Method for extracting collagen peptide through illumination and deep eutectic solvent

Abstract

The invention discloses a method for extracting collagen peptide through cooperation of illumination and a deep eutectic solvent, and belongs to the technical field of collagen peptide preparation. The method comprises the following steps: mixing a collagen raw material and a deep eutectic solvent according to a certain material-liquid ratio, and extracting under illumination with a specific wavelength. The deep eutectic solvent is prepared from choline chloride and hydrogen bond donors such as glycerol, lactic acid, urea or sorbitol according to a certain molar ratio. In the extraction process, blue light of 400-480 nm and / or red light of 630-680 nm are / is adopted for irradiation, the illumination intensity is 10-60 mW / cm < 2 >, the time is 30-90% of the total extraction time, the extraction temperature is 40-65 DEG C, and the time is 1-3 h. And dialyzing, concentrating and drying the extracting solution to obtain a collagen peptide product. The method obviously improves the extraction efficiency, improves the molecular weight distribution and biological activity of the peptide, and has the advantages of mild conditions, simplicity and convenience in operation, greenness, energy conservation and the like.

Description

Technical Field

[0001] This invention belongs to the field of collagen peptide preparation technology, specifically relating to a method for extracting collagen peptides using light-induced synergistic deep eutectic solvent. Background Technology

[0002] Collagen peptides are small molecular weight fragments formed after collagen is hydrolyzed. Due to their excellent biocompatibility, low immunogenicity, and easy absorption by the human body, they are widely used in food, health products, cosmetics, and biomedical materials.

[0003] Traditional collagen extraction methods mainly rely on acid, alkaline, or enzymatic methods. Existing patent CN112341537B discloses a method for preparing and applying fish bone collagen peptides. This involves soaking in a mixed acid solution, followed by ultrasonic treatment, freezing, and then two enzymatic hydrolysis processes and irradiation treatment to obtain the fish bone collagen peptides. While this patent is low-cost and simple to operate, the reaction conditions are harsh, requiring mixed acid soaking, heating, and shaking treatment. This not only consumes a lot of energy but also easily damages the triple helix structure of collagen, resulting in an excessively wide molecular weight distribution, poor functionality, and potentially introducing harmful chemical residues, causing environmental and safety issues. Existing patent CN103783254B discloses a method for preparing yak bone collagen peptides. This method uses a complex enzyme approach—protease, amylase, pectinase, cellulase, and xylanase—to shorten the product preparation time and improve the product's taste. Although the complex enzyme extraction conditions are relatively mild and better preserve the bioactivity of collagen. However, enzymatic methods still significantly increase production costs, and the reactivity of enzymes is easily affected by environmental factors, resulting in insufficient stability.

[0004] Existing research has found that deep eutectic solvents (DES), as a low-melting mixture formed by hydrogen bond donors and acceptors through hydrogen bonding, not only have outstanding advantages such as simple preparation, low cost, biodegradability, low vapor pressure, strong solubility and designable structure, but also can effectively disrupt the cross-linked structure of collagen, dissolve the protein, and may protect the bioactivity of the extract in its unique solvation microenvironment.

[0005] While DES shows promise as a replacement for traditional solvents in collagen extraction, the DES extraction process still relies on external energy sources such as thermal stirring or microwave assistance. Thermal stirring has limited mass transfer efficiency and remains energy-intensive; microwave assistance suffers from expensive equipment and difficulties in scale-up. Therefore, developing a more efficient, energy-saving, and gentler technology that can further enhance the DES extraction process is crucial for improving the extraction rate and quality of collagen peptides and promoting their industrial-scale green production. Summary of the Invention

[0006] To address the problems of insufficient collagen peptide extraction efficiency, easy loss of peptide activity, and imperfections in existing eutectic solvent extraction techniques, there is an urgent need to develop a new method that is more efficient, energy-saving, gentle, and can further enhance the eutectic solvent extraction process. The purpose of this invention is to provide a method for extracting collagen peptides using a light-assisted eutectic solvent extraction process. By adding light of a specific wavelength during the eutectic solvent extraction process, the extraction efficiency of collagen peptides is significantly improved. This method is energy-saving, environmentally friendly, and simple to operate throughout the entire process.

