A collagen peptide composition and preparation method thereof

Abstract

The present invention relates to the field of biotechnology, and in particular to a collagen peptide composition and a preparation method thereof. In the present invention, by preparing a porous caffeic acid starch ester with antioxidant activity, a composite collagen peptide is adsorbed by the porous caffeic acid starch ester, and then a composite collagen peptide inclusion compound is prepared, which can effectively protect the composite collagen peptide, and then, under the synergistic effect of the porous caffeic acid starch ester and the composite collagen peptide, the antioxidant activity of the composite collagen peptide inclusion compound in vivo is improved; through the preparation and addition of the composite emulsion, it is beneficial to stably disperse the composite plant extract in the collagen peptide composition, and promote the composite plant extract to be better absorbed and utilized.

Description

Technical Field

[0001] The present invention relates to the field of biotechnology, and more specifically, to a collagen peptide composition and a preparation method thereof. Background Art

[0002] Collagen peptides are made from collagen-rich animal tissues (including skin, bones, tendons, skeletal muscles, scales, etc.). Under the action of acid, alkali and protease, the macromolecular collagen is hydrolyzed into a small molecule peptide mixture with an average molecular weight of less than 10,000 Da. Studies have shown that collagen peptides are known as "soft gold for the skin" and "skin in skin, bone in bone" because of their small molecular weight, easy absorption, high safety, and biological activity, such as repairing skin damage, antioxidant activity, and anti-hypertensive activity. They are widely used in cosmetics, functional foods, medical materials and pharmaceutical fields.

[0003] Collagen peptides extracted from animal tissues can be divided into fish collagen peptides and terrestrial animal collagen peptides. The structure of fish collagen peptides is similar to that of collagen in human skin, so they are more easily absorbed by the human body. Although fish collagen peptides are more easily absorbed by the human body than intact collagen and terrestrial animal collagen peptides, their bioavailability is still limited. On the one hand, the digestive systems of different people have different absorption capacities for collagen peptides. For people with weak digestive abilities, they may not be able to effectively absorb and utilize collagen peptides. In addition, consuming a large amount of fish collagen peptides in a short period of time can also cause gastrointestinal discomfort, thereby reducing their low bioavailability. In addition, fish collagen peptides still have many problems in actual use, such as fishy taste and poor thermal stability. These problems limit the scope of their processing and consumption applications, and they cannot effectively exert the efficacy of fish collagen peptides.

[0004] Caffeic acid is a functional phenolic acid with significant antioxidant, anti-inflammatory and anti-cancer biological activities. It can scavenge DPPH free radicals, thereby protecting cells from damage and maintaining cell health. However, its molecular structure contains benzene rings and short unsaturated hydrocarbon chains. This structural feature makes it hydrophobic and has low water solubility. This physical and chemical property affects the absorption and distribution of caffeic acid in the body, thereby limiting its bioavailability in the body.

[0005] In addition, plant extracts contain a variety of antioxidant substances, which also have strong antioxidant properties and can scavenge DPPH free radicals. Plant extracts and collagen peptides each have unique health benefits. Combining collagen peptides with plant extracts can produce a synergistic effect and enhance the ability to scavenge DPPH free radicals. However, some plant extracts have poor water solubility and thermal stability, which affects their absorption and utilization in the body, reduces the bioavailability of plant extracts, and also limits the application scope of plant extracts and collagen peptides for co-processing and consumption. Summary of the invention

[0006] In view of the deficiencies in the prior art, the present invention provides a collagen peptide composition and a preparation method thereof.

[0007] A collagen peptide composition comprises the following raw materials:

[0008] A mixed liquid and a composite emulsion, wherein the volume ratio of the mixed liquid to the composite emulsion is (10-30):(1-3), and the mixed liquid is obtained by mixing composite collagen peptide inclusion complex, gamma-aminobutyric acid and distilled water in a mass ratio of (1-10):(0.3-2):(50-100);

[0009] The composite collagen peptide inclusion complex is obtained by adsorbing the composite collagen peptide into caffeic acid porous starch ester, and then encapsulating the caffeic acid porous starch ester adsorbed with the composite collagen peptide using β-cyclodextrin;

[0010] Caffeic acid porous starch ester is obtained by esterification reaction of activated caffeic acid and porous starch;

[0011] The raw materials of the composite emulsion include: konjac glucomannan / quinoa protein complex, composite plant extract and distilled water. The mixing mass ratio of the konjac glucomannan / quinoa protein complex, composite plant extract and distilled water is (1-5): (3-9): (20-70). The konjac glucomannan / quinoa protein complex is obtained by mixing quinoa protein solution and konjac glucomannan solution and then vacuum drying.

[0012] A method for preparing a collagen peptide composition comprises the following steps:

[0013] Step S1: preparation of caffeic acid porous starch ester, after activating caffeic acid, using the activated caffeic acid to react with porous starch for esterification, and then obtaining caffeic acid porous starch ester through precipitation, centrifugal separation, vacuum drying, grinding and sieving;

[0014] Step S2: Preparation of composite collagen peptide inclusion complex, adding caffeic acid porous starch ester to the composite collagen peptide solution, stirring and mixing, and then adding β-cyclodextrin, stirring and mixing, and spray drying to obtain the composite collagen peptide inclusion complex;

[0015] Step S3: preparing a composite emulsion, dissolving the quinoa protein solution and the konjac glucomannan solution to obtain a konjac glucomannan / quinoa protein complex, dissolving the konjac glucomannan / quinoa protein complex in distilled water, adding the composite plant extract, and homogenizing at high speed to obtain a composite emulsion;

[0016] Step S4: Preparation of collagen peptide composition: mixing the composite collagen peptide inclusion complex, γ-aminobutyric acid and distilled water to obtain a mixed solution, adding the composite emulsion of step S3 to the mixed solution, and mixing by magnetic stirring to obtain a collagen peptide composition.

