Bovine bone marrow oil and bovine bone collagen oligopeptide calcium powder as well as preparation method and application thereof

Abstract

The invention discloses bovine bone marrow oil, bovine bone collagen oligopeptide calcium powder and a preparation method and application thereof.The preparation method comprises the following steps that firstly, bovine bones are crushed, then the crushed bovine bones are put into a subcritical ultrasonic extractor to be extracted to obtain bovine bone marrow oil thick paste, then the bovine bone marrow oil thick paste is subjected to supercritical extraction through a supercritical extractor to obtain bovine bone marrow oil, and the bovine bone marrow oil thick paste is obtained; adding the deoiled bone residues into a continuous countercurrent ultrasonic extractor for ultrasonic extraction, removing residues, adjusting the pH value of the obtained extracting solution by using alkali liquor, adding a special enzyme for bone protein hydrolysis for hydrolysis, removing precipitates and insoluble substances after enzyme deactivation, filtering and separating enzymatic hydrolysate by using an ultrafiltration membrane, filtering and separating by using a nanofiltration membrane, and finally concentrating the residual filtrate by using a reverse osmosis membrane and vacuum reduced pressure concentration to obtain the bone protein. And drying the concentrated thick liquid by a drying machine, crushing, and sieving to obtain the bovine bone collagen peptide calcium powder. According to the method, the extraction rate of the bovine bone marrow oil is improved through efficient combination of the subcritical extractor and the ultrasonic extractor, the bovine bone oil and peculiar smell which influence subsequent production are effectively removed for efficient extraction of the bovine bone collagen, and a high-added-value product is obtained.

Description

Technical Field

[0001] The present invention relates to the technical field of food processing, in particular to bovine bone marrow oil, bovine collagen oligopeptide calcium powder and preparation methods and applications thereof. Background Art

[0002] With the advancement and development of the times, calcium supplementation has become a basic health care task in people's minds. Since the loss of calcium in the human body can lead to osteoporosis, which can easily cause fractures and other risks, calcium supplementation has become a major issue of public concern. Our daily calcium supplementation essentially supplements calcium (glucosamine calcium) and collagen. Collagen can be extracted from many organisms, most commonly fish skin, pig skin, and cow bones. Comparing the three materials, cow bones are more suitable than collagen and collagen oligopeptides extracted from fish skin and pig skin.

[0003] Bovine bone has the highest collagen content and density among common biomaterials. Its rich collagen content and stable structure are transformed into small-molecule protein peptides that can be directly absorbed and utilized by the human body through enzymatic hydrolysis with a complex protease. These bovine bone collagen oligopeptides are more suitable for the deposition of bone calcium, promoting rapid bone growth.

[0004] Patent CN109247584A discloses a bovine collagen peptide powder suitable for astronaut nutrition and its preparation method. The bovine collagen peptide powder is prepared through multiple processes including raw material preparation, crushing, cleaning, drying, first crushing, second crushing, high-pressure cooking, high-pressure homogenization, enzymatic hydrolysis, concentration, and spray drying.

[0005] This method separates oil through high-pressure steaming, and its ability to remove oil is weak. The oil produces an odor that affects subsequent normal production. After enzymatic hydrolysis, it will also produce more fatty acid calcium, which is not amino acid chelated calcium or polypeptide chelated calcium that is easily absorbed and utilized by the human body.

[0006] Bovine bone marrow oil is also rich in nutrients, such as acylcarnitines, sphingomyelins, and phosphatidylcholine (lecithin), which has anti-inflammatory and antioxidant effects, including lowering blood lipids, preventing vascular diseases, improving brain and nerve function, improving lipid metabolism, and providing anti-inflammatory and antioxidant benefits. Bovine bone marrow oil is rich in triglycerides (TAG) (93.84%) and a small amount of diglycerides (DAG) (2.89%). Diglycerides, especially 1,3-diglyceride, are an emerging healthy edible oil that can lower serum and liver cholesterol levels, reduce triglyceride levels in the body, and reduce fat accumulation. Therefore, separating the bovine bone marrow oil to the greatest extent possible during the preparation of bovine collagen peptide powder not only helps improve the extraction rate of chelated calcium, but also produces high-value-added bovine bone marrow oil. Summary of the Invention

[0007] The purpose of the present invention is to overcome the shortcomings of the prior art and provide bovine bone marrow oil, bovine collagen oligopeptide calcium powder and their preparation method and application, so as to solve the problems of low bovine bone collagen extraction yield, low bovine bone collagen oligopeptide conversion rate and residual bovine bone oil affecting subsequent production during the production of bovine bone collagen.

