A blood-replenishing and anti-fatigue composition and preparation method thereof

Abstract

The present invention relates to a blood-tonifying and anti-fatigue composition and a preparation method thereof, and belongs to the field of biotechnology. The composition comprises casein phosphopeptide, black fish skin oligopeptide, donkey-hide gelatin peptide, Cistanche deserticola extract, raspberry extract, purple eggplant peel extract, black wolfberry powder, red ginseng powder, American ginseng powder, astragalus powder, Poria powder, cinnamon powder and hemp seed protein powder and other ingredients. The black fish skin oligopeptide is obtained by graded enzymolysis of black fish skin protein by Bacillus subtilis alkaline protease, chymotrypsin and papain; the donkey-hide gelatin peptide is obtained by fermenting donkey-hide gelatin powder by Bacillus subtilis and then enzymolysis by bromelain; the Cistanche deserticola extract is obtained by enzymolysis of Cistanche deserticola by cellulase and pectinase and then by serrapeptidase; the raspberry extract is obtained by enzymolysis of raspberry fruit by ficin and amylase; the purple eggplant peel extract is obtained by enzymolysis of purple eggplant peel by laccase and hemicellulase and then by ficin. The composition has safe and efficient blood-tonifying and anti-fatigue effects.

Description

Technical Field

[0001] The invention belongs to the field of biotechnology, and specifically relates to a blood-enriching and anti-fatigue composition and a preparation method thereof. Background Art

[0002] Anemia refers to a common syndrome in which the volume of red blood cells in the human body's peripheral blood is lower than the lower limit of the normal range. Its causes are relatively complex, among which nutritional anemia is particularly common, mainly including iron deficiency anemia, megaloblastic anemia, etc. Iron deficiency anemia is mostly caused by insufficient iron intake, malabsorption or excessive loss. Megaloblastic anemia is often closely related to a lack of folic acid and / or vitamin B12. In modern fast-paced life, anemia and fatigue have become health problems that plague many people.

[0003] Fatigue is a warning signal from the body. It may be the result of multiple factors such as physical overwork, lack of sleep, and excessive psychological stress. It may also be the external manifestation of underlying diseases such as anemia. Long-term fatigue will not only affect personal work efficiency and quality of life, but may also further cause other health problems, such as decreased immunity and increased risk of cardiovascular disease. In the highly competitive workplace environment, office workers sit at their desks for long periods of time, lack exercise, and are highly focused. They are very prone to fatigue, which is difficult to completely relieve through simple rest.

[0004] Whether due to high-intensity work pressure, bad eating habits, or specific physiological conditions, more and more people are looking for effective blood-replenishing and anti-fatigue solutions, which also makes the research and development of related drugs and health foods an important topic in the field of medicine and health care.

[0005] At present, there are many drugs and health foods on the market for blood tonic and anti-fatigue. Traditional blood tonic drugs such as iron, folic acid and vitamin B12 preparations have clear therapeutic effects in the treatment of nutritional anemia. However, these drugs often have some limitations. For example, iron may cause gastrointestinal discomfort, such as nausea, vomiting, constipation and other side effects, resulting in poor compliance in some patients. In terms of health foods, traditional tonics such as red dates, longan, and donkey-hide gelatin are widely recognized to have blood tonic effects, but the blood tonic and anti-fatigue effects are not obvious. Long-term and large-scale use can cause excessive side effects, including fullness, anorexia, and excessive sugar.

[0006] With the continuous development of modern medicine and biotechnology, the research on blood tonic and anti-fatigue has gradually deepened to the cellular and molecular levels. New studies have found that some bioactive ingredients such as heme iron, certain polysaccharides and flavonoids in Chinese herbal extracts have better bioavailability and fewer side effects, showing potential advantages in blood tonic. At the same time, the research on the mechanism of fatigue has also revealed several key links such as energy metabolism and neurotransmitter regulation, providing new targets for the development of new anti-fatigue drugs or health foods.

[0007] In summary, although the existing blood-tonifying and anti-fatigue drugs and health foods have met the market demand to a certain extent, they still have problems such as high efficiency and safety. Therefore, it is necessary to develop safer, more effective and convenient blood-tonifying and anti-fatigue material ingredients to meet consumers' requirements for fast and safe blood-tonifying and anti-fatigue health improvement. Summary of the invention

[0008] In order to improve the efficiency and safety of blood-tonifying and anti-fatigue material components, the present invention provides a blood-tonifying and anti-fatigue composition and a preparation method thereof, the composition includes casein phosphopeptide, black fish skin oligopeptide, donkey-hide gelatin peptide, Cistanche deserticola extract, raspberry extract, purple eggplant peel extract, black wolfberry powder, red ginseng powder, American ginseng powder, astragalus powder, Poria powder, cinnamon powder, hemp seed protein powder, Patchouli powder and malt powder and other ingredients, and the combination of the ingredients has a rapid, safe and efficient blood-tonifying and anti-fatigue effect. The specific technical scheme is as follows:

[0009] A blood-tonifying and anti-fatigue composition, comprising the following raw materials in parts by mass: 0.5 to 2 parts of casein phosphopeptide, 1.5 to 3 parts of black fish skin oligopeptide, 0.2 to 0.5 parts of donkey-hide gelatin peptide, 0.5 to 1.5 parts of Cistanche deserticola extract, 0.5 to 1.5 parts of raspberry extract, 1.5 to 3 parts of purple eggplant peel extract, 1 to 3 parts of black wolfberry powder, 0.01 to 0.03 parts of red ginseng powder, 0.01 to 0.03 parts of American ginseng powder, 0.5 to 1 parts of astragalus powder, 1 to 3 parts of tuckahoe powder, 0.02 to 0.05 parts of cinnamon powder, 0.01 to 0.03 parts of hemp seed protein powder, 0.05 to 0.08 parts of patchouli powder and 2 to 5 parts of malt powder;

[0010] The black fish skin oligopeptide is a black fish skin oligopeptide with a molecular weight of 1000Da to 3000Da obtained by subjecting black fish skin protein to graded enzymolysis with Bacillus subtilis alkaline protease, chymotrypsin and papain.

[0011] In the above composition, the preparation method of the black fish skin oligopeptide comprises the following steps: dispersing the black fish skin protein in a phosphate buffer with a pH of 7.5 to 8.5 and a concentration of 0.05 mol / L to 0.15 mol / L according to a mass ratio of black fish skin protein to phosphate buffer = 1: (8 to 10), adding 1.5% to 2.5% by mass of Bacillus subtilis alkaline protease of the black fish skin protein, performing enzymolysis for 60 to 90 minutes at 50° C. to 60° C. and stirring at 30 r / min to 80 r / min, heating to 85° C. to 95° C. to inactivate the enzyme for 10 to 15 minutes, cooling to room temperature, and adding 0.5% to 1% by mass of chymoprotein of the black fish skin protein. The method comprises the following steps: enzymolysis for 60 to 90 minutes at 35 to 38 degrees Celsius and 30 to 80 r / min stirring state, heating to 85 to 95 degrees Celsius to inactivate the enzyme for 10 to 15 minutes, cooling to room temperature, adjusting the pH to 5.5 to 6.5, adding papain with 1% to 1.5% by weight of black fish skin protein, enzymolysis for 50 to 80 minutes at 50 to 60 degrees Celsius and 30 to 80 r / min stirring state, heating to 85 to 95 degrees Celsius to inactivate the enzyme for 10 to 15 minutes, cooling to room temperature, filtering with a sterile ultrafiltration membrane, and lyophilizing a filtrate with a molecular weight cutoff of 1000 to 3000 Da to obtain black fish skin oligopeptide.

[0012] In the preparation method of the above-mentioned black fish skin oligopeptide, the preparation method of the black fish skin protein comprises the following steps: washing fresh black fish skin, cutting it into small pieces with a side length range of 0.5 cm to 2 cm, and obtaining black fish skin small pieces; adding the black fish skin small pieces to a sodium hydroxide aqueous solution with a concentration of 0.1 mol / L to 0.5 mol / L according to a mass ratio of black fish skin small pieces: sodium hydroxide aqueous solution = 1: (8 to 12), stirring for 2h to 4h at a temperature of 40° C. to 60° C. and a rotation speed of 200r / min to 400r / min to dissolve the protein and obtain an alkali extract; adjusting the pH value of the alkali extract to 4.0 to 6.0 with a hydrochloric acid aqueous solution with a concentration of 1mol / L to 3mol / L, standing for 30min to 60min, and precipitating the protein; and then centrifuging at a rotation speed of 4000r / min to 6000r / min for 10min to 20min n, obtaining protein precipitate; washing the protein precipitate with distilled water 2 to 3 times to remove residual acid and alkali, and obtaining black fish skin protein.

[0013] In the above composition, the preparation method of the donkey-hide gelatin peptide comprises the following steps: taking donkey-hide gelatin blocks, crushing them into fine powder, passing them through a 80-mesh to 100-mesh sieve, taking the sieve to obtain donkey-hide gelatin powder, preparing the donkey-hide gelatin powder into a 5wt% to 10wt% donkey-hide gelatin liquid with distilled water, stirring at 40°C to 50°C for 30min to 60min until fully dissolved; based on the donkey-hide gelatin liquid, adding 1% to 3% glucose by weight of the donkey-hide gelatin liquid, 0.5% to 1.5% yeast extract by weight of the donkey-hide gelatin liquid, 0.1% to 0.3% potassium dihydrogen phosphate by weight of the donkey-hide gelatin liquid, and 0.0 5% to 0.15% by weight of magnesium sulfate, adjusting the pH to 6.5 to 7.5 to obtain a fermentation medium; inoculating the activated Bacillus subtilis into the fermentation medium, the inoculation amount being 2% to 5% by weight of the fermentation medium, performing aerobic fermentation for 48 hours to 72 hours at a temperature of 35° C. to 38° C. and 150 r / min to 250 r / min, raising the temperature to 120° C. to 125° C., maintaining for 10 minutes to 15 minutes for sterilization, and cooling to room temperature to obtain a fermentation liquid; the fermentation liquid is heated at 6000 r / min to 8000 r / min n Centrifuge for 20min-30min, collect the supernatant, add 0.5%-1.0% bromelain by weight of the supernatant to the supernatant, perform enzymolysis for 40min-60min at 50℃-60℃ and 30r / min-80r / min stirring, heat to 85℃-95℃ to inactivate the enzyme for 10min-15min, cool to room temperature, filter with a sterile ultrafiltration membrane, the ultrafiltration membrane has a molecular weight cutoff of 1000Da-5000Da, and lyophilize to obtain donkey-hide gelatin peptide.

