A collagen tripeptide complex, and a preparation method and application thereof

Abstract

The application belongs to the technical field of biological peptide synthesis, and specifically discloses a collagen tripeptide compound as well as a preparation method and application thereof. The collagen tripeptide compound comprises the following components in mass parts: collagen tripeptide 20-25 parts, black-bone chicken peptide 12-18 parts, elastin peptide 15-20 parts, glycerol 5-8 parts, licorice powder 3-5 parts, centella asiatica powder 1-2 parts, green tea powder 4-9 parts, pomegranate powder 2-3 parts, and blueberry powder 1-3 parts. The collagen tripeptide compound has extremely strong antioxidant capacity, can promote the biosynthesis of collagen, and effectively resists oxidative stress caused by environmental factors such as ultraviolet rays.

Description

Technical Field

[0001] This invention belongs to the field of biopeptide synthesis technology, specifically relating to a collagen tripeptide complex, its preparation method, and its application. Background Technology

[0002] Skin aging is a complex physiological process, mainly manifested as decreased skin elasticity, wrinkle formation, and weakened repair capabilities. One of its core underlying causes is the loss and insufficient synthesis of collagen and elastin in the dermis. Ultraviolet radiation (photoaging) is a major external factor that accelerates this process. Therefore, supplementing with active ingredients that can promote collagen synthesis and possess anti-inflammatory and antioxidant capabilities has become a research hotspot in the field of skin health and beauty.

[0003] Collagen and its hydrolysates (such as collagen peptides) are mainstream skin nutrition supplements on the market. Among them, collagen tripeptides (CTPs) are considered to have better bioavailability due to their small molecular weight. However, single collagen tripeptide products have limitations in application; their function is relatively singular, mainly focusing on supplementing collagen, with limited effects on combating oxidative stress and improving the overall skin microenvironment. Furthermore, the health of human skin depends on the synergistic effects of various structural proteins such as collagen and elastin. To improve efficacy, some compound products have emerged in existing technologies. However, these combinations are mostly simple physical mixtures, lacking synergistic process design between components, and there is still considerable room for improvement in bioavailability and overall efficacy.

[0004] On the other hand, while black-boned chicken, a traditional tonic, is rich in various amino acids and small molecule peptides, its application in skin care is mostly limited to traditional decoction or simple enzymatic hydrolysis. Existing methods for preparing black-boned chicken peptide products are limited, often involving only single enzymatic hydrolysis, which may result in suboptimal peptide distribution, poor flavor, or insufficient activation of functional activities, thus restricting its application value in high-end functional foods and cosmetics. Summary of the Invention

[0005] The present invention aims to provide a collagen tripeptide complex, its preparation method and application. The collagen tripeptide complex has extremely strong antioxidant capacity, can promote the biosynthesis of collagen, and effectively resist oxidative stress caused by environmental factors such as ultraviolet radiation.

[0006] To solve the above-mentioned technical problems, the technical solution adopted by the present invention is as follows: A collagen tripeptide complex comprising the following components in parts by weight: Collagen tripeptide 20-25 parts, black chicken peptide 12-18 parts, elastin peptide 15-20 parts, glycerin 5-8 parts, licorice powder 3-5 parts, centella asiatica powder 1-2 parts, green tea powder 4-9 parts, pomegranate powder 2-3 parts, blueberry powder 1-3 parts.

[0007] This invention also provides a method for preparing the collagen tripeptide complex described above, comprising the following steps: S1. Mix licorice powder, centella asiatica powder and green tea powder, add 8 to 10 times the amount of 60% to 70% ethanol solution, heat and reflux to extract, repeat the extraction 2 to 3 times, combine the extracts, concentrate under reduced pressure, spray dry to obtain the extract powder. S2. Mix pomegranate powder and blueberry powder, add deionized water, inoculate with Lactobacillus plantarum, ferment, heat to stop fermentation, centrifuge, discard the precipitate, and obtain the reactants; S3. Mix the dry powder of the extract obtained in S1 with the reactants obtained in S2, add glycerol, collagen tripeptide, black chicken peptide and elastin peptide, stir evenly and homogenize to obtain collagen tripeptide complex.