[0017] Furthermore, step S1: preparation of caffeic acid porous starch ester specifically comprises the following steps:

[0018] S1.1: 15 to 22 parts by weight of caffeic acid are added to 40 to 60 parts by weight of dichloromethane, and the mixture is stirred evenly by magnetic force to obtain solution A. 8 to 18 parts by weight of thionyl chloride are diluted with 8 to 12 parts by weight of dichloromethane to obtain solution B;

[0019] S1.2: Under nitrogen atmosphere, add solution B dropwise into solution A while stirring. After the addition of solution B is complete, heat to 70-90°C and reflux for 3-5 hours.

[0020] S1.3: After the reaction is completed, distillation under reduced pressure is performed to remove excess thionyl chloride and dichloromethane to obtain caffeoyl chloride;

[0021] S1.4: Disperse 10 to 30 parts by weight of porous starch in 70 to 120 parts by weight of dimethyl sulfoxide, add 3 to 10 parts by weight of caffeoyl chloride and 1 to 5 parts by weight of pyridine, and reflux for reaction at 70 to 80° C. in a nitrogen atmosphere for 2 to 3 hours to obtain a reaction solution;

[0022] S1.5: After the reaction solution is cooled to room temperature, 100 to 200 parts by weight of anhydrous ethanol is slowly added while stirring, and the mixture is centrifuged to remove the supernatant and collect the precipitate. The precipitate is vacuum dried, ground and sieved to obtain caffeic acid porous starch ester.

[0023] Furthermore, step S2: preparation of composite collagen peptide inclusion complex specifically comprises the following steps:

[0024] S2.1: adding the composite collagen peptide to distilled water to prepare a composite collagen peptide solution with a concentration of 4 to 8 mg / mL;

[0025] S2.2: adding caffeic acid porous starch ester to the composite collagen peptide solution having a concentration of 4 to 8 mg / mL, wherein the mass ratio of the composite collagen peptide solution to the caffeic acid porous starch ester is (1 to 5):1, and stirring and mixing at 35 to 45° C. for 30 to 60 minutes to obtain a mixed solution;

[0026] S2.3: Add 15 mg / mL β-cyclodextrin aqueous solution to the mixed solution, stir and mix evenly, and then spray dry to obtain a composite collagen peptide inclusion complex.

[0027] Furthermore, the composite collagen peptide is obtained by mixing fish bone collagen peptide, fish skin collagen peptide and fish scale collagen peptide in a mass ratio of (1-3):(1-3):(1-3).

[0028] Furthermore, the amount of the β-cyclodextrin aqueous solution added is 1 to 3 times the mass of the mixed solution.

[0029] Further, step S3: preparation of the composite emulsion specifically comprises the following steps:

[0030] S3.1: Dissolve konjac glucomannan in distilled water, homogenize at a speed of 8000-10000 rpm for 1-3 min to obtain a konjac glucomannan solution with a concentration of 15-30 mg / mL; dissolve quinoa protein in distilled water, homogenize at a speed of 8000-10000 rpm for 1-3 min, then heat to 32-40° C., and stir magnetically for 2-3 h to fully hydrate the quinoa protein to obtain a quinoa protein solution with a concentration of 15-20 mg / mL;

[0031] S3.2: adding the quinoa protein solution dropwise to the konjac glucomannan solution while stirring; after the quinoa protein solution is added dropwise, stirring is continued for 2 to 4 hours, and vacuum drying is performed to obtain a konjac glucomannan / quinoa protein complex;

[0032] S3.3: Dissolve the konjac glucomannan / quinoa protein complex in distilled water, and then add the composite plant extract, wherein the mixing mass ratio of the konjac glucomannan / quinoa protein complex, the composite plant extract and distilled water is (1-5): (3-9): (20-70), and homogenize at a speed of 10000-15000 rpm for 3-5 minutes to obtain a composite emulsion.

[0033] Furthermore, the mixing volume ratio of the quinoa protein solution to the konjac glucomannan solution is (1-1.5): (1-1.5).

[0034] Furthermore, the composite plant extract contains at least two of lycopene, carotene, blueberry extract, passion fruit extract, sea buckthorn extract, pomegranate extract, flaxseed extract, mangosteen extract, roxburghii extract, grape seed extract and olive fruit extract.

[0035] Further, step S4: preparation of collagen peptide composition, specifically comprising the following steps:

[0036] The composite collagen peptide inclusion complex, γ-aminobutyric acid and distilled water are mixed in a mass ratio of (1-10): (0.3-2): (50-100), and magnetically stirred to obtain a mixed solution. The mixed solution and the composite emulsion are magnetically stirred to obtain a collagen peptide composition in a volume ratio of (10-30): (1-3).

[0037] The present invention has the following advantages:

[0038] 1. In the present invention, caffeic acid is activated to caffeoyl chloride, and then caffeoyl chloride is used to carry out esterification reaction with porous starch, so that the hydroxyl group of the porous starch reacts with the acyl group of the caffeoyl chloride, and caffeic acid is introduced into the surface of the porous starch particles. The porous starch is used as a carrier of caffeic acid, which can effectively improve the bioavailability and water solubility of caffeic acid, and the obtained caffeic acid porous starch ester has strong antioxidant activity and a certain scavenging ability for DPPH free radicals.