[0014] In the above composition, the preparation method of the Cistanche deserticola extract comprises the following steps: taking Cistanche deserticola, washing and then crushing it, adding Cistanche deserticola to deionized water at a mass ratio of Cistanche deserticola: deionized water = 1: (8-12), adjusting the pH to 4.5-5.5, adding 0.5%-1.5% by mass of cellulase and 0.3%-1.0% by mass of pectinase, and heating at 40°C-45°C and 30r / min-80r. / min stirring state for 2h to 3h; then adjust the pH to 7.0-8.0, add 0.5% to 1.5% by weight of serrapeptase from Cistanche deserticola, and perform enzymolysis for 1.5h to 2h at 45°C to 55°C and 30r / min to 80r / min stirring state, heat to 85°C to 95°C to inactivate the enzyme for 10min to 15min, cool to room temperature, filter with a sterile sieve, take the filtrate, and lyophilize to obtain a Cistanche deserticola extract.

[0015] In the above composition, the preparation method of the raspberry extract comprises the following steps: taking ripe raspberry fruits, washing and then crushing them, adding the raspberry fruits into deionized water at a mass ratio of raspberry fruits: deionized water = 1: (8-12), adjusting the pH to 5.0-6.0, adding 1.5%-3% by mass of ficin and 0.2%-0.6% by mass of amylase, performing enzymolysis at 38°C-45°C and 30r / min-80r / min under stirring for 3h-4h; heating to 90°C-95°C to inactivate the enzyme for 10min-15min, cooling to room temperature, filtering with a sterile mesh, taking the filtrate, and freeze-drying to obtain the raspberry extract.

[0016] In the above composition, the preparation method of the purple eggplant peel extract comprises the following steps: taking fresh purple eggplant peel, washing and then crushing, adding the purple eggplant peel to a phosphate buffer with a pH of 5.5 to 6.5 and a concentration of 0.05 mol / L to 0.15 mol / L according to a mass ratio of purple eggplant peel to deionized water = 1: (8 to 12), adding 1.0% to 2.0% by mass of laccase and 0.5% to 1.0% by mass of hemicellulase, and heating at 42° C. to 48° C. and enzymolyze for 2h to 3h under stirring at 30r / min to 80r / min; then adjust the pH to 5.5 to 6.0, add 0.8% to 1.5% by weight of fig protease in the purple eggplant peel, and enzymolyze for 1h to 1.5h under stirring at 60°C to 65°C and 30r / min to 80r / min, raise the temperature to 90°C to 95°C to inactivate the enzyme for 10min to 15min, cool to room temperature, filter with a sterile sieve, take the filtrate, and lyophilize to obtain a purple eggplant peel extract.

[0017] In the above-mentioned method for preparing the extract, the mesh size of the sieve is 200-250 meshes.

[0018] In the above composition, the black wolfberry powder, red ginseng powder, American ginseng powder, astragalus powder, tuckahoe powder, cinnamon powder, hemp seed protein powder, patchouli powder and malt powder are all passed through a 200-250 mesh sieve, and the sieved material is taken.

[0019] The preparation method of the above-mentioned blood-enriching and anti-fatigue composition comprises the following steps:

[0020] S1: Mix casein phosphopeptide, black fish skin oligopeptide, donkey-hide gelatin peptide, Cistanche deserticola extract, raspberry extract and purple eggplant peel extract in proportions by mass to obtain a mixture A;

[0021] S2: Evenly mix black wolfberry powder, red ginseng powder, American ginseng powder, astragalus powder, tuckahoe powder, cinnamon powder, hemp seed protein powder, patchouli powder and malt powder according to parts by mass to obtain a mixture B;

[0022] S3: Mix the mixture A and the mixture B uniformly to obtain a composition.

[0023] The composition is mixed with pharmaceutically acceptable auxiliary materials to prepare a product with blood-enriching and anti-fatigue effects, and the product dosage form is tablets, capsules, powders or emulsions.

[0024] The present invention provides a blood-enriching and anti-fatigue composition and a preparation method thereof, and the beneficial effects are as follows:

[0025] 1. The composition of the present invention includes casein phosphopeptide, black fish skin oligopeptide, donkey-hide gelatin peptide, Cistanche deserticola extract, raspberry extract, purple eggplant peel extract, black wolfberry powder, red ginseng powder, American ginseng powder, astragalus powder, Poria powder, cinnamon powder, hemp seed protein powder, patchouli powder and malt powder; among them, casein phosphopeptide helps to absorb minerals such as calcium and iron, and has a promoting effect on the absorption and utilization of iron in blood-enriching raw materials; black fish skin oligopeptide, donkey-hide gelatin peptide and other peptides can provide easily absorbed nutrients, replenish body energy, and resist fatigue; promote cell activity, promote the absorption and utilization of blood-enriching ingredients, and achieve a good blood-enriching effect. Plant extracts such as Cistanche deserticola extract and raspberry extract contain a variety of biologically active ingredients, which can comprehensively regulate body functions, improve qi and blood circulation, reduce fatigue, and play an auxiliary role in blood-enriching and synergistic effects; the combination of various ingredients has a rapid, safe and efficient blood-enriching and anti-fatigue effect.

[0026] 2. Casein phosphopeptide promotes the absorption of iron. It can form a soluble complex with iron to prevent iron precipitation in the intestine, thereby increasing the concentration of iron on the surface of the small intestinal mucosal cells, which is beneficial to the absorption of iron, and then provides sufficient iron for the synthesis of hemoglobin and assists in blood supplementation. By ensuring the absorption of iron, maintaining normal hematopoietic function, ensuring oxygen transportation, and providing adequate oxygen supply to body tissues, fatigue caused by hypoxia is reduced.

[0027] Black fish skin oligopeptide is a small molecule peptide that is easily absorbed by the human body. It can supplement the body with high-quality protein and other nutrients, help supplement protein and other substances in the blood, provide raw materials for the formation of blood cells, increase hemoglobin levels, stimulate the proliferation and differentiation of hematopoietic stem cells, promote blood formation, and promote hematopoiesis to a certain extent. After being quickly absorbed by the human body, it can provide energy, help repair and nourish tissue cells. It can also regulate the body's metabolism, reduce physical fatigue, and enhance physical endurance.

[0028] Donkey-hide gelatin peptide is more easily absorbed by the human body. Compared with donkey-hide gelatin, it can more effectively increase the number of red blood cells and white blood cells in the blood, improve hemoglobin levels, stimulate the proliferation and differentiation of hematopoietic stem cells, promote blood production, and thus enhance the blood-tonifying effect. It can regulate the body's energy metabolism, provide more energy support for the body, and improve fatigue. At the same time, it can also nourish the body and enhance the body's anti-fatigue ability.

[0029] Cistanche extract contains phenylethanoid glycosides, polypeptides and other ingredients, which can regulate the body's endocrine and metabolism, promote the secretion of hematopoietic hormones, help improve hematopoietic function, and assist in blood supplementation. It can enhance energy and reduce fatigue. By regulating the neuro-endocrine-immune network, it enhances the body's stress resistance and endurance, making the body more resistant to fatigue.

[0030] Raspberry extract contains active ingredients such as flavonoids and peptides, which can improve the hematopoietic microenvironment, promote bone marrow hematopoiesis, and play a certain auxiliary role in blood supplementation. It helps to regulate body functions, enhance the body's antioxidant capacity, remove free radicals, reduce fatigue caused by oxidative stress, and improve the body's anti-fatigue performance.

[0031] Purple eggplant peel extract contains anthocyanins, peptides and other antioxidant ingredients, which can protect hematopoietic stem cells from free radical damage, maintain the normal function of hematopoietic stem cells, and promote blood production. The antioxidant properties of the extract can remove excessive free radicals produced by fatigue, enhance the body's antioxidant defense system, help the body restore energy, and reduce fatigue.

[0032] Black wolfberry powder is rich in anthocyanins and has a strong antioxidant effect. It can protect blood cells from free radical damage, which is beneficial to maintaining the normal structure and function of blood cells to a certain extent, and has a positive significance for blood supplementation. It can remove free radicals in the body, reduce the damage of oxidative stress to the body, improve the body's fatigue state, and enhance the body's anti-fatigue ability.

[0033] Red ginseng powder has the effects of replenishing vital energy, restoring pulse and strengthening deficiency, promoting the circulation of qi and blood, improving the condition of insufficient qi and blood, and helping to replenish blood. It can enhance the vitality of the body, improve endurance, resist fatigue, and provide more energy to the body by regulating body functions, reducing fatigue.

[0034] American ginseng powder can nourish qi and yin, help to harmonize qi and blood, and provide a good internal environment for blood supplementation. It can clear away heat and promote the production of body fluids, which is very helpful for relieving fatigue, improving the body's adaptability, and reducing fatigue.

[0035] Astragalus powder has the effects of replenishing qi and raising yang, can enhance the body's immunity, promote hematopoietic function, and help increase the formed elements in the blood to achieve the effect of replenishing blood. It can improve the body's overall functional state, enhance the body's anti-fatigue ability, and reduce fatigue caused by physical weakness.

[0036] Poria powder has the function of promoting water and moisture, indirectly affecting body functions by regulating the body's water metabolism balance, creating good conditions for the normal production and circulation of blood. It improves fatigue caused by heavy moisture in the body, makes the body feel more refreshed, and reduces fatigue.

[0037] Cinnamon powder has the effects of replenishing fire and yang, promoting blood circulation, helping to transport nutrients to the hematopoietic organs, and providing favorable conditions for blood replenishment. It improves physical coldness, fatigue, etc., warms up the body, promotes metabolism, and relieves fatigue.

[0038] Hemp protein powder provides protein nutrition for the human body, provides raw materials for the formation of blood cells, and assists in blood supplementation to a certain extent. It replenishes energy, reduces fatigue, and maintains the body's normal metabolism and function.

[0039] Patchouli powder is mainly used to eliminate dampness and invigorate the spleen. It can regulate the spleen and stomach functions, promote the absorption of nutrients, and indirectly help to replenish blood. It is helpful to improve fatigue caused by heavy dampness in the body and improve body functions.

[0040] Malt powder can promote qi and digestion, help maintain normal gastrointestinal function, enable the body to better absorb the nutrients needed for hematopoiesis, and indirectly promote blood supplementation. Ensure the body's normal absorption of nutrients, maintain the body's energy supply, and reduce fatigue.

[0041] 3. Black fish skin oligopeptide is a black fish skin oligopeptide with a molecular weight of 1000Da to 3000Da obtained by graded enzymatic hydrolysis of black fish skin protein with Bacillus subtilis alkaline protease, chymotrypsin and papain. Compared with the macromolecular black fish skin protein, the specific enzymatic hydrolysis small molecule peptide of this black fish skin oligopeptide is easier to be absorbed by the human body. In terms of blood supplementation, its efficient absorption characteristics can quickly replenish the body with nutrients needed to synthesize hemoglobin and other blood components. In terms of anti-fatigue, it can enter the blood circulation faster, be transported to the body's tissues, provide energy and nutritional support for cell metabolism, accelerate fatigue recovery, and thus play a synergistic role.

