Composition containing bovine collagen peptide and preparation method thereof

Abstract

The invention belongs to the field of food, and specifically relates to a composition containing bovine bone collagen peptide and a preparation method thereof. The components of the composition include bovine cartilage collagen peptide chelate, a complex of yak bone collagen peptide and chitosan oligosaccharide, enzymatic bone powder, cartilage extract, sucralose, and oligomaltose, and the composition is prepared by mixing the components. The bovine bone collagen peptide composition of the invention has a fresh smell, high market acceptance, high storage stability, strong gastric acid or enzyme resistance, strong stability, high bioavailability, and low-cost and readily available raw materials for preparation, and is easy to industrialize.

Description

Technical Field

[0001] The invention belongs to the field of food, and particularly relates to a composition containing bovine collagen peptide and a preparation method thereof. Background Art

[0002] Collagen peptides are a product obtained from livestock and poultry bones under the action of acid, alkali, heat treatment or enzymatic hydrolysis. Supplementing collagen peptides can increase the regularity and firmness of the body's collagen fiber network, promote the orderly deposition of calcium salts, and increase bone density and bone strength. At least 30 types of collagen have been discovered and confirmed, and the most in-depth research is currently on type I, type II and type III collagen. Bovine bones are rich in collagen, which can be used to prepare collagen peptides. However, during the high-value processing of bovine collagen peptides, there will be a variety of volatile components including butyric acid, acetic acid, propionic acid, etc., or carry sulfhydryls and thiomethyl groups, etc., which produce some unpleasant fishy smells, which seriously restrict the consumption and promotion of bovine collagen peptide products. At the same time, collagen peptides are highly nutritious substances. They are easily contaminated by debris and microorganisms during long-term storage, producing more and heavier odors such as ammonia, which not only affects appetite, but also brings immeasurable health risks. Summary of the invention

[0003] In order to solve the problem in the prior art that bovine collagen peptide products often have peculiar smell and are difficult to store, the present invention mainly provides a composition containing bovine collagen peptides and a preparation method of the composition containing bovine collagen peptides, and the technical scheme thereof is as follows:

[0004] A composition containing bovine collagen peptides comprises the following components by mass: 40-80 parts of bovine cartilage collagen peptide chelates, 1-4 parts of a complex of yak bone collagen peptides and chitosan oligosaccharides, 0.5-2 parts of enzymatic bone powder, 0.5-2 parts of cartilage extracts, 0.01-0.1 parts of sucralose, and 15-25 parts of maltooligosaccharides.

[0005] Furthermore, the bovine cartilage collagen peptide chelate is formed by chelating type II bovine cartilage collagen peptide with ferrous ions.

[0006] A preparation method of the above-mentioned composition containing bovine bone collagen peptides comprises the following steps: removing volatile components in bovine cartilage collagen peptides, completing pretreatment, and then chelating and coating the bovine cartilage collagen peptides with iron ions to obtain bovine cartilage collagen peptide chelates; compounding yak bone collagen peptides with chitosan oligosaccharides and chelating calcium ions to form a complex of yak bone collagen peptides and chitosan oligosaccharides; and mixing modified bovine cartilage collagen peptides, the complex of yak bone collagen peptides and chitosan oligosaccharides, and other components to obtain the composition.

[0007] Furthermore, the preparation of the bovine cartilage collagen peptide chelate comprises the following steps:

[0008] a. Place bovine cartilage collagen peptide in an aqueous solution of ethanol and fully disperse it to form a 1-2 g / mL solution, add calcium bicarbonate to the solution, adjust the pH to 8.5-10, microwave at 100-200w for 1-3h, separate and remove the precipitate, and obtain a pretreated bovine cartilage collagen peptide solution;

[0009] b. Take the pretreated bovine cartilage collagen peptide solution, adjust the pH value to 5-5.5 with sodium acetate buffer solution, add ferrous sulfate, mix thoroughly for 1-2 hours, add ethanol to the system, mix thoroughly until no precipitation is produced, separate and collect the precipitate, and obtain the iron-containing bovine cartilage collagen peptide precursor;

[0010] c. Prepare a dopamine solution, disperse the iron-containing bovine cartilage collagen peptide precursor in the dopamine solution, stir the reaction at 800-1500 rpm for 10-20 hours, filter and wash the filter cake, and obtain the product after freeze-drying.

[0011] Furthermore, the volume concentration of the ethanol aqueous solution in step a is 5-10%; the mass ratio of the ferrous sulfate in step b to the bovine cartilage collagen peptide in step a is 1:2-5; the concentration of the iron-containing bovine cartilage collagen peptide precursor in the dopamine solution in step c is 5-10 mg / mL; and the volume concentration of the ethanol added in step b in the system is 75-99%.

[0012] Furthermore, the freeze-drying in step c is pre-freezing below -18°C, and then vacuum drying at -40 to -80°C and 10 to 16 Pa for 18 to 24 hours.

[0013] Furthermore, in step c, the dopamine solution is prepared by adding dopamine hydrochloride to a 10 mmol / L Tris hydrochloric acid buffer solution at pH=8.5, and dissolving the dopamine hydrochloride to obtain a 2 mg / mL dopamine solution.

