal oil to the herbal extract, stir well, and then add N-acetylneuraminic acid, γ-aminobutyric acid, and vitamin B. 12 Steviol glycosides and concentrated apple juice are mixed evenly, then purified water is added to make up the total volume, the pH is adjusted to 6.0-7.0, filtered to remove impurities, sterilized, cooled and aseptically filled to obtain a brain peptide oral liquid that nourishes the kidneys, strengthens the spleen and improves intelligence.
[0022] Preferably, the sterilization process is carried out at a temperature of 105-110°C for 15-20 minutes.
[0023] In summary, this application has the following beneficial effects:
[0024] In the preparation of peptide components, this application significantly improves the content, targeting, and synergistic efficiency of active peptides by precisely compounding a multi-protease system and scientifically defining the sequence of enzyme and microbial fermentation. This provides a core material guarantee for the kidney-tonifying, spleen-strengthening, and intelligence-enhancing effects of the final oral liquid, while optimizing the stability and controllability of the production process. Regarding enzyme compounding, the preparation of sheep brain peptides utilizes a ternary compound protease system of subtilisin, elastase, and aminopeptidase, achieving complementary enzymatic hydrolysis functions: subtilisin, as a broad-spectrum endopeptidase, lacks strict amino acid specificity and can efficiently cleave peptide bonds formed by various amino acid residues on the sheep brain protein backbone, laying the foundation for subsequent hydrolysis; elastase specifically acts on the carboxyl-side peptide bonds of aliphatic amino acid residues such as alanine, glycine, and valine, precisely targeting and hydrolyzing specific structural domains of large protein molecules; aminopeptidase progressively hydrolyzes small peptide molecules from the amino terminus, generating easily absorbed oligopeptides and free amino acids. The three components work synergistically to form a complete enzymatic hydrolysis chain of "broad-spectrum hydrolysis - specific targeting - terminal modification," resulting in more thorough hydrolysis of sheep brain protein, significantly increased yield of active peptides, and rich in active fragments such as nerve growth factor-promoting peptides and brain function-regulating peptides. This effectively solves the problems of insufficient hydrolysis by single or binary enzyme systems, limited variety of active peptides, and low absorption efficiency. In the preparation of oyster peptides, Bacillus natto fermentation is used first, followed by enzymatic hydrolysis using neutral protease. This sequence allows Bacillus natto to first disrupt the complex spatial structure of oyster meat protein through metabolism, producing small molecule peptide precursors and bioactive substances. Then, neutral protease specifically hydrolyzes these precursors, increasing the content of active peptides and making the peptides more easily absorbed by the human body. At the same time, soybean and walnut peptides strictly follow the enzymatic hydrolysis sequence of alkaline protease first, followed by flavor protease. Alkaline protease first breaks the main chain of large protein molecules, and flavor protease then hydrolyzes the side chains to generate small molecule active peptides, avoiding the defects of low peptide purity and strong off-flavor caused by reversing the sequence. The above-mentioned enzyme system compounding and enzyme-bacterial sequence optimization enable the active ingredients of sheep brain peptide, oyster peptide, and soybean-walnut compound peptide to achieve synergistic effects. This not only improves the overall quality of peptide components, but also complements the traditional Chinese medicine extract, DHA algal oil and other components, ultimately giving the oral liquid significant kidney-tonifying and spleen-strengthening effects. Moreover, the process parameters are controllable and highly repeatable, making it suitable for large-scale industrial production. Detailed Implementation
[0025] The present application will be further described in detail below with reference to the embodiments.
[0026] The DHA algal oil used in the examples and comparative examples of this application was purchased from Hebei Runbu Biotechnology Co., Ltd.; N-acetylneuraminic acid was purchased from Shandong Siyang Biotechnology Co., Ltd.; γ-aminobutyric acid, vitamin B12 and steviol glycosides were all purchased from Chengdu Wanxiang Hongrun Biotechnology Co., Ltd.; concentrated apple juice was purchased from Zhengzhou Daigongbang Network Technology Co., Ltd.; Bacillus subtilis protease (enzyme activity: 100,000 u / g) was purchased from Jiangxi Huihai Biotechnology Co., Ltd.; elastase was purchased from Hefei Nuoerda Biotechnology Co., Ltd.; aminopeptidase was purchased from Shandong Xinxiong Biotechnology Co., Ltd.; Bacillus natto (product number: B90050) was purchased from Ningbo Mingzhou Biotechnology Co., Ltd.; neutral protease (enzyme activity: 100,000 u / g), alkaline protease (enzyme activity: 100,000 u / g) and flavor protease (enzyme activity: 200,000 u / g) were all purchased from Hebei Yulongxiang Biotechnology Co., Ltd.
[0027] Examples 1-3 provide a brain peptide oral liquid for tonifying the kidney, strengthening the spleen, and improving intelligence, and its preparation method.
[0028] Example 1
[0029] A brain peptide oral liquid for tonifying the kidneys, strengthening the spleen, and improving intelligence contains the following raw material components in parts by weight per 100 servings: 3 parts longan pulp, 3 parts wolfberry, 3 parts mulberry, 3 parts red dates, 2 parts poria cocos, 0.5 parts alpinia oxyphylla, 0.05 parts dried ginger, 8 parts peptide components, 0.01 parts DHA algal oil, 0.05 parts N-acetylneuraminic acid, 1 part γ-aminobutyric acid, and 0.002 parts vitamin B. 12 0.05 parts steviol glycosides, 12 parts concentrated apple juice, and the remainder is purified water.
[0030] The preparation method of peptide components includes the following preparation steps:
[0031] Step 1: After homogenizing the sheep brain, add 5 times the volume of deionized water. Adjust the pH to 6.5 with 0.1 mol / L citrate buffer (pH 4.5). Add the complex protease and enzymatically hydrolyze at 50℃ for 2.5 h. Then incubate at 90℃ for 10 min to inactivate the enzyme. Centrifuge at 4000 rpm for 15 min and collect the supernatant to obtain sheep brain peptides. The complex protease is composed of subtilisin, elastase and aminopeptidase in a mass ratio of 10:5:2. The mass ratio of the complex protease to sheep brain is 1:200.
[0032] Step 2: Crush the oyster meat and add 3 times the volume of deionized water to obtain the fermentation substrate. Inoculate the fermentation substrate with Bacillus natto, controlling the inoculation amount of Bacillus natto to be 2% of the mass of the fermentation substrate. Ferment at 37℃ for 18 hours to obtain the fermentation broth. Add neutral protease to the fermentation broth, controlling the mass ratio of fermentation broth to neutral protease to be 100:3. Enzymatically hydrolyze at 40℃ for 2 hours, incubate at 95℃ for 15 minutes to inactivate the enzyme, centrifuge at 4000 rpm for 15 minutes and collect the supernatant to obtain oyster peptides.