[0039] 2. In the present invention, the composite collagen peptide is adsorbed into the pores of caffeic acid porous starch ester, and β-cyclodextrin is used as a wall material to wrap the caffeic acid porous starch ester adsorbed with the composite collagen peptide. The obtained composite collagen peptide inclusion compound can effectively protect the composite collagen peptide, improve the thermal stability of the collagen peptide component, ensure the activity of the composite collagen peptide, and at the same time is beneficial to the slow release of the composite collagen peptide in the gastrointestinal tract, which is beneficial to the absorption and utilization of the composite collagen peptide by people with poor digestive ability, and improves the bioavailability of the composite collagen peptide. Moreover, under the synergistic effect of caffeic acid porous starch ester and the composite collagen peptide, the antioxidant activity of the composite collagen peptide inclusion compound in the body is effectively improved, and the DPPH free radicals are efficiently removed. In addition, the composite collagen peptide can mask the fishy smell of fish collagen peptide after inclusion and improve the flavor.

[0040] 3. In the present invention, the quinoa protein solution and the konjac glucomannan solution are mutually dissolved, and the quinoa protein and the konjac glucomannan are combined into stable composite nanoparticles through the electrostatic adsorption mutual force. The obtained konjac glucomannan / quinoa protein complex can be used as a stabilizer in the process of preparing the composite emulsion, so that the composite emulsion forms a tight and stable droplet polymerization network and covers the particle surface of the composite plant extract, thereby effectively protecting and slow-releasing the composite plant extract, so that the composite plant extract and the composite collagen peptide can be well processed and combined, and it is beneficial for the composite plant extract to be better absorbed and utilized. BRIEF DESCRIPTION OF THE DRAWINGS

[0041] Figure 1 Flow chart of the preparation method of the collagen peptide composition in an embodiment of the present invention.

[0042] Figure 2 The results of the DPPH free radical scavenging ability of the collagen peptide compositions of the examples and comparative examples of the present invention under different conditions are shown in FIG. DETAILED DESCRIPTION

[0043] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. 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.

[0044] Example 1

[0045] A method for preparing a collagen peptide composition, such as Figure 1 As shown, the specific steps include:

[0046] Step S1: Preparation of porous caffeic acid starch ester,

[0047] S1.1: 22 parts by weight of caffeic acid are added to 60 parts by weight of dichloromethane, and stirred evenly by magnetic force to obtain solution A; 18 parts by weight of thionyl chloride are diluted with 12 parts by weight of dichloromethane to obtain solution B;

[0048] S1.2: Under nitrogen atmosphere, add solution B dropwise into solution A while stirring. After the addition of solution B is complete, heat to 80°C and reflux for 5 h.

[0049] S1.3: After the reaction is completed, distillation under reduced pressure is performed to remove excess thionyl chloride and dichloromethane to obtain caffeoyl chloride;

[0050] S1.4: 30 parts by weight of porous starch are dispersed in 120 parts by weight of dimethyl sulfoxide, and then 10 parts by weight of caffeoyl chloride and 5 parts by weight of pyridine are added, and the mixture is refluxed for reaction at 80° C. under a nitrogen atmosphere for 3 h to obtain a reaction solution;

[0051] S1.5: After the reaction solution is cooled to room temperature, 200 parts by weight of anhydrous ethanol is slowly added while stirring, centrifuged, the supernatant is removed, and the precipitate is collected. The precipitate is vacuum dried, ground and sieved to obtain caffeic acid porous starch ester;

[0052] Step S2: Preparation of composite collagen peptide inclusion complex,

[0053] S2.1: fish bone collagen peptide, fish skin collagen peptide and fish scale collagen peptide are mixed in a mass ratio of 1:1:1 to obtain a composite collagen peptide, and the composite collagen peptide is added to distilled water to prepare a composite collagen peptide solution with a concentration of 8 mg / mL;

[0054] S2.2: adding caffeic acid porous starch ester to the composite collagen peptide solution with a concentration of 8 mg / mL, wherein the mass ratio of the composite collagen peptide solution to the caffeic acid porous starch ester is 5:1, and stirring and mixing at 38°C for 60 minutes to obtain a mixed solution;

[0055] S2.3: Add 15 mg / mL β-cyclodextrin aqueous solution to the mixed solution, wherein the amount of β-cyclodextrin aqueous solution added is twice the mass of the mixed solution, stir and mix evenly, and then spray dry to obtain a composite collagen peptide inclusion complex;

[0056] Step S3: Preparation of composite emulsion,

[0057] S3.1: Konjac glucomannan was dissolved in distilled water, and homogenized at 10000 rpm for 3 min to obtain a 30 mg / mL Konjac glucomannan solution; quinoa protein was dissolved in distilled water, and homogenized at 10000 rpm for 3 min, and then heated to 36°C and magnetically stirred for 3 h to fully hydrate the quinoa protein to obtain a 20 mg / mL quinoa protein solution;

[0058] S3.2: adding the quinoa protein solution dropwise into the konjac glucomannan solution while stirring, the volume ratio of the quinoa protein solution to the konjac glucomannan solution is 1:1.5, after the quinoa protein solution is added dropwise, stirring is continued for 4 hours, and the konjac glucomannan / quinoa protein complex is obtained by vacuum drying;

[0059] S3.3: dissolving the konjac glucomannan / quinoa protein complex in distilled water, and then adding a composite plant extract containing lycopene, carotene, blueberry extract, passion fruit extract, sea buckthorn extract, pomegranate extract, flax seed extract, mangosteen extract, roxburghii extract, grape seed extract and olive fruit extract, the mixing mass ratio of the konjac glucomannan / quinoa protein complex, the composite plant extract and distilled water is 3:3:70, and homogenizing at a speed of 15000 rpm for 5 minutes to obtain a composite emulsion;

[0060] Step S4: Preparation of collagen peptide composition,

[0061] The composite collagen peptide inclusion complex, γ-aminobutyric acid and distilled water were mixed at a mass ratio of 5:1:50, and magnetically stirred to obtain a mixed solution. The mixed solution was magnetically stirred and mixed with the composite emulsion at a volume ratio of 30:1 to obtain a collagen peptide composition.