[0008] Preferably, in S1, the heating reflux temperature is 70~80℃ and the time is 1~2h.

[0009] Preferably, in S2, the fermentation temperature is 37~42℃ and the fermentation time is 48~72h.

[0010] Preferably, in step S3, the homogenization pressure is 30-50 MPa, and the homogenization is performed 2-3 times.

[0011] This invention also provides a method for preparing black-boned chicken peptide, used to prepare the collagen tripeptide complex, comprising the following steps: T1. Take fresh black-boned chicken meat, mince it, mix it with deionized water, mix it with deionized water preheated to 50~60℃, stir it evenly, adjust the pH to acidic, sonicate it, centrifuge it, and obtain the supernatant. T2. Adjust the pH of the supernatant extracted in T1 to neutral, let it stand, treat it with variable temperature, inoculate it with yeast, ferment it, sterilize it, filter it through an ultrafiltration membrane, desalinate it by tangential flow filtration, centrifuge it with continuous flow, take the supernatant, spray dry it to obtain black chicken peptide.

[0012] Preferably, in T1, the power of the ultrasonic treatment is 300~500W, and the treatment time is 20~40 minutes.

[0013] Preferably, in T2, the temperature change treatment is to maintain the temperature at 15~20℃ for 8~12h, and then maintain the temperature at 35~38℃ for 10~14h.

[0014] Preferably, in T2, the fermentation temperature is 30~40℃ and the fermentation time is 36~60h.

[0015] The present invention also provides the use of the collagen tripeptide complex as described in the preparation of products for post-sun repair.

[0016] Compared with the prior art, the present invention has the following advantages and technical effects: This invention discloses a collagen tripeptide complex, its preparation method, and its application. The collagen tripeptide complex has extremely strong antioxidant capacity and can promote the biosynthesis of collagen. It can effectively resist oxidative stress caused by environmental factors such as ultraviolet rays, and provide protection for post-sun repair and anti-aging functions.

[0017] The technical solution of the present invention will be further described in detail below with reference to the accompanying drawings and embodiments. Attached Figure Description

[0018] Figure 1 This is a statistical chart showing the DPPH free radical scavenging rate in different groups of the present invention; Figure 2 A statistical chart showing the hydroxyproline content in different groups. Detailed Implementation

[0019] The technical solution of the present invention will be further described below with reference to the accompanying drawings and embodiments.

[0020] Unless otherwise defined, the technical or scientific terms used in this invention shall have the ordinary meaning as understood by one of ordinary skill in the art to which this invention pertains.

[0021] Source of experimental materials: Both Lactobacillus plantarum and yeast were purchased from Weifang Yihao Biotechnology Co., Ltd.

[0022] In this invention, unless otherwise specified, all other test materials and instruments are conventional test materials in the field and can be purchased through commercial channels.

[0023] Example 1 Prepare the ingredients according to the following weight proportions: weigh out 20 parts collagen tripeptide, 18 parts black chicken peptide, 15 parts elastin peptide, 8 parts glycerin, 3 parts licorice powder, 2 parts centella asiatica powder, 4 parts green tea powder, 3 parts pomegranate powder, and 1 part blueberry powder.

[0024] The preparation method of black-boned chicken peptides includes the following steps: T1. Take 1 kg of fresh black-boned chicken meat, mince it, mix it with 3 kg of deionized water preheated to 55℃, adjust the pH to 4.0, sonicate it at 400W power for 30 min, and then centrifuge it at 4℃ and 8000rpm for 20 min. Take the supernatant. T2. Adjust the pH of the supernatant from T1 to 7.0, let it stand at 18℃ for 10 hours, then at 37℃ for 12 hours, inoculate with yeast (inoculation amount 2%), ferment at 35℃ for 48 hours, sterilize at 100℃ for 15 minutes, and then filter through a 10kDa ultrafiltration membrane, desalt by tangential flow, and centrifuge continuously at 4℃ and 10000rpm. Take the supernatant and spray dry it (inlet air temperature 180℃, outlet air temperature 85℃, peristaltic pump frequency 25Hz) to obtain black-boned chicken peptide.

