Collagen peptide composition for repairing joints and preparation method thereof

Abstract

The invention relates to the field of biological medicines, in particular to a collagen peptide composition for repairing joints and a preparation method of the collagen peptide composition. Mainly comprises the following raw materials in parts by weight: 20-25 parts of high-dispersion collagen peptide, 4-6 parts of a lucid ganoderma extract, 4-6 parts of a shiitake mushroom extract, 4-6 parts of composite embedded nano calcium, 2-4 parts of hyaluronic acid, 0.4-0.6 part of vitamin K2, 0.2-0.4 part of tea polyphenol and 5-7 parts of pitaya fermentation puree. The high-dispersion collagen peptide is prepared by specific enzymolysis and ultrafiltration processes. The hydrogel has relatively small molecular weight and good water solubility, and can promote repair and regeneration of articular cartilage; the composite embedded nano-calcium improves the bioavailability of calcium and promotes bone mineral density increase and bone mineralization through the design of a dual embedding structure, the composition can efficiently repair articular cartilage and improve bone mineral density and bone calcium content through the synergistic effect of multiple raw materials, has excellent oxidation resistance and is suitable for joint repair and bone health maintenance.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of biological medicine, in particular to a collagen peptide composition for repairing joints and a preparation method thereof. BACKGROUND

[0002] With the aggravation of population aging, changes in lifestyle (such as sitting for a long time, excessive exercise), the incidence of joint degenerative diseases (such as osteoarthritis) and bone health problems (such as osteoporosis) is increasing year by year, which has become an important health risk affecting the quality of life of the population. The wear and tear of articular cartilage and the decrease of bone density are the core causes of such problems, and the weak self-repairing ability of cartilage and the accelerated calcium loss of bone tissue make it a research hotspot to supplement functional ingredients through diet to assist joint repair and bone health maintenance.

[0003] A Chinese patent with application number CN202011445307.4 discloses a bovine collagen peptide composition and its preparation method and application. The composition includes the following components and their weight fractions: bovine collagen peptide 14-28 parts, chondroitin sulfate 10-16 parts, and plant extract 10-22 parts, which is composed of sinomenium acutum extract and sagittaria sagittifolia extract. A Chinese patent with application number CN202210355647.0 discloses a composition for relieving bone and joint pain and its preparation method and application. The composition includes the following components and their weight fractions: wolfberry 1-12 parts by weight, raspberry 1-12 parts by weight, polygonatum 1-15 parts by weight, yam 1-30 parts by weight, ginseng 1-9 parts by weight, peach kernel 1-10 parts by weight, and pueraria 1-15 parts by weight. The preparation method of the composition includes the following steps: wolfberry, raspberry, polygonatum, yam, ginseng, peach kernel, and pueraria are extracted with water, concentrated, dried, and pulverized, and then mixed with collagen peptide, hydrolyzed egg yolk powder, bovine colostrum basic protein, vitamin C, and auxiliary materials to form an oral preparation. However, the above two schemes lack sufficient calcium source and nutrients that promote calcium absorption and directional deposition. Although the added collagen peptide is an important component of the organic matrix of the skeleton, which helps to maintain the bone weight and toughness, and vitamin C helps to synthesize collagen and has a certain promoting effect on the formation of bone matrix, without the participation of calcium, it is impossible to effectively increase the mineral density and hardness of the skeleton. Therefore, the effects of the above two compositions in improving bone health are limited.

[0004] Therefore, it is a technical problem to be solved in the field to develop a joint repair collagen peptide composition with multi-dimensional synergistic effect, high stability, and high bioavailability, and a preparation method thereof. SUMMARY

[0005] To solve the above problems, the application provides a collagen peptide composition for repairing joints and a preparation method thereof. The high-dispersed collagen peptide is obtained by mixing bovine bone collagen peptide with deionized water, removing impurities through activated carbon, complex protease enzymolysis, and ultrafiltration membrane separation. The composite embedded nano calcium is obtained by reacting casein phosphopeptide and sodium alginate with calcium carbonate, and then embedding with hydroxypropyl-beta-cyclodextrin. Finally, the multi-target and synergistic joint repair composition is obtained by scientifically proportioning the high-dispersed collagen peptide, the composite embedded nano calcium, chondroitin sulfate, hyaluronic acid, vitamin K2, ganoderma lucidum / mushroom extract, and pitaya fermented original pulp, so as to realize the integrated and efficient repair and protection of joint cartilage and bone.

[0006] The technical scheme adopted by the application to achieve the above object is as follows: A collagen peptide composition for repairing joints comprises the following raw materials in parts by weight: 20-25 parts of high-dispersed collagen peptide, 10-15 parts of chondroitin sulfate, 4-6 parts of ganoderma lucidum extract, 4-6 parts of mushroom extract, 4-6 parts of composite embedded nano calcium, 2-4 parts of hyaluronic acid, 0.4-0.6 parts of vitamin K2, 0.2-0.4 parts of tea polyphenol, 5-7 parts of pitaya fermented original pulp, 4-6 parts of malt dextrin, and 0.1-0.2 parts of epsilon-polylysine. The preparation method of the high-dispersed collagen peptide is as follows: Step S1, the bovine bone collagen peptide is added to deionized water, stirred at 40-50 DEG C for 20-25 min, then activated carbon is added, and stirring is continued for 30-40 min. After cooling to room temperature, the filtrate is obtained by filtration. Step S2, the complex protease is added to the filtrate, and the enzyme is reacted at 50-55 DEG C and pH 7.0-7.5 for 2-2.5 h. After the enzyme reaction is completed, the permeate is obtained by ultrafiltration membrane separation after cooling to room temperature. Step S3, the permeate is concentrated, chitosan is added, and the reaction is carried out at 55-65 DEG C and pH 7.8-8.2 for 1.5-2 h. After post-treatment, the high-dispersed collagen peptide is obtained.