[0062] Example 2

[0063] A method for preparing a collagen peptide composition, such as Figure 1 As shown, the specific steps include:

[0064] Step S1: Preparation of porous caffeic acid starch ester,

[0065] S1.1: 22 parts by weight of caffeic acid are added to 60 parts by weight of dichloromethane, and stirred evenly by magnetic force to obtain solution A; 18 parts by weight of thionyl chloride are diluted with 12 parts by weight of dichloromethane to obtain solution B;

[0066] S1.2: Under nitrogen atmosphere, add solution B dropwise into solution A while stirring. After the addition of solution B is complete, heat to 90°C and reflux for 3 h.

[0067] S1.3: After the reaction is completed, distillation under reduced pressure is performed to remove excess thionyl chloride and dichloromethane to obtain caffeoyl chloride;

[0068] S1.4: 30 parts by weight of porous starch are dispersed in 120 parts by weight of dimethyl sulfoxide, and then 10 parts by weight of caffeoyl chloride and 5 parts by weight of pyridine are added, and the mixture is refluxed for reaction at 70° C. under a nitrogen atmosphere for 2 h to obtain a reaction solution;

[0069] S1.5: After the reaction solution is cooled to room temperature, 200 parts by weight of anhydrous ethanol is slowly added while stirring, centrifuged, the supernatant is removed, and the precipitate is collected. The precipitate is vacuum dried, ground and sieved to obtain caffeic acid porous starch ester;

[0070] Step S2: Preparation of composite collagen peptide inclusion complex,

[0071] S2.1: fish bone collagen peptide, fish skin collagen peptide and fish scale collagen peptide are mixed in a mass ratio of 1:1:1 to obtain a composite collagen peptide, and the composite collagen peptide is added to distilled water to prepare a composite collagen peptide solution with a concentration of 8 mg / mL;

[0072] S2.2: adding caffeic acid porous starch ester to the composite collagen peptide solution with a concentration of 8 mg / mL, wherein the mass ratio of the composite collagen peptide solution to the caffeic acid porous starch ester is 5:1, and stirring and mixing at 42° C. for 30 min to obtain a mixed solution;

[0073] S2.3: Add 15 mg / mL β-cyclodextrin aqueous solution to the mixed solution, wherein the amount of β-cyclodextrin aqueous solution added is twice the mass of the mixed solution, stir and mix evenly, and then spray dry to obtain a composite collagen peptide inclusion complex;

[0074] Step S3: Preparation of composite emulsion,

[0075] S3.1: Konjac glucomannan was dissolved in distilled water, and homogenized at 10000 rpm for 1 min to obtain a 30 mg / mL Konjac glucomannan solution; quinoa protein was dissolved in distilled water, and homogenized at 10000 rpm for 1 min, and then heated to 40°C and magnetically stirred for 2 h to fully hydrate the quinoa protein to obtain a 20 mg / mL quinoa protein solution;

[0076] S3.2: adding the quinoa protein solution dropwise into the konjac glucomannan solution while stirring, the volume ratio of the quinoa protein solution to the konjac glucomannan solution is 1:1.5, after the quinoa protein solution is added dropwise, stirring is continued for 2 hours, and the konjac glucomannan / quinoa protein complex is obtained by vacuum drying;

[0077] S3.3: dissolving the konjac glucomannan / quinoa protein complex in distilled water, and then adding a composite plant extract containing lycopene, carotene, blueberry extract, passion fruit extract, sea buckthorn extract, pomegranate extract, flax seed extract, mangosteen extract, roxburghii extract, grape seed extract and olive fruit extract, the mixing mass ratio of the konjac glucomannan / quinoa protein complex, the composite plant extract and distilled water is 3:3:70, and homogenizing at a speed of 15000 rpm for 3 minutes to obtain a composite emulsion;

[0078] Step S4: Preparation of collagen peptide composition,

[0079] The composite collagen peptide inclusion complex, γ-aminobutyric acid and distilled water were mixed at a mass ratio of 5:1:50, and magnetically stirred to obtain a mixed solution. The mixed solution was magnetically stirred and mixed with the composite emulsion at a volume ratio of 30:1 to obtain a collagen peptide composition.

[0080] Example 3

[0081] A method for preparing a collagen peptide composition, such as Figure 1 As shown, the specific steps include:

[0082] Step S1: Preparation of porous caffeic acid starch ester,

[0083] S1.1: 22 parts by weight of caffeic acid are added to 60 parts by weight of dichloromethane, and stirred evenly by magnetic force to obtain solution A; 18 parts by weight of thionyl chloride are diluted with 12 parts by weight of dichloromethane to obtain solution B;

[0084] S1.2: Under nitrogen atmosphere, add solution B dropwise into solution A while stirring. After the addition of solution B is complete, heat to 80°C and reflux for 5 h.