[0014] Furthermore, the preparation of the complex of yak bone collagen peptide and chitosan oligosaccharide includes the following steps: dissolving chitosan oligosaccharide in water, adding yak bone collagen peptide, mixing thoroughly, then adding calcium chloride, adjusting the pH to neutral with ammonia buffer solution, heating to 60-80°C for reaction for 24-48h, using ethanol to precipitate and separate the product from the reaction system, fully washing the precipitate, and drying to obtain the product.

[0015] Furthermore, the mass ratio of the chitosan oligosaccharide to the yak bone collagen peptide is 1:0.8-1.2; the mass ratio of the calcium chloride to the chitosan oligosaccharide is 1:1-2.5.

[0016] Furthermore, the method comprises the following steps: sieving the powdered components through a 40-mesh sieve, weighing the components according to the formula amount, and mixing the components for 25 to 35 minutes to obtain a mixed powder; and quantitatively packaging the mixed powder in a 100,000-grade clean environment, with a loading error within ±3%.

[0017] By adopting the above scheme, the method of the present invention has the following advantages:

[0018] 1. The bovine collagen peptide-containing composition of the present invention has a fresh smell, high market acceptance, high storage stability, strong resistance to gastric acid or enzymes, strong stability, high bioavailability, and low cost and readily available raw materials for preparation, and is easy to industrialize.

[0019] 2. The present invention uses calcium bicarbonate to process bovine cartilage collagen peptides, which can more specifically remove acidic odorous substances. The raw materials are simple, and calcium bicarbonate will not affect the activity of bovine bone collagen peptides. It can also help bovine cartilage collagen peptides increase bone density and bone strength.

[0020] 3. In the composition of the present invention, the content of bovine cartilage collagen peptide accounts for a large proportion. The use of iron and bovine cartilage collagen peptide for competitive chelation, and the difference in solubility between ferrous sulfate and calcium sulfate can partially reduce the large amount of calcium ions introduced by the previous deodorization treatment, thereby avoiding the loss of iron caused by the body's intake of a large amount of calcium ions.

[0021] 4. The bovine cartilage collagen peptide of the present invention is chelated with calcium and iron, which can produce an antibacterial effect, improve the storage time, and improve the stability of the bovine cartilage collagen peptide, reducing the impact of the environment that causes it to continue to produce odorous substances.

[0022] 5. The present invention coats the chelated bovine cartilage collagen peptide with polydopamine to further reduce the contact between the bovine cartilage collagen peptide and the outside world, improve storage stability, and improve the stability of the bovine cartilage collagen peptide chelate in the gastrointestinal tract, thereby improving the bioavailability of the collagen peptide and calcium and iron ions.

[0023] 6. The present invention combines yak bone collagen peptide with chitosan oligosaccharide through calcium ions, which reduces the odor of yak bone collagen peptide and occupies the active groups of yak bone collagen peptide and chitosan oligosaccharide, while increasing the stability of both, saving the cost of processing each substance separately.

[0024] 7. The yak bone collagen peptide and chitosan oligosaccharide of the present invention chelate calcium ions simultaneously, increasing the chelation sites, achieving full utilization of the raw materials, and being able to supplement the calcium element contained in the composition, coordinating and balancing the content of calcium ions and iron ions within the formula amount range of the bovine cartilage collagen peptide chelate. DETAILED DESCRIPTION

[0025] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0026] Example 1: (1) Bovine cartilage collagen peptide was placed in an aqueous solution of 5% ethanol by volume, fully dispersed to form a 1.5 g / mL solution, calcium bicarbonate was added to the solution, the pH was adjusted to 8.5, and microwaved at 150 W for 2 h. After separation and removal of the precipitate, a pretreated bovine cartilage collagen peptide solution was obtained;

[0027] (2) taking the pretreated bovine cartilage collagen peptide solution, adjusting the pH value to 5 with sodium acetate buffer solution, adding ferrous sulfate in an amount 0.2 times the mass of the bovine cartilage collagen peptide added in step (1), and mixing thoroughly for 1 hour, adding ethanol to the system so that the volume concentration of ethanol in the system is 80%, and mixing thoroughly until no precipitation is generated, separating and collecting the precipitate, and obtaining an iron-containing bovine cartilage collagen peptide precursor;

[0028] (3) adding dopamine hydrochloride to a 10 mmol / L Tris hydrochloric acid buffer solution at pH = 8.5, dissolving to obtain a 2 mg / mL dopamine solution, dispersing the iron-containing bovine cartilage collagen peptide precursor in the dopamine solution to obtain a precursor concentration of 5 mg / mL; stirring the reaction at 800 rpm for 15 h, filtering and washing the filter cake, pre-freezing it at below -18°C, and then vacuum drying it at -55°C, 12 Pa for 20 h to obtain a bovine cartilage collagen peptide chelate;