[0007] To achieve the above objectives, the technical solution of the present invention is as follows:

[0008] In a first aspect, the present invention discloses a method for extracting collagen peptides, the method comprising extracting collagen peptides by light synergistic with deep co-solvent extraction.

[0009] Specifically, the illumination is selected from blue light with a wavelength of 400-480nm and / or red light with a wavelength of 630-680nm.

[0010] Specifically, the lighting conditions include:

[0011] Light intensity is 10-60 mW / cm 2 ;

[0012] And / or, the illumination time is 30-90% of the total extraction time;

[0013] And / or, the illumination method is continuous illumination or alternating illumination.

[0014] Specifically, the raw material of the collagen peptide is mixed with the deep co-solvent at a material-to-liquid ratio of 1:5 to 1:12.

[0015] Specifically, the deep co-solution solvent is prepared by mixing hydrogen bond donors and hydrogen bond acceptors in a molar ratio of 1:1–1:10.

[0016] Preferably, the deep co-solution solvent is prepared by mixing a hydrogen bond donor and a hydrogen bond acceptor in a molar ratio of 1:2–1:10.

[0017] Specifically, the hydrogen bond acceptor is selected from at least one of glycerol, lactic acid, urea, and sorbitol; and / or, the hydrogen bond donor is choline chloride.

[0018] Specifically, the extraction conditions include: an extraction temperature of 40-65℃ and a total extraction time of 1-3 hours.

[0019] Specifically, the method also includes dialysis, concentration, and drying processes.

[0020] Specifically, the molecular weight cutoff range of the dialysis is 500-10000 Da; and / or, the concentration temperature is 50-60°C, and the concentration pressure is -0.08 MPa to -0.09 MPa.

[0021] In some embodiments, the method for extracting collagen peptides specifically includes the following steps:

[0022] (1) Raw material pretreatment and deep eutectic solvent preparation:

[0023] Select fresh bone raw materials (preferably pork spine, beef bones or chicken bones), remove fat and muscle tissue, sterilize under high pressure, crush and sieve (preferably sieve mesh of 20-100 mesh); mix the treated bone powder with deep eutectic solvent at a material-to-liquid ratio of 1:5-1:12 (g / mL).

[0024] (2) Photosynergistic deep eutectic solvent extraction:

[0025] Extraction was performed at a constant temperature of 40-65℃ for 1-3 hours, while simultaneously irradiating with blue light at wavelengths of 400-480 nm and / or red light at wavelengths of 630-680 nm, with a light intensity of 10-60 mW / cm². 2 The illumination method can be continuous or alternating (switching every 5–15 minutes), and the total illumination time is 30–90% of the total extraction time;

[0026] (3) Separation and purification:

[0027] The polypeptide extract obtained in step (2) was dialyzed to remove impurities. The molecular weight cutoff range of the dialysis bag was 500–10000 Da, so as to obtain polypeptide components with different molecular weight ranges.

[0028] (4) Concentration and Drying

[0029] The polypeptide solution obtained in step (3) was vacuum concentrated at 50–60°C and a pressure of -0.08 MPa to -0.09 MPa until the solid mass fraction was 15–30%. The concentrate was dried by spray drying at an inlet air temperature of 150–170°C and an outlet air temperature of 80–90°C to obtain a light yellow powdered collagen peptide product.

[0030] Secondly, the present invention discloses a collagen peptide prepared by the aforementioned method.

[0031] The collagen peptide products prepared by the aforementioned method can have a total peptide content of over 80%, of which the proportion of small molecule peptides with a molecular weight of less than 1000 Da can reach over 75%, and they maintain high functional activity, making them suitable as functional ingredients in food, health products, and cosmetics.

