A method for preparing high-purity and high-concentration collagen product

Abstract

The invention discloses a method for preparing a high-purity and high-concentration collagen product, which belongs to the technical field of medical materials, and specifically relates to using bovine Achilles tendon as a biomass material for preparing collagen products, pre-treating the bovine Achilles tendon to prepare a pre-treated bovine Achilles tendon slice, preparing a collagen crude extract from the pre-treated bovine Achilles tendon slice under the action of a protease and / or an extraction aid, and then preparing a collagen product through high-pressure homogenization, ultrafiltration treatment, and adjusting ionic strength. The collagen prepared by the method of the invention has high purity, high collagen yield, low collagen loss, and the method of the invention has a good recovery rate for collagen.

Description

Technical Field

[0001] The invention belongs to the technical field of medical materials, and specifically relates to a method for preparing a high-purity and high-concentration collagen product. Background Art

[0002] Collagen is mainly found in connective tissues such as skin, bones, and tendons of animals, among which the collagen content in tendon tissue is the highest. Collagen is an important component of the extracellular matrix and a natural nanomaterial with high biocompatibility, biodegradability, film-forming property, and low antigenicity. Collagen interacts with other protein family members in the signaling pathway through a triple helix structure region composed of three α-type peptide chains, and mediates cytological regulatory behaviors, such as cell adhesion and migration, cell differentiation and proliferation, hemostasis, wound healing, and regulation of immune function. Therefore, it plays an important role in food, cosmetics, biomedicine, and other fields, especially in the preparation of biomedical materials.

[0003] CN105063148A discloses a "collagen preparation method", which uses cowhide as raw material, and prepares collagen stock solution through adsorption filtration, ultrafiltration concentration for buffer solution, ion exchange chromatography, ultrafiltration concentration for storage solution and filtration sterilization. CN114452444A discloses a "low endotoxin collagen preparation method", which uses cation exchange chromatography, and prepares low endotoxin collagen stock solution by rebalancing, washing, eluting and ultrafiltration concentration of the cation exchange chromatography column. CN115124613A discloses "a high-fluidity high-concentration collagen solution and its preparation method", which is to prepare a collagen stock solution with a concentration of 10~20 mg / mL. CN107155305A discloses a "high-concentration collagen preparation method for use as a medical material", which uses enzymatic hydrolysis, purified water dissolution, filtration, ultrafiltration and concentration to prepare collagen with a concentration of 120 mg / mL.

[0004] Currently, the collagen products on the market are prepared from animal skin tissue (such as pig skin and cow skin). However, due to the high content of adipose tissue in the skin tissue, production process technology difficulties are prone to occur, the production process is complicated, and protein loss is high during the production process. The collagen degradation time is short, which is not conducive to the mass production of collagen products and corporate cost control. Summary of the invention

[0005] The object of the present invention is to provide a method for preparing a high-purity and high-concentration collagen product with high collagen purity, high collagen yield, low collagen loss and high collagen recovery rate.

[0006] The technical solution adopted by the present invention to achieve the above-mentioned purpose is:

[0007] A method for preparing a collagen product comprises preparing a collagen product from a biomass raw material through an extraction process, wherein the biomass raw material is firstly used to prepare a collagen crude extract, and then the collagen crude extract is subjected to high-pressure homogenization and ultrafiltration to prepare an ultrafiltered collagen solution, and the ultrafiltered collagen solution is subjected to ionic strength adjustment and separation to obtain a collagen product. The present invention performs enzymatic hydrolysis on a biomass raw material containing collagen, extracts the collagen into a solution, and then separates the collagen through high-pressure homogenization, ultrafiltration and ionic strength adjustment. The high-pressure homogenization method can break the incompletely enzymatically hydrolyzed fine tissue particles and crudely extracted collagen fibers in the collagen crude extract into nanometer levels, so that the collagen in the fine tissue can be fully extracted during the homogenization process. At the same time, the interaction and isoelectric point between collagen molecules are adjusted to achieve the effect of being easily induced to assemble into fibers and precipitate. The ultrafiltration membrane separation and purification technology abandons the complicated purification operation steps, removes pepsin, terminal peptides, residual reagents and other small molecule impurities, and controls the endotoxin level to obtain high-purity collagen precipitation. By adjusting the ionic strength and pH, high-concentration collagen is prepared, and the pH range of collagen is weakly acidic and weakly alkaline. Protein precipitation with a high recovery rate can be obtained by isoelectric precipitation, without the need to add neutral salt salting-out process, which simplifies the production operation steps and saves production costs. While obtaining high-concentration, high-purity medical-grade collagen, the natural biological activity of its triple helix structure is maintained, and it has both biodegradability, tissue absorbability and biocompatibility. It can be formulated into collagen products of different concentrations, which can achieve the wide application of collagen in the biomedical field.