[0029] (4) Dissolve chitosan oligosaccharide in water, add yak bone collagen peptide in an amount of 0.8 times the mass of chitosan oligosaccharide, mix thoroughly, then add calcium chloride in an amount of 0.4 times the mass of chitosan oligosaccharide, adjust the pH to neutral using an ammonia buffer solution, heat to 70° C. and react for 24 hours, use ethanol to precipitate and separate the product from the reaction system, fully wash the precipitate, and dry to obtain a complex of yak bone collagen peptide and chitosan oligosaccharide;

[0030] (5) The powdered components are sieved with a 40-mesh sieve, and the following components are added by weight: 55 parts of bovine cartilage collagen peptide chelate, 2 parts of yak bone collagen peptide and chitosan oligosaccharide complex, 1 part of enzymatic bone powder, 1 part of cartilage extract, 0.04 parts of sucralose, and 20.96 parts of oligomaltose. After weighing, the mixture is mixed for 25 to 35 minutes to obtain a mixed powder. The mixed powder is quantitatively packaged at 8 g / bag under a cleanliness level of 100,000, and the filling error is within ±3%.

[0031] Example 2: (1) Bovine cartilage collagen peptide was placed in an aqueous solution of 10% ethanol by volume, fully dispersed to form a 1.5 g / mL solution, calcium bicarbonate was added to the solution, the pH was adjusted to 8.5, and microwaved at 150 W for 2 h. After separation and removal of the precipitate, a pretreated bovine cartilage collagen peptide solution was obtained;

[0032] (2) taking the pretreated bovine cartilage collagen peptide solution, adjusting the pH value to 5 with sodium acetate buffer solution, adding ferrous sulfate in an amount 0.2 times the mass of the bovine cartilage collagen peptide added in step (1), and mixing thoroughly for 1 hour, adding ethanol to the system so that the volume concentration of ethanol in the system is 80%, and mixing thoroughly until no precipitation is generated, separating and collecting the precipitate, and obtaining an iron-containing bovine cartilage collagen peptide precursor;

[0033] (3) adding dopamine hydrochloride to a 10 mmol / L Tris hydrochloric acid buffer solution at pH = 8.5, dissolving to obtain a 2 mg / mL dopamine solution, dispersing the iron-containing bovine cartilage collagen peptide precursor in the dopamine solution to obtain a precursor concentration of 5 mg / mL; stirring the reaction at 800 rpm for 15 h, filtering and washing the filter cake, pre-freezing it at below -18°C, and then vacuum drying it at -55°C, 12 Pa for 20 h to obtain a bovine cartilage collagen peptide chelate;

[0034] (4) Dissolve chitosan oligosaccharide in water, add yak bone collagen peptide in an amount of 0.8 times the mass of chitosan oligosaccharide, mix thoroughly, then add calcium chloride in an amount of 0.4 times the mass of chitosan oligosaccharide, adjust the pH to neutral using an ammonia buffer solution, heat to 70° C. and react for 24 hours, use ethanol to precipitate and separate the product from the reaction system, fully wash the precipitate, and dry to obtain a complex of yak bone collagen peptide and chitosan oligosaccharide;

[0035] (5) The powdered components are sieved with a 40-mesh sieve, and the following components are added by weight: 55 parts of bovine cartilage collagen peptide chelate, 2 parts of yak bone collagen peptide and chitosan oligosaccharide complex, 1 part of enzymatic bone powder, 1 part of cartilage extract, 0.04 parts of sucralose, and 20.96 parts of oligomaltose. After weighing, the mixture is mixed for 25 to 35 minutes to obtain a mixed powder. The mixed powder is quantitatively packaged at 8 g / bag under a cleanliness level of 100,000, and the filling error is within ±3%.

[0036] Example 3: (1) The bovine cartilage collagen peptide was placed in an aqueous solution of 5% ethanol by volume, and fully dispersed to form a 1.5 g / mL solution. Calcium bicarbonate was added to the solution, and the pH was adjusted to 10. The solution was microwaved at 150 W for 2 h, and the precipitate was separated and removed to obtain a pretreated bovine cartilage collagen peptide solution;

[0037] (2) taking the pretreated bovine cartilage collagen peptide solution, adjusting the pH value to 5 with sodium acetate buffer solution, adding ferrous sulfate in an amount 0.2 times the mass of the bovine cartilage collagen peptide added in step (1), and mixing thoroughly for 1 hour, adding ethanol to the system so that the volume concentration of ethanol in the system is 80%, and mixing thoroughly until no precipitation is generated, separating and collecting the precipitate, and obtaining an iron-containing bovine cartilage collagen peptide precursor;

[0038] (3) adding dopamine hydrochloride to a 10 mmol / L Tris hydrochloric acid buffer solution at pH = 8.5, dissolving to obtain a 2 mg / mL dopamine solution, dispersing the iron-containing bovine cartilage collagen peptide precursor in the dopamine solution to obtain a precursor concentration of 5 mg / mL; stirring the reaction at 800 rpm for 15 h, filtering and washing the filter cake, pre-freezing it at below -18°C, and then vacuum drying it at -55°C, 12 Pa for 20 h to obtain a bovine cartilage collagen peptide chelate;