[0032] Existing research indicates that blue light (400–480 nm) can enhance the activity of active enzymes (such as peroxidase and proteolytic enzymes) in plants, promoting the degradation of macromolecules; while red light (620–680 nm) can enhance free radical scavenging, protect unsaturated bond structures, and maintain biological activity. This suggests that light has potential value in regulating molecular degradation rates and maintaining product activity.

[0033] By incorporating the technical solution of this invention, introducing specific wavelengths of light into the DES system may have the following mechanisms of action:

[0034] (1) Photoexcitation effect: Blue light can excite trace amounts of peroxides or hydroxyl radicals in the DES system, generating mild oxidative stress, promoting the breakage of collagen macromolecular chains, thereby increasing the proportion of small molecule peptides generated.

[0035] (2) Structural protection: Red light can induce the scavenging of free radicals in the solution, slow down the destruction of active groups during the oxidative breakage of peptide chains, and prevent the degradation of key amino acid residues such as hydroxyproline, thereby improving the functional activity of the product;

[0036] (3) Synergistic effect: Alternating red and blue light irradiation can form a dynamic balance between “promoting breakage (blue light)” and “protecting activity (red light)”, making collagen decomposition more complete while maintaining high biological activity.

[0037] Some literature reports that blue light can enhance the conformational flexibility of proteases and improve substrate binding efficiency; while red light helps reduce enzyme inactivation during the reaction, thus synergistically improving enzymatic hydrolysis efficiency. Therefore, selecting red and blue light as specific wavelength light sources can both promote the cleavage of large collagen chains and maintain the functional activity of small peptides.

[0038] Compared with the prior art, the present invention has the following beneficial effects:

[0039] (1) Significantly improved extraction efficiency: By combining specific wavelengths of light (400-480nm blue light and / or 630-680nm red light) with deep eutectic solvent (DES) extraction technology, a significant synergistic effect was achieved. This combined strategy can more effectively disrupt the cross-linked structure of collagen, promote its dissolution and hydrolysis, thereby greatly improving the extraction rate and yield of collagen peptides;

[0040] (2) High product activity and excellent quality: The method of this invention is carried out under mild conditions (40-65℃) and utilizes the free radical scavenging and protective effects of red light, effectively reducing the damage to heat-sensitive amino acids (such as hydroxyproline) and peptide chain functional structures. The prepared collagen peptides have a more concentrated molecular weight distribution, a high proportion of small molecule peptides (<1000Da) (up to 75% or more), and better retain their biological activity;

[0041] (3) Energy saving and cost reduction in the process: Light irradiation, as a clean energy input method, significantly reduces energy consumption compared to traditional simple thermal stirring or microwave assistance. At the same time, this method reduces dependence on expensive enzyme preparations, and the DES solvent used is simple to prepare and inexpensive, which helps to reduce the overall production cost;

[0042] (4) Green and environmentally friendly with good safety: The entire extraction process does not require the use of large amounts of strong acids, strong alkalis or toxic chemical reagents. The DES solvent used is usually biodegradable and low in toxicity, avoiding harmful chemical residues and meeting the development requirements of green chemistry and clean production. Detailed Implementation

[0043] The following embodiments are merely illustrative of the present invention and do not limit the scope of protection of the present invention in any way. For those skilled in the art, all equivalent implementations or modifications made without departing from the spirit of the present invention are within the scope of protection of the present invention.

[0044] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to embodiments.

[0045] Example 1

[0046] (1) Take fresh pig spine, remove fat and muscle tissue, sterilize under high pressure (121℃, 15min), and then pulverize through a 40-mesh sieve. Mix bone meal with eutectic solvent (choline chloride: glycerol = 1:2 molar ratio) at a material-to-liquid ratio of 1:10 and stir evenly.

[0047] (2) Extraction was performed at 40℃ for 3 hours, while alternating irradiation with blue light (450nm) and red light (660nm) at an intensity of 40mW / cm². 2 Irradiation time accounts for 70% of the total extraction time.