[0007] Further, the solution prepared by adding the bovine bone collagen peptide to deionized water in step S1 has a mass concentration of 7-9%, and the mass ratio of bovine bone collagen peptide to activated carbon is 1:0.0015-0.0025.

[0008] Further, the complex protease in step S2 is composed of alkaline protease and papain in a mass ratio of 2.5-3.5:1, and the mass-volume ratio of complex protease to filtrate is 0.09-0.1 g / 100 mL.

[0009] Further, the concentration of the permeate in step S3 is to 9-11% solid content, and the mass / volume ratio of chitosan to the permeate is 0.035-0.045 g / 100 mL.

[0010] The preparation method of the composite embedded nano-calcium is as follows: Step a, first add casein phosphopeptide and sodium alginate into deionized water, stir at 55-60℃ and pH 6.5-7.0 for 30-40 min to obtain a composite embedding solution, then add calcium carbonate, stir at 40-45℃ for 60-70 min, and obtain primary embedded nano-calcium after post-processing; Step b, add hydroxypropyl-β-cyclodextrin into deionized water, stir at 60-65℃ for 20-30 min to obtain a hydroxypropyl-β-cyclodextrin solution, then cool to 35-40℃, add the primary embedded nano-calcium, and stir at constant temperature for 90-100 min, and obtain the composite embedded nano-calcium after post-processing.

[0011] Further, in step a, the mass ratio of casein phosphopeptide to sodium alginate is 1.5-2.5:1, the amount of sodium alginate added in deionized water is 1-1.5 g / 100 mL, and the mass ratio of calcium carbonate to the composite embedding solution is 1:5-6.

[0012] Further, in step b, the mass concentration of the hydroxypropyl-β-cyclodextrin solution is 9.5-10.5%, and the mass ratio of the primary embedded nano-calcium to hydroxypropyl-β-cyclodextrin is 2.5-3.5:7.

[0013] The dragon fruit fermented original pulp is prepared by blending red heart dragon fruit fermented liquid and galactooligosaccharide at a mass ratio of 10:3-6.

[0014] A preparation method of a collagen peptide composition for repairing joints, comprising the following steps: (1) Screen high-dispersed collagen peptide, chondroitin sulfate, ganoderma extract, lentinus edodes extract, composite embedded nano-calcium, hyaluronic acid, vitamin K2, tea polyphenol, and ε-polylysine, respectively, and reserve; (2) Add malt dextrin into deionized water, heat to 60-65℃ and stir to dissolve, cool to 30-35℃, then add tea polyphenol, ε-polylysine, high-dispersed collagen peptide, chondroitin sulfate in sequence, stir at a rotation speed of 500-600 r / min for 20-30 min, add dragon fruit fermented original pulp and continue to stir for 10-15 min to form a mixed solution A; (3) Add ganoderma extract, lentinus edodes extract, and hyaluronic acid into the mixed solution A, continue to stir for 20-30 min to form a mixed solution B, mix vitamin K2 and composite embedded nano-calcium uniformly, and add into the mixed solution B, and then perform homogenization treatment to obtain the product.

[0015] The present application has the following advantages: The collagen peptide composition for repairing joints prepared by the present application realizes integrated repair of joints and bones through the mutual synergy between raw materials. The core is that the high-dispersed collagen peptide and the complex-embedded nano-calcium form a "harmony of rigidity and flexibility" bone repair cornerstone: the collagen peptide as a high-quality raw material of an organic framework ensures efficient absorption and targeted delivery due to its high dispersibility; and the double-embedded nano-calcium as a calcium source significantly improves the absorption rate and stability, and the two synergistically enhance each other under the precise regulation of vitamin K2, vitamin K2 activates osteocalcin to guide calcium ions to be efficiently deposited in the collagen network, thereby significantly improving bone density, bone calcium content and bone mass.

[0016] Meanwhile, chondroitin sulfate and hyaluronic acid directly supplement the cartilage matrix and lubricate the joints; ganoderma lucidum and lentinus edodes extract and tea polyphenol form a powerful antioxidant network to scavenge free radicals and reduce joint inflammatory damage, and this network is more stable due to the carrier protection effect of collagen peptide; pitaya fermented raw pulp has antioxidant and prebiotic functions, and together with ε-polylysine, it regulates the intestinal microenvironment and indirectly promotes nutrient absorption. These ingredients provide multiple supports required for joint repair through synergistic effect, not only repairing cartilage and bones, but also protecting joints from oxidative stress damage, significantly improving the repair effect. Therefore, the collagen peptide composition of the present application has significant advantages in joint repair and bone health. BRIEF DESCRIPTION OF DRAWINGS

[0017] Fig. 1 is a DPPH free radical scavenging capacity line graph; Fig. 2 is a superoxide anion free radical scavenging capacity line graph; Fig. 3 is a hydroxyl radical scavenging capacity line graph. DETAILED DESCRIPTION

[0018] The technical solutions in the embodiments of the present application will be described below in conjunction with the embodiments of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, not all embodiments. Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art without creative labor fall within the scope of the present application.