[0039] (4) Dissolve chitosan oligosaccharide in water, add yak bone collagen peptide in an amount of 0.8 times the mass of chitosan oligosaccharide, mix thoroughly, then add calcium chloride in an amount of 0.4 times the mass of chitosan oligosaccharide, adjust the pH to neutral using an ammonia buffer solution, heat to 70° C. and react for 24 hours, use ethanol to precipitate and separate the product from the reaction system, fully wash the precipitate, and dry to obtain a complex of yak bone collagen peptide and chitosan oligosaccharide;

[0040] (5) The powdered components are sieved with a 40-mesh sieve, and the following components are added by weight: 55 parts of bovine cartilage collagen peptide chelate, 2 parts of yak bone collagen peptide and chitosan oligosaccharide complex, 1 part of enzymatic bone powder, 1 part of cartilage extract, 0.04 parts of sucralose, and 20.96 parts of oligomaltose. After weighing, the mixture is mixed for 25 to 35 minutes to obtain a mixed powder. The mixed powder is quantitatively packaged at 8 g / bag under a cleanliness level of 100,000, and the filling error is within ±3%.

[0041] Example 4: (1) The bovine cartilage collagen peptide was placed in an aqueous solution of 5% ethanol by volume, and fully dispersed to form a 1.5 g / mL solution. Calcium bicarbonate was added to the solution, and the pH was adjusted to 8.5. The solution was microwaved at 150 W for 2 h, and the precipitate was separated and removed to obtain a pretreated bovine cartilage collagen peptide solution;

[0042] (2) taking the pretreated bovine cartilage collagen peptide solution, adjusting the pH value to 5 with sodium acetate buffer solution, adding ferrous sulfate in an amount 0.5 times the mass of the bovine cartilage collagen peptide added in step (1), and mixing thoroughly for 1 hour, adding ethanol to the system so that the volume concentration of ethanol in the system is 80%, and mixing thoroughly until no precipitation is generated, separating and collecting the precipitate, and obtaining an iron-containing bovine cartilage collagen peptide precursor;

[0043] (3) adding dopamine hydrochloride to a 10 mmol / L Tris hydrochloric acid buffer solution at pH = 8.5, dissolving to obtain a 2 mg / mL dopamine solution, dispersing the iron-containing bovine cartilage collagen peptide precursor in the dopamine solution to obtain a precursor concentration of 5 mg / mL; stirring the reaction at 800 rpm for 15 h, filtering and washing the filter cake, pre-freezing it at below -18°C, and then vacuum drying it at -55°C, 12 Pa for 20 h to obtain a bovine cartilage collagen peptide chelate;

[0044] (4) Dissolve chitosan oligosaccharide in water, add yak bone collagen peptide in an amount of 0.8 times the mass of chitosan oligosaccharide, mix thoroughly, then add calcium chloride in an amount of 0.4 times the mass of chitosan oligosaccharide, adjust the pH to neutral using an ammonia buffer solution, heat to 70° C. and react for 24 hours, use ethanol to precipitate and separate the product from the reaction system, fully wash the precipitate, and dry to obtain a complex of yak bone collagen peptide and chitosan oligosaccharide;

[0045] (5) The powdered components are sieved with a 40-mesh sieve, and the following components are added by weight: 55 parts of bovine cartilage collagen peptide chelate, 2 parts of yak bone collagen peptide and chitosan oligosaccharide complex, 1 part of enzymatic bone powder, 1 part of cartilage extract, 0.04 parts of sucralose, and 20.96 parts of oligomaltose. After weighing, the mixture is mixed for 25 to 35 minutes to obtain a mixed powder. The mixed powder is quantitatively packaged at 8 g / bag under a cleanliness level of 100,000, and the filling error is within ±3%.

[0046] Example 5: (1) The bovine cartilage collagen peptide was placed in an aqueous solution of 5% ethanol by volume, and fully dispersed to form a 1.5 g / mL solution. Calcium bicarbonate was added to the solution, and the pH was adjusted to 8.5. The solution was microwaved at 150 W for 2 h, and the precipitate was separated and removed to obtain a pretreated bovine cartilage collagen peptide solution;

[0047] (2) taking the pretreated bovine cartilage collagen peptide solution, adjusting the pH value to 5 with sodium acetate buffer solution, adding ferrous sulfate in an amount 0.2 times the mass of the bovine cartilage collagen peptide added in step (1), and mixing thoroughly for 1 hour, adding ethanol to the system so that the volume concentration of ethanol in the system is 80%, and mixing thoroughly until no precipitation is generated, separating and collecting the precipitate, and obtaining an iron-containing bovine cartilage collagen peptide precursor;

[0048] (3) adding dopamine hydrochloride to a 10 mmol / L Tris hydrochloric acid buffer solution at pH = 8.5, dissolving to obtain a 2 mg / mL dopamine solution, dispersing the iron-containing bovine cartilage collagen peptide precursor in the dopamine solution to make the precursor concentration 10 mg / mL; stirring the reaction at 800 rpm for 15 h, filtering and washing the filter cake, pre-freezing it at below -18°C, and then vacuum drying it at -55°C, 12 Pa for 20 h to obtain the bovine cartilage collagen peptide chelate;