[0008] The purpose of the present invention is achieved through the following technical solution: A method for preparing bovine bone marrow oil and bovine collagen oligopeptide calcium powder comprises the following steps:

[0009] S1. Wash and remove the meat from fresh beef leg bones, then crush them into bone fragments using a bone crusher.

[0010] S2. The crushed cattle bones were put into a subcritical ultrasonic extractor to obtain a bovine bone marrow oily paste;

[0011] S3. The thick paste of bovine bone marrow oil was extracted by supercritical extraction to obtain bovine bone marrow oil;

[0012] S4. The de-oiled bone residue was put into a continuous countercurrent ultrasonic extractor for ultrasonic extraction, and the extract was obtained after removing the residue;

[0013] S5. The extract was adjusted with alkali solution to pH, and a bone protein hydrolyzing enzyme was added to hydrolyze the extract. After the enzyme was inactivated, the precipitate and insoluble matter were removed.

[0014] S6. The enzymatic hydrolyzate is first separated by ultrafiltration membrane filtration, and then separated by nanofiltration membrane filtration;

[0015] S7. The filtrate is concentrated by reverse osmosis membrane, vacuum decompression concentration, and the concentrated thick liquid is dried in a dryer, crushed, and sieved to obtain bovine collagen peptide calcium powder.

[0016] Preferably, in step S1, the particle size of the bovine bone fragments is 1-5 mm.

[0017] Preferably, in step S2, subcritical ultrasonic extraction is performed for 30 minutes.

[0018] Preferably, in step S4, ultrasonic extraction is performed at 50-60° C. for 60 min.

[0019] Preferably, in step S5, the pH is adjusted to 6.0-8.0 with 0.5-2.0 mol / L dilute alkali solution, the enzymatic hydrolysis temperature is controlled at 55-65° C., the amount of bone protein hydrolysis enzyme is 0.1-0.3%, the enzymatic hydrolysis time is controlled at 2-6 h, and the enzyme is inactivated by boiling water for 10-15 min after enzymatic hydrolysis.

[0020] Preferably, in step S6, the ultrafiltration membrane is 3000D and the nanofiltration membrane is 200D.

[0021] A bovine bone marrow oil prepared by any of the above methods.

[0022] The bovine collagen oligopeptide calcium powder obtained by any of the above methods has a bovine collagen peptide recovery rate of more than 65%, a peptide content with a molecular weight less than 1000Da of more than 85%, and an organic calcium content of more than 45 mg / g.

[0023] The invention discloses an application of bovine bone collagen oligopeptide calcium powder in eucommia ulmoides bovine bone collagen oligopeptide beauty and skin care nutrient powder, which comprises eucommia ulmoides leaves, cordyceps militaris, kudzu root, white poria, ganoderma lucidum, liquorice and bovine collagen oligopeptide.

[0024] The invention discloses an application of bovine collagen oligopeptide calcium powder in kudzu root bovine collagen oligopeptide calcium nutritional powder for relieving fatigue and improving bone density, comprising kudzu root, cordyceps militaris, eucommia leaf, white poria, ginseng, wolfberry fruit and bovine collagen oligopeptide.