[0019] The raw materials used in the following examples are all ordinary commercially available products. Chondroitin sulfate with a particle size of 80-100 mesh and an active ingredient content of 99% was purchased from Shandong Pingju Biological Technology Co., Ltd.; Ganoderma extract with a specification of 10:1 and a mesh size of 80-100 was purchased from Shanyang Lianfeng Biological Technology Co., Ltd.; Shiitake mushroom extract with a particle size of 80 mesh and an active ingredient content of 99% was purchased from Sichuan Huanxu Biological Technology Co., Ltd.; Hyaluronic acid with an active ingredient content of 99% was purchased from Hebei Jiuxing Chemical Product Co., Ltd.; Vitamin K2 with an active ingredient content of 99% and a particle size of 120 mesh was purchased from Hubei Haijia Biological Technology Co., Ltd.; Tea polyphenols with a particle size of 100 mesh, a specification of 10:1 and an active ingredient content of 98% were purchased from Lanzhou Waterless Biological Technology Co., Ltd.; Malt dextrin with an active ingredient content of 99.9% and a density of 1.3-1.6 was purchased from Shandong Yingsheng Chemical Co., Ltd.; ε-Polylysine with an active ingredient content of 97.2% was purchased from Sichuan Huanxu Biological Technology Co., Ltd.; Bovine bone collagen peptide with an active ingredient content of 99.2% was purchased from Jiangsu Caiwei Biological Technology Co., Ltd.; Activated carbon with a particle size of 325 mesh was purchased from Shijiazhuang Hongsen Activated Carbon Co., Ltd.; Chitosan with an active ingredient content of 99% was purchased from Xi'an Hongyao Pharmaceutical Auxiliary Material Co., Ltd.; Alkaline protease was purchased from Taian Xindeli Biological Engineering Co., Ltd.; Papain was purchased from Nanning Dongheng Huadao Biological Technology Co., Ltd.; Casein phosphopeptide with a peptide content of 37.98% was purchased from Hebei Hongtao Biological Engineering Co., Ltd.; Sodium alginate with an active ingredient content of 99% was purchased from Xi'an Hongyao Pharmaceutical Auxiliary Material Co., Ltd.; Calcium carbonate with a particle size of 1250 mesh and an active ingredient content of 99% and a density of 1.7 was purchased from Henan Tuohong Chemical Co., Ltd.; Hydroxypropyl-β-cyclodextrin with an active ingredient content of 99.5% was purchased from Shandong Tongwang Biological Technology Co., Ltd.; Galacto-oligosaccharide with an active ingredient content of 99% was purchased from Sichuan Kangbaier Biological Technology Co., Ltd.

[0020] Example 1 A collagen peptide composition for repairing joints comprises the following raw materials in parts by weight: high-dispersion collagen peptide 20 parts, chondroitin sulfate 10 parts, Ganoderma extract 4 parts, Shiitake mushroom extract 4 parts, composite embedded nano-calcium 4 parts, hyaluronic acid 2 parts, vitamin K2 0.4 parts, tea polyphenols 0.2 parts, dragon fruit fermentation original paste 5 parts, malt dextrin 4 parts, ε-polylysine 0.1 part; The preparation method of the high-dispersion collagen peptide is as follows: Step S1, the bovine bone collagen peptide is added into deionized water, stirred at 45℃ and 300rpm for 22min, then activated carbon is added and stirred for another 35min, and then filtered through a filter membrane with a pore size of 0.22μm after natural cooling to room temperature to obtain a filtrate, wherein the bovine bone collagen peptide is prepared into a solution with a mass concentration of 8% in deionized water, and the mass ratio of bovine bone collagen peptide to activated carbon is 1:0.002; Step S2, the protease is added into the filtrate, and enzymolysis is carried out at 52℃ and pH 7.0-7.5 for 2.2h, and then the enzyme is inactivated at 90℃ for 15min, and then the filtrate is separated through an ultrafiltration membrane with a molecular weight cut-off of 3000Da after natural cooling to room temperature, wherein the protease is composed of alkaline protease and papain with a mass ratio of 3:1, and the mass / volume ratio of protease to filtrate is 0.095g / 100mL; Step S3, the permeate is concentrated, and chitosan is added, and reacted at 60℃ and pH 7.8-8.2 for 1.7h, and then natural cooling to room temperature, and then pre-frozen at-40℃ for 3h, and then sublimation dried at a vacuum degree of 0.01-0.02MPa and sublimation temperature of-10 to-5℃ for 10h to obtain the high dispersion collagen peptide, wherein the permeate is concentrated to a solid content of 10%, and the mass / volume ratio of chitosan to permeate is 0.04g / 100mL.

[0021] The preparation method of the composite embedded nano calcium is as follows: Step a, first, casein phosphopeptide and sodium alginate are added into deionized water, stirred at 57℃ and pH 6.5-7.0 for 35min to obtain a composite embedding liquid, then calcium carbonate is added, stirred at 42℃ and 400rpm for 65min, and then spray dried to obtain the primary embedded nano calcium, wherein the mass ratio of casein phosphopeptide to sodium alginate is 2:1, the amount of sodium alginate added in deionized water is 1.2g / 100mL, and the mass ratio of calcium carbonate to composite embedding liquid is 1:5.5; Step b, hydroxypropyl-β-cyclodextrin is added into deionized water, stirred at 62℃ for 25min to obtain a hydroxypropyl-β-cyclodextrin solution, then cooled to 37℃, and the primary embedded nano calcium is added, stirred at 300rpm for 95min, and then natural cooling to room temperature, and then pre-frozen at-40℃ for 3h, and then sublimation dried at a vacuum degree of 0.01-0.02MPa and sublimation temperature of-15 to-10℃ for 15h to obtain the composite embedded nano calcium, wherein the mass concentration of the hydroxypropyl-β-cyclodextrin solution is 10%, and the mass ratio of the primary embedded nano calcium to hydroxypropyl-β-cyclodextrin is 3:7.

[0022] The pitaya fermentation original pulp is prepared by red heart pitaya fermentation liquid and galacto-oligosaccharide at a mass ratio of 10:3.

[0023] The red heart pitaya fermentation liquid and the preparation method thereof have been disclosed in a patent document with the application number CN201810157407.3.