[0049] (4) Dissolve chitosan oligosaccharide in water, add yak bone collagen peptide in an amount of 0.8 times the mass of chitosan oligosaccharide, mix thoroughly, then add calcium chloride in an amount of 0.4 times the mass of chitosan oligosaccharide, adjust the pH to neutral using an ammonia buffer solution, heat to 70° C. and react for 24 hours, use ethanol to precipitate and separate the product from the reaction system, fully wash the precipitate, and dry to obtain a complex of yak bone collagen peptide and chitosan oligosaccharide;

[0050] (5) The powdered components are sieved with a 40-mesh sieve, and the following components are added by weight: 55 parts of bovine cartilage collagen peptide chelate, 2 parts of yak bone collagen peptide and chitosan oligosaccharide complex, 1 part of enzymatic bone powder, 1 part of cartilage extract, 0.04 parts of sucralose, and 20.96 parts of oligomaltose. After weighing, the mixture is mixed for 25 to 35 minutes to obtain a mixed powder. The mixed powder is quantitatively packaged at 8 g / bag under a cleanliness level of 100,000, and the filling error is within ±3%.

[0051] Example 6: (1) The bovine cartilage collagen peptide was placed in an aqueous solution of 5% ethanol by volume, and fully dispersed to form a 1.5 g / mL solution. Calcium bicarbonate was added to the solution, and the pH was adjusted to 8.5. The solution was microwaved at 150 W for 2 h, and the precipitate was separated and removed to obtain a pretreated bovine cartilage collagen peptide solution;

[0052] (2) taking the pretreated bovine cartilage collagen peptide solution, adjusting the pH value to 5 with sodium acetate buffer solution, adding ferrous sulfate in an amount 0.2 times the mass of the bovine cartilage collagen peptide added in step (1), and mixing thoroughly for 1 hour, adding ethanol to the system so that the volume concentration of ethanol in the system is 80%, and mixing thoroughly until no precipitation is generated, separating and collecting the precipitate, and obtaining an iron-containing bovine cartilage collagen peptide precursor;

[0053] (3) adding dopamine hydrochloride to a 10 mmol / L Tris hydrochloric acid buffer solution at pH = 8.5, dissolving to obtain a 2 mg / mL dopamine solution, dispersing the iron-containing bovine cartilage collagen peptide precursor in the dopamine solution to make the precursor concentration 5 mg / mL; stirring the reaction at 1500 rpm for 15 h, filtering and washing the filter cake, pre-freezing it at below -18°C, and then vacuum drying it at -55°C, 12 Pa for 20 h to obtain the bovine cartilage collagen peptide chelate;

[0054] (4) Dissolve chitosan oligosaccharide in water, add yak bone collagen peptide in an amount of 0.8 times the mass of chitosan oligosaccharide, mix thoroughly, then add calcium chloride in an amount of 0.4 times the mass of chitosan oligosaccharide, adjust the pH to neutral using an ammonia buffer solution, heat to 70° C. and react for 24 hours, use ethanol to precipitate and separate the product from the reaction system, fully wash the precipitate, and dry to obtain a complex of yak bone collagen peptide and chitosan oligosaccharide;

[0055] (5) The powdered components are sieved with a 40-mesh sieve, and the following components are added by weight: 55 parts of bovine cartilage collagen peptide chelate, 2 parts of yak bone collagen peptide and chitosan oligosaccharide complex, 1 part of enzymatic bone powder, 1 part of cartilage extract, 0.04 parts of sucralose, and 20.96 parts of oligomaltose. After weighing, the mixture is mixed for 25 to 35 minutes to obtain a mixed powder. The mixed powder is quantitatively packaged at 8 g / bag under a cleanliness level of 100,000, and the filling error is within ±3%.

[0056] Example 7: (1) The bovine cartilage collagen peptide was placed in an aqueous solution of 5% ethanol by volume, and fully dispersed to form a 1.5 g / mL solution. Calcium bicarbonate was added to the solution, and the pH was adjusted to 8.5. The solution was microwaved at 150 W for 2 h, and the precipitate was separated and removed to obtain a pretreated bovine cartilage collagen peptide solution;

[0057] (2) taking the pretreated bovine cartilage collagen peptide solution, adjusting the pH value to 5 with sodium acetate buffer solution, adding ferrous sulfate in an amount 0.2 times the mass of the bovine cartilage collagen peptide added in step (1), and mixing thoroughly for 1 hour, adding ethanol to the system so that the volume concentration of ethanol in the system is 80%, and mixing thoroughly until no precipitation is generated, separating and collecting the precipitate, and obtaining an iron-containing bovine cartilage collagen peptide precursor;

[0058] (3) adding dopamine hydrochloride to a 10 mmol / L Tris hydrochloric acid buffer solution at pH = 8.5, dissolving to obtain a 2 mg / mL dopamine solution, dispersing the iron-containing bovine cartilage collagen peptide precursor in the dopamine solution to obtain a precursor concentration of 5 mg / mL; stirring the reaction at 800 rpm for 15 h, filtering and washing the filter cake, pre-freezing it at below -18°C, and then vacuum drying it at -55°C, 12 Pa for 20 h to obtain a bovine cartilage collagen peptide chelate;