[0085] S1.3: After the reaction is completed, distillation under reduced pressure is performed to remove excess thionyl chloride and dichloromethane to obtain caffeoyl chloride;

[0086] S1.4: 30 parts by weight of porous starch are dispersed in 120 parts by weight of dimethyl sulfoxide, and then 10 parts by weight of caffeoyl chloride and 5 parts by weight of pyridine are added, and the mixture is refluxed for reaction at 80° C. under a nitrogen atmosphere for 3 h to obtain a reaction solution;

[0087] S1.5: After the reaction solution is cooled to room temperature, 200 parts by weight of anhydrous ethanol is slowly added while stirring, centrifuged, the supernatant is removed, and the precipitate is collected. The precipitate is vacuum dried, ground and sieved to obtain caffeic acid porous starch ester;

[0088] Step S2: Preparation of composite collagen peptide inclusion complex,

[0089] S2.1: fish bone collagen peptide, fish skin collagen peptide and fish scale collagen peptide are mixed in a mass ratio of 1:1:1 to obtain a composite collagen peptide, and the composite collagen peptide is added to distilled water to prepare a composite collagen peptide solution with a concentration of 4 mg / mL;

[0090] S2.2: adding caffeic acid porous starch ester to the composite collagen peptide solution with a concentration of 4 mg / mL, wherein the mass ratio of the composite collagen peptide solution to the caffeic acid porous starch ester is 3:1, and stirring and mixing at 38°C for 60 minutes to obtain a mixed solution;

[0091] S2.3: Add 15 mg / mL β-cyclodextrin aqueous solution to the mixed solution, wherein the amount of β-cyclodextrin aqueous solution added is 3 times the mass of the mixed solution, stir and mix evenly, and then spray dry to obtain a composite collagen peptide inclusion complex;

[0092] Step S3: Preparation of composite emulsion,

[0093] S3.1: Konjac glucomannan was dissolved in distilled water, and homogenized at 10000 rpm for 3 min to obtain a 15 mg / mL Konjac glucomannan solution; quinoa protein was dissolved in distilled water, and homogenized at 10000 rpm for 3 min, and then heated to 36°C and magnetically stirred for 3 h to fully hydrate the quinoa protein to obtain a 15 mg / mL quinoa protein solution;

[0094] S3.2: adding the quinoa protein solution dropwise into the konjac glucomannan solution while stirring, the volume ratio of the quinoa protein solution to the konjac glucomannan solution is 1:1, after the quinoa protein solution is added dropwise, stirring is continued for 4 hours, and vacuum drying is performed to obtain a konjac glucomannan / quinoa protein complex;

[0095] S3.3: dissolving the konjac glucomannan / quinoa protein complex in distilled water, and then adding a composite plant extract containing lycopene, carotene, blueberry extract, passion fruit extract, sea buckthorn extract, pomegranate extract, flax seed extract, mangosteen extract, roxburghii extract, grape seed extract and olive fruit extract, the mixing mass ratio of the konjac glucomannan / quinoa protein complex, the composite plant extract and distilled water is 5:9:70, and homogenizing at a speed of 15000 rpm for 5 minutes to obtain a composite emulsion;

[0096] Step S4: Preparation of collagen peptide composition,

[0097] The composite collagen peptide inclusion complex, γ-aminobutyric acid and distilled water were mixed at a mass ratio of 5:1:50, and magnetically stirred to obtain a mixed solution. The mixed solution was magnetically stirred and mixed with the composite emulsion at a volume ratio of 30:1 to obtain a collagen peptide composition.

[0098] Comparative Example 1

[0099] Compared with Example 1, the difference of Comparative Example 1 is that step S1 is removed, the caffeic acid porous starch ester used in step S2 is replaced with porous starch, the porous starch is used to adsorb the composite collagen peptide, and then β-cyclodextrin is used as a wall material to wrap the porous starch adsorbed with the composite collagen peptide to prepare a composite collagen peptide inclusion complex, and the remaining steps and components remain unchanged to prepare a collagen peptide composition, which is recorded as Comparative Example 1.

[0100] Comparative Example 2

[0101] Compared with Example 1, the difference of Comparative Example 2 is that the composite collagen peptide inclusion complex in step S4 is replaced by composite collagen peptide, and the other steps and components remain unchanged to prepare a collagen peptide composition, which is recorded as Comparative Example 2.

[0102] Comparative Example 3

[0103] Compared with Example 1, the difference of Comparative Example 3 is that the composite collagen peptide inclusion complex in step S4 is replaced by caffeic acid porous starch ester, and the other steps and ingredients remain unchanged to prepare a collagen peptide composition, which is recorded as Comparative Example 3.

[0104] Comparative Example 4

[0105] Compared with Example 1, the difference of Comparative Example 4 is that the composite emulsion in step S4 is removed, and the remaining steps and components remain unchanged to prepare a collagen peptide composition, which is recorded as Comparative Example 4.

[0106] Comparative Example 5

[0107] Compared with Example 1, the difference of Comparative Example 5 is that steps S3.1-S3.2 are removed, the konjac glucomannan / quinoa protein complex in step S3.3 is replaced with konjac glucomannan, and the other steps and ingredients remain unchanged to prepare a collagen peptide composition, which is recorded as Comparative Example 5.

[0108] Comparative Example 6

[0109] Compared with Example 1, the difference of Comparative Example 6 is that steps S3.1-S3.2 are removed, the konjac glucomannan / quinoa protein complex in step S3.3 is replaced with quinoa protein, and the remaining steps and ingredients remain unchanged to prepare a collagen peptide composition, which is recorded as Comparative Example 6.