[0025] The preparation method of collagen tripeptide complex includes the following steps: S1. Mix licorice powder, centella asiatica powder and green tea powder, add 9 times the amount of 65% ethanol solution, heat and reflux at 75℃ for 1.5h, repeat the extraction twice, combine the extracts, concentrate under reduced pressure at 60℃ and -0.08MPa for 20min, and spray dry (inlet air temperature 175℃, outlet air temperature 80℃, peristaltic pump frequency 20Hz) to obtain plant extract powder. S2. Mix pomegranate powder and blueberry powder, add deionized water (material-to-liquid ratio 1:8), inoculate with 4% Lactobacillus plantarum, ferment at 40℃ for 60h, heat at 80℃ for 15min to terminate fermentation, then centrifuge at 4℃ and 8000rpm for 15min, discard the precipitate, and obtain the supernatant as the fermentation reaction product. S3. Mix the dry powder obtained in S1 with the reactants obtained in S2, add glycerol, collagen tripeptide, homemade black chicken peptide and elastin peptide, stir evenly, and homogenize twice under 40MPa pressure to obtain collagen tripeptide complex.

[0026] Example 2 Prepare the ingredients according to the following weight proportions: weigh out 23 parts collagen tripeptide, 15 parts black chicken peptide, 18 parts elastin peptide, 6 parts glycerin, 4 parts licorice powder, 1.5 parts centella asiatica powder, 7 parts green tea powder, 2.5 parts pomegranate powder, and 2 parts blueberry powder.

[0027] The preparation method of black-boned chicken peptides includes the following steps: T1. Take 1 kg of fresh black-boned chicken meat, mince it, mix it with 3 kg of deionized water preheated to 55℃, adjust the pH to 4.0, sonicate it at 350W for 40 min, and then centrifuge it at 4℃ and 8500rpm for 25 min. Take the supernatant. T2. Adjust the pH of the supernatant from T1 to 7.0, let it stand at 16℃ for 12 hours, then at 36℃ for 14 hours, inoculate with yeast (inoculation amount 2%), ferment at 38℃ for 42 hours, sterilize at 100℃ for 15 minutes, and then filter through a 10kDa ultrafiltration membrane, desalt by tangential flow, and centrifuge continuously at 4℃ and 10000rpm. Take the supernatant and spray dry it (inlet air temperature 185℃, outlet air temperature 85℃, peristaltic pump frequency 25Hz) to obtain black-boned chicken peptide.

[0028] The preparation method of collagen tripeptide complex includes the following steps: S1. Mix licorice powder, centella asiatica powder and green tea powder, add 9 times the amount of 65% ethanol solution, heat and reflux at 78℃ for 1 hour, repeat the extraction twice, combine the extracts, concentrate under reduced pressure at 65℃ and -0.09MPa for 20 minutes, and spray dry (inlet air temperature 170℃, outlet air temperature 80℃, peristaltic pump frequency 20Hz) to obtain plant extract powder. S2. Mix pomegranate powder and blueberry powder, add deionized water (material-to-liquid ratio 1:8), inoculate with 4% Lactobacillus plantarum, ferment at 38℃ for 72h, heat at 80℃ for 15min to terminate fermentation, then centrifuge at 4℃ and 7500rpm for 20min, discard the precipitate, and obtain the supernatant as the fermentation product. S3. Mix the dry powder obtained in S1 with the reactants obtained in S2, add glycerol, collagen tripeptide, homemade black chicken peptide and elastin peptide, stir evenly, and homogenize 3 times under 35 MPa pressure to obtain collagen tripeptide complex.

[0029] Example 3 Prepare the ingredients according to the following weight proportions: weigh out 25 parts collagen tripeptide, 12 parts black chicken peptide, 20 parts elastin peptide, 5 parts glycerin, 5 parts licorice powder, 1 part centella asiatica powder, 9 parts green tea powder, 2 parts pomegranate powder, and 3 parts blueberry powder.