[0042] 4. Donkey-hide gelatin peptide is a 1000Da to 5000Da molecular weight donkey-hide gelatin peptide obtained by fermenting donkey-hide gelatin powder with Bacillus subtilis and then enzymolyzing it with bromelain. This donkey-hide gelatin peptide is easier to be absorbed by the human body than traditional donkey-hide gelatin. In terms of blood tonic, its specific enzymatic small molecule form can make the effective ingredients in donkey-hide gelatin play a better role, promote the proliferation and differentiation of bone marrow hematopoietic stem cells, quickly increase blood formed elements, and improve blood quality. In terms of anti-fatigue, it can more effectively regulate the body's energy metabolism, quickly replenish the body's energy, and reduce the accumulation of fatigue substances. It has more advantages than donkey-hide gelatin itself in blood tonic and anti-fatigue.

[0043] Bacillus subtilis fermentation is a complex microbial metabolic process. During the fermentation process, in addition to protease, Bacillus subtilis will also secrete a variety of other enzymes (such as amylase, lipase, etc.) and some metabolites. These enzymes and metabolites will interact with donkey-hide gelatin to produce synergistic and antagonistic effects. For example, in addition to protein hydrolysis reactions, other modification reactions such as glycosylation reactions will also occur, which will affect the composition of the final peptide. Due to the combined effect of multiple factors in the fermentation process, the size range of the peptides produced is wider. Some peptides will be further modified, polymerized or decomposed due to the metabolic action of other enzymes, resulting in more complex and diverse lengths and compositions of the peptides. During the fermentation process, the metabolic activities of microorganisms will introduce new chemical groups, such as sugar groups, phosphate groups, etc., causing the peptides to be chemically modified and exhibit different characteristics in terms of solubility, stability, and biological activity.

[0044] 5. Cistanche deserticola extract is obtained by enzymolysis of Cistanche deserticola with cellulase and pectinase, and then enzymolysis with serrapeptase. The extract can more fully release the active ingredients in Cistanche deserticola. In terms of blood supplementation, it is beneficial to extract and retain more components that can promote hematopoiesis, such as phenylethanol glycosides and polypeptides. In terms of anti-fatigue, these components can better regulate the body's neuro-endocrine-immune network, improve the body's stress resistance and endurance, and enhance the anti-fatigue and blood supplement functions of Cistanche deserticola.

[0045] 6. Raspberry extract is obtained by enzymolysis of raspberry fruit with ficin and amylase. It can better release the effective ingredients such as flavonoids and peptides in raspberries. In terms of blood supplementation, these ingredients will promote the improvement of the hematopoietic microenvironment. In terms of anti-fatigue, the extract can regulate the body's metabolism and physiological functions, improve the body's antioxidant capacity, remove free radicals, more effectively reduce fatigue, and play a synergistic role.

[0046] 7. Purple eggplant peel extract is obtained by enzymolysis of purple eggplant peel with laccase and hemicellulase, and then with ficin, which can better extract anthocyanins, polypeptides and other effective ingredients in purple eggplant peel. In terms of blood supplementation, anthocyanins, polypeptides and other ingredients can protect hematopoietic stem cells and maintain their normal functions. In terms of anti-fatigue, it can remove excessive free radicals produced by fatigue, enhance the body's antioxidant defense system, and produce a synergistic effect in blood supplementation and anti-fatigue. DETAILED DESCRIPTION

[0047] The present invention is further described below in conjunction with specific implementation cases, but the present invention is not limited to these embodiments.

[0048] Example 1

[0049] A blood-enriching and anti-fatigue composition comprises the following raw materials in parts by mass: 1.2 parts of casein phosphopeptide, 2.0 parts of black fish skin oligopeptide, 0.3 parts of donkey-hide gelatin peptide, 1.0 parts of Cistanche deserticola extract, 1.0 parts of raspberry extract, 2.2 parts of purple eggplant peel extract, 2 parts of black wolfberry powder, 0.02 parts of red ginseng powder, 0.02 parts of American ginseng powder, 0.8 parts of astragalus powder, 2 parts of tuckahoe powder, 0.03 parts of cinnamon powder, 0.02 parts of hemp seed protein powder, 0.06 parts of patchouli powder and 3 parts of malt powder.

[0050] In the composition, the preparation method of black fish skin oligopeptide comprises the following steps:

[0051] S1: Wash fresh snakehead skin and cut it into small pieces with a side length of 0.5 cm to 1 cm to obtain snakehead skin small pieces; add the snakehead skin small pieces to a sodium hydroxide aqueous solution with a concentration of 0.3 mol / L according to the mass ratio of the snakehead skin small pieces: sodium hydroxide aqueous solution=1:10, stir for 3 hours at a temperature of 50°C and a speed of 300 r / min to dissolve the protein and obtain an alkali extract; adjust the pH value of the alkali extract to 5.0 with a hydrochloric acid aqueous solution with a concentration of 2 mol / L, leave it to stand for 40 minutes, and precipitate the protein; then centrifuge at a speed of 5000 r / min for 15 minutes to obtain a protein precipitate; wash the protein precipitate with distilled water 2 times (add distilled water and mix, and centrifuge at a speed of 5000 r / min for 15 minutes), remove the residual acid and alkali, and obtain snakehead skin protein;

[0052] S2: According to the mass ratio of black fish skin protein: phosphate buffer = 1:9, black fish skin protein is dispersed in a phosphate buffer with a pH of 8.0 and a concentration of 0.1 mol / L, 2.0% of the mass of Bacillus subtilis alkaline protease of black fish skin protein is added, and enzymolysis is carried out at 55°C and 60r / min stirring for 80min, the temperature is raised to 90°C and the enzyme is inactivated for 12min, and after cooling to room temperature, 0.8% of the mass of chymotrypsin of black fish skin protein is added, and enzymolysis is carried out at 37°C and 60r / min stirring for 80min, the temperature is raised to 90°C and the enzyme is inactivated for 12min, and after cooling to room temperature, pH is adjusted to 6.0, 1.2% of the mass of papain of black fish skin protein is added, and enzymolysis is carried out at 55°C and 60r / min stirring for 60min, the temperature is raised to 90°C and the enzyme is inactivated for 12min, and after cooling to room temperature, a sterile ultrafiltration membrane is used for filtration, and the filtrate with a molecular weight cutoff of the ultrafiltration membrane is 1000Da to 3000Da, and freeze-dried to obtain black fish skin oligopeptide.

[0053] In the composition, the preparation method of donkey-hide gelatin peptide comprises the following steps: taking donkey-hide gelatin blocks, crushing them into fine powder, passing through an 80-mesh sieve, taking the sieve to obtain donkey-hide gelatin powder, preparing the donkey-hide gelatin powder into 8wt% donkey-hide gelatin liquid with distilled water, stirring at 45°C for 50 minutes until fully dissolved; adding 2% glucose by weight of the donkey-hide gelatin liquid, 1% yeast extract by weight of the donkey-hide gelatin liquid, 0.2% potassium dihydrogen phosphate by weight of the donkey-hide gelatin liquid, and 0.1% magnesium sulfate by weight of the donkey-hide gelatin liquid as the basis, adjusting the pH to 7.0, and obtaining a fermentation medium; inoculating the activated Bacillus subtilis into the fermentation medium, and the inoculation amount is 3% of the mass of the fermentation medium %, aerobic fermentation was carried out at 37°C and 200r / min for 60h, the temperature was raised to 121°C, maintained for 12min for sterilization, and then cooled to room temperature to obtain a fermentation liquid; the fermentation liquid was centrifuged at 7000r / min for 25min, the supernatant was collected, 0.8% mass of bromelain of the supernatant was added to the supernatant, enzymolysis was carried out at 55°C and 60r / min for 50min under stirring, the temperature was raised to 90°C for 12min for enzyme inactivation, and then cooled to room temperature, and then filtered through a sterile ultrafiltration membrane, the ultrafiltration membrane had a molecular weight cutoff of 1000Da to 5000Da, and the filtrate was freeze-dried to obtain donkey-hide gelatin.

[0054] In the composition, the preparation method of the Cistanche deserticola extract comprises the following steps: taking Cistanche deserticola, washing and then crushing it, adding Cistanche deserticola to deionized water at a mass ratio of Cistanche deserticola to deionized water = 1:10, adjusting the pH to 5.0, adding 1.0% by mass of cellulase and 0.6% by mass of pectinase of Cistanche deserticola, and performing enzymolysis at 42°C and 60r / min stirring for 2.5h; then adjusting the pH to 7.5, adding 1.0% by mass of serrapeptase of Cistanche deserticola, performing enzymolysis at 50°C and 60r / min stirring for 1.5h, heating to 90°C to inactivate the enzyme for 12min, cooling to room temperature, filtering with a 200-mesh sterile sieve, taking the filtrate, and freeze-drying to obtain the Cistanche deserticola extract.

[0055] In the composition, the preparation method of the raspberry extract comprises the following steps: taking ripe raspberry fruits, washing and then crushing them, adding the raspberry fruits into deionized water at a mass ratio of raspberry fruits: deionized water = 1:10, adjusting the pH to 5.5, adding 2.2% by mass of fig protease and 0.5% by mass of amylase of raspberry fruits, performing enzymolysis for 3.5 hours at 42° C. and 60 r / min stirring; heating to 92° C. to inactivate the enzyme for 12 minutes, cooling to room temperature, filtering with a 200-mesh sterile sieve, taking the filtrate, and freeze-drying to obtain the raspberry extract.

[0056] In the composition, the preparation method of the purple eggplant peel extract comprises the following steps: taking fresh purple eggplant peel, washing and then crushing, adding the purple eggplant peel into a phosphate buffer with a pH of 6.0 and a concentration of 0.1 mol / L according to a mass ratio of purple eggplant peel: deionized water = 1:10, adding 1.5% by mass of laccase and 0.8% by mass of hemicellulase of the purple eggplant peel, and enzymolysis for 2.5 hours at 45°C and 60 r / min stirring; then adjusting the pH to 5.8, adding 1.2% by mass of fig protease of the purple eggplant peel, and enzymolysis for 1 hour at 62°C and 60 r / min stirring, heating to 92°C to inactivate the enzyme for 12 minutes, cooling to room temperature, filtering with a 200-mesh sterile sieve, taking the filtrate, and freeze-drying to obtain the purple eggplant peel extract.

[0057] In the composition, black wolfberry powder, red ginseng powder, American ginseng powder, astragalus powder, tuckahoe powder, cinnamon powder, hemp seed protein powder, patchouli powder and malt powder are all passed through a 200-mesh sieve, and the sieved material is taken.