[0048] (3) The extract was dialyzed (molecular weight cutoff 500-10000 Da) to remove impurities.

[0049] (4) Collagen peptide powder was obtained by vacuum concentration (50℃, -0.08MPa, 60min) and spray drying (inlet air temperature 160℃, outlet air temperature 85℃) of the dialysate.

[0050] Example 2

[0051] (1) Take fresh beef tibia, defatted, sterilized by high pressure (121℃, 15min), and then crushed through a 60-mesh sieve. Mix bone meal with deep eutectic solvent (choline chloride: lactic acid = 1:3 molar ratio) at a material-to-liquid ratio of 1:8.

[0052] (2) Extraction was performed at 55℃ for 1.5 hours, while simultaneously applying alternating pulsed irradiation of blue light (460nm) and red light (660nm) (5 seconds of irradiation followed by 15 seconds of irradiation), with a light intensity of 35mW / cm². 2 Irradiation time accounts for 80% of the total extraction time.

[0053] (3) The extract was dialyzed (molecular weight cutoff 500-10000 Da) to remove impurities.

[0054] (4) Collagen peptide powder was obtained by vacuum concentration (55℃, -0.09MPa, 70min) and spray drying (inlet air temperature 165℃, outlet air temperature 88℃) of the dialysate.

[0055] Example 3

[0056] (1) Take chicken breastbone, remove fat, autoclave at 121℃ for 20 minutes, and pulverize through an 80-mesh sieve. Mix bone meal with eutectic solvent (choline chloride: urea = 1:4 molar ratio) at a material-to-liquid ratio of 1:12.

[0057] (2) Extraction was performed at 50℃ for 2 hours, while continuous blue light (445nm) was applied at an intensity of 50mW / cm². 2 Irradiation time accounts for 60% of the extraction time.

[0058] (3) Impurities were removed by dialysis (molecular weight cutoff 500-10000 Da), and the dialysate was concentrated under vacuum (50℃, -0.08MPa, 60min) and spray dried (inlet air temperature 160℃, outlet air temperature 85℃) to obtain collagen peptide powder.

[0059] Example 4

[0060] (1) Take sheep bone powder (pass through a 40-mesh sieve), and mix bone powder with deep eutectic solvent (choline chloride: sorbitol = 1:5 molar ratio) at a material-to-liquid ratio of 1:9.

[0061] (2) Extraction was performed at 65℃ for 1.5 hours, followed by continuous irradiation with red light (650nm) at an intensity of 30mW / cm². 2 The irradiation time is 75% of the extraction time.

[0062] (3) Impurities were removed by dialysis (molecular weight cutoff 500-10000 Da), and the dialysate was concentrated under vacuum (50℃, -0.08MPa, 60min) and spray dried (inlet air temperature 160℃, outlet air temperature 85℃) to obtain collagen peptide powder.

[0063] Comparative Example 1

[0064] The difference from Example 1 is that only deep eutectic solvent extraction was used, and no light was applied.

[0065] (1) Take fresh pig spine, remove fat and muscle tissue, sterilize under high pressure (121℃, 15min), and then pulverize through a 40-mesh sieve. Mix bone meal with eutectic solvent (choline chloride: glycerol = 1:2 molar ratio) at a material-to-liquid ratio of 1:10.

[0066] (2) Extracted at 40℃ for 3 hours without light exposure.

[0067] (3) The extract was dialyzed (molecular weight cutoff 500-10000 Da) to remove impurities.

[0068] (4) Collagen peptide powder was obtained by vacuum concentration (50℃, -0.08MPa, 60min) and spray drying (inlet air temperature 160℃, outlet air temperature 85℃) of the dialysate.

[0069] Comparative Example 2

[0070] The difference from Example 1 is that only light extraction was used, without a deep eutectic solvent.

[0071] (1) Take fresh pig spine, remove fat and muscle tissue, sterilize under high pressure (121℃, 15min), and then pulverize through a 40-mesh sieve. Mix bone meal with deionized water at a material-to-liquid ratio of 1:10 and stir evenly.