[0025] The present invention has the following advantages:

[0026] 1. The efficient combination of a subcritical extractor and an ultrasonic extractor solves the low extraction rate problem caused by the existing high-temperature cooking method for removing beef bone oil. It also effectively removes beef bone oil and odor that affect subsequent production for efficient extraction of beef bone collagen, while turning waste into valuable, highly nutritious, high-value-added beef bone marrow oil. Subcritical extraction features low temperature, low pressure, and high efficiency. While efficiently extracting beef bone marrow oil, it effectively protects the heat-sensitive active ingredients in the beef bones, ensuring the effective extraction of subsequent active ingredients.

[0027] 2. Using de-oiled bone residue, a continuous countercurrent ultrasonic extractor is used to ultrasonically extract bovine bone collagen. An ultrasonic generator is used to emit ultrasonic waves into the extraction solvent. The cavitation effect and mechanical action of the ultrasonic waves in the extraction solvent can effectively break up the bovine bone cells, freeing the active ingredients and dissolving them into the extraction solvent. It can also accelerate the molecular movement of the extraction solvent, allowing the active ingredients in the extraction solvent and the bovine bones to quickly contact, dissolve, and mix with each other, which is beneficial to the leaching of bovine bone collagen. At the same time, the continuous countercurrent ultrasonic extractor is a modern high-tech means of extraction that achieves high efficiency, energy saving, and environmental protection.

[0028] 3. Utilizing a scientifically screened enzyme specifically designed to hydrolyze bone protein and an enzymatic hydrolysis process determined through single-factor, orthogonal experiments, the conversion rate of bovine bone collagen peptides is increased, the off-flavor of the product is effectively reduced, and the product meets the quality requirements for use as a food additive. This patent converts large amounts of bovine bone collagen, which is beneficial to the human body but difficult to absorb, into collagen oligopeptides (small molecule peptides) that are beneficial to human absorption. These collagen oligopeptides have excellent solubility, emulsification, and biological activity (physiological activity, transport physiological activity, carrier physiological activity, dynamic physiological activity, and transmitter physiological activity, etc.) (for example, bovine bone collagen oligopeptides bind to metal ions of calcium to promote the body's absorption of mineral calcium). This improves the functional properties of the protein and enhances the efficacy of all functional elements of bovine bone, enabling its wider application in functional foods.

[0029] 4. Use subcritical ultrasonic extractors, continuous countercurrent ultrasonic extractors for efficient extraction, horizontal screw centrifuges, butterfly centrifuges for efficient separation, special enzyme directional enzymatic cutting, ultrafiltration membrane separation, nanofiltration membrane desalination separation and concentration, vacuum low-temperature belt continuous dryers and other advanced equipment for seamless connection to achieve the automation and scale of bovine bone marrow oil and bovine collagen oligopeptide calcium production.

[0030] 5. Leveraging the outstanding benefits of bovine collagen oligopeptide calcium, combined with extracts from medicinal and edible plants, we formulate formulations such as Eucommia ulmoides bovine collagen oligopeptide for skincare and Pueraria lobata bovine collagen oligopeptide calcium for fatigue relief and bone density improvement. Bovine collagen oligopeptide fully unleashes the benefits of bovine collagen oligopeptide, while the mineral calcium, carried by bovine collagen oligopeptide, is absorbed and fully utilized. The rational organic compatibility of these formulations fully demonstrates the unique efficacy of each of these ingredients. DETAILED DESCRIPTION

[0031] In order to make the purpose, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below in combination with the specific experimental process of the present invention.

[0032] 1Experiment on optimization of preparation process of bovine collagen oligopeptide

[0033] 1.1 Single factor investigation

[0034] 1.1.1 Effect of oil yield of fresh beef bones

[0035] 200 g of fresh beef bones were extracted using solvent method, ultrasound-assisted method, cooking method, steam method, enzyme method and subcritical-supercritical method. The oil yield was calculated and the results are shown in Table 1.