[0008] Preferably, in high-pressure homogenization, the homogenization pressure is 100-400 bar and the feed rate is 10-150 L / h; or, acidic reagents and alkaline solutions are used to adjust the ionic strength; or, proteases are used to enzymatically hydrolyze the biomass raw materials in the preparation of the crude collagen extract.

[0009] Preferably, the acidic reagent is at least one of acetic acid, hydrochloric acid, citric acid and phosphate; or, the amount of the acidic reagent used is in a ratio of 0.1-0.5 mol / L to the ultrafiltered collagen solution; or, the alkaline medium in the alkaline solution is at least one of sodium hydroxide, potassium hydroxide and sodium bicarbonate; or, the alkaline solution contains 5-20 wt% of the alkaline medium; or, pepsin is added during the enzymatic hydrolysis; or, an extraction aid is added during the enzymatic hydrolysis.

[0010] More preferably, the extraction aid is a functionalized aminopolysiloxane; or, the amount of the extraction aid used is 2-8wt% of the biomass raw material. In the method for extracting collagen of the present invention, aminopolysiloxane can be firstly prepared by polymerizing γ-aminopropylmethyldiethoxysilane and diethyldiethoxysilane, and then the aminopolysiloxane is functionalized by at least a functional agent including cinnamaldehyde to prepare a functionalized aminopolysiloxane, which can keep the structure of collagen stable in the enzymatic hydrolysis solution and the subsequent high-pressure homogenization treatment, and make the collagen easier to extract during the enzymatic hydrolysis process. The use of functionalized aminopolysiloxane can increase the extraction amount of collagen and stabilize the structure of collagen. Functionalized aminopolysiloxane will not promote the dissolution of non-collagenous proteins, and the purity of the obtained collagen is higher.

[0011] More preferably, the functionalized aminopolysiloxane contains at least a cinnamaldehyde group; or, the functionalized aminopolysiloxane is prepared by reacting an aminopolysiloxane and a functional agent, wherein the aminopolysiloxane is polymerized by γ-aminopropylmethyldiethoxysilane and diethyldiethoxysilane and capped by a capping agent, and the functional agent includes cinnamaldehyde.

[0012] More preferably, the usage amount of diethyldiethoxysilane is 100-300wt% of γ-aminopropylmethyldiethoxysilane; or, the end-capping agent is hexamethyldisiloxane, and the usage amount of hexamethyldisiloxane is 5-20wt% of γ-aminopropylmethyldiethoxysilane.

[0013] Preferably, the collagen product includes solid collagen and liquid collagen.

[0014] Preferably, the liquid collagen is solid collagen dispersed in a dispersion system, and the dispersion system includes at least one of a phosphate buffer, a sodium hyaluronate system, a hydroxyapatite system and a silicone gel system.

[0015] Preferably, the solvent of the dispersion system is water, alcohol or other medical solvents.

[0016] Preferably, the biomass raw material is bovine Achilles tendon.

[0017] Preferably, in the preparation of the pretreated bovine Achilles tendon slice, foreign matter on the surface of the bovine Achilles tendon is removed, and then the bovine Achilles tendon with the foreign matter removed from the surface is cryogenically frozen, sliced, the fascia is removed, and finally decellularized to obtain the pretreated bovine Achilles tendon slice.

[0018] More preferably, the foreign matter is hair, adipose tissue or the like.