[0072] (2) Extraction was performed at 40℃ for 3 hours, while alternating irradiation with blue light (450nm) and red light (660nm) at an intensity of 40mW / cm². 2 Irradiation time accounts for 70% of the total extraction time.

[0073] (3) The extract was dialyzed (molecular weight cutoff 500-10000 Da) to remove impurities.

[0074] (4) Collagen peptide powder was obtained by vacuum concentration (50℃, -0.08MPa, 60min) and spray drying (inlet air temperature 160℃, outlet air temperature 85℃) of the dialysate.

[0075] Comparative Example 3

[0076] The difference from Example 2 is that the sampling method involves conventional alkaline solution and light extraction.

[0077] (1) Take fresh beef tibia, defatted, sterilized by high pressure (121℃, 15min), and then crushed through a 60-mesh sieve. Mix the bone meal with 1% sodium hydroxide solution at a material-to-liquid ratio of 1:8.

[0078] (2) Extraction was performed at 55℃ for 1.5 hours, while alternating irradiation with blue light (460nm) and red light (660nm) (5 seconds of irradiation followed by 15 seconds of irradiation), with a light intensity of 35mW / cm². 2 Irradiation time accounts for 80% of the total extraction time.

[0079] (3) The extract was dialyzed (molecular weight cutoff 500-10000 Da) to remove impurities.

[0080] (4) Collagen peptide powder was obtained by vacuum concentration (55℃, -0.09MPa, 70min) and spray drying (inlet air temperature 165℃, outlet air temperature 88℃) of the dialysate.

[0081] Comparative Example 4

[0082] The difference from Example 3 is that only deep eutectic solvent was used for extraction, and no light was applied.

[0083] (1) Take chicken breastbone, remove fat, autoclave at 121℃ for 20 minutes, and pulverize through an 80-mesh sieve. Mix bone meal with eutectic solvent (choline chloride: urea = 1:4 molar ratio) at a material-to-liquid ratio of 1:12.

[0084] (2) Extract at 50℃ for 2 hours without light.

[0085] (3) Impurities were removed by dialysis (molecular weight cutoff 500-10000 Da), and the dialysate was concentrated under vacuum (50℃, -0.08MPa, 60min) and spray dried (inlet air temperature 160℃, outlet air temperature 85℃) to obtain collagen peptide powder.

[0086] Comparative Example 5

[0087] (1) Take fish scales, crush them and pass them through an 80-mesh sieve. Mix bone meal with deep eutectic solvent (choline chloride: glycerol = 1:3 molar ratio) at a material-to-liquid ratio of 1:15.

[0088] (2) Extraction at 45℃ for 2 hours, while simultaneously applying continuous irradiation with a mixture of blue (450nm), red (660nm), and green (530nm) light at an intensity of 40mW / cm². 2 Irradiation time accounts for 70% of the total extraction time.

[0089] (3) Impurities were removed by dialysis (molecular weight cutoff 500-10000 Da), and the dialysate was concentrated under vacuum (50℃, -0.08MPa, 60min) and spray dried (inlet air temperature 160℃, outlet air temperature 85℃) to obtain collagen peptide powder.

[0090] Example of effect

[0091] The total peptide content, small molecule peptide ratio, hydroxyproline content, and antioxidant properties of the collagen peptide powders prepared in the above examples and comparative examples were tested.

[0092] Total peptide content was determined using the biuret method.

[0093] (1) Preparation of standard curve: Prepare a series of peptide standard solutions with concentrations (e.g., 0, 1, 2, 3, 4, 5 mg / mL) precisely. Take 6 test tubes and add 1.0 mL of each concentration of standard solution to each tube.