[0036] Table 1 Effect of oil yield of fresh beef bones (n=3)

[0037]

[0038] Table 1 shows that the oil yield from fresh beef bones ranks in the order of subcritical-supercritical extraction, ultrasonic-assisted extraction, solvent extraction, steam extraction, and cooking extraction. The solvent extraction method uses solvents such as light gasoline and is suitable for industrial production. However, its disadvantages are that it is flammable and explosive, requiring strict production environment requirements. Consumers are also concerned about the presence of small amounts of solvent residue in the product. The ultrasonic-assisted extraction method uses solvents such as light gasoline and is suitable for industrial production. However, its disadvantages are that it is flammable and explosive, requiring strict production environment requirements, and consumer concerns about the presence of small amounts of solvent residue in the product. The steam extraction method and cooking extraction method are not suitable for industrial production due to their low yields. The subcritical-supercritical extraction method offers high extraction yields, eliminates solvent residues, and is suitable for large-scale and industrial production. However, its disadvantage is that the equipment is expensive. It is suitable for use in the comprehensive utilization of beef bones.

[0039] 1.1.2 Effect of oil extraction method on protein content in raw materials

[0040] 200 g of fresh beef bones were extracted using solvent method, ultrasound-assisted method, cooking method, steam method, enzyme method and subcritical-supercritical method. Protein was extracted from the residue after oil extraction. The extraction rate results are shown in Table 2.

[0041] Table 2 Effect of oil extraction method on protein extraction effect in raw materials (n=3)

[0042]

[0043] Table 2 shows that the protein extraction efficiency of the residual residue after subcritical-supercritical extraction of bovine bone marrow oil from fresh bovine bones follows the order of subcritical-supercritical extraction > ultrasound-assisted extraction > solvent extraction > steam extraction > cooking extraction. The subcritical-supercritical extraction and ultrasound-assisted extraction methods facilitate protein extraction from the residual bovine bone due to the ultrasonic effect. However, the steam extraction and cooking extraction methods are hampered by the high temperature damage to the protein. The solvent extraction and ultrasound-assisted extraction methods require the complete removal of residual solvent, otherwise the protein extraction will be affected.

[0044] 1.1.3 Effect of temperature on enzymatic hydrolysis

[0045] The residue after extracting bovine bone marrow oil was extracted by subcritical-supercritical method, the enzyme dosage was 2.5 mg / g, the pH value was about 6.5, the enzymatic hydrolysis time was 4 h, and the enzymatic hydrolysis temperature was 50℃, 55℃, 60℃, 65℃, and 70℃ (enzymatic hydrolysis was performed in parallel for 3 times, the same below). The results are shown in Table 3.

[0046] Table 3 Effect of temperature on enzymatic hydrolysis effect (n=3)

[0047]

[0048] Table 3 shows that the peptide extraction rate from the hydrolyzate increases with increasing temperature. This is because the enzyme activity gradually increases with temperature, leading to an increase in the hydrolysis rate. As the temperature continues to rise, the peptide extraction rate from the hydrolyzate decreases, meaning the hydrolysis rate decreases. This is because high temperature denatures the enzyme, weakening its activity and reducing the hydrolysis rate. Enzyme activity is maximum at 60°C, resulting in the highest hydrolysis rate.

[0049] 1.1.4 Effect of pH on enzymatic hydrolysis

[0050] The residue after extracting bovine bone marrow oil was extracted by subcritical-supercritical method. The water bath temperature was 60℃, the enzyme dosage was 2.5 mg / g, the enzymatic hydrolysis time was 4 h, the pH value was adjusted to 5.0, 6.0, 7.0, 8.0, and 9.0, and the operation was performed. The results are shown in Table 4.

[0051] Table 4 Effect of pH value on enzymatic hydrolysis effect (n=3)

[0052]

[0053] Table 4 shows that the peptide extraction rate in the enzymatic hydrolysis solution increases with increasing pH. This is because changes in pH have a significant impact on the enzymatic hydrolysis process. At lower pH values, enzymatic activity increases with increasing pH. At pH 7.0, the enzyme has the highest activity and enzymatic hydrolysis rate. As the pH value continues to rise, the peptide extraction rate in the enzymatic hydrolysis solution decreases, that is, the enzymatic hydrolysis rate decreases. This is because the high pH value denatures the enzyme, weakening its activity and reducing the enzymatic hydrolysis rate.