[0059] (4) Dissolve chitosan oligosaccharide in water, add yak bone collagen peptide in an amount of 1.2 times the mass of chitosan oligosaccharide, mix thoroughly, then add calcium chloride in an amount of 0.4 times the mass of chitosan oligosaccharide, adjust the pH to neutral using an ammonia buffer solution, heat to 70° C. and react for 24 hours, use ethanol to precipitate and separate the product from the reaction system, fully wash the precipitate, and dry to obtain a complex of yak bone collagen peptide and chitosan oligosaccharide;

[0060] (5) The powdered components are sieved with a 40-mesh sieve, and the following components are added by weight: 55 parts of bovine cartilage collagen peptide chelate, 2 parts of yak bone collagen peptide and chitosan oligosaccharide complex, 1 part of enzymatic bone powder, 1 part of cartilage extract, 0.04 parts of sucralose, and 20.96 parts of oligomaltose. After weighing, the mixture is mixed for 25 to 35 minutes to obtain a mixed powder. The mixed powder is quantitatively packaged at 8 g / bag under a cleanliness level of 100,000, and the filling error is within ±3%.

[0061] Example 8: (1) The bovine cartilage collagen peptide was placed in an aqueous solution of 5% ethanol by volume, and fully dispersed to form a 1.5 g / mL solution. Calcium bicarbonate was added to the solution, and the pH was adjusted to 8.5. The solution was microwaved at 150 W for 2 h, and the precipitate was separated and removed to obtain a pretreated bovine cartilage collagen peptide solution;

[0062] (2) taking the pretreated bovine cartilage collagen peptide solution, adjusting the pH value to 5 with sodium acetate buffer solution, adding ferrous sulfate in an amount 0.2 times the mass of the bovine cartilage collagen peptide added in step (1), and mixing thoroughly for 1 hour, adding ethanol to the system so that the volume concentration of ethanol in the system is 80%, and mixing thoroughly until no precipitation is generated, separating and collecting the precipitate, and obtaining an iron-containing bovine cartilage collagen peptide precursor;

[0063] (3) adding dopamine hydrochloride to a 10 mmol / L Tris hydrochloric acid buffer solution at pH = 8.5, dissolving to obtain a 2 mg / mL dopamine solution, dispersing the iron-containing bovine cartilage collagen peptide precursor in the dopamine solution to obtain a precursor concentration of 5 mg / mL; stirring the reaction at 800 rpm for 15 h, filtering and washing the filter cake, pre-freezing it at below -18°C, and then vacuum drying it at -55°C, 12 Pa for 20 h to obtain a bovine cartilage collagen peptide chelate;

[0064] (4) Dissolve chitosan oligosaccharide in water, add yak bone collagen peptide in an amount of 0.8 times the mass of chitosan oligosaccharide, mix thoroughly, then add calcium chloride in an amount of 1 times the mass of chitosan oligosaccharide, adjust the pH to neutral using an ammonia buffer solution, heat to 70° C. and react for 24 hours, use ethanol to precipitate and separate the product from the reaction system, fully wash the precipitate, and dry to obtain a complex of yak bone collagen peptide and chitosan oligosaccharide;

[0065] (5) The powdered components are sieved with a 40-mesh sieve, and the following components are added by weight: 55 parts of bovine cartilage collagen peptide chelate, 2 parts of yak bone collagen peptide and chitosan oligosaccharide complex, 1 part of enzymatic bone powder, 1 part of cartilage extract, 0.04 parts of sucralose, and 20.96 parts of oligomaltose. After weighing, the mixture is mixed for 25 to 35 minutes to obtain a mixed powder. The mixed powder is quantitatively packaged at 8 g / bag under a cleanliness level of 100,000, and the filling error is within ±3%.

[0066] Comparative Example 1: (1) dopamine hydrochloride was added to a 10 mmol / L Tris hydrochloric acid buffer solution at pH = 8.5, and a 2 mg / mL dopamine solution was obtained after dissolving, and bovine cartilage collagen peptide was dispersed in the dopamine solution to make the concentration of the precursor 5 mg / mL; the reaction was stirred at 800 rpm for 15 h, and the filter cake was filtered and washed, pre-frozen at below -18°C, and then vacuum dried at -55°C, 12 Pa for 20 h to obtain the product;

[0067] (2) Dissolve chitosan oligosaccharide in water, add yak bone collagen peptide in an amount of 0.8 times the mass of chitosan oligosaccharide, mix thoroughly, then add calcium chloride in an amount of 0.4 times the mass of chitosan oligosaccharide, adjust the pH to neutral using an ammonia buffer solution, heat to 70° C. and react for 24 hours, use ethanol to precipitate and separate the product from the reaction system, fully wash the precipitate, and dry to obtain a complex of yak bone collagen peptide and chitosan oligosaccharide;

[0068] (3) Sieve the powdered components through a 40-mesh sieve, and add the following components by mass: 55 parts of bovine cartilage collagen peptide obtained in step (1), 2 parts of a complex of yak bone collagen peptide and chitosan oligosaccharide, 1 part of enzymatic bone powder, 1 part of cartilage extract, 0.04 parts of sucralose, and 20.96 parts of oligomaltose; weigh and mix for 25 to 35 minutes to obtain a mixed powder; and quantitatively package the mixed powder in a 100,000-grade cleanliness level at 8 g / bag, with a filling error within ±3%.