[0094] (2) Dilute the sample to be tested appropriately so that its estimated concentration falls within the range of the standard curve. Take 1.0 mL of the diluted sample solution into a test tube. Add 4.0 mL of biuret reagent to all standard tubes and sample tubes, and vortex thoroughly to mix. Let the mixture stand at room temperature (20-25℃) for 30 minutes. Afterward, use a spectrophotometer at a wavelength of 540 nm, zeroing the instrument with a blank tube (standard tube with a concentration of 0), and measure the absorbance value (OD value) of each tube.

[0095] A standard curve was plotted with the concentration of the standard on the x-axis and the absorbance value on the y-axis. The OD value of the sample tube was substituted into the regression equation of the standard curve to calculate the total peptide concentration in the sample dilution. This concentration was then multiplied by the dilution factor to obtain the total peptide content of the original sample.

[0096] Small molecule peptide percentage detection:

[0097] The trichloroacetic acid (TCA) precipitation method is used because large protein and peptide molecules can be precipitated by a certain concentration of TCA, while small peptide molecules are soluble in TCA solution. After centrifugation, the contents of the supernatant (small peptide molecules) and the precipitate (large protein molecules) are measured separately, allowing the calculation of the proportion of small peptide molecules.

[0098] (1) Pipette a certain volume (V0, e.g., 2.0 mL) of peptide solution (the total peptide content C0 must be known) into a centrifuge tube, add an equal volume of 15% TCA solution (2.0 mL), so that the final TCA concentration is 7.5%. Vortex to mix, and let it stand in a refrigerator at 4°C for 1 hour to precipitate. Centrifuge at 10,000 rpm for 15 minutes at 4°C, completely dissolve the precipitate with a small amount of 0.5 M NaOH solution, and make up to a certain volume (V1, e.g., 2.0 mL).

[0099] (2) The concentrations of small molecule peptides in the supernatant were determined using the biuret method (C0). small ) and the concentration of macromolecular proteins in the dissolved precipitate (C large )

[0100] Small molecule peptide percentage (%) = [C small ×(V0+V TCA [(C0×V0)]×100%; where V TCA For the volume of TCA added, V0 is usually equal to V. TCA .

[0101] Hydroxyproline content was determined by acid hydrolysis-colorimetric method.

[0102] (1) Weigh an appropriate amount of sample (containing approximately 1-10 mg of hydroxyproline) into a hydrolysis tube, add 10 mL of 6M HCl, seal the tube under vacuum or fill it with nitrogen and then tighten the cap. Hydrolyze the solution in an oven at 110°C for 16-24 hours. After cooling, filter and transfer the hydrolysate, adjust the pH to neutral with NaOH solution, and bring the volume to a certain level (V) with distilled water.

[0103] (2) Take an appropriate amount of diluted hydrolysate or standard solution into a test tube, add chloramine-T solution, and oxidize at room temperature for 5 minutes. Then add DMAB colorimetric reagent, mix well, and heat in a 60°C water bath for 20 minutes to ensure complete colorimetric reaction. Remove and cool to room temperature.

[0104] (3) Use a spectrophotometer at a wavelength of 560 nm, zero the instrument with a blank reagent, and measure the absorbance value.

[0105] First, plot the hydroxyproline standard curve. Calculate the hydroxyproline concentration in the test solution based on the sample absorbance. Then, multiply the concentration by the dilution factor and the total volume, and divide by the sample mass to obtain the hydroxyproline content in the sample.

[0106] Antioxidant performance testing: DPPH free radical scavenging capacity determination.

[0107] Prepare a series of solutions of different concentrations using distilled water or buffer solution. Take 2.0 mL of each solution and add 2.0 mL of DPPH ethanol working solution, then vortex to mix. Incubate at room temperature in the dark for 30 minutes. Measure the absorbance (A) of the reaction solution at 517 nm, using anhydrous ethanol as a blank. 样本组 Simultaneously, a control group (with 2.0 mL of water or anhydrous ethanol and 2.0 mL of DPPH ethanol working solution added) and a blank group (with the absorbance of 2.0 mL of sample solution + 2.0 mL of anhydrous ethanol) were set up, and A was measured. 对照组 and A 空白组 ;

[0108] DPPH free radical scavenging rate (%) = [1-(A 样本组 -A 空白组 ) / A 对照组 ×100%.