[0054] 1.1.5 Effect of enzyme dosage on enzymatic hydrolysis effect

[0055] The residue after extracting bovine bone marrow oil was extracted by subcritical-supercritical method. The operation was carried out under the conditions of water bath temperature of 60℃, pH value of 6.5, enzymatic hydrolysis time of 4h, and bone protein hydrolysis enzyme dosage of 1.5, 2.0, 2.5, 3.0, and 3.5 mg / g. The results are shown in Table 5.

[0056] Table 5 Effect of enzyme dosage on enzymatic hydrolysis effect (n=3)

[0057]

[0058] Table 5 shows that the peptide extraction rate from the hydrolyzate increases with increasing enzyme dosage, i.e., the enzymatic hydrolysis rate increases, reaching its highest level at an enzyme dosage of 2.5 mg / g. As the enzyme dosage continues to increase, the peptide extraction rate from the hydrolyzate changes little, i.e., the enzymatic hydrolysis rate remains unchanged. Considering production costs, an enzyme dosage of 2.5 mg / g is recommended.

[0059] 1.1.6 Effect of time on enzymatic hydrolysis effect

[0060] The residue after extracting bovine bone marrow oil was extracted by subcritical-supercritical method, with the operation under the conditions of water bath temperature of 60℃, pH value of 6.5, enzyme dosage of 2.5mg / g, and enzymatic hydrolysis time of 1, 2, 4, 6, and 8h. The results are shown in Table 6.

[0061] Table 6 Effect of time on enzymatic hydrolysis effect (n=3)

[0062]

[0063] Table 6 shows that as the enzymatic hydrolysis time increases, the polypeptide extraction rate in the hydrolyzate gradually increases. That is, the enzymatic hydrolysis rate increases, reaching its highest level after 4 hours of hydrolysis. Further extension of the hydrolysis time results in essentially no change in the polypeptide extraction rate in the hydrolyzate. This is because the enzyme requires a certain amount of time to interact with the raw material. If the time is too short, the enzyme cannot fully interact with the raw material, and the optimal enzymatic hydrolysis rate cannot be achieved. If the time is too long, the enzyme and raw material have already fully reacted, and further extension of the time will not increase the enzymatic hydrolysis rate. Therefore, considering production costs, an enzymatic hydrolysis time of 4-6 hours is appropriate.

[0064] 1.2 Orthogonal test

[0065] The residue after extracting bovine bone marrow oil using the subcritical-supercritical method was used as raw material, and bovine bone protein was extracted by ultrasound. The orthogonal experimental results of protein hydrolysis using a special enzyme for bone protein hydrolysis are shown in Table 7; the visual analysis results of protein hydrolysis using the special enzyme for bone protein hydrolysis are shown in Table 8; the variance analysis results of protein hydrolysis using the special enzyme for bone protein hydrolysis are shown in Table 9.

[0066] Table 7 Orthogonal table of protein hydrolysis by special enzymes for bone protein hydrolysis L9 (3 4 ) and results

[0067]

[0068] Table 8 Results of visual analysis of protein hydrolysis by special enzyme for bone protein hydrolysis

[0069]

[0070]

[0071] Note: a K is the sum of the results of a certain level of each factor; corner numbers 1-3 are the experimental levels of each factor;

[0072] bk is the average value of the sum of the results of a certain level of each factor;

[0073] c G is the sum of the experimental yields of the designed 9 groups;

[0074] d CT = G2 / 9;

[0075] e S is the sum of squared deviations.

[0076] Analysis of Variance Results of Protein Hydrolysis by Special Enzyme for Bone Protein Hydrolysis

[0077]

[0078] Note: F0.01(2, 2) = 99.0, F0.05(2, 2) = 19.0, F0.1(2, 2) = 9.0. e is the error, and * indicates no significance.