[0110] Comparative Example 7

[0111] Compared with Example 1, the difference of Comparative Example 7 is that the composite emulsion in step S4 is replaced by a composite plant extract, so that the mixed liquid and the composite plant extract are mixed in a volume ratio of 150:1, and the other steps and ingredients remain unchanged to prepare a collagen peptide composition, which is recorded as Comparative Example 7.

[0112] Test Example 1

[0113] Antioxidant performance test

[0114] The ability of the collagen peptide compositions of Examples 1-3 and Comparative Examples 1-7 to scavenge DPPH free radicals under normal conditions was determined. The specific steps are as follows:

[0115] The collagen peptide compositions of Examples 1-3 and Comparative Examples 1-7 were diluted 100 times with distilled water as test solutions, and then a solution with a concentration of 1×10 -4 mol / L DPPH anhydrous ethanol solution, store in dark for later use. Take 2mL of each test solution as a sample, add 2mL of DPPH anhydrous ethanol solution to each sample, shake vigorously, react at room temperature for 30min, and then measure the absorbance at 517nm In addition, the blank group used an equal volume of anhydrous ethanol solution instead of the DPPH anhydrous ethanol solution. Similarly, 2 mL of the test solution was taken from each group of test solutions as a sample, and 2 mL of anhydrous ethanol solution was added to each sample, shaken vigorously, reacted at room temperature for 30 minutes, and then the absorbance was measured at 517 nm. The control group used an equal volume of distilled water instead of DPPH anhydrous ethanol solution. Similarly, 2 mL of the test solution was taken from each group as a sample, and 2 mL of distilled water was added to each sample. The mixture was shaken vigorously and reacted at room temperature for 30 minutes. Then, the absorbance was measured at 517 nm. Calculate the scavenging rate of DPPH free radicals, the result is as follows Figure 2 As shown in Figure 2, the calculation formula for the scavenging rate of DPPH free radicals is:

[0116] DPPH free radical scavenging rate (%)

[0117] in, is the absorbance value of the control group, is the absorbance value of each test group, is the absorbance value of the blank group.

[0118] like Figure 2 It can be seen that the scavenging rate of the collagen peptide composition of Example 1-3 to DPPH free radicals is higher than that of Comparative Example 1-7, and has excellent antioxidant properties. It can be seen from Example 1-3 compared with Comparative Example 1-4 that the composite collagen peptide inclusion compound and the composite emulsion component in the collagen peptide composition have antioxidant properties, wherein the caffeic acid porous starch ester and the composite collagen peptide used in the composite collagen peptide inclusion compound have antioxidant activity, and can play a certain scavenging ability to DPPH free radicals, and, under the synergistic effect of caffeic acid porous starch ester and composite collagen peptide, the antioxidant activity can be further improved, and the scavenging ability to DPPH free radicals is strengthened; it can be seen from Example 1-3 compared with Comparative Example 5-7 that compared with the use of konjac glucomannan or quinoa protein alone, the konjac glucomannan / quinoa protein complex can be used as a stabilizer in the process of preparing a composite emulsion, which is conducive to the antioxidant activity of the composite plant extract, and ensures the ability to scavenge DPPH free radicals.

[0119] Test Example 2

[0120] Gastrointestinal simulated digestion stability test

[0121] The ability of the collagen peptide compositions of Examples 1-3 and Comparative Examples 1-7 to scavenge DPPH free radicals after simulated gastrointestinal digestion in vitro was determined. The specific steps are as follows:

[0122] The collagen peptide compositions of Examples 1-3 and Comparative Examples 1-7 were added to a potassium chloride-hydrochloric acid buffer solution with a concentration of 0.1 mol / L and a pH of 2, respectively, and the volume ratio of the collagen peptide composition to the potassium chloride-hydrochloric acid buffer solution was 1:100. After stirring and mixing, a simulated test solution was obtained. Pepsin was added to each group of simulated test solutions, and the amount of pepsin added was 1% of the mass of each group of simulated test solutions. Subsequently, each group of simulated test solutions was stirred in a water bath at 37° C. for 4 h, and then in a boiling water bath. The enzyme was inactivated for 5 minutes, and the pH value of each group of simulated test liquid after the enzyme inactivation was adjusted to 8 with sodium hydroxide solution to obtain digestion liquid. Trypsin was added to each group of digestion liquid, and the amount of trypsin added was 1% of the mass of each group of digestion liquid. Then, the mixture was stirred in a water bath at 37°C for 4 hours. Then, the enzyme was inactivated in a boiling water bath for 5 minutes. The digestion liquid of each group after the enzyme inactivation was centrifuged at 10,000 rpm for 30 minutes, and the supernatant was taken to determine the scavenging rate of each group of supernatant for DPPH free radicals.

[0123] Take 2 mL of the supernatant from each group as a sample, and add 2 mL of 1×10 - 4 mol / L DPPH anhydrous ethanol solution, shake vigorously, react at room temperature for 30 minutes, and then measure the absorbance at 517nm The blank group was replaced with an equal volume of anhydrous ethanol solution instead of DPPH anhydrous ethanol solution. 2 mL of the supernatant from each group was taken as a sample. 2 mL of anhydrous ethanol solution was added to each sample, and the mixture was shaken vigorously. The mixture was reacted at room temperature for 30 min, and then the absorbance was measured at 517 nm. The control group used an equal volume of distilled water instead of DPPH anhydrous ethanol solution. Similarly, 2 mL of the supernatant from each group was taken as a sample, and 2 mL of distilled water was added to each sample, and the mixture was shaken vigorously and reacted at room temperature for 30 min. Then, the absorbance was measured at 517 nm. Calculate the scavenging rate of DPPH free radicals, the result is as follows Figure 2 shown.