[0019] More preferably, the low-temperature freezing is to freeze the raw material bovine Achilles tendon at -20°C to -80°C for 12-48h.

[0020] More preferably, the low-temperature frozen bovine Achilles tendon is placed at 20-40° C. for 20-120 seconds for surface thawing before slicing.

[0021] More preferably, during the removal of the fascia, a slicing device is used, the thickness of the bovine Achilles tendon slice is set at 0.1-1.0 mm, the spindle speed is set at 150-300 r / min, and after slicing, the fascia is peeled off from the blade.

[0022] More preferably, the decellularization treatment includes at least one of a hypertonic salt treatment, an organic solution treatment, and an oxidation treatment.

[0023] More preferably, in the hypertonic saline treatment, the bovine Achilles tendon slice with the fascia removed is immersed in a hypertonic saline solution for 10-24 hours, then washed with water and squeezed dry, the initial temperature of the hypertonic saline solution is 0-10°C, and the temperature is not maintained constant when left naturally, and the hypertonic saline solution is 15-25wt% sodium chloride solution.

[0024] More preferably, in the organic solution treatment, the bovine Achilles tendon slices with the fascia removed or the bovine Achilles tendon slices treated with hypertonic salt are immersed in an organic solution for 20-60 minutes, and then washed with water and squeezed dry, wherein the organic solution contains 0.05-1wt% TritonX-100 and 0.1-0.5wt% TNBP.

[0025] More preferably, during the oxidation treatment, the bovine Achilles tendon slices with the fascia removed or the bovine Achilles tendon slices treated with an organic solution are immersed in a hydrogen peroxide solution for 5-15 minutes, and then washed with water and squeezed dry, wherein the hydrogen peroxide solution contains 3-5wt% hydrogen peroxide.

[0026] Preferably, in the preparation of the crude collagen extract, a protease is added to an acidic solution, and the pretreated bovine Achilles tendon slices are added thereto for enzymatic hydrolysis for 40-72 hours at a temperature of 2-8° C., and the solution is centrifuged to obtain the supernatant to obtain the crude collagen extract.

[0027] More preferably, the acidic medium in the acidic solution is at least one of acetic acid, hydrochloric acid, citric acid, or tartaric acid and tartaric acid, and the concentration of the acidic medium in the acidic solution is 0.01-1 mol / L. When the concentration of the acidic medium meets the requirements, the pH is adjusted to 2.0-3.0.

[0028] More preferably, the protease is pepsin, and the usage ratio of the protease to the acidic solution is 1 g: 1-40 L.

[0029] More preferably, the usage ratio of the pretreated bovine Achilles tendon slice and the acidic solution is 1g:0.01-0.08L.

[0030] More preferably, during centrifugation, the centrifugal speed is 8000-12000 rpm / min, and the centrifugal time is 45-60 min.

[0031] Preferably, functionalized aminopolysiloxane may be added in the preparation of the crude collagen extract, and the amount of the functionalized aminopolysiloxane used is 2-8wt% of the pretreated bovine Achilles tendon slice.

[0032] Preferably, the functionalized aminopolysiloxane is prepared by reacting aminopolysiloxane with a functional agent.

[0033] Preferably, in the preparation of aminopolysiloxane, γ-aminopropylmethyldiethoxysilane and diethyldiethoxysilane are mixed with water, and then an alkaline reagent and DMSO are added and stirred, and the mixture is reacted at 40-60° C. for 1-4 hours. After the reaction is completed, the mixture is distilled under reduced pressure for 0.5-2 hours, and then hexamethyldisiloxane is added to react for 2-6 hours. After the reaction is completed, the pH is adjusted to neutral to obtain aminopolysiloxane.

[0034] More preferably, the usage amount of γ-aminopropylmethyldiethoxysilane is 100-200 wt % of water.

[0035] More preferably, the usage amount of diethyldiethoxysilane is 100-300 wt % of γ-aminopropylmethyldiethoxysilane.

[0036] More preferably, the alkaline agent is at least one of sodium hydroxide or potassium hydroxide, and the amount of the alkaline agent used is 0.1-0.4 wt % of the γ-aminopropylmethyldiethoxysilane.