[0069] Comparative Example 2: (1) The bovine cartilage collagen peptide was placed in an aqueous solution of 5% ethanol by volume, and fully dispersed to form a 1.5 g / mL solution. Calcium bicarbonate was added to the solution, and the pH was adjusted to 8.5. The solution was microwaved at 150 W for 2 h, and the precipitate was separated and removed to obtain a pretreated bovine cartilage collagen peptide solution;

[0070] (2) taking the pretreated bovine cartilage collagen peptide solution, adjusting the pH value to 5 with sodium acetate buffer solution, adding ferrous sulfate in an amount 0.2 times the mass of the bovine cartilage collagen peptide added in step (1), and mixing thoroughly for 1 hour, adding ethanol to the system so that the volume concentration of ethanol in the system is 80%, and mixing thoroughly until no precipitation is generated, separating and collecting the precipitate, and obtaining an iron-containing bovine cartilage collagen peptide precursor;

[0071] (3) adding dopamine hydrochloride to a 10 mmol / L Tris hydrochloric acid buffer solution at pH = 8.5, dissolving to obtain a 2 mg / mL dopamine solution, dispersing the iron-containing bovine cartilage collagen peptide precursor in the dopamine solution to obtain a precursor concentration of 5 mg / mL; stirring the reaction at 800 rpm for 15 h, filtering and washing the filter cake, pre-freezing it at below -18°C, and then vacuum drying it at -55°C, 12 Pa for 20 h to obtain a bovine cartilage collagen peptide chelate;

[0072] (4) The powdered components are sieved with a 40-mesh sieve and the following components are mixed by weight: 55 parts of bovine cartilage collagen peptide chelate, 1 part of yak bone collagen peptide, 1 part of chitosan oligosaccharide, 1 part of enzymatic bone powder, 1 part of cartilage extract, 0.04 parts of sucralose, and 20.96 parts of maltooligosaccharide; the mixed components are weighed and mixed for 25 to 35 minutes to obtain a mixed powder; the mixed powder is quantitatively packaged at 8 g / bag under a cleanliness level of 100,000, and the filling error is within ±3%.

[0073] Example sample test:

[0074] Simulated gastric fluid: NaCl 1.0g, pepsin 1.6g dissolved in 500mL 0.084M hydrochloric acid solution, pH 2.0 adjusted; simulated intestinal fluid: bile salt 250mg, lipase 80mg and CaCl21.39 g, NaCl 10.95g dissolved in 50mL distilled water, pH 7.0 adjusted. The products of each embodiment and comparative example were placed open for 0d and 30d, and then the samples placed for 0d and 30d were placed in simulated gastric fluid at 37℃ for 2h, 10000r / min, centrifuged for 10min, and the precipitate was placed in simulated intestinal fluid at 37℃ for 2h, the enzyme was inactivated in a boiling water bath for 10min, 10000r / min, centrifuged for 10min, and the precipitate was freeze-dried. The content of volatile substances in the samples placed open for 0d, 30d and 0d, 30d after simulated gastric fluid and simulated intestinal fluid were measured by headspace solid phase microextraction and gastrometry-mass spectrometry technology. The total iron content in the simulated gastric fluid and simulated intestinal fluid after 0d and 30d was determined by the o-phenanthroline method.

[0075] Results: The main volatile substances in the samples left open for 0 days and 30 days were not detected, while some volatile substances that can produce odor were detected in the samples exposed to simulated gastric fluid and simulated intestinal fluid. The results after treatment with simulated gastrointestinal fluid are as follows:

[0076]

[0077]

[0078] As can be seen from the above table, after being treated with simulated gastrointestinal fluid, the volatile substances inside the embodiments of the present invention are exposed, and the low content of the substances indicates that the preparation method of the present invention can greatly reduce the peculiar smell in the bovine collagen peptide during the oral stage. In the process of sodium bicarbonate treatment, compared with Example 1, the concentration of ethanol in the system of Example 2 is greater, and the content of each main volatile substance is significantly reduced, indicating that more ethanol is conducive to the removal of such substances, but more ethanol will affect the solubility of calcium bicarbonate, and the cost needs to be considered comprehensively; the pH of Example 3 is 10, indicating that the amount of calcium bicarbonate added is increased, and the content of volatile substances is significantly increased. When polydopamine is coated, the dopamine content of Example 5 is relatively less than that of Example 1, and the content of peculiar smell substances is increased. It may be that less dopamine causes the coating integrity to decrease; the stirring speed of Example 6 is faster, and the content of volatile substances measured by the sample is reduced, indicating that a faster stirring speed is conducive to the complete coating of bovine cartilage collagen peptides. The volatile substance content of Comparative Example 1, which was directly coated with dopamine without calcium and iron treatment of bovine cartilage collagen peptide, increased significantly, indicating that it was more likely to produce peculiar smell, and the smell was not as pleasant as the smell of each embodiment. In the process of treating yak bone collagen peptide, compared with Example 1, the yak bone collagen peptide involved in the treatment of Example 7 had a higher content, and the volatile substance content of the sample increased, while the volatile substance content of Example 8 with a higher amount of calcium chloride added decreased; the volatile substance content of Comparative Example 2, which was not treated with simultaneous chelation, was significantly higher, indicating that chelating yak bone collagen peptide and chitosan oligosaccharide with calcium ions together can significantly reduce the volatilization of peculiar smell substances and increase the acceptance of the composition.