[0033] Step 3: Mix walnuts and soybeans at a mass ratio of 3:2, grind them through a 100-mesh sieve, add 8 times the volume of deionized water, stir at 200 rpm until well mixed, adjust the pH to 8.0 with 0.05 mol / L sodium bicarbonate-sodium carbonate buffer (pH 9.0) to obtain soybean-walnut extract; first add alkaline protease to the soybean-walnut extract, and enzymatically hydrolyze at 50℃ for 2 hours, then add flavor protease, and enzymatically hydrolyze at 45℃ for 1 hour, then inactivate the enzyme at 100℃ for 10 minutes, centrifuge at 4000 rpm for 15 minutes and collect the supernatant to obtain soybean-walnut complex peptides;
[0034] Step 4: Mix sheep brain peptide, oyster peptide and soybean-walnut compound peptide in a mass ratio of 3:2:1, concentrate under vacuum at 0.08 MPa and 50°C until the solid content is 20%, and then freeze-dry at -40°C for 10 hours to obtain peptide components.
[0035] A method for preparing a brain peptide oral liquid that tonifies the kidneys, strengthens the spleen, and improves intelligence includes the following preparation steps:
[0036] Mix longan pulp, goji berries, mulberries, red dates, poria cocos, alpinia oxyphylla, and dried ginger. Add 12 times the volume of purified water to the longan pulp and decoct twice, 1 hour each time. Combine the decoctions, filter through a 100-mesh filter bag, and concentrate to 1 / 4 of the original volume to obtain a traditional Chinese medicine extract. Add peptide components and DHA algal oil to the extract, stir well, and then add N-acetylneuraminic acid, γ-aminobutyric acid, and vitamin B. 12 Stevioside and concentrated apple juice were stirred at 200 rpm until homogeneous. Then, purified water was added to bring the total volume up to 6.0. The pH was adjusted to 6.0 using 0.1 mol / L citrate buffer solution (pH 4.5). The mixture was filtered through a 100-mesh filter bag to remove impurities. It was then sterilized at 105°C for 15 minutes, cooled, and aseptically filled to obtain a brain peptide oral liquid that tonifies the kidneys, strengthens the spleen, and improves intelligence.
[0037] Example 2
[0038] A brain peptide oral liquid for tonifying the kidneys, strengthening the spleen, and improving intelligence contains the following raw material components in parts by weight per 100 servings: 4 parts longan pulp, 5 parts wolfberry, 5 parts mulberry, 5 parts red dates, 4 parts poria cocos, 1 part alpinia oxyphylla, 0.08 parts dried ginger, 10 parts peptide components, 0.03 parts DHA algal oil, 0.08 parts N-acetylneuraminic acid, 2 parts γ-aminobutyric acid, and 0.003 parts vitamin B. 12 0.08 parts steviol glycosides, 14 parts concentrated apple juice, and the remainder is purified water.
[0039] The preparation method of peptide components includes the following preparation steps:
[0040] Step 1: After homogenizing the sheep brain, add 8 times the volume of deionized water. Adjust the pH to 6.8 with 0.1 mol / L citrate buffer (pH 4.5). Add the complex protease and enzymatically hydrolyze at 52℃ for 2.8 h. Then incubate at 92℃ for 12 min to inactivate the enzyme. Centrifuge at 4500 rpm for 18 min and collect the supernatant to obtain sheep brain peptides. The complex protease is composed of subtilisin, elastase and aminopeptidase in a mass ratio of 11:7:4. The mass ratio of the complex protease to sheep brain is 2:250.
[0041] Step 2: After crushing the oyster meat, add 4 times the volume of deionized water to obtain the fermentation substrate. Inoculate the fermentation substrate with Bacillus natto, controlling the inoculation amount of Bacillus natto to be 4% of the mass of the fermentation substrate. Ferment at 38℃ for 21 hours to obtain the fermentation broth. Add neutral protease to the fermentation broth, controlling the mass ratio of fermentation broth to neutral protease to be 150:4. Enzymatically hydrolyze at 45℃ for 3.5 hours, incubate at 98℃ for 18 minutes to inactivate the enzyme, centrifuge at 4500 rpm for 18 minutes, and collect the supernatant to obtain oyster peptides.
[0042] Step 3: Mix walnuts and soybeans at a mass ratio of 4:3, grind them through a 150-mesh sieve, add 10 times the volume of deionized water, mix evenly at a stirring speed of 300 rpm, and adjust the pH to 8.2 with a 0.05 mol / L sodium bicarbonate-sodium carbonate buffer solution (pH 9.0) to obtain soybean-walnut extract; first add alkaline protease to the soybean-walnut extract and hydrolyze it at 55℃ for 2.5 h, then add flavor protease and hydrolyze it at 50℃ for 2 h, followed by inactivation at 105℃ for 12 min, centrifuge at 4500 rpm for 18 min, and collect the supernatant to obtain soybean-walnut complex peptides;
[0043] Step 4: Mix sheep brain peptide, oyster peptide and soybean-walnut compound peptide in a mass ratio of 4.5:3.5:1.5, concentrate under vacuum at 0.085 MPa and 52°C until the solid content is 22%, and then freeze-dry at -30°C for 11 hours to obtain peptide components.
[0044] A method for preparing a brain peptide oral liquid that tonifies the kidneys, strengthens the spleen, and improves intelligence includes the following preparation steps:
[0045] Mix longan pulp, goji berries, mulberries, red dates, poria cocos, alpinia oxyphylla, and dried ginger. Add 15 times the volume of purified water to the longan pulp and decoct three times, 1.5 hours each time. Combine the decoctions, filter through a 150-mesh filter bag, and concentrate to 1 / 3 of the original volume to obtain a traditional Chinese medicine extract. Add peptide components and DHA algal oil to the extract, stir well, and then add N-acetylneuraminic acid, γ-aminobutyric acid, and vitamin B. 12 Stevioside and concentrated apple juice were stirred at 300 rpm until homogeneous. Then, purified water was added to bring the total volume up to 6.5. The pH was adjusted to 6.5 using 0.1 mol / L citrate buffer solution (pH 4.5). The mixture was filtered through a 110-mesh filter bag to remove impurities. It was then sterilized at 108°C for 18 minutes, cooled, and aseptically filled to obtain a brain peptide oral liquid that tonifies the kidneys, strengthens the spleen, and improves intelligence.