[0037] More preferably, the amount of DMSO used is 0.5-2 wt % of γ-aminopropylmethyldiethoxysilane.

[0038] More preferably, the amount of hexamethyldisiloxane used is 5-20 wt % of γ-aminopropylmethyldiethoxysilane.

[0039] Preferably, in the preparation of the functionalized aminopolysiloxane, the aminopolysiloxane and the functional agent are added to methanol and mixed under an inert gas atmosphere, and refluxed at 60-90° C. for 12-36 hours. After the reaction is completed, the methanol is removed and dried to obtain the functionalized aminopolysiloxane.

[0040] More preferably, the inert gas is nitrogen.

[0041] More preferably, the aminopolysiloxane is used in an amount of 3-12 wt % of methanol.

[0042] More preferably, the functional agent is cinnamaldehyde, and the usage amount of the functional agent is 2-10wt% of the aminopolysiloxane.

[0043] More preferably, the functional agent may also be glycidyl methacrylate.

[0044] More preferably, the functional agent can also be cinnamaldehyde and methacrylic acid glycidyl ether, and cinnamaldehyde and methacrylic acid glycidyl ether are used in combination at a mass ratio of 1:0.5-10. In the present invention, when aminopolysiloxane is functionalized, methacrylic acid glycidyl ether can also be used when cinnamaldehyde is used. After aminopolysiloxane is functionalized with cinnamaldehyde and methacrylic acid glycidyl ether at the same time, it is applied to the method for preparing collagen of the present invention, and the obtained collagen has high purity, high yield of collagen, and can better maintain its helical structure.

[0045] Preferably, in the preparation of ultrafiltered collagen solution, the crude collagen extract is subjected to high-pressure homogenization at a temperature of 4-15°C, and the solution after high-pressure homogenization is ultrafiltered using an ultrafiltration membrane to recover collagen with a larger molecular weight at the cutoff end to obtain an ultrafiltered collagen solution.

[0046] More preferably, the solution treated by high pressure homogenization can be circulated, and the number of cycles of high pressure homogenization is 2-15 times.

[0047] More preferably, in the high-pressure homogenization process, the homogenization pressure is 100-400 bar, and the feed rate is 10-150 L / h.

[0048] More preferably, the ultrafiltration membrane is an ultrafiltration membrane with a relative molecular mass of 100-150KD. During the ultrafiltration treatment, the ultrafiltration flow rate is 80-1500 cubic meters / (hour×square meters), the membrane pressure of the ultrafiltration membrane is 0.1-0.6MPa, the ultrafiltration temperature is 2-15°C, the ultrafiltration replacement fluid is a 0.01-0.1mol / L dilute acid solution, the acidic medium in the dilute acid solution is at least one of hydrochloric acid, acetic acid, and citric acid, the volume of the ultrafiltration replacement fluid used is 3-9 times the volume of the crude collagen extract to ensure that the system composition before ultrafiltration is fully replaced, the solution pH is maintained in the range of 1-4 during the ultrafiltration treatment, the collagen with a larger molecular weight at the retention end is recovered, and the small molecular impurities such as pepsin, terminal peptide, endotoxin, antigen and inorganic salt in the permeation end are removed.

[0049] Preferably, in the preparation of collagen, the ionic strength of the ultrafiltered collagen solution is adjusted, an acidic reagent is first added, and then an alkaline solution is added to adjust the pH to 3-9.5, the solution is stirred for 6-24 hours, centrifuged, and the precipitate is taken to obtain collagen.

[0050] More preferably, the acidic agent is at least one of acetic acid, hydrochloric acid, citric acid and phosphate.

[0051] More preferably, the usage ratio of the acidic reagent to the ultrafiltered collagen solution is 0.1-0.5 mol / L.

[0052] More preferably, the alkaline medium in the alkaline solution is at least one of sodium hydroxide, potassium hydroxide and sodium bicarbonate, and the alkaline solution contains 5-20 wt % of the alkaline medium.

[0053] More preferably, during centrifugation, the centrifugal speed is 8000-12000 rpm / min, and the centrifugal time is 25-60 min.