[0058] The preparation method of the above-mentioned blood-enriching and anti-fatigue composition comprises the following steps:

[0059] S1: Mix casein phosphopeptide, black fish skin oligopeptide, donkey-hide gelatin peptide, Cistanche deserticola extract, raspberry extract and purple eggplant peel extract in proportions by mass to obtain a mixture A;

[0060] S2: Evenly mix black wolfberry powder, red ginseng powder, American ginseng powder, astragalus powder, tuckahoe powder, cinnamon powder, hemp seed protein powder, patchouli powder and malt powder according to parts by mass to obtain a mixture B;

[0061] S3: Mix the mixture A and the mixture B uniformly to obtain a composition.

[0062] The composition is mixed with pharmaceutically acceptable auxiliary materials to prepare a product with blood-enriching and anti-fatigue effects, and the product dosage form is tablets, capsules, powders or emulsions.

[0063] Example 2

[0064] A blood-enriching and anti-fatigue composition comprises the following raw materials in parts by mass: 0.5 parts of casein phosphopeptide, 3 parts of black fish skin oligopeptide, 0.2 parts of donkey-hide gelatin peptide, 1.5 parts of Cistanche deserticola extract, 0.5 parts of raspberry extract, 3 parts of purple eggplant peel extract, 1 part of black wolfberry powder, 0.03 parts of red ginseng powder, 0.01 parts of American ginseng powder, 1 part of astragalus powder, 1 part of poria powder, 0.05 parts of cinnamon powder, 0.01 parts of hemp seed protein powder, 0.08 parts of patchouli powder and 2 parts of malt powder.

[0065] In the composition, the preparation method of black fish skin oligopeptide comprises the following steps:

[0066] S1: Wash fresh snakehead skin and cut it into small pieces with a side length of 1 cm to 2 cm to obtain snakehead skin small pieces; add the snakehead skin small pieces to a sodium hydroxide aqueous solution with a concentration of 0.1 mol / L according to a mass ratio of the snakehead skin small pieces: sodium hydroxide aqueous solution = 1:8, stir for 2 hours at a temperature of 40°C and a rotation speed of 200 r / min to dissolve the protein and obtain an alkali extract; adjust the pH value of the alkali extract to 4.0 with a hydrochloric acid aqueous solution with a concentration of 1 mol / L, let it stand for 30 minutes, and precipitate the protein; then centrifuge at a rotation speed of 4000 r / min for 10 minutes to obtain a protein precipitate; wash the protein precipitate with distilled water twice (add distilled water and mix, and centrifuge at a rotation speed of 4000 r / min for 10 minutes), remove the residual acid and alkali, and obtain snakehead skin protein;

[0067] S2: According to the mass ratio of black fish skin protein: phosphate buffer = 1:8, black fish skin protein is dispersed in a phosphate buffer with a pH of 7.5 and a concentration of 0.05 mol / L, 1.5% of the mass of Bacillus subtilis alkaline protease of black fish skin protein is added, and enzymolysis is carried out at 50°C and 30r / min stirring for 60min, the temperature is raised to 85°C and the enzyme is inactivated for 10min, and after cooling to room temperature, 0.5% of the mass of chymotrypsin of black fish skin protein is added, and enzymolysis is carried out at 35°C and 30r / min stirring for 60min, the temperature is raised to 85°C and the enzyme is inactivated for 10min, and after cooling to room temperature, the pH is adjusted to 5.5, and 1% of the mass of papain of black fish skin protein is added, and enzymolysis is carried out at 50°C and 30r / min stirring for 50min, the temperature is raised to 85°C and the enzyme is inactivated for 10min, and after cooling to room temperature, a sterile ultrafiltration membrane is used for filtering, and the filtrate with a molecular weight cutoff of the ultrafiltration membrane is 1000Da to 3000Da, and freeze-dried to obtain black fish skin oligopeptide.

[0068] In the composition, the preparation method of donkey-hide gelatin peptide comprises the following steps: taking donkey-hide gelatin blocks, crushing them into fine powder, passing through an 80-mesh sieve, taking the sieve to obtain donkey-hide gelatin powder, preparing the donkey-hide gelatin powder into a 5wt% donkey-hide gelatin liquid with distilled water, stirring at 40°C for 30 minutes until fully dissolved; adding 1% glucose by weight of the donkey-hide gelatin liquid, 0.5% yeast extract by weight of the donkey-hide gelatin liquid, 0.1% potassium dihydrogen phosphate by weight of the donkey-hide gelatin liquid, and 0.05% magnesium sulfate by weight of the donkey-hide gelatin liquid as the basis, adjusting the pH to 6.5 to obtain a fermentation medium; inoculating the activated Bacillus subtilis into the fermentation medium, and the inoculation amount is 1% by weight of the fermentation medium. 2%, aerobic fermentation was carried out at 35°C and 150r / min for 48h, the temperature was raised to 120°C, kept for 10min for sterilization, and then cooled to room temperature to obtain a fermentation liquid; the fermentation liquid was centrifuged at 6000r / min for 20min, the supernatant was collected, 0.5% bromelain by weight of the supernatant was added to the supernatant, enzymolysis was carried out at 50°C and 30r / min for 40min under stirring, the temperature was raised to 85°C for 10min for enzyme inactivation, and then cooled to room temperature, and filtered through a sterile ultrafiltration membrane, the ultrafiltration membrane had a molecular weight cutoff of 1000Da to 5000Da, and the filtrate was freeze-dried to obtain donkey-hide gelatin.

[0069] In the composition, the preparation method of the Cistanche deserticola extract comprises the following steps: taking Cistanche deserticola, washing and then crushing it, adding Cistanche deserticola to deionized water at a mass ratio of Cistanche deserticola to deionized water = 1:8, adjusting the pH to 4.5, adding 0.5% by mass of cellulase and 0.3% by mass of pectinase of Cistanche deserticola, and performing enzymolysis at 40°C and 30r / min stirring for 2h; then adjusting the pH to 7.0, adding 0.5% by mass of serrapeptase of Cistanche deserticola, performing enzymolysis at 45°C and 30r / min stirring for 1.5h, heating to 85°C to inactivate the enzyme for 10min, cooling to room temperature, filtering with a 200-mesh sterile sieve, taking the filtrate, and freeze-drying to obtain the Cistanche deserticola extract.

[0070] In the composition, the preparation method of the raspberry extract comprises the following steps: taking ripe raspberry fruits, washing and then crushing them, adding the raspberry fruits into deionized water at a mass ratio of raspberry fruits: deionized water = 1:8, adjusting the pH to 5.0, adding 1.5% by mass of fig protease and 0.2% by mass of amylase of raspberry fruits, and performing enzymolysis for 3 hours at 38°C and 30r / min stirring; heating to 90°C to inactivate the enzyme for 10 minutes, cooling to room temperature, filtering with a 200-mesh sterile sieve, taking the filtrate, and freeze-drying to obtain the raspberry extract.

[0071] In the composition, the preparation method of the purple eggplant peel extract comprises the following steps: taking fresh purple eggplant peel, washing and then crushing, adding the purple eggplant peel into a phosphate buffer with a pH of 5.5 and a concentration of 0.05 mol / L according to a mass ratio of purple eggplant peel: deionized water = 1:8, adding 1.0% by mass of laccase and 0.5% by mass of hemicellulase of the purple eggplant peel, and enzymolysis at 42°C and 30 r / min stirring for 2 hours; then adjusting the pH to 5.5, adding 0.8% by mass of fig protease of the purple eggplant peel, enzymolysis at 60°C and 30 r / min stirring for 1 hour, heating to 90°C to inactivate the enzyme for 10 minutes, cooling to room temperature, filtering with a 200-mesh sterile sieve, taking the filtrate, and freeze-drying to obtain the purple eggplant peel extract.

[0072] In the composition, black wolfberry powder, red ginseng powder, American ginseng powder, astragalus powder, tuckahoe powder, cinnamon powder, hemp seed protein powder, patchouli powder and malt powder are all passed through a 200-mesh sieve, and the sieved material is taken.

[0073] The preparation method of the above-mentioned blood-enriching and anti-fatigue composition comprises the following steps:

[0074] S1: Mix casein phosphopeptide, black fish skin oligopeptide, donkey-hide gelatin peptide, Cistanche deserticola extract, raspberry extract and purple eggplant peel extract in proportions by mass to obtain a mixture A;

[0075] S2: Evenly mix black wolfberry powder, red ginseng powder, American ginseng powder, astragalus powder, tuckahoe powder, cinnamon powder, hemp seed protein powder, patchouli powder and malt powder according to parts by mass to obtain a mixture B;

[0076] S3: Mix the mixture A and the mixture B uniformly to obtain a composition.

[0077] The composition is mixed with pharmaceutically acceptable auxiliary materials to prepare a product with blood-enriching and anti-fatigue effects, and the product dosage form is tablets, capsules, powders or emulsions.

[0078] Example 3

[0079] A blood-enriching and anti-fatigue composition comprises the following raw materials in parts by mass: 0.5 parts of casein phosphopeptide, 1.5 parts of black fish skin oligopeptide, 0.2 parts of donkey-hide gelatin peptide, 0.5 parts of Cistanche deserticola extract, 0.5 parts of raspberry extract, 1.5 parts of purple eggplant peel extract, 1 part of black wolfberry powder, 0.01 parts of red ginseng powder, 0.01 parts of American ginseng powder, 0.5 parts of astragalus powder, 1 part of tuckahoe powder, 0.02 parts of cinnamon powder, 0.01 parts of hemp seed protein powder, 0.05 parts of patchouli powder and 2 parts of malt powder.