[0024] A preparation method of a collagen peptide composition for repairing joints, comprising the following steps: (1) Screen high-dispersed collagen peptide, chondroitin sulfate, ganoderma lucidum extract, lentinus edodes extract, composite embedded nano calcium, hyaluronic acid, vitamin K2, tea polyphenol, and ε-polylysine through a 60-mesh screen, and reserve for use; (2) Add malt dextrin into deionized water, heat to 60 DEG C and stir to dissolve, cool to 30 DEG C, then add tea polyphenol, ε-polylysine, high-dispersed collagen peptide, and chondroitin sulfate in sequence, stir for 20 min under the condition of 500 rpm, add pitaya fermentation original pulp and continue to stir for 10 min, to form a mixed solution A; (3) Add ganoderma lucidum extract, lentinus edodes extract, and hyaluronic acid into the mixed solution A, continue to stir for 20 min, to form a mixed solution B, mix vitamin K2 and composite embedded nano calcium uniformly, add into the mixed solution B, and homogenize under the homogenization pressure of 30-35 MPa, 3 times, to obtain the product.

[0025] Example 2 A collagen peptide composition for repairing joints, comprising the following raw materials in parts by weight: high-dispersed collagen peptide 25 parts, chondroitin sulfate 15 parts, ganoderma lucidum extract 6 parts, lentinus edodes extract 6 parts, composite embedded nano calcium 6 parts, hyaluronic acid 4 parts, vitamin K2 0.6 parts, tea polyphenol 0.4 parts, pitaya fermentation original pulp 7 parts, malt dextrin 6 parts, and ε-polylysine 0.2 parts. The preparation method of the high-dispersed collagen peptide and the preparation method of the composite embedded nano calcium are the same as those in Example 1. The pitaya fermentation original pulp is prepared by red heart pitaya fermentation liquid and galacto-oligosaccharide at a mass ratio of 10:6.

[0026] The red heart pitaya fermentation liquid and the preparation method thereof have been disclosed in a patent document with the application number CN201810157407.3.

[0027] A preparation method of a collagen peptide composition for repairing joints, comprising the following steps: (1) Screen high-dispersed collagen peptide, chondroitin sulfate, ganoderma lucidum extract, lentinus edodes extract, composite embedded nano calcium, hyaluronic acid, vitamin K2, tea polyphenol, and ε-polylysine through a 60-mesh screen, and reserve for use; (2) malt dextrin is added to deionized water, heated to 65℃ and stirred to dissolve, cooled to 35℃, then tea polyphenols, ε-polylysine, high dispersion collagen peptide, chondroitin sulfate are added in sequence, stirred at 600 rpm for 30 min, pitaya fermentation original syrup is added and stirred for 15 min to form a mixed solution A; (3) ganoderma extract, lentinus edodes extract, hyaluronic acid are added to the mixed solution A, continue to stir for 30 min to form a mixed solution B, vitamin K2 and composite embedded nano calcium are mixed uniformly and added to the mixed solution B, homogenized at a pressure of 30-35 MPa, 3 times, to obtain the collagen peptide composition.

[0028] Example 3 A collagen peptide composition for repairing joints, comprising the following raw materials in parts by weight: high dispersion collagen peptide 23 parts, chondroitin sulfate 12 parts, ganoderma extract 5 parts, lentinus edodes extract 5 parts, composite embedded nano calcium 5 parts, hyaluronic acid 3 parts, vitamin K2 0.5 parts, tea polyphenols 0.3 parts, pitaya fermentation original syrup 6 parts, malt dextrin 5 parts, ε-polylysine 0.15 parts. The preparation method of the high dispersion collagen peptide and the preparation method of the composite embedded nano calcium are the same as those in Example 1. The pitaya fermentation original syrup is prepared by blending red heart pitaya fermentation broth and galactooligosaccharide at a mass ratio of 10:5.

[0029] The red heart pitaya fermentation broth and its preparation method have been disclosed in the patent document with application number CN201810157407.3.

[0030] A preparation method of a collagen peptide composition for repairing joints, comprising the following steps: (1) high dispersion collagen peptide, chondroitin sulfate, ganoderma extract, lentinus edodes extract, composite embedded nano calcium, hyaluronic acid, vitamin K2, tea polyphenols, and ε-polylysine are sieved through a 60-mesh sieve and prepared for use; (2) malt dextrin is added to deionized water, heated to 62℃ and stirred to dissolve, cooled to 32℃, then tea polyphenols, ε-polylysine, high dispersion collagen peptide, and chondroitin sulfate are added in sequence, stirred at 550 rpm for 25 min, pitaya fermentation original syrup is added and stirred for 13 min to form a mixed solution A; (3) ganoderma extract, lentinus edodes extract, and hyaluronic acid are added to the mixed solution A, continue to stir for 25 min to form a mixed solution B, vitamin K2 and composite embedded nano calcium are mixed uniformly and added to the mixed solution B, homogenized at a pressure of 30-35 MPa, 3 times, to obtain the collagen peptide composition.

[0031] Example 4 A collagen peptide composition for repairing joints comprises the following raw materials in parts by weight: 23 parts of high-dispersion collagen peptide, 12 parts of chondroitin sulfate, 5 parts of ganoderma extract, 5 parts of lentinus edodes extract, 5 parts of composite-embedded nano calcium, 3 parts of hyaluronic acid, 0.5 part of vitamin K2, 0.3 part of tea polyphenol, 6 parts of pitaya fermented raw pulp, 5 parts of malt dextrin, and 0.15 part of epsilon-polylysine; The preparation method of the high-dispersion collagen peptide is as follows: In step S1, the collagen peptide of bovine bone is added to deionized water, stirred at 50 DEG C and 300 rpm for 25 min, then activated carbon is added and stirred for another 40 min, and then the mixture is naturally cooled to room temperature and filtered through a filter membrane with a pore size of 0.22 microns to obtain a filtrate, wherein the solution prepared by adding the collagen peptide of bovine bone to deionized water has a mass concentration of 9%, and the mass ratio of the collagen peptide of bovine bone to activated carbon is 1:0.0025. In step S2, complex protease is added to the filtrate, and enzymolysis is carried out at 55 DEG C and pH 7.0-7.5 for 2.5 h, then the enzyme is inactivated at 90 DEG C for 15 min, and then the mixture is naturally cooled to room temperature and separated through an ultrafiltration membrane with a molecular weight cut-off of 3000 Da to obtain a permeate, wherein the complex protease is composed of alkaline protease and papain at a mass ratio of 3.5:1, and the mass-volume ratio of the complex protease to the filtrate is 0.1 g / 100 mL. In step S3, the permeate is concentrated, chitosan is added, and reaction is carried out at 65 DEG C and pH 7.8-8.2 for 2 h, then the mixture is naturally cooled to room temperature, pre-frozen at -40 DEG C for 3 h, and then sublimation dried at a vacuum degree of 0.01-0.02 MPa and a sublimation temperature of -10 to -5 DEG C for 10 h to obtain the high-dispersion collagen peptide, wherein the permeate is concentrated to a solid content of 11%, and the mass-volume ratio of chitosan to the permeate is 0.045 g / 100 mL.