[0054] Preferably, in the preparation of the collagen solution, the collagen precipitate is added into the dispersion system to prepare the collagen solution.

[0055] More preferably, the dispersion system includes at least one of a phosphate buffer solution, a sodium hyaluronate solution, a hydroxyapatite solution and a silicone gel solution.

[0056] More preferably, the pH of the phosphate buffer is 5.5-10, and the molar concentration of the substance in the phosphate buffer is 0.01-0.1 mol / L.

[0057] More preferably, the concentration of the collagen solution is 0.5-230 mg / mL.

[0058] The invention discloses use of an extraction aid in preparing a collagen product and / or enzymatic hydrolysis. The extraction aid is a functionalized aminopolysiloxane, and the functionalized aminopolysiloxane has at least a cinnamaldehyde group.

[0059] The present invention uses bovine Achilles tendon as a biomass material for preparing collagen products, and pre-treats the bovine Achilles tendon to prepare pre-treated bovine Achilles tendon slices. The pre-treated bovine Achilles tendon slices are used to prepare a collagen crude extract under the action of a protease and / or an extraction aid, and then the collagen product is prepared by high-pressure homogenization, ultrafiltration, and ion strength adjustment. Therefore, the present invention has the following beneficial effects: high collagen purity, high collagen yield, low collagen loss, and the method of the present invention has a good recovery rate for collagen. Therefore, the present invention is a method for preparing a high-purity, high-concentration collagen product with high collagen purity, high collagen yield, low collagen loss, and high collagen recovery rate. BRIEF DESCRIPTION OF THE DRAWINGS

[0060] Figure 1 It is an infrared spectrum.

[0061] Figure 2 This is a graph showing the purity of collagen.

[0062] Figure 3 Collagen concentration graph.

[0063] Figure 4 Collagen electrophoresis diagram. DETAILED DESCRIPTION

[0064] The technical solution of the present invention is further described in detail below in conjunction with specific implementations and drawings:

[0065] Example 1: A method for preparing solid collagen

[0066] Preparation of pretreated bovine Achilles tendon slices: remove foreign matter on the surface of the bovine Achilles tendon, then cryo-freeze the bovine Achilles tendon with foreign matter removed from the surface, slice it, remove the fascia, and finally perform decellularization to obtain pretreated bovine Achilles tendon slices. Foreign matter includes hair, adipose tissue, etc. Cryo-freezing is to freeze the raw material bovine Achilles tendon at minus 20°C for 24 hours; before slicing, the low-temperature frozen bovine Achilles tendon is placed at 30°C for 30 seconds to defrost the surface; in the removal of fascia, a slicing device is used to operate, and the thickness of the bovine Achilles tendon slice is set at 0.5mm and the spindle speed is 200r / min. After slicing, the fascia is peeled off from the blade; the decellularization treatment includes hypertonic salt treatment, organic solution treatment and oxidation treatment; in the hypertonic salt treatment, the bovine Achilles tendon slice with fascia removed is immersed in a hypertonic salt solution for 16 hours, then washed with water and squeezed dry. The initial temperature of the hypertonic salt solution is 0°C, and it is naturally placed. The temperature is not maintained constant, and the hypertonic saline solution is a 15wt% sodium chloride solution; in the organic solution treatment, the bovine Achilles tendon slices with the fascia removed or the bovine Achilles tendon slices treated with hypertonic saline are immersed in the organic solution for 20 minutes, then washed with water and squeezed dry, and the organic solution contains 0.1wt% TritonX-100 and 0.1wt% TNBP; in the oxidation treatment, the bovine Achilles tendon slices with the fascia removed or the bovine Achilles tendon slices treated with the organic solution are immersed in a hydrogen peroxide solution for 15 minutes, then washed with water and squeezed dry, and the hydrogen peroxide solution contains 3wt% hydrogen peroxide.