[0046] Example 3
[0047] A brain peptide oral liquid for tonifying the kidneys, strengthening the spleen, and improving intelligence contains the following raw material components in parts by weight per 100 servings: 5 parts longan pulp, 6 parts wolfberry, 8 parts mulberry, 6 parts red dates, 6 parts poria cocos, 2 parts alpinia oxyphylla, 0.1 parts dried ginger, 12 parts peptide components, 0.05 parts DHA algal oil, 0.1 parts N-acetylneuraminic acid, 3 parts γ-aminobutyric acid, and 0.004 parts vitamin B. 12 0.1 parts steviol glycosides, 15 parts concentrated apple juice, and the remainder is purified water.
[0048] The preparation method of peptide components includes the following preparation steps:
[0049] Step 1: After homogenizing the sheep brain, add 10 times the volume of deionized water to the sheep brain homogenate. Adjust the pH to 7.0 with 0.1 mol / L citrate buffer (pH 4.5). Add the complex protease and hydrolyze at 55℃ for 3 hours. Then incubate at 95℃ for 15 minutes to inactivate the enzyme. Centrifuge at 5000 rpm for 20 minutes and collect the supernatant to obtain sheep brain peptides. The complex protease is composed of subtilisin, elastase and aminopeptidase in a mass ratio of 12:8:5. The mass ratio of the complex protease to sheep brain is 3:300.
[0050] Step 2: Crush the oyster meat and add 5 times the volume of deionized water to obtain the fermentation substrate. Inoculate the fermentation substrate with Bacillus natto, controlling the inoculation amount of Bacillus natto to be 5% of the mass of the fermentation substrate. Ferment at 40℃ for 24 hours to obtain the fermentation broth. Add neutral protease to the fermentation broth, controlling the mass ratio of fermentation broth to neutral protease to be 200:5. Enzymatically hydrolyze at 50℃ for 5 hours, incubate at 100℃ for 20 minutes to inactivate the enzyme, centrifuge at 5000 rpm for 20 minutes and collect the supernatant to obtain oyster peptides.
[0051] Step 3: Mix walnuts and soybeans at a mass ratio of 5:4, grind them through a 200-mesh sieve, add 12 times the volume of deionized water, mix evenly at a stirring speed of 400 rpm, and adjust the pH to 8.5 with a 0.05 mol / L sodium bicarbonate-sodium carbonate buffer solution (pH 9.0) to obtain soybean-walnut extract; first add alkaline protease to the soybean-walnut extract and hydrolyze it at 60℃ for 3 hours, then add flavor protease and hydrolyze it at 55℃ for 3 hours, then inactivate the enzyme at 110℃ for 15 minutes, centrifuge at 5000 rpm for 20 minutes and collect the supernatant to obtain soybean-walnut complex peptides;
[0052] Step 4: Mix sheep brain peptide, oyster peptide and soybean-walnut compound peptide in a mass ratio of 6:5:2, concentrate under vacuum at 0.09 MPa and 55°C until the solid content is 25%, and then freeze-dry at -20°C for 12 hours to obtain peptide components.
[0053] A method for preparing a brain peptide oral liquid that tonifies the kidneys, strengthens the spleen, and improves intelligence includes the following preparation steps:
[0054] Mix longan pulp, goji berries, mulberries, red dates, poria cocos, alpinia oxyphylla, and dried ginger. Add 20 times the volume of purified water to the longan pulp and decoct three times, 2 hours each time. Combine the decoctions, filter through a 200-mesh filter bag, and concentrate to 1 / 3 of the original volume to obtain a traditional Chinese medicine extract. Add peptide components and DHA algal oil to the extract, stir well, and then add N-acetylneuraminic acid, γ-aminobutyric acid, and vitamin B. 12 Stevioside and concentrated apple juice were stirred at 400 rpm until homogeneous. Then, purified water was added to bring the total volume up to 7.0. The pH was adjusted to 7.0 using 0.1 mol / L citrate buffer solution with a pH of 4.5. The mixture was filtered through a 120-mesh filter bag to remove impurities. It was then sterilized at 110°C for 20 minutes, cooled, and aseptically filled to obtain a brain peptide oral liquid that tonifies the kidneys, strengthens the spleen, and improves intelligence.
[0055] Comparative Example 1
[0056] A brain peptide oral liquid for tonifying the kidneys, strengthening the spleen, and improving intelligence contains the following raw material components in parts by weight per 100 servings: 3 parts longan pulp, 3 parts wolfberry, 3 parts mulberry, 3 parts red dates, 2 parts poria cocos, 0.5 parts alpinia oxyphylla, 0.05 parts dried ginger, 8 parts peptide components, 0.01 parts DHA algal oil, 0.05 parts N-acetylneuraminic acid, 1 part γ-aminobutyric acid, and 0.002 parts vitamin B. 12 0.05 parts steviol glycosides, 12 parts concentrated apple juice, and the remainder is purified water.
[0057] The preparation method of peptide components includes the following preparation steps:
[0058] Step 1: After homogenizing the sheep brain, add 5 times the volume of deionized water to the sheep brain homogenate. Adjust the pH to 6.5 with 0.1 mol / L citrate buffer (pH 4.5). Add the complex protease and enzymatically hydrolyze at 50℃ for 2.5 h. Then incubate at 90℃ for 10 min to inactivate the enzyme. Centrifuge at 4000 rpm for 15 min and collect the supernatant to obtain sheep brain peptides. The complex protease is composed of Bacillus subtilis protease and aminopeptidase in a mass ratio of 10:2. The mass ratio of the complex protease to sheep brain is 1:200.
[0059] Step 2: Crush the oyster meat and add 3 times the volume of deionized water to obtain the fermentation substrate. Inoculate the fermentation substrate with Bacillus natto, controlling the inoculation amount of Bacillus natto to be 2% of the mass of the fermentation substrate. Ferment at 37℃ for 18 hours to obtain the fermentation broth. Add neutral protease to the fermentation broth, controlling the mass ratio of fermentation broth to neutral protease to be 100:3. Enzymatically hydrolyze at 40℃ for 2 hours, incubate at 95℃ for 15 minutes to inactivate the enzyme, centrifuge at 4000 rpm for 15 minutes and collect the supernatant to obtain oyster peptides.