[0080] In the composition, the preparation method of black fish skin oligopeptide comprises the following steps:

[0081] S1: Wash fresh snakehead skin and cut it into small pieces with a side length of 0.8 cm to 1.5 cm to obtain small pieces of snakehead skin; add the small pieces of snakehead skin to a sodium hydroxide aqueous solution with a concentration of 0.5 mol / L according to a mass ratio of small pieces of snakehead skin: sodium hydroxide aqueous solution = 1:8, stir for 2 h at a temperature of 40°C and a rotation speed of 400 r / min to dissolve the protein and obtain an alkali extract; adjust the pH value of the alkali extract to 4.0 with a hydrochloric acid aqueous solution with a concentration of 3 mol / L, leave it to stand for 60 min, and precipitate the protein; then centrifuge at a rotation speed of 4000 r / min for 20 min to obtain a protein precipitate; wash the protein precipitate with distilled water twice (add distilled water and mix, and centrifuge at a rotation speed of 4000 r / min for 20 min), remove residual acid and alkali, and obtain snakehead skin protein;

[0082] S2: According to the mass ratio of black fish skin protein: phosphate buffer = 1:8, black fish skin protein is dispersed in a phosphate buffer with a pH of 8.5 and a concentration of 0.05 mol / L, 2.5% of the mass of Bacillus subtilis alkaline protease of black fish skin protein is added, and enzymolysis is carried out at 50°C and 80r / min stirring for 60min, the temperature is raised to 95°C and the enzyme is inactivated for 10min, and after cooling to room temperature, chymotrypsin with a mass of 1% of the black fish skin protein is added, and enzymolysis is carried out at 35°C and 80r / min stirring for 60min, the temperature is raised to 95°C and the enzyme is inactivated for 10min, and after cooling to room temperature, the pH is adjusted to 6.5, papain with a mass of 1% of the black fish skin protein is added, and enzymolysis is carried out at 60°C and 30r / min stirring for 80min, the temperature is raised to 85°C and the enzyme is inactivated for 15min, and after cooling to room temperature, a sterile ultrafiltration membrane is used for filtration, and the filtrate with a molecular weight cutoff of the ultrafiltration membrane is 1000Da to 3000Da, and freeze-dried to obtain black fish skin oligopeptide.

[0083] In the composition, the preparation method of donkey-hide gelatin peptide comprises the following steps: taking donkey-hide gelatin blocks, crushing them into fine powder, passing through an 80-mesh sieve, taking the sieve to obtain donkey-hide gelatin powder, preparing the donkey-hide gelatin powder into a 10wt% donkey-hide gelatin liquid with distilled water, stirring at 40°C for 60 minutes until fully dissolved; adding 1% glucose by weight of the donkey-hide gelatin liquid, 1.5% yeast extract by weight of the donkey-hide gelatin liquid, 0.1% potassium dihydrogen phosphate by weight of the donkey-hide gelatin liquid, and 0.15% magnesium sulfate by weight of the donkey-hide gelatin liquid as the basis, adjusting the pH to 6.5, and obtaining a fermentation medium; inoculating the activated Bacillus subtilis into the fermentation medium, and the inoculation amount is 10wt% of the fermentation medium. 5%, aerobic fermentation was carried out at 35°C and 250r / min for 48h, the temperature was raised to 125°C, kept for 10min for sterilization, and cooled to room temperature to obtain a fermentation liquid; the fermentation liquid was centrifuged at 8000r / min for 20min, the supernatant was collected, 1.0% bromelain of the supernatant was added to the supernatant, enzymolysis was carried out at 50°C and 80r / min for 40min under stirring, the temperature was raised to 95°C for 10min for enzyme inactivation, and the temperature was cooled to room temperature, and the filtrate was filtered with a sterile ultrafiltration membrane, the ultrafiltration membrane had a molecular weight cutoff of 1000Da to 5000Da, and freeze-dried to obtain donkey-hide gelatin.

[0084] In the composition, the preparation method of the Cistanche deserticola extract comprises the following steps: taking Cistanche deserticola, washing and then crushing it, adding Cistanche deserticola to deionized water at a mass ratio of Cistanche deserticola to deionized water = 1:12, adjusting the pH to 4.5, adding 1.5% by mass of cellulase and 0.3% by mass of pectinase of Cistanche deserticola, and performing enzymolysis at 45°C and 30r / min stirring for 3h; then adjusting the pH to 7.0, adding 1.5% by mass of serrapeptase of Cistanche deserticola, performing enzymolysis at 45°C and 80r / min stirring for 1.5h, heating to 95°C to inactivate the enzyme for 10min, cooling to room temperature, filtering with a 250-mesh sterile sieve, taking the filtrate, and freeze-drying to obtain the Cistanche deserticola extract.

[0085] In the composition, the preparation method of the raspberry extract comprises the following steps: taking ripe raspberry fruits, washing and then crushing them, adding the raspberry fruits into deionized water at a mass ratio of raspberry fruits: deionized water = 1:8, adjusting the pH to 6.0, adding 1.5% by mass of fig protease and 0.6% by mass of amylase of raspberry fruits, performing enzymolysis at 38°C and 80r / min stirring for 3h; heating to 95°C to inactivate the enzyme for 10min, cooling to room temperature, filtering with a 250-mesh sterile sieve, taking the filtrate, and freeze-drying to obtain the raspberry extract.

[0086] In the composition, the preparation method of the purple eggplant peel extract comprises the following steps: taking fresh purple eggplant peel, washing and then crushing, adding the purple eggplant peel into a phosphate buffer with a pH of 6.5 and a concentration of 0.05 mol / L according to a mass ratio of purple eggplant peel: deionized water = 1:8, adding 2.0% by mass of laccase and 0.5% by mass of hemicellulase of the purple eggplant peel, and enzymolysis for 3 hours at 48°C and 30 r / min stirring; then adjusting the pH to 5.5, adding 1.5% by mass of fig protease of the purple eggplant peel, enzymolysis for 1 hour at 60°C and 80 r / min stirring, heating to 95°C to inactivate the enzyme for 10 minutes, cooling to room temperature, filtering with a 250-mesh sterile sieve, taking the filtrate, and freeze-drying to obtain the purple eggplant peel extract.

[0087] In the composition, black wolfberry powder, red ginseng powder, American ginseng powder, astragalus powder, tuckahoe powder, cinnamon powder, hemp seed protein powder, patchouli powder and malt powder are all passed through a 200-mesh sieve, and the sieved material is taken.

[0088] The preparation method of the above-mentioned blood-enriching and anti-fatigue composition comprises the following steps:

[0089] S1: Mix casein phosphopeptide, black fish skin oligopeptide, donkey-hide gelatin peptide, Cistanche deserticola extract, raspberry extract and purple eggplant peel extract in proportions by mass to obtain a mixture A;

[0090] S2: Evenly mix black wolfberry powder, red ginseng powder, American ginseng powder, astragalus powder, tuckahoe powder, cinnamon powder, hemp seed protein powder, patchouli powder and malt powder according to parts by mass to obtain a mixture B;

[0091] S3: Mix the mixture A and the mixture B uniformly to obtain a composition.

[0092] The composition is mixed with pharmaceutically acceptable auxiliary materials to prepare a product with blood-enriching and anti-fatigue effects, and the product dosage form is tablets, capsules, powders or emulsions.

[0093] Example 4

[0094] A blood-enriching and anti-fatigue composition comprises the following raw materials in parts by mass: 2 parts of casein phosphopeptide, 1.5 parts of black fish skin oligopeptide, 0.5 parts of donkey-hide gelatin peptide, 0.5 parts of Cistanche deserticola extract, 1.5 parts of raspberry extract, 1.5 parts of purple eggplant peel extract, 3 parts of black wolfberry powder, 0.01 parts of red ginseng powder, 0.03 parts of American ginseng powder, 0.5 parts of astragalus powder, 3 parts of tuckahoe powder, 0.02 parts of cinnamon powder, 0.03 parts of hemp seed protein powder, 0.05 parts of patchouli powder and 5 parts of malt powder.

[0095] In the composition, the preparation method of black fish skin oligopeptide comprises the following steps:

[0096] S1: fresh snakehead skin is cleaned and cut into small pieces with a side length of 0.5 cm to 2 cm to obtain snakehead skin small pieces; according to the mass ratio of snakehead skin small pieces: sodium hydroxide aqueous solution=1:12, the snakehead skin small pieces are added to a sodium hydroxide aqueous solution with a concentration of 0.5 mol / L, stirred for 4 hours at a temperature of 60°C and a rotation speed of 400 r / min, so that the protein is dissolved to obtain an alkali extract; the pH value of the alkali extract is adjusted to 6.0 with a hydrochloric acid aqueous solution with a concentration of 3 mol / L, and the alkali extract is allowed to stand for 60 minutes to precipitate the protein; then centrifuged at a rotation speed of 6000 r / min for 20 minutes to obtain a protein precipitate; the protein precipitate is washed 3 times with distilled water (after adding distilled water and mixing, centrifuged at a rotation speed of 6000 r / min for 20 minutes), and residual acid and alkali are removed to obtain snakehead skin protein;

[0097] S2: According to the mass ratio of black fish skin protein: phosphate buffer = 1:10, black fish skin protein is dispersed in a phosphate buffer with a pH of 8.5 and a concentration of 0.15 mol / L, 2.5% of the mass of Bacillus subtilis alkaline protease of black fish skin protein is added, and enzymolysis is carried out at 60°C and 80r / min stirring for 90min, the temperature is raised to 95°C and the enzyme is inactivated for 15min, and after cooling to room temperature, chymotrypsin with a mass of 1% of the black fish skin protein is added, and enzymolysis is carried out at 38°C and 80r / min stirring for 90min, the temperature is raised to 95°C and the enzyme is inactivated for 15min, and after cooling to room temperature, the pH is adjusted to 6.5, papain with a mass of 1.5% of the black fish skin protein is added, and enzymolysis is carried out at 60°C and 80r / min stirring for 80min, the temperature is raised to 95°C and the enzyme is inactivated for 15min, and after cooling to room temperature, a sterile ultrafiltration membrane is used for filtration, and the filtrate with a molecular weight cutoff of the ultrafiltration membrane is 1000Da to 3000Da, and freeze-dried to obtain black fish skin oligopeptide.

[0098] In the composition, the preparation method of donkey-hide gelatin peptide comprises the following steps: taking donkey-hide gelatin blocks, crushing them into fine powder, passing through a 100-mesh sieve, taking the sieve to obtain donkey-hide gelatin powder, preparing the donkey-hide gelatin powder into a 10wt% donkey-hide gelatin liquid with distilled water, stirring at 50°C for 60 minutes until fully dissolved; adding 3% glucose by weight of the donkey-hide gelatin liquid, 1.5% yeast extract by weight of the donkey-hide gelatin liquid, 0.3% potassium dihydrogen phosphate by weight of the donkey-hide gelatin liquid, and 0.15% magnesium sulfate by weight of the donkey-hide gelatin liquid as the basis, adjusting the pH to 7.5, and obtaining a fermentation medium; inoculating the activated Bacillus subtilis into the fermentation medium, and the inoculation amount is 10wt% of the fermentation medium medium. The mixture was centrifuged at 8000 r / min for 30 min, and the supernatant was collected. 1.0% bromelain of the supernatant was added to the supernatant, and the mixture was enzymolyzed at 60°C and 80 r / min for 60 min. The mixture was heated to 95°C and the enzyme was inactivated for 15 min. The mixture was cooled to room temperature and filtered through a sterile ultrafiltration membrane. The filtrate had a molecular weight cutoff of 1000 Da to 5000 Da. The mixture was freeze-dried to obtain donkey-hide gelatin.