[0067] Preparation of crude collagen extract: Add protease to the acidic solution, add pretreated bovine Achilles tendon slices at 5°C for enzymatic hydrolysis for 40 hours, centrifuge, and take the supernatant to obtain crude collagen extract. The acidic medium in the acidic solution is acetic acid, and the concentration of the acidic medium in the acidic solution is 0.5 mol / L. When the concentration of the acidic medium meets the requirements, adjust the pH to 2.0; the protease is pepsin, and the amount of protease used is 0.5g:2L; the amount of pretreated bovine Achilles tendon slices and acidic solution is 50g:2L. During centrifugation, the centrifugal speed is 10000rpm / min, and the centrifugal time is 60min.

[0068] Preparation of ultrafiltration collagen solution: The crude collagen extract is subjected to high pressure homogenization at a temperature of 4°C, and the solution after high pressure homogenization is subjected to ultrafiltration treatment using an ultrafiltration membrane to recover collagen with a larger molecular weight at the cutoff end to obtain an ultrafiltration collagen solution. The solution after high pressure homogenization treatment can be circulated, and the number of cycles of high pressure homogenization treatment is 5 times. During the high-pressure homogenization treatment, the homogenization pressure is 200 bar and the feed rate is 15 L / h. The ultrafiltration membrane has a relative molecular mass of 100 KD. During the ultrafiltration treatment, the ultrafiltration flow rate is 80 cubic meters / (hour × square meters). The membrane pressure of the ultrafiltration membrane is 0.1 MPa, the ultrafiltration temperature is 5°C, and the ultrafiltration replacement fluid is a 0.01 mol / L dilute acid solution. The acidic medium in the dilute acid solution is hydrochloric acid. The volume of the ultrafiltration replacement fluid used is 6 times the volume of the collagen crude extract to ensure that the system composition before ultrafiltration is fully replaced. During the ultrafiltration treatment, the solution pH is maintained at 2, the collagen with a larger molecular weight at the retention end is recovered, and small molecular impurities such as pepsin, terminal peptide, endotoxin, antigen and inorganic salt in the permeation end are removed.

[0069] Preparation of collagen: Adjust the ionic strength of the ultrafiltered collagen solution, first add an acidic reagent, then add an alkaline solution, adjust the pH to 7.5, stir for 6 hours, centrifuge, take the precipitate, and obtain collagen, i.e., solid collagen. The acidic reagent is hydrochloric acid. The amount of the acidic reagent used is 0.3 mol / L to the ultrafiltered collagen solution, the alkaline medium in the alkaline solution is sodium hydroxide, and the alkaline solution contains 20 wt% of the alkaline medium. During centrifugation, the centrifugal speed is 10000 rpm / min, and the centrifugal time is 30 min.

[0070] Example 2: A method for preparing solid collagen

[0071] Preparation of pretreated bovine Achilles tendon slices: remove foreign matter on the surface of the bovine Achilles tendon, then cryo-freeze the bovine Achilles tendon with foreign matter removed from the surface, slice it, remove the fascia, and finally perform decellularization to obtain pretreated bovine Achilles tendon slices. Foreign matter includes hair, adipose tissue, etc. Cryo-freezing is to freeze the raw material bovine Achilles tendon at minus 40°C for 48 hours; before slicing, the bovine Achilles tendon after cryo-freezing is placed at 30°C for 60 seconds to defrost the surface; in the removal of fascia, a slicing device is used to operate, and the thickness of the bovine Achilles tendon slice is set at 1mm and the spindle speed is 200r / min. After slicing, the fascia is peeled off from the blade; the decellularization treatment includes hypertonic salt treatment, organic solution treatment and oxidation treatment; in the hypertonic salt treatment, the bovine Achilles tendon slice with fascia removed is immersed in a hypertonic salt solution for 16 hours, then washed with water and squeezed dry. The initial temperature of the hypertonic salt solution is 0°C, and it is naturally placed. The temperature is not maintained constant, and the hypertonic saline solution is a 20wt% sodium chloride solution. In the organic solution treatment, the bovine Achilles tendon slices with the fascia removed or the bovine Achilles tendon slices treated with hypertonic salt are immersed in the organic solution for 20 minutes, then washed with water and squeezed dry, and the organic solution contains 0.1wt% TritonX-100 and 0.3wt% TNBP. In the oxidation treatment, the bovine Achilles tendon slices with the fascia removed or the bovine Achilles tendon slices treated with organic solution are immersed in a hydrogen peroxide solution for 15 minutes, then washed with water and squeezed dry, and the hydrogen peroxide solution contains 5wt% hydrogen peroxide.