[0109] Table 1. Results of total peptide content, small molecule peptide ratio, hydroxyproline content, and antioxidant properties of the collagen peptides of this invention.

[0110]

[0111] As shown in Table 1, compared with Comparative Examples 1-2, the actual effect of Example 1 is far greater than the theoretical sum of the comparative examples. This proves that the combined action of light and DES produces a synergistic effect. DES can effectively disrupt the tight structure of collagen, allowing it to be more fully exposed to light; while light of a specific wavelength can excite the DES system and promote the breaking of peptide bonds, thereby generating small molecule peptides more efficiently. Hydroxyproline is a characteristic amino acid of collagen, and its content directly reflects the purity and structural retention of the product. DPPH scavenging rate represents the antioxidant activity of the product. Example 1 is significantly superior to either single method in both indicators, indicating that the "light-DES" system not only improves extraction efficiency but also creates a milder and more protective extraction environment. DES may reduce oxidative damage, while the introduction of red light (660nm) is presumably able to scavenge free radicals, jointly protecting the active groups and characteristic structures of collagen peptides, thus better preserving their functionality. Examples 1-4 employed different DES systems and illumination modes (alternating / continuous / monochromatic light), but their results were consistently and significantly superior to the corresponding unilluminated comparative examples (Comparative Examples 1 and 4). This demonstrates that "illumination-assisted DES" is a universal technical solution, rather than a coincidence under specific formulations. While the effect of collagen peptides in Comparative Example 5 was better than the traditional method, it was slightly inferior to Examples 1 and 2. This indicates that the introduction of green light did not bring any gain, but may have diluted the effective blue and red light energy. This also reveals the scientific basis and necessity of the present invention's selection of 400-480nm blue light and 630-680nm red light, further highlighting the subtlety of the synergy between specific wavelength illumination and DES.

[0112] In summary, the "specific wavelength light synergistic deep eutectic solvent" technology provided by this invention is not a simple combination of two technologies, but rather produces a powerful synergistic effect. This synergistic effect significantly improves extraction efficiency and small molecule peptide yield while effectively protecting the functional activity of collagen peptides. Its overall effect is far superior to any single technology (DES only or light irradiation only) or traditional chemical methods.

Claims

1. A method for extracting collagen peptides, characterized in that, The method includes photo-assisted deep co-solvent extraction of collagen peptides.

2. The method according to claim 1, characterized in that, The illumination is selected from blue light with a wavelength of 400-480nm and / or red light with a wavelength of 630-680nm.

3. The method according to claim 2, characterized in that, The lighting conditions include: a light intensity of 10-60 mW / cm². 2 ; and / or, the illumination time is 30-90% of the total extraction time; and / or, the illumination method is continuous irradiation or alternating irradiation.

4. The method according to claim 1, characterized in that, The collagen peptide raw material is mixed with a deep co-solvent at a material-to-liquid ratio of 1:5 to 1:

12.

5. The method according to claim 4, characterized in that, The deep co-solution solvent is prepared by mixing hydrogen bond donors and hydrogen bond acceptors in a molar ratio of 1:1–1:

10.

6. The method according to claim 5, characterized in that, The hydrogen bond acceptor is selected from at least one of glycerol, lactic acid, urea, and sorbitol; and / or, the hydrogen bond donor is choline chloride.

7. The method according to claim 1, characterized in that, The extraction conditions include: an extraction temperature of 40-65℃ and a total extraction time of 1-3 hours.

8. The method according to claim 1, characterized in that, The method also includes dialysis, concentration, and drying processes.

9. The method according to claim 8, characterized in that, The molecular weight cutoff range of the dialysis is 500-10000 Da; and / or the concentration temperature is 50-60°C, and the concentration pressure is -0.08 MPa to -0.09 MPa.

10. A collagen peptide prepared by the method according to any one of claims 1-9.

Images