[0099] In the composition, the preparation method of the Cistanche deserticola extract comprises the following steps: taking Cistanche deserticola, washing and then crushing it, adding Cistanche deserticola to deionized water at a mass ratio of Cistanche deserticola to deionized water = 1:12, adjusting the pH to 5.5, adding 1.5% by mass of cellulase and 1.0% by mass of pectinase of Cistanche deserticola, and performing enzymolysis at 45°C and 80r / min stirring for 3h; then adjusting the pH to 8.0, adding 1.5% by mass of serrapeptase of Cistanche deserticola, performing enzymolysis at 55°C and 80r / min stirring for 2h, heating to 95°C to inactivate the enzyme for 15min, cooling to room temperature, filtering with a 250-mesh sterile sieve, taking the filtrate, and freeze-drying to obtain the Cistanche deserticola extract.

[0100] In the composition, the preparation method of the raspberry extract comprises the following steps: taking ripe raspberry fruits, washing and then crushing them, adding the raspberry fruits into deionized water at a mass ratio of raspberry fruits: deionized water = 1:12, adjusting the pH to 6.0, adding 3% of the mass of fig protease and 0.6% of the mass of amylase of raspberry fruits, and performing enzymolysis for 4 hours at 45°C and 80r / min stirring; heating to 95°C to inactivate the enzyme for 15 minutes, cooling to room temperature, filtering with a 250-mesh sterile sieve, taking the filtrate, and freeze-drying to obtain the raspberry extract.

[0101] In the composition, the preparation method of the purple eggplant peel extract comprises the following steps: taking fresh purple eggplant peel, washing and then crushing, adding the purple eggplant peel into a phosphate buffer with a pH of 6.5 and a concentration of 0.15 mol / L according to a mass ratio of purple eggplant peel: deionized water = 1:12, adding 2.0% by mass of laccase and 1.0% by mass of hemicellulase of the purple eggplant peel, and enzymolysis for 3 hours at 48°C and 80 r / min stirring; then adjusting the pH to 6.0, adding 1.5% by mass of fig protease of the purple eggplant peel, and enzymolysis for 1.5 hours at 65°C and 80 r / min stirring, heating to 95°C to inactivate the enzyme for 15 minutes, cooling to room temperature, filtering with a 250-mesh sterile sieve, taking the filtrate, and freeze-drying to obtain the purple eggplant peel extract.

[0102] In the composition, black wolfberry powder, red ginseng powder, American ginseng powder, astragalus powder, tuckahoe powder, cinnamon powder, hemp seed protein powder, patchouli powder and malt powder are all passed through a 250-mesh sieve, and the sieved material is taken.

[0103] The preparation method of the above-mentioned blood-enriching and anti-fatigue composition comprises the following steps:

[0104] S1: Mix casein phosphopeptide, black fish skin oligopeptide, donkey-hide gelatin peptide, Cistanche deserticola extract, raspberry extract and purple eggplant peel extract in proportions by mass to obtain a mixture A;

[0105] S2: Evenly mix black wolfberry powder, red ginseng powder, American ginseng powder, astragalus powder, tuckahoe powder, cinnamon powder, hemp seed protein powder, patchouli powder and malt powder according to parts by mass to obtain a mixture B;

[0106] S3: Mix the mixture A and the mixture B uniformly to obtain a composition.

[0107] The composition is mixed with pharmaceutically acceptable auxiliary materials to prepare a product with blood-enriching and anti-fatigue effects, and the product dosage form is tablets, capsules, powders or emulsions.

[0108] Example 5

[0109] A blood-enriching and anti-fatigue composition comprises the following raw materials in parts by mass: 2 parts of casein phosphopeptide, 3 parts of black fish skin oligopeptide, 0.5 parts of donkey-hide gelatin peptide, 1.5 parts of Cistanche deserticola extract, 1.5 parts of raspberry extract, 3 parts of purple eggplant peel extract, 3 parts of black wolfberry powder, 0.03 parts of red ginseng powder, 0.03 parts of American ginseng powder, 1 part of astragalus powder, 3 parts of poria powder, 0.05 parts of cinnamon powder, 0.03 parts of hemp seed protein powder, 0.08 parts of patchouli powder and 5 parts of malt powder.

[0110] In the composition, the preparation method of black fish skin oligopeptide comprises the following steps:

[0111] S1: fresh snakehead skin is cleaned and cut into small pieces with a side length of 0.6 cm to 1.8 cm to obtain snakehead skin small pieces; according to the mass ratio of snakehead skin small pieces: sodium hydroxide aqueous solution=1:12, the snakehead skin small pieces are added to a sodium hydroxide aqueous solution with a concentration of 0.1 mol / L, stirred for 4 h at a temperature of 60°C and a rotation speed of 200 r / min, so that the protein is dissolved to obtain an alkali extract; the pH value of the alkali extract is adjusted to 6.0 with a hydrochloric acid aqueous solution with a concentration of 1 mol / L, and the alkali extract is allowed to stand for 30 min to precipitate the protein; then centrifuged at a rotation speed of 6000 r / min for 10 min to obtain a protein precipitate; the protein precipitate is washed 3 times with distilled water (after adding distilled water and mixing, centrifuged at a rotation speed of 6000 r / min for 20 min), and residual acid and alkali are removed to obtain snakehead skin protein;

[0112] S2: According to the mass ratio of black fish skin protein: phosphate buffer = 1:10, black fish skin protein is dispersed in a phosphate buffer with a pH of 7.5 and a concentration of 0.15 mol / L, 1.5% of the mass of Bacillus subtilis alkaline protease of black fish skin protein is added, and enzymolysis is carried out at 60°C and 30r / min stirring for 90min, the temperature is increased to 85°C and the enzyme is inactivated for 15min, and after being cooled to room temperature, 0.5% of the mass of chymotrypsin of black fish skin protein is added, and enzymolysis is carried out at 38°C and 30r / min stirring for 90min, the temperature is increased to 85°C and the enzyme is inactivated for 15min, and after being cooled to room temperature, the pH is adjusted to 5.5, and 1.5% of the mass of papain of black fish skin protein is added, and enzymolysis is carried out at 50°C and 80r / min stirring for 50min, the temperature is increased to 95°C and the enzyme is inactivated for 10min, and after being cooled to room temperature, a sterile ultrafiltration membrane is used for filtering, and the filtrate with a molecular weight cutoff of the ultrafiltration membrane is 1000Da to 3000Da, and freeze-dried to obtain black fish skin oligopeptide.

[0113] In the composition, the preparation method of donkey-hide gelatin peptide comprises the following steps: taking donkey-hide gelatin blocks, crushing them into fine powder, passing through a 100-mesh sieve, taking the sieve to obtain donkey-hide gelatin powder, preparing the donkey-hide gelatin powder into a 5wt% donkey-hide gelatin liquid with distilled water, stirring at 50°C for 30 minutes until fully dissolved; adding 3% glucose by weight of the donkey-hide gelatin liquid, 0.5% yeast extract by weight of the donkey-hide gelatin liquid, 0.3% potassium dihydrogen phosphate by weight of the donkey-hide gelatin liquid, and 0.05% magnesium sulfate by weight of the donkey-hide gelatin liquid as the basis, adjusting the pH to 7.5 to obtain a fermentation medium; inoculating the activated Bacillus subtilis into the fermentation medium, and the inoculation amount is 100% by weight of the fermentation medium. 2%, aerobic fermentation was carried out at 38°C and 150r / min for 72h, the temperature was raised to 120°C, sterilized for 15min, and cooled to room temperature to obtain a fermentation liquid; the fermentation liquid was centrifuged at 6000r / min for 30min, the supernatant was collected, 0.5% bromelain by weight of the supernatant was added to the supernatant, enzymolysis was carried out at 60°C and 30r / min with stirring for 60min, the temperature was raised to 85°C for 15min to inactivate the enzyme, and the temperature was cooled to room temperature, and the filtrate was filtered with a sterile ultrafiltration membrane, the ultrafiltration membrane had a molecular weight cutoff of 1000Da to 5000Da, and freeze-dried to obtain donkey-hide gelatin.

[0114] In the composition, the preparation method of the Cistanche deserticola extract comprises the following steps: taking Cistanche deserticola, washing and then crushing it, adding Cistanche deserticola to deionized water at a mass ratio of Cistanche deserticola to deionized water = 1:8, adjusting the pH to 5.5, adding 0.5% by mass of cellulase and 1.0% by mass of pectinase of Cistanche deserticola, and performing enzymolysis at 40°C and 80r / min stirring for 2h; then adjusting the pH to 8.0, adding 0.5% by mass of serrapeptase of Cistanche deserticola, performing enzymolysis at 55°C and 30r / min stirring for 2h, heating to 85°C to inactivate the enzyme for 15min, cooling to room temperature, filtering with a 200-mesh sterile sieve, taking the filtrate, and freeze-drying to obtain the Cistanche deserticola extract.

[0115] In the composition, the preparation method of the raspberry extract comprises the following steps: taking ripe raspberry fruits, washing and then crushing them, adding the raspberry fruits into deionized water at a mass ratio of raspberry fruits: deionized water = 1:12, adjusting the pH to 5.0, adding 3% of the mass of fig protease and 0.2% of the mass of amylase of raspberry fruits, and performing enzymolysis at 45°C and 30r / min stirring for 4h; heating to 90°C to inactivate the enzyme for 15min, cooling to room temperature, filtering with a 200-mesh sterile sieve, taking the filtrate, and freeze-drying to obtain the raspberry extract.

[0116] In the composition, the preparation method of the purple eggplant peel extract comprises the following steps: taking fresh purple eggplant peel, washing and then crushing, adding the purple eggplant peel into a phosphate buffer with a pH of 5.5 and a concentration of 0.15 mol / L according to a mass ratio of purple eggplant peel: deionized water = 1:12, adding 1.0% by mass of laccase and 1.0% by mass of hemicellulase of the purple eggplant peel, and enzymolysis at 42°C and 80 r / min stirring for 2 hours; then adjusting the pH to 6.0, adding 0.8% by mass of fig protease of the purple eggplant peel, enzymolysis at 65°C and 30 r / min stirring for 1.5 hours, heating to 90°C to inactivate the enzyme for 15 minutes, cooling to room temperature, filtering with a 200-mesh sterile sieve, taking the filtrate, and freeze-drying to obtain the purple eggplant peel extract.

[0117] In the composition, black wolfberry powder, red ginseng powder, American ginseng powder, astragalus powder, tuckahoe powder, cinnamon powder, hemp seed protein powder, patchouli powder and malt powder are all passed through a 250-mesh sieve, and the sieved material is taken.