[0072] Preparation of crude collagen extract: Add protease to the acidic solution, add pretreated bovine Achilles tendon slices at 5°C for enzymatic hydrolysis for 48 hours, centrifuge, and take the supernatant to obtain crude collagen extract. The acidic medium in the acidic solution is acetic acid, and the concentration of the acidic medium in the acidic solution is 0.5 mol / L. When the concentration of the acidic medium meets the requirements, adjust the pH to 2.0; the protease is pepsin, and the amount of protease used is 0.5g:3L; the amount of pretreated bovine Achilles tendon slices and acidic solution is 50g:3L. During centrifugation, the centrifugal speed is 10000rpm / min, and the centrifugal time is 60min.

[0073] Preparation of ultrafiltration collagen solution: The crude collagen extract is subjected to high pressure homogenization at a temperature of 10°C, and the solution after high pressure homogenization is subjected to ultrafiltration treatment using an ultrafiltration membrane to recover collagen with a larger molecular weight at the cutoff end to obtain an ultrafiltration collagen solution. The solution after high pressure homogenization treatment can be recycled, and the number of cycles of high pressure homogenization treatment is 3 times. During the high-pressure homogenization treatment, the homogenization pressure is 250 bar and the feed rate is 15 L / h. The ultrafiltration membrane has a relative molecular mass of 100 KD. During the ultrafiltration treatment, the ultrafiltration flow rate is 80 cubic meters / (hour × square meters). The membrane pressure of the ultrafiltration membrane is 0.1 MPa, the ultrafiltration temperature is 5°C, and the ultrafiltration replacement fluid is a 0.02 mol / L dilute acid solution. The acidic medium in the dilute acid solution is citric acid. The volume of the ultrafiltration replacement fluid used is 6 times the volume of the collagen crude extract to ensure that the system composition before ultrafiltration is fully replaced. During the ultrafiltration treatment, the solution pH is maintained at 2, the collagen with a larger molecular weight at the retention end is recovered, and small molecular impurities such as pepsin, terminal peptide, endotoxin, antigen and inorganic salt in the permeation end are removed.

[0074] Preparation of collagen: adjust the ionic strength of the ultrafiltered collagen solution, first add an acidic reagent, then add an alkaline solution, adjust the pH to 4.5, stir for 24 hours, centrifuge, take the precipitate, and obtain collagen, i.e., solid collagen. The acidic reagent is hydrochloric acid. The amount of the acidic reagent used is 0.4 mol / L to the ultrafiltered collagen solution, the alkaline medium in the alkaline solution is sodium hydroxide, and the alkaline solution contains 10 wt% of the alkaline medium. During centrifugation, the centrifugal speed is 12000 rpm / min, and the centrifugal time is 30 min.

[0075] Example 3: A method for preparing solid collagen

[0076] Preparation of pretreated bovine Achilles tendon slices: remove foreign matter on the surface of the bovine Achilles tendon, then cryo-freeze the bovine Achilles tendon with foreign matter removed from the surface, slice it, remove the fascia, and finally perform decellularization to obtain pretreated bovine Achilles tendon slices. Foreign matter includes hair, adipose tissue, etc. Cryo-freezing is to freeze the raw material bovine Achilles tendon at minus 80°C for 48 hours; before slicing, the low-temperature frozen bovine Achilles tendon is placed at 30°C for 60 seconds to defrost the surface; in the removal of fascia, a slicing device is used to operate, and the thickness of the bovine Achilles tendon slice is set at 1mm and the spindle speed is 200r / min. After slicing, the fascia is peeled off from the blade; the decellularization treatment includes hypertonic salt treatment, organic solution treatment and oxidation treatment; in the hypertonic salt treatment, the bovine Achilles tendon slice with fascia removed is immersed in a hypertonic salt solution for 20 hours, then washed with water and squeezed dry. The initial temperature of the hypertonic salt solution is 0°C, and it is naturally placed. The temperature is not maintained constant, and the hypertonic saline solution is a 20wt% sodium chloride solution. In the organic solution treatment, the bovine Achilles tendon slices with the fascia removed or the bovine Achilles tendon slices treated with hypertonic salt are immersed in the organic solution for 20 minutes, then washed with water and squeezed dry, and the organic solution contains 0.1wt% TritonX-100 and 0.3wt% TNBP. In the oxidation treatment, the bovine Achilles tendon slices with the fascia removed or the bovine Achilles tendon slices treated with organic solution are immersed in a hydrogen peroxide solution for 15 minutes, then washed with water and squeezed dry, and the hydrogen peroxide solution contains 5wt% hydrogen peroxide.