[0032] The preparation method of the composite-embedded nano calcium is as follows: In step a, casein phosphopeptide and sodium alginate are first added to deionized water, stirred at 60 DEG C and pH 6.5-7.0 for 40 min to obtain a composite embedding solution, then calcium carbonate is added, stirred at 45 DEG C and 400 rpm for 70 min, and then spray dried to obtain the primary embedded nano calcium, wherein the mass ratio of casein phosphopeptide to sodium alginate is 2.5:1, the amount of sodium alginate added to deionized water is 1.5 g / 100 mL, and the mass ratio of calcium carbonate to the composite embedding solution is 1:6. Step b, add hydroxypropyl-β-cyclodextrin into deionized water, stir at 65℃ for 30 min to obtain a hydroxypropyl-β-cyclodextrin solution, then cool to 40℃, add the primary embedded nano-calcium, stir at a rotation speed of 300 rpm for 100 min, naturally cool to room temperature, pre-freeze at -40℃ for 3 h, then sublimate dry under the conditions of a vacuum degree of 0.01-0.02 MPa and a sublimation temperature of -15 to -10℃ for 15 h to obtain the composite embedded nano-calcium, wherein the mass concentration of the hydroxypropyl-β-cyclodextrin solution is 10.5%, and the mass ratio of the primary embedded nano-calcium to the hydroxypropyl-β-cyclodextrin is 3.5:7.

[0033] The pitaya fermented original pulp is prepared by blending red heart pitaya fermented liquid and galactooligosaccharide at a mass ratio of 10:5.

[0034] The red heart pitaya fermented liquid and the preparation method thereof have been disclosed in a patent document with the application number CN201810157407.3.

[0035] A preparation method of a collagen peptide composition for repairing joints, comprising the following steps: (1) Screen high-dispersed collagen peptide, chondroitin sulfate, ganoderma lucidum extract, lentinus edodes extract, composite embedded nano-calcium, hyaluronic acid, vitamin K2, tea polyphenol, and ε-polylysine through a 60-mesh screen, and reserve for use; (2) Add malt dextrin into deionized water, heat to 62℃ and stir to dissolve, cool to 32℃, then add tea polyphenol, ε-polylysine, high-dispersed collagen peptide, and chondroitin sulfate in sequence, stir at a rotation speed of 550 rpm for 25 min, add pitaya fermented original pulp and continue to stir for 13 min to form a mixed solution A; (3) Add ganoderma lucidum extract, lentinus edodes extract, and hyaluronic acid into the mixed solution A, continue to stir for 25 min to form a mixed solution B, mix vitamin K2 and composite embedded nano-calcium uniformly, add into the mixed solution B, and homogenize under a homogenization pressure of 30-35 MPa for 3 times to obtain the collagen peptide composition.

[0036] Comparative Example 1 A collagen peptide composition for repairing joints, comprising the following raw materials in parts by weight: high-dispersed collagen peptide 10 parts, chondroitin sulfate 12 parts, ganoderma lucidum extract 5 parts, lentinus edodes extract 5 parts, composite embedded nano-calcium 1 part, hyaluronic acid 3 parts, vitamin K2 0.5 part, tea polyphenol 0.3 part, pitaya fermented original pulp 1 part, malt dextrin 5 parts, and ε-polylysine 0.15 part. The preparation method of the high-dispersed collagen peptide is as follows: Step S1, the bovine bone collagen peptide is added into deionized water, stirred at 45℃ and 300rpm for 10min, then activated carbon is added and stirred for 35min, and then cooled to room temperature, filtered with a filter membrane with a pore size of 0.22μm to obtain a filtrate, wherein the bovine bone collagen peptide is prepared into a solution with a mass concentration of 8% in deionized water, and the mass ratio of bovine bone collagen peptide to activated carbon is 1:0.001; Step S2, the protease is added into the filtrate, and enzymolysis is carried out at 70℃ and pH 4.0-4.5 for 2.2h, then the enzyme is inactivated at 90℃ for 15min, and then cooled to room temperature, and then separated by ultrafiltration membrane with a molecular weight cut-off of 3000Da to obtain a permeate, wherein the protease is composed of alkaline protease and papain with a mass ratio of 3:1, and the mass / volume ratio of protease to filtrate is 0.095g / 100mL; Step S3, the permeate is concentrated, chitosan is added, and reacted at 60℃ and pH 7.8-8.2 for 1.7h, then cooled to room temperature, pre-frozen at-40℃ for 3h, and then sublimed and dried at a vacuum degree of 0.01-0.02MPa and a sublimation temperature of-10 to-5℃ for 10h to obtain the high-dispersion collagen peptide, wherein the permeate is concentrated to a solid content of 10%, and the mass / volume ratio of chitosan to permeate is 0.01g / 100mL.