[0060] Step 3: Mix walnuts and soybeans at a mass ratio of 3:2, grind them through a 100-mesh sieve, add 8 times the volume of deionized water, stir at 200 rpm until well mixed, adjust the pH to 8.0 with 0.05 mol / L sodium bicarbonate-sodium carbonate buffer (pH 9.0) to obtain soybean-walnut extract; first add alkaline protease to the soybean-walnut extract, and enzymatically hydrolyze at 50℃ for 2 hours, then add flavor protease, and enzymatically hydrolyze at 45℃ for 1 hour, then inactivate the enzyme at 100℃ for 10 minutes, centrifuge at 4000 rpm for 15 minutes and collect the supernatant to obtain soybean-walnut complex peptides;
[0061] Step 4: Mix sheep brain peptide, oyster peptide and soybean-walnut compound peptide in a mass ratio of 3:2:1, concentrate under vacuum at 0.08 MPa and 50°C until the solid content is 20%, and then freeze-dry at -40°C for 10 hours to obtain peptide components.
[0062] A method for preparing a brain peptide oral liquid that tonifies the kidneys, strengthens the spleen, and improves intelligence includes the following preparation steps:
[0063] Mix longan pulp, goji berries, mulberries, red dates, poria cocos, alpinia oxyphylla, and dried ginger. Add 12 times the volume of purified water to the longan pulp and decoct twice, 1 hour each time. Combine the decoctions, filter through a 100-mesh filter bag, and concentrate to 1 / 4 of the original volume to obtain a traditional Chinese medicine extract. Add peptide components and DHA algal oil to the extract, stir well, and then add N-acetylneuraminic acid, γ-aminobutyric acid, and vitamin B. 12Stevioside and concentrated apple juice were stirred at 200 rpm until homogeneous. Then, purified water was added to bring the total volume up to 6.0. The pH was adjusted to 6.0 using 0.1 mol / L citrate buffer solution (pH 4.5). The mixture was filtered through a 100-mesh filter bag to remove impurities. It was then sterilized at 105°C for 15 minutes, cooled, and aseptically filled to obtain a brain peptide oral liquid that tonifies the kidneys, strengthens the spleen, and improves intelligence.
[0064] Comparative Example 2
[0065] A brain peptide oral liquid for tonifying the kidneys, strengthening the spleen, and improving intelligence contains the following raw material components in parts by weight per 100 servings: 3 parts longan pulp, 3 parts wolfberry, 3 parts mulberry, 3 parts red dates, 2 parts poria cocos, 0.5 parts alpinia oxyphylla, 0.05 parts dried ginger, 8 parts peptide components, 0.01 parts DHA algal oil, 0.05 parts N-acetylneuraminic acid, 1 part γ-aminobutyric acid, and 0.002 parts vitamin B. 12 0.05 parts steviol glycosides, 12 parts concentrated apple juice, and the remainder is purified water.
[0066] The preparation method of peptide components includes the following preparation steps:
[0067] Step 1: After homogenizing the sheep brain, add 5 times the volume of deionized water to the sheep brain homogenate. Adjust the pH to 6.5 with 0.1 mol / L citrate buffer (pH 4.5). Add the complex protease and enzymatically hydrolyze at 50℃ for 2.5 h. Then incubate at 90℃ for 10 min to inactivate the enzyme. Centrifuge at 4000 rpm for 15 min and collect the supernatant to obtain sheep brain peptides. The complex protease is composed of Bacillus subtilis protease and elastase in a mass ratio of 10:5, and the mass ratio of the complex protease to sheep brain is 1:200.
[0068] Step 2: Crush the oyster meat and add 3 times the volume of deionized water to obtain the fermentation substrate. Inoculate the fermentation substrate with Bacillus natto, controlling the inoculation amount of Bacillus natto to be 2% of the mass of the fermentation substrate. Ferment at 37℃ for 18 hours to obtain the fermentation broth. Add neutral protease to the fermentation broth, controlling the mass ratio of fermentation broth to neutral protease to be 100:3. Enzymatically hydrolyze at 40℃ for 2 hours, incubate at 95℃ for 15 minutes to inactivate the enzyme, centrifuge at 4000 rpm for 15 minutes and collect the supernatant to obtain oyster peptides.
[0069] Step 3: Mix walnuts and soybeans at a mass ratio of 3:2, grind them through a 100-mesh sieve, add 8 times the volume of deionized water, stir at 200 rpm until well mixed, adjust the pH to 8.0 with 0.05 mol / L sodium bicarbonate-sodium carbonate buffer (pH 9.0) to obtain soybean-walnut extract; first add alkaline protease to the soybean-walnut extract, and enzymatically hydrolyze at 50℃ for 2 hours, then add flavor protease, and enzymatically hydrolyze at 45℃ for 1 hour, then inactivate the enzyme at 100℃ for 10 minutes, centrifuge at 4000 rpm for 15 minutes and collect the supernatant to obtain soybean-walnut complex peptides;
[0070] Step 4: Mix sheep brain peptide, oyster peptide and soybean-walnut compound peptide in a mass ratio of 3:2:1, concentrate under vacuum at 0.08 MPa and 50°C until the solid content is 20%, and then freeze-dry at -40°C for 10 hours to obtain peptide components.
[0071] A method for preparing a brain peptide oral liquid that tonifies the kidneys, strengthens the spleen, and improves intelligence includes the following preparation steps:
[0072] Mix longan pulp, goji berries, mulberries, red dates, poria cocos, alpinia oxyphylla, and dried ginger. Add 12 times the volume of purified water to the longan pulp and decoct twice, 1 hour each time. Combine the decoctions, filter through a 100-mesh filter bag, and concentrate to 1 / 4 of the original volume to obtain a traditional Chinese medicine extract. Add peptide components and DHA algal oil to the extract, stir well, and then add N-acetylneuraminic acid, γ-aminobutyric acid, and vitamin B. 12 Stevioside and concentrated apple juice were stirred at 200 rpm until homogeneous. Then, purified water was added to bring the total volume up to 6.0. The pH was adjusted to 6.0 using 0.1 mol / L citrate buffer solution (pH 4.5). The mixture was filtered through a 100-mesh filter bag to remove impurities. It was then sterilized at 105°C for 15 minutes, cooled, and aseptically filled to obtain a brain peptide oral liquid that tonifies the kidneys, strengthens the spleen, and improves intelligence.
[0073] Comparative Example 3
[0074] A brain peptide oral liquid for tonifying the kidneys, strengthening the spleen, and improving intelligence contains the following raw material components in parts by weight per 100 servings: 3 parts longan pulp, 3 parts wolfberry, 3 parts mulberry, 3 parts red dates, 2 parts poria cocos, 0.5 parts alpinia oxyphylla, 0.05 parts dried ginger, 8 parts peptide components, 0.01 parts DHA algal oil, 0.05 parts N-acetylneuraminic acid, 1 part γ-aminobutyric acid, and 0.002 parts vitamin B. 12 0.05 parts steviol glycosides, 12 parts concentrated apple juice, and the remainder is purified water.