[0124] like Figure 2It can be seen that after simulated gastrointestinal digestion, the scavenging rate of the collagen peptide composition of Examples 1-3 for DPPH radicals can be maintained at about 85%, indicating that the DPPH antioxidant activity of the collagen peptide composition in the digestive system can be effectively maintained and has good digestive stability. The inclusion composition of the composite collagen peptide inclusion complex and the droplet polymerization composition of the composite emulsion are both conducive to the slow release of the antioxidant active substances in the collagen peptide composition in the body, thereby improving the bioavailability of the collagen peptide composition in the body.

[0125] Test Example 3

[0126] Thermal stability test

[0127] The ability of the collagen peptide compositions of Examples 1-3 and Comparative Examples 1-7 to scavenge DPPH free radicals after heating was measured. The specific steps are as follows:

[0128] 20 mL of each collagen peptide composition of Examples 1-3 and Comparative Examples 1-8 was taken as a test sample, and each test sample was placed in a 65°C water bath and heated for 20 min. Subsequently, each group of test samples after heating were diluted 100 times with distilled water as a test solution, and 2 mL of each group of test solutions was taken as a sample, and 2 mL of a 1×10 -4 mol / L DPPH anhydrous ethanol solution, shake vigorously, react at room temperature for 30 minutes, and then measure the absorbance at 517nm In addition, the blank group used an equal volume of anhydrous ethanol solution instead of the DPPH anhydrous ethanol solution. Similarly, 2 mL of the test solution was taken from each group of test solutions as a sample, and 2 mL of anhydrous ethanol solution was added to each sample, shaken vigorously, reacted at room temperature for 30 minutes, and then the absorbance was measured at 517 nm. The control group used an equal volume of distilled water instead of DPPH anhydrous ethanol solution. Similarly, 2 mL of the test solution was taken from each group as a sample, and 2 mL of distilled water was added to each sample. The mixture was shaken vigorously and reacted at room temperature for 30 minutes. Then, the absorbance was measured at 517 nm. . Calculate the scavenging rate of DPPH free radicals. Figure 2 shown.

[0129] like Figure 2 It can be seen that after the heat treatment, although the antioxidant activity of the collagen peptide composition of Examples 1-3 is reduced, the scavenging rate of DPPH free radicals is maintained at about 80%, indicating that the antioxidant activity of the collagen peptide composition remains relatively stable after the heat treatment, and can effectively ensure that the collagen peptide component and the composite plant extract component therein will not be oxidized due to high temperature.

[0130] Test Example 4

[0131] Sensory evaluation

[0132] A sensory evaluation panel was formed by 10 sensory assessors to evaluate the collagen peptide compositions of Examples 1-3 and Comparative Examples 1-7 according to smell and taste. Each sensory assessor put 5 mL of the collagen peptide composition in the mouth each time for sensory evaluation, so that the collagen peptide composition fully filled the entire oral cavity, stayed for 10 seconds, and rinsed the mouth three times before and after each sensory evaluation. The total score of the sensory evaluation was 10 points. The sensory evaluation criteria are shown in Table 1 below.

[0133] Table 1:

[0134]

[0135] According to the sensory evaluation standard, the collagen peptide compositions of Examples 1-3 and Comparative Examples 1-7 were subjected to sensory evaluation, and the average sensory score of the assessors was finally used as the result. The results are shown in Table 2 below.

[0136] Table 2:

[0137]

[0138] As can be seen in Table 2, it is obvious after sensory evaluation that directly adding the composite collagen peptide that has not been included in the collagen peptide composition will make the collagen peptide composition have a fishy taste and a certain bitterness, indicating that the composite collagen peptide can cover up the original fishy smell and improve the flavor after inclusion.

[0139] It should be understood that those skilled in the art can make improvements or changes based on the above description, and all such improvements and changes should fall within the scope of protection of the appended claims of the present invention. Parts not described in detail in this specification belong to the prior art known to those skilled in the art.

Claims

1. A collagen peptide composition, characterized in that: Including the following raw materials: A mixed liquid and a composite emulsion, wherein the volume ratio of the mixed liquid to the composite emulsion is (10-30):(1-3), and the mixed liquid is obtained by mixing composite collagen peptide inclusion complex, gamma-aminobutyric acid and distilled water in a mass ratio of (1-10):(0.3-2):(50-100); The composite collagen peptide inclusion complex is obtained by adsorbing the composite collagen peptide into caffeic acid porous starch ester, and then encapsulating the caffeic acid porous starch ester adsorbed with the composite collagen peptide using β-cyclodextrin; Caffeic acid porous starch ester is obtained by esterification reaction of activated caffeic acid and porous starch; The raw materials of the composite emulsion include: konjac glucomannan / quinoa protein complex, composite plant extract and distilled water. The mixing mass ratio of the konjac glucomannan / quinoa protein complex, composite plant extract and distilled water is (1-5): (3-9): (20-70). The konjac glucomannan / quinoa protein complex is obtained by mixing quinoa protein solution and konjac glucomannan solution and then vacuum drying.