[0037] The preparation method of the composite embedded nano-calcium is as follows: Step a, first, casein phosphopeptide and sodium alginate are added into deionized water, stirred at 57℃ and pH 6.5-7.0 for 35min to obtain a composite embedding liquid, then calcium carbonate is added, stirred at 42℃ and 400rpm for 65min, and then spray dried to obtain the primary embedded nano-calcium, wherein the mass ratio of casein phosphopeptide to sodium alginate is 2:1, the amount of sodium alginate added in deionized water is 1.2g / 100mL, and the mass ratio of calcium carbonate to composite embedding liquid is 1:1; Step b, hydroxypropyl-β-cyclodextrin is added into deionized water, stirred at 62℃ for 25min to obtain a hydroxypropyl-β-cyclodextrin solution, then cooled to 37℃, the primary embedded nano-calcium is added, stirred at 300rpm for 10min, then cooled to room temperature, pre-frozen at-40℃ for 3h, and then sublimed and dried at a vacuum degree of 0.01-0.02MPa and a sublimation temperature of-15 to-10℃ for 15h to obtain the composite embedded nano-calcium, wherein the mass concentration of the hydroxypropyl-β-cyclodextrin solution is 10%, and the mass ratio of the primary embedded nano-calcium to hydroxypropyl-β-cyclodextrin is 1:7.

[0038] The dragon fruit fermented pulp is made by mixing red dragon fruit fermentation liquid and galactooligosaccharides in a mass ratio of 10:1.

[0039] The fermented liquid of red dragon fruit and its preparation method have been disclosed in the patent document with application number CN201810157407.3.

[0040] A method for preparing a collagen peptide composition for repairing joints includes the following steps: (1) Highly dispersed collagen peptides, chondroitin sulfate, Ganoderma lucidum extract, shiitake mushroom extract, composite encapsulated nano-calcium, hyaluronic acid, vitamin K2, tea polyphenols, and ε-polylysine were sieved through a 60-mesh sieve and set aside for later use; (2) Add maltodextrin to deionized water and stir. Then add tea polyphenols, ε-polylysine, highly dispersed collagen peptides and chondroitin sulfate in sequence. Stir for 25 minutes at 550 rpm. Add dragon fruit fermentation pulp and continue stirring for 13 minutes to form mixture A. (3) Add Ganoderma lucidum extract, shiitake mushroom extract and hyaluronic acid to mixture A, and continue stirring for 25 minutes to form mixture B. Mix vitamin K2 and composite embedded nano calcium evenly and add it to mixture B. Homogenize under a homogenization pressure of 30-35 MPa for 3 times to obtain the final product.

[0041] Comparative Example 2 In this comparative example, commercially available bovine bone collagen peptides were used instead of highly dispersed collagen peptides, and everything else was the same as in Example 1.

[0042] Comparative Example 3 In this comparative example, commercially available calcium carbonate was used instead of composite-encapsulated nano-calcium, and all other aspects were the same as in Example 1.

[0043] Comparative Example 4 In this comparative example, commercially available collagen peptides were used instead of highly dispersed collagen peptides, and commercially available calcium carbonate was used instead of composite encapsulated nano-calcium. All other aspects were the same as in Example 1.

[0044] (1) Antioxidant test DPPH free radical scavenging ability: Samples from Examples 1-4 and Comparative Examples 1-4 were prepared into 0.5 mg / mL solutions, with concentrations of [missing information]. 2 mL of DPPH anhydrous ethanol solution was prepared and stored protected from light. 2 mL of the sample solution was mixed with 2 mL of DPPH anhydrous ethanol solution and shaken vigorously. The mixture was allowed to react at room temperature for 30 min, and the absorbance (Ai) was measured at 517 nm. The blank group was prepared using an equal volume of anhydrous ethanol solution instead of DPPH solution, and the control group was prepared using an equal volume of distilled water instead of the sample solution. The DPPH free radical scavenging rate was calculated using the following formula: In the formula, A0 represents the absorbance of the control group, Ai represents the absorbance of the sample group, and Aj represents the absorbance of the blank group.

[0045] Superoxide anion radical scavenging capacity: Under constant temperature of 25℃, take 3 mL of 50 mmol / L pH 8.2 Tris-HCl buffer solution (containing 1 mmol / L EDTA) and 10 μL of 50 mmol / L pyrogallol, mix rapidly, place in a 1 cm quartz cuvette, and measure the absorbance at 325 nm every 30 s. The reaction is stopped after 4.5 min. Plot absorbance against time, and the slope is the pyrogallol self-oxidation rate, A0. Add collagen peptide composition at a ratio of 0.5 mg / mL to 3 mL of pH 8.2, 50 mmol / L Tris-HCl buffer, and measure the pyrogallol oxidation rate As at 325 nm using the above method. The superoxide anion radical scavenging rate is calculated using the following formula: .

[0046] Hydroxyl radical scavenging ability: Take 0.1 mL of a solution with a concentration of 10 mmol / mL. The mixture was prepared by adding 0.3 mL of deoxyribose (10 mmol / mL), followed by 0.2 mL of the collagen peptide composition sample solution prepared in Examples 1-4 and Comparative Examples 1-4 with a concentration of 0.5 mg / mL. The volume was adjusted to 1.9 mL with 0.1 mol / L phosphate buffer (pH 7.4), and then 0.1 mL of H2O2 (10 mmol / mL) was added. After mixing, the mixture was placed in a 37°C water bath for 1 h. Then, 1 mL of 2.8% (w / w) trichloroacetic acid (TCA) solution and 1 mL of 1.0% (w / w) thiobarbituric acid (TBA) solution were added and mixed. The mixture was then reacted in a boiling water bath for 15 min. After cooling, the absorbance was measured at 532 nm. The absorbance without the addition of a scavenging agent is Ac. If the sample is added and has a scavenging effect on ·OH, it can inhibit the formation of oxidation products, resulting in a decrease in absorbance, which is measured as As. The actual blank absorbance is represented by A0. The formula for calculating the scavenging rate of hydroxyl radicals is: .

[0047] The results are shown in Table 1 and Figs. 1-3 As shown.

[0048] Table 1. Antioxidant performance test of collagen peptide compositions used for joint repair.