[0075] The preparation method of peptide components includes the following preparation steps:
[0076] Step 1: After homogenizing the sheep brain, add 5 times the volume of deionized water. Adjust the pH to 6.5 with 0.1 mol / L citrate buffer (pH 4.5). Add the complex protease and enzymatically hydrolyze at 50℃ for 2.5 h. Then incubate at 90℃ for 10 min to inactivate the enzyme. Centrifuge at 4000 rpm for 15 min and collect the supernatant to obtain sheep brain peptides. The complex protease is composed of subtilisin, elastase and aminopeptidase in a mass ratio of 10:5:2. The mass ratio of the complex protease to sheep brain is 1:200.
[0077] Step 2: After crushing the oyster meat, add 3 times the volume of deionized water to obtain a matrix. Add neutral protease to the matrix and enzymatically hydrolyze at 40℃ for 2 hours, controlling the mass ratio of matrix to neutral protease to be 100:3 to obtain the enzymatic hydrolysate. Inoculate the enzymatic hydrolysate with Bacillus natto, controlling the inoculation amount of Bacillus natto to be 2% of the mass of the enzymatic hydrolysate. Ferment at 37℃ for 18 hours, then sterilize by incubating at 95℃ for 15 minutes. Centrifuge at 4000 rpm for 15 minutes and collect the supernatant to obtain oyster peptides.
[0078] Step 3: Mix walnuts and soybeans at a mass ratio of 3:2, grind them through a 100-mesh sieve, add 8 times the volume of deionized water, stir at 200 rpm until well mixed, adjust the pH to 8.0 with 0.05 mol / L sodium bicarbonate-sodium carbonate buffer (pH 9.0) to obtain soybean-walnut extract; first add alkaline protease to the soybean-walnut extract, and enzymatically hydrolyze at 50℃ for 2 hours, then add flavor protease, and enzymatically hydrolyze at 45℃ for 1 hour, then inactivate the enzyme at 100℃ for 10 minutes, centrifuge at 4000 rpm for 15 minutes and collect the supernatant to obtain soybean-walnut complex peptides;
[0079] Step 4: Mix sheep brain peptide, oyster peptide and soybean-walnut compound peptide in a mass ratio of 3:2:1, concentrate under vacuum at 0.08 MPa and 50°C until the solid content is 20%, and then freeze-dry at -40°C for 10 hours to obtain peptide components.
[0080] A method for preparing a brain peptide oral liquid that tonifies the kidneys, strengthens the spleen, and improves intelligence includes the following preparation steps:
[0081] Mix longan pulp, goji berries, mulberries, red dates, poria cocos, alpinia oxyphylla, and dried ginger. Add 12 times the volume of purified water to the longan pulp and decoct twice, 1 hour each time. Combine the decoctions, filter through a 100-mesh filter bag, and concentrate to 1 / 4 of the original volume to obtain a traditional Chinese medicine extract. Add peptide components and DHA algal oil to the extract, stir well, and then add N-acetylneuraminic acid, γ-aminobutyric acid, and vitamin B. 12Stevioside and concentrated apple juice were stirred at 200 rpm until homogeneous. Then, purified water was added to bring the total volume up to 6.0. The pH was adjusted to 6.0 using 0.1 mol / L citrate buffer solution (pH 4.5). The mixture was filtered through a 100-mesh filter bag to remove impurities. It was then sterilized at 105°C for 15 minutes, cooled, and aseptically filled to obtain a brain peptide oral liquid that tonifies the kidneys, strengthens the spleen, and improves intelligence.
[0082] Comparative Example 4
[0083] A brain peptide oral liquid for tonifying the kidneys, strengthening the spleen, and improving intelligence contains the following raw material components in parts by weight per 100 servings: 3 parts longan pulp, 3 parts wolfberry, 3 parts mulberry, 3 parts red dates, 2 parts poria cocos, 0.5 parts alpinia oxyphylla, 0.05 parts dried ginger, 8 parts peptide components, 0.01 parts DHA algal oil, 0.05 parts N-acetylneuraminic acid, 1 part γ-aminobutyric acid, and 0.002 parts vitamin B. 12 0.05 parts steviol glycosides, 12 parts concentrated apple juice, and the remainder is purified water.
[0084] The preparation method of peptide components includes the following preparation steps:
[0085] Step 1: After homogenizing the sheep brain, add 5 times the volume of deionized water. Adjust the pH to 6.5 with 0.1 mol / L citrate buffer (pH 4.5). Add the complex protease and enzymatically hydrolyze at 50℃ for 2.5 h. Then incubate at 90℃ for 10 min to inactivate the enzyme. Centrifuge at 4000 rpm for 15 min and collect the supernatant to obtain sheep brain peptides. The complex protease is composed of subtilisin, elastase and aminopeptidase in a mass ratio of 10:5:2. The mass ratio of the complex protease to sheep brain is 1:200.
[0086] Step 2: After crushing the oyster meat, add 3 times the volume of deionized water to obtain the fermentation substrate. Inoculate the fermentation substrate with Bacillus natto, controlling the inoculation amount of Bacillus natto to be 2% of the mass of the fermentation substrate. Ferment at 37℃ for 18 hours to obtain the fermentation broth. Sterilize by incubating at 95℃ for 15 minutes, centrifuge at 4000rpm for 15 minutes, and collect the supernatant to obtain oyster peptides.
[0087] Step 3: Mix walnuts and soybeans at a mass ratio of 3:2, grind them through a 100-mesh sieve, add 8 times the volume of deionized water, stir at 200 rpm until well mixed, adjust the pH to 8.0 with 0.05 mol / L sodium bicarbonate-sodium carbonate buffer (pH 9.0) to obtain soybean-walnut extract; first add alkaline protease to the soybean-walnut extract, and enzymatically hydrolyze at 50℃ for 2 hours, then add flavor protease, and enzymatically hydrolyze at 45℃ for 1 hour, then inactivate the enzyme at 100℃ for 10 minutes, centrifuge at 4000 rpm for 15 minutes and collect the supernatant to obtain soybean-walnut complex peptides;
[0088] Step 4: Mix sheep brain peptide, oyster peptide and soybean-walnut compound peptide in a mass ratio of 3:2:1, concentrate under vacuum at 0.08 MPa and 50°C until the solid content is 20%, and then freeze-dry at -40°C for 10 hours to obtain peptide components.