[0079] The results are as follows:

[0080]

[0081]

[0082] As can be seen from the above table, the iron ion release rate of Example 3 is reduced, indicating that more calcium bicarbonate is conducive to the stable chelation of iron ions. Example 4 adds more ferrous ions compared to Example 1, and the release rate of iron ions increases. The release rate of Example 5 increases, while that of Example 6 decreases, indicating that more dopamine content and stirring speed are conducive to the integrity and uniformity of the coating, and the protection ability of the iron ions inside is also stronger.

[0083] For those skilled in the art, various other corresponding changes and deformations can be made according to the technical solutions and concepts described above, and all of these changes and deformations should fall within the protection scope of the claims of the present invention.

Claims

1. A method for preparing a composition containing bovine collagen peptide, characterized in that: The following components are included by mass: 40-80 parts of bovine cartilage collagen peptide chelate, 1-4 parts of yak bone collagen peptide and chitosan oligosaccharide complex, 0.5-2 parts of enzymatic bone powder, 0.5-2 parts of cartilage extract, 0.01-0.1 parts of sucralose, and 15-25 parts of oligomaltose; The preparation of the bovine cartilage collagen peptide chelate comprises the following steps: a. Place bovine cartilage collagen peptide in an aqueous solution of ethanol with a volume concentration of 5-10%, fully disperse it to form a 1-2 g / mL solution, add calcium bicarbonate to the solution, adjust the pH to 8.5-10, microwave at 100-200w for 1-3h, separate and remove the precipitate, and obtain a pretreated bovine cartilage collagen peptide solution; b. Take the pretreated bovine cartilage collagen peptide solution, adjust the pH value to 5-5.5 with sodium acetate buffer solution, add ferrous sulfate, mix thoroughly for 1-2 hours, add ethanol to the system so that the volume concentration of ethanol in the system is 75-99%, mix thoroughly until no precipitation is generated, separate and collect the precipitate, and obtain the iron-containing bovine cartilage collagen peptide precursor; the mass ratio of the ferrous sulfate in step b to the bovine cartilage collagen peptide in step a is 1:2-5; c. Prepare a dopamine solution, disperse the iron-containing bovine cartilage collagen peptide precursor in the dopamine solution to make the concentration of the iron-containing bovine cartilage collagen peptide precursor in the dopamine solution be 5-10 mg / mL, stir at 800-1500 rpm for 10-20 hours, filter and wash the filter cake, and lyophilize to obtain the product; The preparation of the complex of yak bone collagen peptide and chitosan oligosaccharide comprises the following steps: dissolving chitosan oligosaccharide in water, adding yak bone collagen peptide, mixing thoroughly, adding calcium chloride, adjusting the pH to neutral with ammonia buffer solution, heating to 60-80° C. for reaction for 24-48 h, separating the product from the reaction system with ethanol, washing the precipitate thoroughly, and drying to obtain the complex; the mass ratio of chitosan oligosaccharide to yak bone collagen peptide is 1:0.8-1.2; the mass ratio of calcium chloride to chitosan oligosaccharide is 1:1-2.

5.

2. The method for preparing the composition containing bovine collagen peptide according to claim 1, characterized in that: The bovine cartilage collagen peptide is type II bovine cartilage collagen peptide.

3. The method for preparing the composition containing bovine collagen peptide according to claim 1, characterized in that: The following steps are involved: The bovine cartilage collagen peptide chelate, the yak bone collagen peptide and chitosan oligosaccharide complex and other components are mixed to obtain the product.

4. The method for preparing the composition containing bovine collagen peptide according to claim 1, characterized in that: The freeze-drying in step c is pre-freezing below -18°C and then vacuum drying at -40~-80°C and 10~16Pa for 18~24h.

5. The method for preparing the composition containing bovine collagen peptide according to claim 1, characterized in that: In step c, the dopamine solution is prepared by adding dopamine hydrochloride to a 10 mmol / L Tris hydrochloric acid buffer solution at pH=8.5, and dissolving the dopamine hydrochloride to obtain a 2 mg / mL dopamine solution.

6. The method for preparing the composition containing bovine collagen peptide according to claim 1, characterized in that: The following steps are involved: The powdered components are sieved with a 40-mesh sieve, weighed according to the formula amount, and mixed for 25 to 35 minutes to obtain a mixed powder; In a 100,000-grade clean environment, the mixed powder is quantitatively packaged, and the filling error is within ±3%.

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