[0049] (2) Rat experiment Select the weight of 180g-230g female clean level Wistar rats, the rats were divided into 9 groups (Examples 1-4, Comparative Examples 1-4, solvent control group each corresponding to a group of rats), each group of 10, according to the recommended dose of 20g / day, adult body weight of 60kg, then the recommended dose of rats per day is 0.33g / kg·bw. According to 0.1mL / 10g·bw body weight, rats were gavaged once a day, the solvent control group with distilled water gavage. The test period is 12 weeks, during which the rats were single caged, fed with estrogen-free active substances of the configuration of the feed, free to drink deionized water, the end of the femoral artery bloodletting animals. The left femur of rats was dried in 105℃ oven to constant weight, then weighed, placed in a flask for digestion, and the bone calcium content was determined by atomic absorption method, and the bone density was determined by bone density meter. The results are shown in Table 2.

[0050] Table 2 femur bone weight, bone density and bone calcium content performance test of collagen peptide composition for repairing joints

[0051] As shown in Tables 1-2 and Figs. 1-3 The collagen peptide composition for repairing joints prepared in Examples 1-4 is significantly better than Comparative Example 1 in terms of antioxidant performance (DPPH free radical scavenging ability, superoxide anion radical scavenging ability, hydroxyl radical scavenging ability), femur bone weight, bone density and bone calcium content, etc. It can be seen that the collagen peptide composition prepared in the examples can repair joints, improve bone density and maintain bone health, so it can be concluded that the ratio of the collagen peptide composition proposed in the present application is the best, and the femur bone weight, bone density and bone calcium content are better than the solvent control group, which shows that the compositions prepared in Examples 1-3 have the effect of improving bone health.

[0052] As shown in Tables 1-2 and Figs. 1-3As shown, compared with the examples, the antioxidant performance and femur-related data of the collagen peptide composition prepared in Comparative Example 2 are all decreased, because the commercially available bovine bone collagen peptide is used to replace the high-dispersion collagen peptide, which provides the most direct and essential building block for the repair of articular cartilage, bone and connective tissue, i.e., collagen, and through a special preparation process, the molecular weight is smaller, the distribution is more concentrated, and the surface properties are improved, so that it can be highly dispersed in aqueous solution and body fluid and is not easy to aggregate, so that it can be more quickly and in larger amounts absorbed by the intestinal tract, and more effectively targeted to the joint and bone tissue that needs to be repaired, and the special preparation process makes the high-dispersion collagen peptide have better biocompatibility, so that it can fully play a synergistic effect with other active ingredients (such as chondroitin sulfate, calcium, etc.); the complex embedding of nano-calcium can improve the stability and absorption rate of calcium, and the high-dispersion collagen peptide provides a precise anchoring site for the absorbed calcium, and the two synergize to ensure that calcium can be efficiently transported to the bone and deposited, thereby significantly increasing the bone calcium content and bone density. The high-dispersion collagen peptide itself has a certain antioxidant activity, the histidine, proline and other amino acids in the collagen peptide chain and the terminal amino acid residues have the ability to scavenge free radicals, and after the complex enzymolysis of step S2, more active groups are exposed, enhancing its direct antioxidant capacity; in addition, tea polyphenols, ganoderma extract, shiitake mushroom extract and other antioxidants in the composition are strong but may be unstable, and the high-dispersion collagen peptide can combine with these ingredients through intermolecular interactions such as hydrogen bonds and hydrophobic interactions, achieve a protective effect, reduce their oxidation inactivation during processing and storage, and ensure that they can fully play a role when they reach the body.

[0053] As shown in Tables 1-2 and Figs. 1-3As shown, compared with the examples, the antioxidant properties and femoral-related data of the collagen peptide composition prepared in Comparative Example 3 still decreased. This is because commercially available calcium carbonate was used instead of composite-encapsulated nano-calcium. Composite-encapsulated nano-calcium mainly achieves easier absorption and utilization in vivo by encapsulating the calcium source at the nanoscale. Through its combination with collagen peptides, composite-encapsulated nano-calcium can play a role in joint repair, enhancing bone mineralization and improving bone structure stability. Oxidative stress is an important cause of articular cartilage degeneration and bone matrix decomposition. Although composite-encapsulated nano-calcium is not a direct strong antioxidant, its encapsulation layer components (casein phosphopeptide, sodium alginate, hydroxypropyl-β-cyclodextrin) can form an antioxidant synergistic effect with highly dispersed collagen peptides and tea polyphenols through their own structure and activity, further reducing the damage of oxidative stress to articular cartilage and bone matrix, and providing a better microenvironment for joint repair. The calcium carbonate raw material used in the preparation of composite-encapsulated nano-calcium is nanoscale, with a huge specific surface area. Its contact area with digestive juices in the intestine is much larger than that of ordinary calcium supplements, making it easier to dissolve and ionize. This is the physical basis for its high absorption rate. The composite-encapsulated nano-calcium employs a double-encapsulation structure. The first layer consists of casein phosphopeptide (CPP) and sodium alginate. CPP is an excellent calcium absorption promoter; it can bind with calcium ions to form a soluble complex, effectively preventing calcium from binding with phosphate and phytate ions to form precipitates in the neutral or alkaline environment of the intestine. This prolongs the absorption time of calcium in the intestine and promotes its absorption through the small intestinal mucosal cells. The second layer consists of hydroxypropyl-β-cyclodextrin. Cyclodextrin has a special structure of being hydrophilic on the outside and hydrophobic on the inside, which can form inclusion complexes, further improving the stability and dispersibility of the initially encapsulated nano-calcium. It also helps it penetrate the intestinal mucus layer and make more thorough contact with the absorptive cells. Therefore, this double-encapsulation design makes the bioavailability of calcium much higher than that of ordinary calcium supplements, providing an ample source of calcium for increasing bone calcium content.