[0089] A method for preparing a brain peptide oral liquid that tonifies the kidneys, strengthens the spleen, and improves intelligence includes the following preparation steps:
[0090] Mix longan pulp, goji berries, mulberries, red dates, poria cocos, alpinia oxyphylla, and dried ginger. Add 12 times the volume of purified water to the longan pulp and decoct twice, 1 hour each time. Combine the decoctions, filter through a 100-mesh filter bag, and concentrate to 1 / 4 of the original volume to obtain a traditional Chinese medicine extract. Add peptide components and DHA algal oil to the extract, stir well, and then add N-acetylneuraminic acid, γ-aminobutyric acid, and vitamin B. 12 Stevioside and concentrated apple juice were stirred at 200 rpm until homogeneous. Then, purified water was added to bring the total volume up to 6.0. The pH was adjusted to 6.0 using 0.1 mol / L citrate buffer solution (pH 4.5). The mixture was filtered through a 100-mesh filter bag to remove impurities. It was then sterilized at 105°C for 15 minutes, cooled, and aseptically filled to obtain a brain peptide oral liquid that tonifies the kidneys, strengthens the spleen, and improves intelligence.
[0091] Comparative Example 5
[0092] A brain peptide oral liquid for tonifying the kidneys, strengthening the spleen, and improving intelligence contains the following raw material components in parts by weight per 100 servings: 3 parts longan pulp, 3 parts wolfberry, 3 parts mulberry, 3 parts red dates, 2 parts poria cocos, 0.5 parts alpinia oxyphylla, 0.05 parts dried ginger, 8 parts peptide components, 0.01 parts DHA algal oil, 0.05 parts N-acetylneuraminic acid, 1 part γ-aminobutyric acid, and 0.002 parts vitamin B. 12 0.05 parts steviol glycosides, 12 parts concentrated apple juice, and the remainder is purified water.
[0093] The preparation method of peptide components includes the following preparation steps:
[0094] Step 1: After homogenizing the sheep brain, add 5 times the volume of deionized water. Adjust the pH to 6.5 with 0.1 mol / L citrate buffer (pH 4.5). Add the complex protease and enzymatically hydrolyze at 50℃ for 2.5 h. Then incubate at 90℃ for 10 min to inactivate the enzyme. Centrifuge at 4000 rpm for 15 min and collect the supernatant to obtain sheep brain peptides. The complex protease is composed of subtilisin, elastase and aminopeptidase in a mass ratio of 10:5:2. The mass ratio of the complex protease to sheep brain is 1:200.
[0095] Step 2: After crushing the oyster meat, add 3 times the volume of deionized water to obtain a matrix. Add neutral protease to the matrix, and control the mass ratio of matrix to neutral protease to 100:3. Enzymatically hydrolyze at 40℃ for 20h, incubate at 95℃ for 15min to inactivate the enzyme, centrifuge at 4000rpm for 15min and collect the supernatant to obtain oyster peptides.
[0096] Step 3: Mix walnuts and soybeans at a mass ratio of 3:2, grind them through a 100-mesh sieve, add 8 times the volume of deionized water, stir at 200 rpm until well mixed, adjust the pH to 8.0 with 0.05 mol / L sodium bicarbonate-sodium carbonate buffer (pH 9.0) to obtain soybean-walnut extract; first add alkaline protease to the soybean-walnut extract, and enzymatically hydrolyze at 50℃ for 2 hours, then add flavor protease, and enzymatically hydrolyze at 45℃ for 1 hour, then inactivate the enzyme at 100℃ for 10 minutes, centrifuge at 4000 rpm for 15 minutes and collect the supernatant to obtain soybean-walnut complex peptides;
[0097] Step 4: Mix sheep brain peptide, oyster peptide and soybean-walnut compound peptide in a mass ratio of 3:2:1, concentrate under vacuum at 0.08 MPa and 50°C until the solid content is 20%, and then freeze-dry at -40°C for 10 hours to obtain peptide components.
[0098] A method for preparing a brain peptide oral liquid that tonifies the kidneys, strengthens the spleen, and improves intelligence includes the following preparation steps:
[0099] Mix longan pulp, goji berries, mulberries, red dates, poria cocos, alpinia oxyphylla, and dried ginger. Add 12 times the volume of purified water to the longan pulp and decoct twice, 1 hour each time. Combine the decoctions, filter through a 100-mesh filter bag, and concentrate to 1 / 4 of the original volume to obtain a traditional Chinese medicine extract. Add peptide components and DHA algal oil to the extract, stir well, and then add N-acetylneuraminic acid, γ-aminobutyric acid, and vitamin B. 12 Stevioside and concentrated apple juice were stirred at 200 rpm until homogeneous. Then, purified water was added to bring the total volume up to 6.0. The pH was adjusted to 6.0 using 0.1 mol / L citrate buffer solution (pH 4.5). The mixture was filtered through a 100-mesh filter bag to remove impurities. It was then sterilized at 105°C for 15 minutes, cooled, and aseptically filled to obtain a brain peptide oral liquid that tonifies the kidneys, strengthens the spleen, and improves intelligence.
[0100] Comparative Example 6
[0101] A brain peptide oral liquid for tonifying the kidneys, strengthening the spleen, and improving intelligence contains the following raw material components in parts by weight per 100 servings: 3 parts longan pulp, 3 parts wolfberry, 3 parts mulberry, 3 parts red dates, 2 parts poria cocos, 0.5 parts alpinia oxyphylla, 0.05 parts dried ginger, 8 parts peptide components, 0.01 parts DHA algal oil, 0.05 parts N-acetylneuraminic acid, 1 part γ-aminobutyric acid, and 0.002 parts vitamin B. 120.05 parts steviol glycosides, 12 parts concentrated apple juice, and the remainder is purified water.
[0102] The preparation method of peptide components includes the following preparation steps:
[0103] Step 1: After homogenizing the sheep brain, add 5 times the volume of deionized water. Adjust the pH to 6.5 with 0.1 mol / L citrate buffer (pH 4.5). Add the complex protease and enzymatically hydrolyze at 50℃ for 2.5 h. Then incubate at 90℃ for 10 min to inactivate the enzyme. Centrifuge at 4000 rpm for 15 min and collect the supernatant to obtain sheep brain peptides. The complex protease is composed of subtilisin, elastase and aminopeptidase in a mass ratio of 10:5:2. The mass ratio of the complex protease to sheep brain is 1:200.
[0104] Step 2: Crush the oyster meat and add 3 times the volume of deionized water to obtain the fermentation substrate. Inoculate the fermentation substrate with Bacillus natto, controlling the inoculation amount of Bacillus natto to be 2% of the mass of the fermentation substrate. Ferment at 37℃ for 18 hours to obtain the fermentation broth. Add neutral protease to the fermentation broth, controlling the mass ratio of fermentation broth to neutral protease to be 100:3. Enzymatically hydrolyze at 40℃ for 2 hours, incubate at 95℃ for 15 minutes to inactivate the enzyme, centrifuge at 4000 rpm for 15 minutes and collect the supernatant to obtain oyster peptides.