[0030] The preparation method of black-boned chicken peptides includes the following steps: T1. Take 1 kg of fresh black-boned chicken meat, mince it, mix it with 3 kg of deionized water preheated to 55℃, adjust the pH to 4.0, sonicate it at 450W for 20 min, and then centrifuge it at 4℃ and 9000rpm for 15 min. Take the supernatant. T2. Adjust the pH of the supernatant from T1 to 7.0, let it stand at 20℃ for 8 hours, then at 38℃ for 10 hours, inoculate with yeast (inoculation amount 2%), ferment at 32℃ for 60 hours, sterilize at 100℃ for 15 minutes, and then filter through a 10kDa ultrafiltration membrane, desalt by tangential flow, and centrifuge continuously at 4℃ and 10000rpm. Take the supernatant and spray dry it (inlet air temperature 185℃, outlet air temperature 85℃, peristaltic pump frequency 25Hz) to obtain black-boned chicken peptide.

[0031] The preparation method of collagen tripeptide complex includes the following steps: S1. Mix licorice powder, centella asiatica powder and green tea powder, add 9 times the amount of 65% ethanol solution, heat and reflux at 72℃ for 2 hours, repeat the extraction twice, combine the extracts, concentrate under reduced pressure at 58℃ and -0.09MPa for 20 minutes, and spray dry (inlet air temperature 170℃, outlet air temperature 85℃, peristaltic pump frequency 20Hz) to obtain plant extract powder. S2. Mix pomegranate powder and blueberry powder, add deionized water (material-to-liquid ratio 1:8), inoculate with 4% Lactobacillus plantarum, ferment at 38℃ for 72h, heat at 80℃ for 15min to terminate fermentation, then centrifuge at 4℃ and 7500rpm for 20min, discard the precipitate, and obtain the supernatant as the fermentation product. S3. Mix the dry powder obtained in S1 with the reactants obtained in S2, add glycerol, collagen tripeptide, homemade black chicken peptide and elastin peptide, stir evenly, and homogenize twice under 45 MPa pressure to obtain collagen tripeptide complex.

[0032] Comparative Example 1 The preparation method is the same as in Example 1, except that the black-boned chicken peptide is commercially available.

[0033] Comparative Example 2 The preparation method is the same as in Example 1, except that the S2 fermentation step is omitted, and the pomegranate powder and blueberry powder of the formula amount are directly mixed with other powder raw materials (S1 dry powder, collagen tripeptide, black chicken peptide, etc.) in the S3 step by simple physical mixing.

[0034] Comparative Example 3 The preparation method is the same as in Example 1, except that pomegranate powder and blueberry powder are not added to the raw materials.

[0035] Comparative Example 4 The preparation method is the same as in Example 1, except that licorice powder, centella asiatica powder and green tea powder are not added to the raw materials.

[0036] The effects of the collagen tripeptide complexes prepared in Examples 1-3 and Comparative Examples 1-4 were verified.

[0037] In vitro antioxidant activity assay (DPPH free radical scavenging rate): Accurately weigh DPPH powder and prepare a 0.1 mmol solution with anhydrous ethanol. Solution L, freshly prepared and used immediately, has an initial absorbance of 1.0 at a wavelength of 517 nm. 0.05.

[0038] Sample solutions: Dissolve each sample in distilled water and dilute to a series of concentration gradients (0.01, 0.05, 0.1, 0.5, 1.0, 5.0 mg). mL).

[0039] Positive control: Prepare aqueous solutions of vitamin C (Vc) with the same concentration gradient.

[0040] Solvent control: Distilled water with the same solvent percentage as the highest concentration point of the sample was used as a control.

[0041] Examples 1-3 and Comparative Examples 1-4 were respectively set as groups A, B, C, D, E, F, and G.

[0042] Take 100 L of sample solutions (or control solutions) of different concentrations, and 100 L DPPH solution was mixed in 96-well plates. Distilled water was used as a background control instead of DPPH solution. The reaction mixture was incubated at 25°C for 30 min, and the absorbance was immediately measured at 517 nm using a microplate reader. Each concentration was tested in triplicate, and the experiment was independently repeated three times. The results are shown below. Figure 1 As shown.

[0043] Depend on Figure 1 It can be seen that the collagen tripeptide complexes of Examples 1-3 exhibited strong DPPH free radical scavenging ability, which was better than that of the comparative examples, indicating that they have excellent in vitro antioxidant activity and are superior to commercially available black chicken peptide products.