[0054] As shown in Table 1-2 and Figs. 1-3 As shown, compared with the examples, the collagen peptide composition prepared in Comparative Example 4 had the worst overall performance. This is because commercially available bovine bone collagen peptides replaced highly dispersed collagen peptides, and commercially available calcium carbonate replaced composite-encapsulated nano-calcium. Highly dispersed collagen peptides provide structural support for joints and bones and promote repair, while composite-encapsulated nano-calcium promotes bone mineralization and enhances bone structural strength by providing an effective calcium source. The combination of the two can effectively promote bone matrix repair, improve bone density, enhance antioxidant capacity, and enhance bone calcium deposition. Therefore, highly dispersed collagen peptides and composite-encapsulated nano-calcium are indispensable synergistic core components in the preparation of this collagen composition.

[0055] While embodiments of the application have been shown and described, it is to be understood that the embodiments described are merely exemplary and that changes can be made in detail without departing from the principles and spirit of the application. The scope of the application is therefore defined by the appended claims and their equivalents.

Claims

1. A collagen peptide composition for repairing joints, characterized by, The raw materials include the following by weight: 20-25 parts of high-dispersion collagen peptide, 10-15 parts of chondroitin sulfate, 4-6 parts of ganoderma extract, 4-6 parts of lentinus edodes extract, 4-6 parts of composite embedded nano calcium, 2-4 parts of hyaluronic acid, 0.4-0.6 parts of vitamin K2, 0.2-0.4 parts of tea polyphenol, 5-7 parts of pitaya fermentation original paste, 4-6 parts of malt dextrin, and 0.1-0.2 parts of epsilon-polylysine. The preparation method of the high-dispersion collagen peptide is as follows: In step S1, the bovine bone collagen peptide is added to deionized water, stirred at 40-50 DEG C for 20-25 min, then activated carbon is added and stirred for another 30-40 min, and the filtrate is obtained after cooling to room temperature. In step S2, the composite protease is added to the filtrate, and enzymolysis is carried out at 50-55 DEG C and pH 7.0-7.5 for 2-2.5 h, then the reaction is terminated, the mixture is cooled to room temperature, and the permeate is obtained by ultrafiltration membrane separation. In step S3, the permeate is concentrated, chitosan is added, and reaction is carried out at 55-65 DEG C and pH 7.8-8.2 for 1.5-2 h, and the high-dispersion collagen peptide is obtained after post-treatment.

2. The collagen peptide composition for repairing joints according to claim 1, characterized in that, In step S1, the solution prepared by adding the bovine bone collagen peptide to deionized water has a mass concentration of 7-9%, and the mass ratio of the bovine bone collagen peptide to activated carbon is 1:0.0015-0.0025.

3. The collagen peptide composition for repairing joints according to claim 1, characterized in that, In step S2, the composite protease is composed of alkaline protease and papain at a mass ratio of 2.5-3.5:1, and the mass-volume ratio of the composite protease to the filtrate is 0.09-0.1 g / 100 mL.

4. The collagen peptide composition for repairing joints according to claim 1, characterized in that, In step S3, the permeate is concentrated to a solid content of 9-11%, and the mass-volume ratio of chitosan to the permeate is 0.035-0.045 g / 100 mL.

5. The collagen peptide composition for repairing joints according to claim 1, characterized in that, The preparation method of the composite embedded nano calcium is as follows: In step a, casein phosphopeptide and sodium alginate are first added to deionized water, stirred at 55-60 DEG C and pH 6.5-7.0 for 30-40 min to obtain a composite embedding solution, then calcium carbonate is added, stirred at 40-45 DEG C for 60-70 min, and the primary embedded nano calcium is obtained after post-treatment. In step b, hydroxypropyl-beta-cyclodextrin is added to deionized water, stirred at 60-65 DEG C for 20-30 min to obtain a hydroxypropyl-beta-cyclodextrin solution, then the temperature is lowered to 35-40 DEG C, the primary embedded nano calcium is added, and constant-temperature stirring is carried out for 90-100 min, and the composite embedded nano calcium is obtained after post-treatment.

6. The collagen peptide composition for repairing joints according to claim 5, characterized in that, In step a, the mass ratio of casein phosphopeptide to sodium alginate is 1.5-2.5:1, the amount of sodium alginate added to deionized water is 1-1.5 g / 100 mL, and the mass ratio of calcium carbonate to the composite embedding solution is 1:5-6.

7. The collagen peptide composition for repairing joints according to claim 5, characterized in that, In step b, the mass concentration of the hydroxypropyl-beta-cyclodextrin solution is 9.5-10.5%, and the mass ratio of the primary embedded nano calcium to hydroxypropyl-beta-cyclodextrin is 2.5-3.5:

7.

8. The collagen peptide composition for repairing joints according to claim 1, wherein The pitaya fermentation original paste is prepared by blending red heart pitaya fermentation broth and galactooligosaccharide at a mass ratio of 10:3-6.

9. The method for preparing a collagen peptide composition for repairing joints according to any one of claims 1 to 8, characterized in that, The method comprises the following steps: (1) high dispersion collagen peptide, chondroitin sulfate, ganoderma extract, lentinus edodes extract, composite embedding nano calcium, hyaluronic acid, vitamin K2, tea polyphenol, epsilon-polylysine are sieved respectively and prepared for use; (2) malt dextrin is added to deionized water, heated to 60-65 DEG C and stirred to dissolve, cooled to 30-35 DEG C, then tea polyphenol, epsilon-polylysine, high dispersion collagen peptide, chondroitin sulfate are added in sequence, stirred at 500-600 r / min for 20-30 min, pitaya fermentation original slurry is added and stirred for 10-15 min to form a mixed solution A; (3) ganoderma extract, lentinus edodes extract, hyaluronic acid are added to the mixed solution A, continue to stir for 20-30 min to form a mixed solution B, vitamin K2 and composite embedding nano calcium are mixed uniformly and added to the mixed solution B, and then homogenized to obtain.

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