[0105] Step 3: Add 8 times the volume of deionized water to the walnuts, mix thoroughly at 200 rpm, and adjust the pH to 8.0 with 0.05 mol / L sodium bicarbonate-sodium carbonate buffer solution (pH 9.0) to obtain soybean-walnut extract. First, add flavor protease to the soybean-walnut extract and hydrolyze for 2 hours at 50℃. Then, add alkaline protease and hydrolyze for 1 hour at 45℃. After that, inactivate the enzyme at 100℃ for 10 minutes, centrifuge at 4000 rpm for 15 minutes, and collect the supernatant to obtain soybean-walnut complex peptides.
[0106] Step 4: Mix sheep brain peptide, oyster peptide and soybean-walnut compound peptide in a mass ratio of 3:2:1, concentrate under vacuum at 0.08 MPa and 50°C until the solid content is 20%, and then freeze-dry at -40°C for 10 hours to obtain peptide components.
[0107] A method for preparing a brain peptide oral liquid that tonifies the kidneys, strengthens the spleen, and improves intelligence includes the following preparation steps:
[0108] Mix longan pulp, goji berries, mulberries, red dates, poria cocos, alpinia oxyphylla, and dried ginger. Add 12 times the volume of purified water to the longan pulp and decoct twice, 1 hour each time. Combine the decoctions, filter through a 100-mesh filter bag, and concentrate to 1 / 4 of the original volume to obtain a traditional Chinese medicine extract. Add peptide components and DHA algal oil to the extract, stir well, and then add N-acetylneuraminic acid, γ-aminobutyric acid, and vitamin B. 12 Stevioside and concentrated apple juice were stirred at 200 rpm until homogeneous. Then, purified water was added to bring the total volume up to 6.0. The pH was adjusted to 6.0 using 0.1 mol / L citrate buffer solution (pH 4.5). The mixture was filtered through a 100-mesh filter bag to remove impurities. It was then sterilized at 105°C for 15 minutes, cooled, and aseptically filled to obtain a brain peptide oral liquid that tonifies the kidneys, strengthens the spleen, and improves intelligence.
[0109] Performance testing
[0110] This application was formulated in accordance with the "Methodology of Pharmacological Research of Traditional Chinese Medicine" and the "Technical Specifications for Inspection and Evaluation of Health Foods". It systematically verified the kidney-tonifying and spleen-strengthening effects of the Kidney-tonifying, Spleen-strengthening, and Intelligence-enhancing Brain Peptide Oral Liquid by establishing animal models of two TCM syndrome types: kidney-yang deficiency and spleen deficiency, combined with macroscopic behavioral, biochemical, and pathological morphological tests. SPF-grade ICR mice were used as experimental animals, and all procedures were approved by the animal ethics committee and followed the 3R principle.
[0111] Model establishment of kidney-yang deficiency and detection of kidney-tonifying indicators:
[0112] Experimental animals: SPF-grade ICR male mice, 6-8 weeks old, weighing 20-22g, were selected. They were kept at a temperature of 22-25℃, relative humidity of 50-60%, and a 12-hour light-dark cycle (8:00-20:00 light). They were allowed free access to standard feed and sterile water. The experiment began after 7 days of acclimatization.
[0113] Modeling method: 110 SPF-grade ICR male mice were randomly divided into 11 groups of 10 mice each, namely blank control group, model control group, Example 1-3 groups and Comparative Example 1-6 groups;
[0114] Modeling procedures: The model control group, Examples 1-3 groups, and Comparative Examples 1-6 groups were intraperitoneally injected with hydrocortisone solution (20 mg / kg, diluted with physiological saline to 0.1 mL / 10 g body weight) daily, while the blank control group was intraperitoneally injected with an equal volume of physiological saline, for 10 consecutive days. From the first day of modeling, Examples 1-3 groups and Comparative Examples 1-6 groups were administered the kidney-tonifying, spleen-strengthening, and intelligence-enhancing brain peptide oral solution prepared in Examples 1-3 and Comparative Examples 1-6 (0.4 mL / 10 g body weight) by gavage, while the blank control group and model group were administered an equal volume of physiological saline by gavage, once daily for 14 consecutive days.
[0115] After successful modeling, mice were fasted for 12 hours, and blood was collected from their orbital sinuses into coagulation tubes. After standing for 30 minutes, the blood was centrifuged at 3000 rpm for 15 minutes to collect serum. The serum levels of testosterone (T), cortisol (CORT), triiodothyronine (T3), and thyroxine (T4) were measured. 0.1 g of kidney tissue was taken, added to physiological saline to make a 10% homogenate, centrifuged at 3000 rpm for 15 minutes, and the supernatant was collected to measure SOD activity (unit: U / mg prot) and MDA content (unit: nmol / mg prot).
[0116] Specific data on kidney-tonifying indicators are shown in Table 1.
[0117] Table 1. Detection results of kidney-tonifying indicators of the brain peptide oral liquids prepared in Examples 1-3 and Comparative Examples 1-6 (x±s, n=10)
[0118] Model establishment of spleen deficiency and detection of spleen-strengthening indicators:
[0119] Experimental animals: SPF-grade ICR male mice, 6-8 weeks old, weighing 20-22g, were selected and fed under the same conditions as the kidney yang deficiency model.
[0120] Modeling method: 110 SPF-grade ICR male mice were randomly divided into 11 groups of 10 mice each, namely blank control group, model control group, Example 1-3 groups and Comparative Example 1-6 groups;
[0121] Preparation of rhubarb decoction: Add 10 times the amount of distilled water to raw rhubarb, soak for 30 minutes, boil for 30 minutes, filter, concentrate to 1 g / mL (crude drug concentration), and store at 4℃ for later use.
[0122] Modeling procedure: The model control group, Examples 1-3 and Comparative Examples 1-6 were administered rhubarb decoction (10g / kg, i.e., 0.1mL / 10g body weight) by gavage daily, while the blank control group was administered an equal volume of physiological saline by gavage, for 7 consecutive days. From the first day of modeling, Examples 1-3 and Comparative Examples 1-6 were administered kidney-tonifying, spleen-strengthening, and brain-boosting peptide oral liquid (0.4mL / 10g body weight) prepared in Examples 1-3 and Comparative Examples 1-6 by gavage, while the blank control group and model control group were administered an equal volume of physiological saline by gavage once daily for 10 consecutive days.