[0079] Results of Orthogonal Experiment

[0080] From the intuitive analysis of Table 8, it can be seen that the factor with the largest range is Factor A, followed by Factor B, Factor D, and Factor C. This indicates that the factor having the greatest impact on the yield of protein hydrolysis by the special enzyme for bone protein hydrolysis is A, followed by B, D, and C. From the analysis of variance results in Table 9: Factors A, B, D, and C have no significant meaning (F < F0.1). Therefore, according to the change rate of each factor k, Factor A is selected at level 2, Factor B is selected at level 2, Factor C is selected at level 2, and Factor D is selected at level 2. Referring to the results of single-factor experiments, the determined conditions are A2B2C2D2.

[0081] 1.3 Verification of the Optimal Process Conditions of the Enzyme Pretreatment Method

[0082] Through the above single-factor experiments and orthogonal experiments, the process conditions for extracting bovine bone marrow oil, extracting protein, and the optimal enzyme hydrolysis treatment from fresh bovine bones have been determined. That is, using the residue after extracting bovine bone marrow oil by the subcritical-supercritical method as the raw material, ultrasonic extraction of bovine bone protein, hydrolysis of protein with the special enzyme for bone protein hydrolysis, pH value of 6.5, enzyme dosage of 2.5 mg / g, water bath temperature of 60 °C, and enzyme hydrolysis time of 4 h. According to this process, 100.00 g of raw material of crushed bovine bone powder was weighed, enzymatically hydrolyzed according to the method, and at the same time, a blank control of the raw material of crushed bovine bone powder without enzymatic hydrolysis was made. Each enzymatically hydrolyzed solution and non-enzymatically hydrolyzed solution were measured according to the method, and the results are shown in Table 10.

[0083] Table 10 Verification Results of the Optimal Process Conditions of the Enzyme Pretreatment Method

[0084]

[0085] As can be seen from Table 10, under the same enzymatic hydrolysis conditions, the extraction rate of protein peptides from enzymatically hydrolyzed crushed bovine bone powder is 4.9 times that of the raw material without enzymatic hydrolysis. This shows that enzymatic hydrolysis treatment is beneficial to the extraction of protein peptides from bovine bones.

[0086] 2 Conclusions

[0087] 2.1 Taking the oil yield of fresh bovine bones as the investigation index, the research results show that the subcritical-supercritical method > ultrasonic-assisted method > solvent method > steam method > cooking method. The subcritical-supercritical method has a high extraction rate, no solvent residue, and can be produced on a large scale and industrially. The disadvantage is that the equipment is expensive, and it is suitable for use in the comprehensive utilization production of bovine bones.

[0088] 2.2 Using the protein extraction rate from fresh bovine bone as an evaluation indicator, the results showed that the protein extraction rate of the residue after subcritical-supercritical extraction of bovine bone marrow oil from fresh bovine bone was in the following order: subcritical-supercritical method > ultrasound-assisted method > solvent method > steam method > cooking method. The subcritical-supercritical method facilitates the extraction of protein from the bovine bone residue due to the effect of ultrasound.

[0089] 2.3 Taking the polypeptide extraction rate in bovine bone hydrolysate as the evaluation index, the research results show that the optimal enzymatic hydrolysis process conditions are: enzymatic hydrolysis temperature 60℃, enzymatic hydrolysis pH 6.5, enzymatic hydrolysis dosage 2.5mg / g, enzymatic hydrolysis time 4h, and solid-liquid ratio 1:15.

[0090] 2.4 Compared with non-enzymatically hydrolyzed raw materials, the protein peptide extraction rate of bovine bone after enzymatic hydrolysis was 4.9 times that of non-enzymatically hydrolyzed raw materials. This shows that the enzyme specifically for hydrolyzing bone protein can be used in the enzymatic hydrolysis process to extract protein peptides from bovine bone.

[0091] 3. Obtain the best preparation method based on the optimized selected conditions

[0092] A bovine bone marrow oil and bovine collagen oligopeptide calcium powder is prepared according to the following method, comprising the following steps:

[0093] S1. Wash and remove the meat from fresh beef shanks. Chop the bones into small pieces of appropriate size and crush them into small pieces with a bone crusher to a particle size of 1 to 5 mm.