[0118] The preparation method of the above-mentioned blood-enriching and anti-fatigue composition comprises the following steps:

[0119] S1: Mix casein phosphopeptide, black fish skin oligopeptide, donkey-hide gelatin peptide, Cistanche deserticola extract, raspberry extract and purple eggplant peel extract in proportions by mass to obtain a mixture A;

[0120] S2: Evenly mix black wolfberry powder, red ginseng powder, American ginseng powder, astragalus powder, tuckahoe powder, cinnamon powder, hemp seed protein powder, patchouli powder and malt powder according to parts by mass to obtain a mixture B;

[0121] S3: Mix the mixture A and the mixture B uniformly to obtain a composition.

[0122] The composition is mixed with pharmaceutically acceptable auxiliary materials to prepare a product with blood-enriching and anti-fatigue effects, and the product dosage form is tablets, capsules, powders or emulsions.

[0123] The sources of raw materials used in the above embodiments are as follows: casein phosphopeptide comes from Shanxi Hengtian Biotechnology Co., Ltd.; black wolfberry powder comes from Xi'an Furuida Biotechnology Co., Ltd.; red ginseng powder comes from Shanxi Bohong Biotechnology Co., Ltd.; American ginseng powder comes from Shaanxi Shengheng Biotechnology Co., Ltd.; astragalus powder comes from Shaanxi Shengheng Biotechnology Co., Ltd.; Poria powder comes from Shaanxi Shengheng Biotechnology Co., Ltd.; cinnamon powder comes from Shaanxi Shengheng Biotechnology Co., Ltd.; hemp seed protein powder comes from Shanxi Tongling Biotechnology Co., Ltd.; patchouli powder comes from Shaanxi Shengheng Biotechnology Co., Ltd.; malt powder comes from Shaanxi Shengheng Biotechnology Co., Ltd.

[0124] The sources of enzymes, bacteria and auxiliary materials used in the above embodiments are as follows: the enzyme activity of Bacillus subtilis alkaline protease is 200,000 U / g, which is sourced from Dongguan Varisi Chemical Co., Ltd.; the enzyme activity of papain is 200,000 U / g, which is sourced from Shaanxi Xulin Borui Biotechnology Co., Ltd.; the enzyme activity of bromelain is 100,000 U / g, which is sourced from Shaanxi Xulin Borui Biotechnology Co., Ltd.; the enzyme activity of cellulase is 100,000 U / g, which is sourced from Shandong Jurong Bioengineering Co., Ltd.; the enzyme activity of pectinase is 100,000 U / g, which is sourced from Shandong Jurong Bioengineering Co., Ltd.; the enzyme activity of serrapeptidase is 100,000 U / g, sourced from Jiangsu Aofu Biotechnology Co., Ltd.; the enzyme activity of ficin is 100,000 U / g, sourced from Changsha Shenghe Bioengineering Co., Ltd.; the enzyme activity of amylase is 2000 U / g, sourced from Jiangsu Aofu Biotechnology Co., Ltd.; the enzyme activity of laccase is 10,000 U / g, sourced from Henan Anrui Biotechnology Co., Ltd.; the enzyme activity of hemicellulase is 100,000 U / g, sourced from Henan Anrui Biotechnology Co., Ltd.; the bacterial activity of Bacillus subtilis is 100 billion CFU / g, sourced from Jinan Jinyuyuan Biotechnology Co., Ltd.; the yeast extract is sourced from Henan Yiduoyuan Biotechnology Co., Ltd.

[0125] Comparative Example 1

[0126] No black fish skin oligopeptide is added to the composition, and the content of black fish skin oligopeptide is replaced by malt powder; other parameters and methods are the same as in Example 1.

[0127] Comparative Example 2

[0128] The donkey-hide gelatin peptide in the composition is replaced by donkey-hide gelatin; other parameters and methods are the same as in Example 1.

[0129] Comparative Example 3

[0130] The purple eggplant peel extract is not added to the composition, and the content of the purple eggplant peel extract is replaced by malt powder; other parameters and methods are the same as in Example 1.

[0131] Comparative Example 4

[0132] In the method for preparing black fish skin oligopeptide, chymotrypsin and papain are not used for enzymatic hydrolysis; other parameters and methods are the same as in Example 1.

[0133] Comparative Example 5

[0134] In the method for preparing black fish skin oligopeptide, Bacillus subtilis alkaline protease and chymotrypsin are not used for enzymatic hydrolysis; other parameters and methods are the same as in Example 1.

[0135] Comparative Example 6

[0136] In the preparation method of donkey-hide gelatin peptide, bromelain is not used for enzymatic hydrolysis; other parameters and methods are the same as those in Example 1.

[0137] Comparative Example 7

[0138] In the preparation method of donkey-hide gelatin peptide, Bacillus subtilis is not used for fermentation, and bromelain is directly used to enzymolyze the donkey-hide gelatin liquid; other parameters and methods are the same as in Example 1.

[0139] Comparative Example 8

[0140] In the preparation method of the Cistanche deserticola extract, serrapeptase is not used for enzymatic hydrolysis; other parameters and methods are the same as those in Example 1.

[0141] Comparative Example 9

[0142] In the preparation method of raspberry extract, ficin is not used for enzymatic hydrolysis; other parameters and methods are the same as in Example 1.

[0143] Comparative Example 10

[0144] In the preparation method of purple eggplant peel extract, laccase and ficin are not used for enzymolysis; other parameters and methods are the same as in Example 1.

[0145] The compositions of the above-mentioned embodiments and comparative examples were tested for their efficacy.

[0146] 1. Safety Testing

[0147] (I) Acute toxicity test

[0148] Experimental animals: Healthy adult SPF Kunming mice weighing 18-22 g were selected, half male and half female, with 10 mice in each group.

[0149] Dosage and route of administration: 5 mg / 20 g body weight of the composition was administered by oral gavage, and a blank control group was set up to be administered with an equal mass of physiological saline.

[0150] Observation indicators and time: The mice were observed for 14 consecutive days after administration, and the symptoms of toxic reactions were recorded. Results: The activities, diet, fur gloss, mental state, breathing, feces, etc. were normal, and there was no death; each composition had good safety.

[0151] (II) Long-term toxicity test

[0152] Experimental animals: Healthy SD rats weighing 120-150 g, half male and half female, were selected and divided into a high-dose group (administered 6 mg / 20 g body weight of the composition), a medium-dose group (administered 4 mg / 20 g body weight of the composition), a low-dose group (administered 2 mg / 20 g body weight of the composition) and a blank control group (no composition was administered), 20 rats in each group.

[0153] Dosage and route of administration: Different doses of the composition were administered by oral gavage for 90 consecutive days.

[0154] Observation index and time: During the administration period, the general condition of the rats was regularly observed: body weight, food intake, water intake, physical signs, and behavioral activities; Histopathological examination: At the end of the experiment, the rats were killed, and the main organs were fixed, sliced, and stained to observe pathological changes. Results: The daily behavioral activities were normal, and there were no obvious specific lesions in the histopathological examination. Each composition has good safety.

[0155] 2. Efficacy Testing

[0156] (I) Blood-enriching efficacy test

[0157] Establishment of animal model and grouping: Female Kunming mice were selected, and an anemia model was established by intraperitoneal injection of cyclophosphamide 100 mg / kg. The anemic mice were randomly divided into a model control group, a positive control group (given ferrous sulfate 60 mg / kg body weight) and a test group (given the composition 50 mg / kg body weight), with 10 mice in each group; a normal control group (10 mice) was also set up.

[0158] Administration and detection indexes: The test group was given oral administration of the corresponding dose of each composition, the positive control group was given oral administration of ferrous sulfate, and the normal control group and the model control group were given normal saline, and the administration was continued for 14 days. Blood was collected from the retro-orbital venous plexus of mice on the day before administration and on the 14th day after administration for detection of red blood cell count (RBC), hemoglobin content (Hb), hematocrit (HCT), serum ferritin (SF), and serum erythropoietin (EPO). The test results are shown in Table 1 below:

[0159] Table 1 Blood-enriching effect test results (average value of 10 animals)

[0160]

[0161]

[0162] It can be seen from the above results that the compositions of Examples 1 to 5 have an efficient and stable blood-enriching effect. It can be seen from the results of Comparative Examples 1, 4 and 5 that black fish skin oligopeptide has a good blood-enriching effect. Changing the preparation method of black fish skin oligopeptide affects the components of the polypeptide product and reduces the blood-enriching effect. It can be seen from the results of Comparative Examples 2, 6 and 7 that the blood-enriching effect can be greatly improved after donkey-hide gelatin is prepared into donkey-hide gelatin peptide. Changing the preparation method of donkey-hide gelatin peptide affects the components of the polypeptide product and reduces the blood-enriching effect. It can be seen from the results of Comparative Examples 3 and 10 that purple eggplant peel extract has a good blood-enriching effect. Changing the preparation method of purple eggplant peel extract affects different product components and reduces the blood-enriching effect. It can be seen from the results of Comparative Example 8 that changing the preparation method of Cistanche deserticola extract has different products, which affects the blood-enriching effect. It can be seen from the results of Comparative Example 9 that changing the preparation method of raspberry extract has different products, which affects the blood-enriching effect.

[0163] (II) Anti-fatigue efficacy test

[0164] Animal model establishment and grouping: Male SD rats were selected, and a fatigue model was constructed by a weighted swimming experiment. The swimming time of rats to exhaustion was 300s-600s as the fatigue model. Rats in the fatigue model were randomly divided into a model control group, a positive control group (administered ginsenoside 80 mg / kg body weight) and a test group (administered the composition 80 mg / kg body weight), with 10 rats in each group; a normal control group (rats without modeling) was also set up.

[0165] Administration and testing indicators: The test group was gavaged with each composition, the positive control group was gavaged with ginsenosides, and the model control group and the normal control group were given normal saline, and the administration was continued for 28 days. 30 minutes after the last administration, the rats were placed in a swimming box for weighted swimming (a lead block of 5% of their body weight was attached to the tail of the rats), and the swimming exhaustion time of the rats was recorded (i.e., the rats sank to the bottom of the water for more than 10 seconds and could not float to the surface). At the same time, the blood of the rats was collected immediately after the end of swimming to detect fatigue-related indicators such as blood lactate (BLA), serum urea nitrogen (SUN), liver glycogen (LG), and muscle glycogen (MG). The test results are shown in Table 2 below:

[0166] Table 2 Anti-fatigue efficacy test results (average value of 10 animals)

[0167]

[0168] It can be seen from the above results that the compositions of Examples 1 to 5 have excellent anti-fatigue effects. It can be seen from the results of Comparative Examples 1, 4 and 5 that black fish skin oligopeptides have a good anti-fatigue effect. Changing the preparation method of black fish skin oligopeptides affects the components of the polypeptide product and reduces the anti-fatigue effect. It can be seen from the results of Comparative Examples 2, 6 and 7 that the anti-fatigue effect can be greatly improved after donkey-hide gelatin is prepared into donkey-hide gelatin peptides. Changing the preparation method of donkey-hide gelatin peptides affects the components of the polypeptide product and reduces the anti-fatigue effect. It can be seen from the results of Comparative Examples 3 and 10 that purple eggplant peel extract has a good anti-fatigue effect. Changing the preparation method of purple eggplant peel extract affects different product components and reduces the anti-fatigue effect. It can be seen from the results of Comparative Example 8 that changing the preparation method of Cistanche deserticola extract has different products, which affects the reduction of anti-fatigue effect. It can be seen from the results of Comparative Example 9 that changing the preparation method of raspberry extract has different products, which affects the reduction of anti-fatigue effect.