[0044] The specific experimental protocol for determining the hydroxyproline content is as follows: Human skin fibroblasts (HSF) were seeded in 6-well plates at a density of 5 × 10⁶ cells / well. 4 cells The wells were incubated at 37℃ in a 5% CO2 incubator for 24 hours until fully adherent. The old culture medium was discarded, and different concentrations of the sample (0.01, 0.1, 1.0, 10, 50 g) were added. Fresh, complete culture medium was used. A blank control (without sample) and a positive control (containing 0.5 g of sample) were set up. Retinol) and solvent controls (Example 1, Comparative Example 1, Comparative Example 2, Comparative Example 3 and Comparative Example 4). Each group had 3 replicates and was cultured for another 48 hours.

[0045] After culturing, collect the cell supernatant. Follow the instructions strictly according to the hydroxyproline assay kit. Results are as follows: Figure 2 As shown.

[0046] Depend on Figure 2 It can be seen that the hydroxyproline content in human skin fibroblasts treated in Examples 1-3 was significantly higher than that in each pair, indicating that the complex can effectively promote collagen synthesis.

[0047] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and not to limit them. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can still be made to the technical solutions of the present invention, and these modifications or equivalent substitutions cannot cause the modified technical solutions to deviate from the spirit and scope of the technical solutions of the present invention.

Claims

1. A collagen tripeptide complex, characterized in that, It contains the following components by weight: Collagen tripeptide 20-25 parts, black chicken peptide 12-18 parts, elastin peptide 15-20 parts, glycerin 5-8 parts, licorice powder 3-5 parts, centella asiatica powder 1-2 parts, green tea powder 4-9 parts, pomegranate powder 2-3 parts, blueberry powder 1-3 parts.

2. The method for preparing the collagen tripeptide complex as described in claim 1, characterized in that, Includes the following steps: S1. Mix licorice powder, centella asiatica powder and green tea powder, add 8 to 10 times the amount of 60% to 70% ethanol solution, heat and reflux to extract, repeat the extraction 2 to 3 times, combine the extracts, concentrate under reduced pressure, spray dry to obtain the extract powder. S2. Mix pomegranate powder and blueberry powder, add deionized water, inoculate with Lactobacillus plantarum, ferment, heat to stop fermentation, centrifuge, discard the precipitate, and obtain the reactants; S3. Mix the dry powder of the extract obtained in S1 with the reactants obtained in S2, add glycerol, collagen tripeptide, black chicken peptide and elastin peptide, stir evenly and homogenize to obtain collagen tripeptide complex.

3. The preparation method according to claim 2, characterized in that, In S1, the heating reflux temperature is 70~80℃ and the time is 1~2h.

4. The preparation method according to claim 2, characterized in that, In S2, the fermentation temperature is 37~42℃ and the fermentation time is 48~72h.

5. The preparation method according to claim 2, characterized in that, In S3, the homogenization pressure is 30~50MPa, and the homogenization is performed 2~3 times.

6. A method for preparing black-boned chicken peptide, used to prepare the collagen tripeptide complex of claim 1, characterized in that, Includes the following steps: T1. Take fresh black-boned chicken meat, mince it, mix it with deionized water, mix it with deionized water preheated to 50~60℃, stir it evenly, adjust the pH to acidic, sonicate it, centrifuge it, and obtain the supernatant. T2. Adjust the pH of the supernatant extracted in T1 to neutral, let it stand, treat it with variable temperature, inoculate it with yeast, ferment it, sterilize it, filter it through an ultrafiltration membrane, desalinate it by tangential flow filtration, centrifuge it with continuous flow, take the supernatant, spray dry it to obtain black chicken peptide.

7. The preparation method according to claim 6, characterized in that, In T1, the power of the ultrasonic treatment is 300~500W, and the treatment time is 20~40 minutes.

8. The preparation method according to claim 6, characterized in that, In T2, the temperature change treatment is to maintain the temperature at 15~20℃ for 8~12h, and then maintain the temperature at 35~38℃ for 10~14h.

9. The preparation method according to claim 6, characterized in that, In T2, the fermentation temperature is 30~40℃ and the fermentation time is 36~60h.

10. The use of the collagen tripeptide complex as described in claim 1 in the preparation of products for post-sun repair.

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