[0123] After successful modeling, mice were fasted for 12 hours and then administered D-xylose solution (2 g / kg, 0.2 mL / 10 g body weight) by gavage. Two hours later, blood was collected from the orbital sinus, serum was separated, and the absorbance was measured at 540 nm using the phloroglucinol colorimetric method to calculate the serum D-xylose concentration. 0.1 g of gastric tissue was taken, added to physiological saline to prepare a 10% homogenate, centrifuged at 3000 rpm for 15 min, and the supernatant was collected to determine pepsin activity (unit: U / mg prot). 0.1 g of pancreatic tissue was taken, prepared into a 10% homogenate, centrifuged, and the supernatant was collected to determine pancreatic amylase activity (unit: U / mg prot). ALB content was determined using the bromocresol green method, and TP content was determined using the biuret method. The albumin / total protein ratio (A / G) was calculated.
[0124] Specific data on spleen-strengthening indicators are shown in Table 2.
[0125] Table 2. Detection results of spleen-strengthening indicators of the brain peptide oral liquid prepared in Examples 1-3 and Comparative Examples 1-6 (x±s, n=10)
[0126] As shown in Tables 1 and 2, the brain peptide oral liquid prepared in this application, which tonifies the kidney, strengthens the spleen, and improves intelligence, has significant kidney-tonifying and spleen-strengthening effects. Regarding kidney tonification, it can significantly increase serum testosterone, T3, and T4 levels in mice with kidney-yang deficiency, reduce cortisol levels, enhance SOD activity in kidney tissue, reduce MDA accumulation, and improve kidney-yang deficiency-related indicators. Regarding spleen strengthening, it can increase serum D-xylose concentration in mice with spleen deficiency, enhance pepsin and pancreatic amylase activity, increase ALB content and A / G ratio, and improve digestive and absorptive functions, providing a highly efficient and stable oral liquid preparation for tonifying the kidney and strengthening the spleen.
[0127] This specific embodiment is merely an explanation of this application and is not intended to limit it. After reading this specification, those skilled in the art can make modifications to this embodiment without contributing any inventive step, but such modifications are protected by patent law as long as they fall within the scope of the claims of this application.
Claims
1. A brain peptide oral liquid for tonifying the kidneys, strengthening the spleen, and improving intelligence, characterized in that, Each 100 servings of oral liquid contains the following raw material components by weight: 3-5 parts longan pulp, 3-6 parts wolfberry, 3-8 parts mulberry, 3-6 parts red dates, 2-6 parts poria cocos, 0.5-2 parts alpinia oxyphylla, 0.05-0.1 parts dried ginger, 8-12 parts peptide components, 0.01-0.05 parts DHA algal oil, 0.05-0.1 parts N-acetylneuraminic acid, 1-3 parts γ-aminobutyric acid, and 0.002-0.004 parts vitamin B. 12 0.05-0.1 parts steviol glycosides, 12-15 parts concentrated apple juice, and the remainder is purified water.
2. The brain peptide oral liquid for tonifying the kidney, strengthening the spleen, and improving intelligence according to claim 1, characterized in that, The preparation method of the peptide component includes the following preparation steps: Step 1: Homogenize sheep brain and add deionized water to adjust the pH to 6.5-7.
0. Add complex protease and hydrolyze for 2.5-3 hours. After inactivating the enzyme, centrifuge and collect the supernatant to obtain sheep brain peptides. Step 2: Crush the oyster meat and add deionized water to obtain a fermentation substrate. Inoculate the fermentation substrate with Bacillus natto and ferment for 18-24 hours to obtain a fermentation broth. Add neutral protease to the fermentation broth and hydrolyze for 2-5 hours. Centrifuge to remove the supernatant and obtain oyster peptides. Step 3: Mix walnuts and soybeans, grind and sieve them, add deionized water and mix evenly, adjust the pH to 8.0-8.5 to obtain soybean-walnut liquid; first add alkaline protease to soybean-walnut liquid and hydrolyze for 2-3 hours, then add flavor protease and hydrolyze for 1-3 hours, inactivate the enzyme, centrifuge and take the supernatant to obtain soybean-walnut complex peptide. Step 4: Mix sheep brain peptides, oyster peptides and soybean-walnut complex peptides, vacuum concentrate to a solid content of 20-25%, and then freeze-dry to obtain peptide components.
3. The brain peptide oral liquid for tonifying the kidney, strengthening the spleen, and improving intelligence according to claim 2, characterized in that, In step 1, the complex protease is composed of subtilisin, elastase and aminopeptidase in a mass ratio of 10-12:5-8:2-5.
4. The brain peptide oral liquid for tonifying the kidney, strengthening the spleen, and improving intelligence according to claim 2, characterized in that, In step 1, the mass ratio of the complex protease to sheep brain is 1-3:200-300.
5. The brain peptide oral liquid for tonifying the kidney, strengthening the spleen, and improving intelligence according to claim 2, characterized in that, In step 2, the inoculation amount of Bacillus natto is 2-5% of the mass of the fermentation substrate.
6. The brain peptide oral liquid for tonifying the kidney, strengthening the spleen, and improving intelligence according to claim 2, characterized in that, In step 2, the mass ratio of fermentation broth to neutral protease is 100-200:3-5.
7. The brain peptide oral liquid for tonifying the kidney, strengthening the spleen, and improving intelligence according to claim 2, characterized in that, In step 3, the mass ratio of soybean-walnut liquid, alkaline protease, and flavor protease is 200-300:3-5:1-3.
8. The brain peptide oral liquid for tonifying the kidney, strengthening the spleen, and improving intelligence according to claim 2, characterized in that, In step 4, the mass ratio of sheep brain peptide, oyster peptide, and soybean-walnut complex peptide is 3-6:2-5:1-2.
9. A method for preparing a brain peptide oral liquid for tonifying the kidney, strengthening the spleen, and improving intelligence as described in any one of claims 1-8, characterized in that, Specifically, the preparation steps include the following: Mix longan pulp, goji berries, mulberries, red dates, poria cocos, alpinia oxyphylla, dried ginger, and purified water, and decoct 2-3 times, 1-2 hours each time. Combine the decoctions, filter and concentrate to obtain a herbal extract. Add peptide components and DHA algal oil to the herbal extract, stir well, and then add N-acetylneuraminic acid, γ-aminobutyric acid, and vitamin B. 12 Steviol glycosides and concentrated apple juice are mixed evenly, then purified water is added to make up the total volume, the pH is adjusted to 6.0-7.0, filtered, sterilized, cooled and aseptically filled to obtain a brain peptide oral liquid that nourishes the kidneys, strengthens the spleen and improves intelligence.