2. A method for preparing the collagen peptide composition according to claim 1, characterized in that: The steps include: Step S1: preparation of caffeic acid porous starch ester, after activating caffeic acid, using the activated caffeic acid to react with porous starch for esterification, and then obtaining caffeic acid porous starch ester through precipitation, centrifugal separation, vacuum drying, grinding and sieving; Step S2: Preparation of composite collagen peptide inclusion complex, adding caffeic acid porous starch ester to the composite collagen peptide solution, stirring and mixing, and then adding β-cyclodextrin, stirring and mixing, and spray drying to obtain the composite collagen peptide inclusion complex; Step S3: preparing a composite emulsion, dissolving the quinoa protein solution and the konjac glucomannan solution to obtain a konjac glucomannan / quinoa protein complex, dissolving the konjac glucomannan / quinoa protein complex in distilled water, adding the composite plant extract, and homogenizing at high speed to obtain a composite emulsion; Step S4: Preparation of collagen peptide composition: mixing the composite collagen peptide inclusion complex, γ-aminobutyric acid and distilled water to obtain a mixed solution, adding the composite emulsion of step S3 to the mixed solution, and mixing by magnetic stirring to obtain a collagen peptide composition.

3. The method for preparing the collagen peptide composition according to claim 2, characterized in that: Step S1: Preparation of porous caffeic acid starch ester, specifically comprising the following steps: S1.1: 15 to 22 parts by weight of caffeic acid are added to 40 to 60 parts by weight of dichloromethane, and the mixture is stirred evenly by magnetic force to obtain solution A. 8 to 18 parts by weight of thionyl chloride are diluted with 8 to 12 parts by weight of dichloromethane to obtain solution B; S1.2: Under nitrogen atmosphere, add solution B dropwise into solution A while stirring. After the addition of solution B is complete, heat to 70-90°C and reflux for 3-5 hours. S1.3: After the reaction is completed, distillation under reduced pressure is performed to remove excess thionyl chloride and dichloromethane to obtain caffeoyl chloride; S1.4: Disperse 10 to 30 parts by weight of porous starch in 70 to 120 parts by weight of dimethyl sulfoxide, add 3 to 10 parts by weight of caffeoyl chloride and 1 to 5 parts by weight of pyridine, and reflux for reaction at 70 to 80° C. in a nitrogen atmosphere for 2 to 3 hours to obtain a reaction solution; S1.5: After the reaction solution is cooled to room temperature, 100 to 200 parts by weight of anhydrous ethanol is slowly added while stirring, and the mixture is centrifuged to remove the supernatant and collect the precipitate. The precipitate is vacuum dried, ground and sieved to obtain caffeic acid porous starch ester.

4. The method for preparing the collagen peptide composition according to claim 3, characterized in that: Step S2: Preparation of composite collagen peptide inclusion complex, specifically comprising the following steps: S2.1: adding the composite collagen peptide to distilled water to prepare a composite collagen peptide solution with a concentration of 4 to 8 mg / mL; S2.2: adding caffeic acid porous starch ester to the composite collagen peptide solution having a concentration of 4 to 8 mg / mL, wherein the mass ratio of the composite collagen peptide solution to the caffeic acid porous starch ester is (1 to 5):1, and stirring and mixing at 35 to 45° C. for 30 to 60 minutes to obtain a mixed solution; S2.3: Add 15 mg / mL β-cyclodextrin aqueous solution to the mixed solution, stir and mix evenly, and then spray dry to obtain a composite collagen peptide inclusion complex.

5. The method for preparing the collagen peptide composition according to claim 4, characterized in that: The composite collagen peptide is obtained by mixing fish bone collagen peptide, fish skin collagen peptide and fish scale collagen peptide in a mass ratio of (1-3):(1-3):(1-3).

6. The method for preparing the collagen peptide composition according to claim 5, characterized in that: The amount of β-cyclodextrin aqueous solution added is 1 to 3 times the mass of the mixed solution.

7. The method for preparing the collagen peptide composition according to claim 6, characterized in that: Step S3: Preparation of composite emulsion, specifically comprising the following steps: S3.1: Dissolve konjac glucomannan in distilled water, homogenize at a speed of 8000-10000 rpm for 1-3 min to obtain a konjac glucomannan solution with a concentration of 15-30 mg / mL; dissolve quinoa protein in distilled water, homogenize at a speed of 8000-10000 rpm for 1-3 min, then heat to 32-40° C., and stir magnetically for 2-3 h to fully hydrate the quinoa protein to obtain a quinoa protein solution with a concentration of 15-20 mg / mL; S3.2: adding the quinoa protein solution dropwise to the konjac glucomannan solution while stirring; after the quinoa protein solution is added dropwise, stirring is continued for 2 to 4 hours, and vacuum drying is performed to obtain a konjac glucomannan / quinoa protein complex; S3.3: Dissolve the konjac glucomannan / quinoa protein complex in distilled water, and then add the composite plant extract, wherein the mixing mass ratio of the konjac glucomannan / quinoa protein complex, the composite plant extract and distilled water is (1-5): (3-9): (20-70), and homogenize at a speed of 10000-15000 rpm for 3-5 minutes to obtain a composite emulsion.

8. The method for preparing the collagen peptide composition according to claim 7, characterized in that: The mixing volume ratio of the quinoa protein solution to the konjac glucomannan solution is (1-1.5): (1-1.5).

9. The method for preparing the collagen peptide composition according to claim 8, characterized in that: The composite plant extract contains at least two of lycopene, carotene, blueberry extract, passion fruit extract, sea buckthorn extract, pomegranate extract, flaxseed extract, mangosteen extract, prickly pear extract, grape seed extract and olive fruit extract.

10. The method for preparing the collagen peptide composition according to claim 9, characterized in that: Step S4: Preparation of collagen peptide composition, specifically comprising the following steps: The composite collagen peptide inclusion complex, γ-aminobutyric acid and distilled water are mixed in a mass ratio of (1-10): (0.3-2): (50-100), and magnetically stirred to obtain a mixed solution. The mixed solution and the composite emulsion are magnetically stirred to obtain a collagen peptide composition in a volume ratio of (10-30): (1-3).

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