[0094] S2. The crushed cattle bones were put into a subcritical ultrasonic extractor and extracted with subcritical ultrasonic technology for 30 minutes to obtain a thick oily paste of bovine bone marrow;

[0095] S3. The thick paste of bovine bone marrow oil was extracted by supercritical extraction to obtain bovine bone marrow oil;

[0096] S4. The de-oiled bone residue was put into a continuous countercurrent ultrasonic extractor for ultrasonic extraction at 50-60°C for 60 min, and the extract was removed by centrifugation in a decanter centrifuge to obtain an extract;

[0097] S5. The extract was poured into the enzymatic hydrolysis tank and the pH was adjusted to 6.0-8.0 with a dilute alkali solution of 0.5-2.0 mol / L. The enzymatic hydrolysis temperature was controlled at 55-65°C, the amount of bone protein hydrolysis enzyme was 0.1-0.3%, the enzymatic hydrolysis time was controlled at 2-6h, and the enzyme was inactivated by boiling water for 10-15min after enzymatic hydrolysis. The precipitate and insoluble matter were removed by centrifugation in a butterfly centrifuge.

[0098] S6. The enzymatic hydrolysate was separated using a 3000D ultrafiltration membrane to remove macromolecular components, and the filtrate was separated using a 200D nanofiltration membrane to remove small molecular components and inorganic salts;

[0099] S7. The filtrate is concentrated by reverse osmosis membrane and vacuum decompression concentration. The concentrated thick liquid is dried by low-temperature vacuum belt dryer, crushed, and sieved to obtain bovine collagen peptide calcium powder.

[0100] The obtained bovine collagen peptide calcium powder was measured, and the results showed that the recovery rate of bovine collagen peptide was over 65%, the content of peptides with a molecular weight of less than 1000 Da was over 85%, and the organic calcium content was over 45 mg / g.

[0101] 4 Applications

[0102] 4.1 The obtained bovine bone collagen peptide calcium powder is used to prepare Eucommia bovine bone collagen oligopeptide beauty and skin care nutritional powder. The preparation method is as follows:

[0103] First, weigh the following components in proportion: 160-240g of eucommia leaves, 40-60g of cordyceps militaris, 80-120g of kudzu root, 40-60g of white poria, 40-60g of ganoderma lucidum, 40-60g of licorice, 10g of bovine collagen oligopeptide, 9.6g of inulin, 5g of maltodextrin, 0.3g of citric acid, and 0.1g of stevioside. Put the weighed eucommia leaves, cordyceps militaris, kudzu root, white poria, ganoderma lucidum, and licorice into a multifunctional extraction tank, add 8 times the amount of purified water, heat reflux extraction for 120min, filter, use 6 times the amount of purified water to extract the residue, heat reflux extraction for 120min, filter, combine the two extracts, clarify with chitosan clarifier, filter, concentrate the filtrate under reduced pressure, spray dry, add bovine collagen oligopeptide, inulin, maltodextrin, citric acid, and stevioside into the extract powder, mix thoroughly, bag, sterilize, and produce the finished product.

[0104] 4.2 The obtained bovine collagen peptide calcium powder is used to prepare Pueraria bovine collagen oligopeptide calcium nutritional powder for relieving fatigue and improving bone density. The preparation method is as follows:

[0105] First, weigh the following components according to the proportion: 120-180g of kudzu root, 40-60g of cordyceps militaris, 80-120g of eucommia leaf, 40-60g of white poria, 40-60g of ginseng, 80-120g of wolfberry, 10g of bovine collagen oligopeptide, 9.6g of inulin, 5g of maltodextrin, 0.3g of citric acid, and 0.1g of stevioside. Put the kudzu root, cordyceps militaris, eucommia leaf, white poria, ginseng, and wolfberry weighed according to the formula into a multifunctional extraction tank, add 8 times the amount of pure water, heat reflux extraction for 120min, filter, use 6 times the amount of pure water to extract the residue, heat reflux extraction for 120min, filter, combine the two extracts, clarify with chitosan clarifier, filter, concentrate the filtrate under reduced pressure, spray dry, add bovine collagen oligopeptide, inulin, maltodextrin, citric acid, and stevioside into the extract powder, mix thoroughly, bag, sterilize, and produce the finished product.