Claims

1. A blood-enriching and anti-fatigue composition, characterized in that: The composition comprises the following raw materials in parts by weight: 0.5 to 2 parts of casein phosphopeptide, 1.5 to 3 parts of black fish skin oligopeptide, 0.2 to 0.5 parts of donkey-hide gelatin peptide, 0.5 to 1.5 parts of Cistanche deserticola extract, 0.5 to 1.5 parts of raspberry extract, 1.5 to 3 parts of purple eggplant peel extract, 1 to 3 parts of black wolfberry powder, 0.01 to 0.03 parts of red ginseng powder, 0.01 to 0.03 parts of American ginseng powder, 0.5 to 1 parts of astragalus powder, 1 to 3 parts of tuckahoe powder, 0.02 to 0.05 parts of cinnamon powder, 0.01 to 0.03 parts of hemp seed protein powder, 0.05 to 0.08 parts of patchouli powder and 2 to 5 parts of malt powder; The black fish skin oligopeptide is a black fish skin oligopeptide with a molecular weight of 1000Da to 3000Da obtained by subjecting black fish skin protein to graded enzymolysis with Bacillus subtilis alkaline protease, chymotrypsin and papain.

2. A blood-enriching and anti-fatigue composition according to claim 1, characterized in that: The preparation method of the black fish skin oligopeptide comprises the following steps: dispersing the black fish skin protein in a phosphate buffer solution with a pH value of 7.5 to 8.5 and a concentration of 0.05 mol / L to 0.15 mol / L according to a mass ratio of black fish skin protein to phosphate buffer solution = 1: (8 to 10), adding 1.5% to 2.5% by mass of Bacillus subtilis alkaline protease of the black fish skin protein, performing enzymolysis for 60 to 90 minutes at 50 to 60° C. and 30 to 80 r / min under stirring, heating to 85 to 95° C. to inactivate the enzyme for 10 to 15 minutes, cooling to room temperature, adding 0.5% to 1% by mass of chymotrypsin of the black fish skin protein, and The method comprises the following steps: performing enzymolysis for 60 to 90 minutes at 35 to 38 degrees Celsius and 30 to 80 r / min under stirring conditions, heating the temperature to 85 to 95 degrees Celsius to inactivate the enzyme for 10 to 15 minutes, cooling the temperature to room temperature, adjusting the pH to 5.5 to 6.5, adding papain in an amount of 1% to 1.5% by weight of black fish skin protein, performing enzymolysis for 50 to 80 minutes at 50 to 60 degrees Celsius and 30 to 80 r / min under stirring conditions, heating the temperature to 85 to 95 degrees Celsius to inactivate the enzyme for 10 to 15 minutes, cooling the temperature to room temperature, filtering the mixture with a sterile ultrafiltration membrane, and performing freeze-drying to obtain a black fish skin oligopeptide.

3. A blood-enriching and anti-fatigue composition according to claim 2, characterized in that: The method for preparing black fish skin protein comprises the following steps: washing fresh black fish skin, cutting it into small pieces with a side length ranging from 0.5 cm to 2 cm, so as to obtain small pieces of black fish skin; adding the small pieces of black fish skin to a sodium hydroxide aqueous solution with a concentration of 0.1 mol / L to 0.5 mol / L according to a mass ratio of small pieces of black fish skin to sodium hydroxide aqueous solution = 1: (8 to 12), stirring for 2 h to 4 h at a temperature of 40° C. to 60° C. and a rotation speed of 200 r / min to 400 r / min to dissolve the protein, so as to obtain an alkali extract; adjusting the pH value of the alkali extract to 4.0 to 6.0 with a hydrochloric acid aqueous solution with a concentration of 1 mol / L to 3 mol / L, standing for 30 min to 60 min, so as to precipitate the protein; and then centrifuging for 10 min to 20 min at a rotation speed of 4000 r / min to 6000 r / min to obtain a protein precipitate; The protein precipitate was washed with distilled water for 2 to 3 times to remove the residual acid and alkali to obtain black fish skin protein.

4. The blood-enriching and anti-fatigue composition according to claim 1, characterized in that: The preparation method of donkey-hide gelatin peptide comprises the following steps: taking donkey-hide gelatin blocks, crushing them into fine powder, passing through a 80-100 mesh sieve, taking the sieve to obtain donkey-hide gelatin powder, preparing the donkey-hide gelatin powder into a 5wt%-10wt% donkey-hide gelatin liquid with distilled water, stirring at 40°C-50°C for 30min-60min until fully dissolved; adding 1%-3% glucose by weight of the donkey-hide gelatin liquid, 0.5%-1.5% yeast extract by weight of the donkey-hide gelatin liquid, 0.1%-0.3% potassium dihydrogen phosphate by weight of the donkey-hide gelatin liquid, and 0.05%-0.15% magnesium sulfate by weight of the donkey-hide gelatin liquid as a basis, adjusting the pH to 6.5-7.5, and obtaining a fermentation medium; inoculating the activated Bacillus subtilis into the fermentation medium in an inoculation amount of 2%-5% of the mass of the fermentation medium, and culturing the culture medium at a temperature of 35°C-38°C and 15 The method comprises the following steps: aerobic fermentation is performed at 0 r / min to 250 r / min for 48 h to 72 h, the temperature is raised to 120° C. to 125° C., sterilized for 10 min to 15 min, and cooled to room temperature to obtain a fermentation liquid; the fermentation liquid is centrifuged at 6000 r / min to 8000 r / min for 20 min to 30 min, the supernatant is collected, 0.5% to 1.0% by weight of bromelain is added to the supernatant, enzymolysis is performed at 50° C. to 60° C. and 30 r / min to 80 r / min under stirring for 40 min to 60 min, the temperature is raised to 85° C. to 95° C. to inactivate the enzyme for 10 min to 15 min, the temperature is lowered to room temperature, the filtrate is filtered through a sterile ultrafiltration membrane, the ultrafiltration membrane has a molecular weight cutoff of 1000 Da to 5000 Da, and the filtrate is freeze-dried to obtain donkey-hide gelatin peptide.

5. The blood-enriching and anti-fatigue composition according to claim 1, characterized in that: The preparation method of the Cistanche deserticola extract comprises the following steps: taking Cistanche deserticola, washing it and then crushing it, adding Cistanche deserticola to deionized water in a mass ratio of Cistanche deserticola to deionized water = 1: (8-12), adjusting the pH to 4.5-5.5, adding 0.5%-1.5% by mass of cellulase and 0.3%-1.0% by mass of pectinase to Cistanche deserticola, and performing enzymolysis for 2h-3h at 40-45°C and 30r / min-80r / min under stirring; then adjusting the pH to 7.0-8.0, adding 0.5%-1.5% by mass of serrapeptase to Cistanche deserticola, and stirring at 45-55°C and 30r / min-80r / min n The enzyme was hydrolyzed for 1.5 h to 2 h under stirring, and the temperature was raised to 85°C to 95°C for 10 min to 15 min to inactivate the enzyme. After cooling to room temperature, the enzyme was filtered through a sterile sieve, and the filtrate was taken and freeze-dried to obtain the Cistanche deserticola extract.

6. The blood-enriching and anti-fatigue composition according to claim 1, characterized in that: The preparation method of the raspberry extract comprises the following steps: taking ripe raspberry fruits, washing and then crushing them, adding the raspberry fruits into deionized water according to the mass ratio of raspberry fruits: deionized water=1:(8-12), adjusting the pH to 5.0-6.0, adding 1.5%-3% by mass of fig protease and 0.2%-0.6% by mass of amylase of the raspberry fruits, performing enzymolysis for 3h-4h at 38-45℃ and 30r / min-80r / min stirring; heating to 90-95℃ to inactivate the enzyme for 10-15min, cooling to room temperature, filtering with a sterile mesh, taking the filtrate, and freeze-drying to obtain the raspberry extract.

7. The blood-enriching and anti-fatigue composition according to claim 1, characterized in that: The preparation method of the purple eggplant peel extract comprises the following steps: taking fresh purple eggplant peel, washing and then crushing, adding the purple eggplant peel to a phosphate buffer solution with a pH of 5.5 to 6.5 and a concentration of 0.05 mol / L to 0.15 mol / L according to a mass ratio of purple eggplant peel to deionized water = 1: (8 to 12), adding 1.0% to 2.0% by mass of laccase and 0.5% to 1.0% by mass of hemicellulase, and heating at 42° C. to 48° C. and 30 r. / min~80r / min stirring state, enzymolysis for 2h~3h; then adjust the pH to 5.5~6.0, add 0.8%~1.5% of the mass of purple eggplant peel ficin, enzymolysis for 1h~1.5h at 60℃~65℃ and 30r / min~80r / min stirring state, heat to 90℃~95℃ to inactivate the enzyme for 10min~15min, cool to room temperature, filter with a sterile sieve, take the filtrate, and lyophilize to obtain the purple eggplant peel extract.

8. A blood-enriching and anti-fatigue composition according to claim 5, 6 or 7, characterized in that: The mesh number of the sieve is 200-250 meshes.

9. The blood-enriching and anti-fatigue composition according to claim 1, characterized in that: The black wolfberry powder, red ginseng powder, American ginseng powder, astragalus powder, tuckahoe powder, cinnamon powder, hemp seed protein powder, patchouli powder and malt powder are all passed through a 200-250 mesh sieve, and the sieved material is taken.

10. A method for preparing a blood-enriching and anti-fatigue composition, used for preparing the blood-enriching and anti-fatigue composition according to claim 1, characterized in that: The preparation method comprises the following steps: S1: Mix casein phosphopeptide, black fish skin oligopeptide, donkey-hide gelatin peptide, Cistanche deserticola extract, raspberry extract and purple eggplant peel extract in proportions by mass to obtain a mixture A; S2: Evenly mix black wolfberry powder, red ginseng powder, American ginseng powder, astragalus powder, tuckahoe powder, cinnamon powder, hemp seed protein powder, patchouli powder and malt powder according to parts by mass to obtain a mixture B; S3: Mix the mixture A and the mixture B uniformly to obtain a composition.

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