[0106] Although the present invention has been described in detail with reference to the aforementioned embodiments, it is still possible for those skilled in the art to modify the technical solutions described in the aforementioned embodiments, or to make equivalent substitutions for some of the technical features therein. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A method for preparing bovine bone marrow oil and bovine collagen oligopeptide calcium powder, characterized in that: The steps include: S1. Wash and remove the meat from fresh beef leg bones, then crush them into bone fragments using a bone crusher. S2. The crushed cattle bones were put into a subcritical ultrasonic extractor to obtain a bovine bone marrow oily paste; S3. The thick paste of bovine bone marrow oil was extracted by supercritical extraction to obtain bovine bone marrow oil; S4. The de-oiled bone residue was put into a continuous countercurrent ultrasonic extractor for ultrasonic extraction, and the extract was obtained after removing the residue; S5. The extract was adjusted with alkali solution to pH, and a bone protein hydrolyzing enzyme was added to hydrolyze the extract. After the enzyme was inactivated, the precipitate and insoluble matter were removed. S6. The enzymatic hydrolyzate is first separated by ultrafiltration membrane filtration, and then separated by nanofiltration membrane filtration; S7. The filtrate is concentrated by reverse osmosis membrane, vacuum decompression concentration, and the concentrated thick liquid is dried in a dryer, crushed, and sieved to obtain bovine collagen peptide calcium powder.

2. The method for preparing bovine bone marrow oil and bovine collagen oligopeptide calcium powder according to claim 1, wherein: In step S1, the particle size of the bovine bone is 1-5 mm.

3. The method for preparing bovine bone marrow oil and bovine collagen oligopeptide calcium powder according to claim 1, wherein: In step S2, subcritical ultrasonic extraction is performed for 30 minutes.

4. The method for preparing bovine bone marrow oil and bovine collagen oligopeptide calcium powder according to claim 1, wherein: In step S4, ultrasonic extraction is performed at 50-60°C for 60 minutes.

5. The method for preparing bovine bone marrow oil and bovine collagen oligopeptide calcium powder according to claim 1, wherein: In step S5, the pH is adjusted to 6.0-8.0 with 0.5-2.0 mol / L dilute alkali solution, the enzymatic hydrolysis temperature is controlled at 55-65°C, the amount of bone protein hydrolysis enzyme is 0.1-0.3%, the enzymatic hydrolysis time is controlled at 2-6 hours, and the enzyme is inactivated by boiling water for 10-15 minutes after enzymatic hydrolysis.

6. The method for preparing bovine bone marrow oil and bovine collagen oligopeptide calcium powder according to claim 1, wherein: In step S6, the ultrafiltration membrane is selected as 3000D, and the nanofiltration membrane is selected as 200D.

7. A bovine bone marrow oil obtained according to any one of claims 1 to 3.

8. A bovine collagen oligopeptide calcium powder obtained according to any one of claims 1 to 6, characterized in that: The recovery rate of bovine collagen peptides is over 65%, the content of peptides with a molecular weight of less than 1000Da is over 85%, and the organic calcium content is over 45mg / g.

9. The use of a bovine bone collagen oligopeptide calcium powder according to claim 8 in the Eucommia ulmoides bovine bone collagen oligopeptide beauty and skin care nutritional powder, characterized in that: Including Eucommia ulmoides leaves, Cordyceps militaris, Pueraria root, White Poria cocos, Ganoderma lucidum, Licorice, and bovine collagen oligopeptides.

10. The use of the bovine collagen oligopeptide calcium powder according to claim 8 in the kudzu root bovine collagen oligopeptide calcium nutritional powder for relieving fatigue and improving bone density, characterized in that: Including Pueraria root, Cordyceps militaris, Eucommia ulmoides leaves, white Poria cocos, ginseng, wolfberry, and bovine collagen oligopeptide.

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