[0077] Preparation of crude collagen extract: Add protease to the acidic solution, add pretreated bovine Achilles tendon slices at 5°C for enzymatic hydrolysis for 72 hours, centrifuge, and take the supernatant to obtain crude collagen extract. The acidic medium in the acidic solution is hydrochloric acid, and the concentration of the acidic medium in the acidic solution is 0.01 mol / L. When the concentration of the acidic medium meets the requirements, adjust the pH to 2.0; the protease is pepsin, and the usage ratio of the protease to the acidic solution is 1g:4L; the usage ratio of the pretreated bovine Achilles tendon slices to the acidic solution is 100g:4L. During centrifugation, the centrifugal speed is 10000rpm / min, and the centrifugal time is 60min.

[0078] Preparation of ultrafiltration collagen solution: The crude collagen extract is subjected to high pressure homogenization at a temperature of 10°C, and the solution after high pressure homogenization is subjected to ultrafiltration treatment using an ultrafiltration membrane to recover collagen with a larger molecular weight at the cutoff end to obtain an ultrafiltration collagen solution. The solution after high pressure homogenization treatment can be recycled, and the number of cycles of high pressure homogenization treatment is 3 times. During the high-pressure homogenization treatment, the homogenization pressure is 300 bar and the feed rate is 20 L / h. The ultrafiltration membrane has a relative molecular mass of 100 KD. During the ultrafiltration treatment, the ultrafiltration flow rate is 80 cubic meters / (hour × square meters). The membrane pressure of the ultrafiltration membrane is 0.1 MPa, the ultrafiltration temperature is 5°C, and the ultrafiltration replacement fluid is a 0.02 mol / L dilute acid solution. The acidic medium in the dilute acid solution is citric acid. The volume of the ultrafiltration replacement fluid used is 3 times the volume of the collagen crude extract to ensure that the system composition before ultrafiltration is fully replaced. During the ultrafiltration treatment, the solution pH is maintained at 2 to recover collagen with a larger molecular weight at the retention end and remove small molecular impurities such as pepsin, terminal peptides, endotoxins, antigens and inorganic salts at the permeation end.

[0079] Preparation of collagen: Adjust the ionic strength of the ultrafiltered collagen solution, first add an acidic reagent, then add an alkaline solution, adjust the pH to 9.2, stir for 16 hours, centrifuge, take the precipitate, and obtain collagen, i.e., solid collagen. The acidic reagent is citric acid. The amount of the acidic reagent used is 0.4 mol / L to the ultrafiltered collagen solution, the alkaline medium in the alkaline solution is sodium hydroxide, and the alkaline solution contains 20 wt% of the alkaline medium. During centrifugation, the centrifugal speed is 12000 rpm / min, and the centrifugal time is 30 min.

[0080] Example 4: A method for preparing solid collagen

[0081] The difference between this embodiment and embodiment 3 lies in the preparation of the crude collagen extract.

[0082] Preparation of aminopolysiloxane: γ-aminopropylmethyldiethoxysilane and diethyldiethoxysilane are mixed with water, and then an alkaline agent and DMSO are added and stirred and mixed, and the mixture is reacted at 50°C for 2 hours. After the reaction is completed, the mixture is distilled under reduced pressure for 1 hour, and then hexamethyldisiloxane is added to react for 4 hours. After the reaction is completed, the pH is adjusted to neutral to obtain aminopolysiloxane. The amount of γ-aminopropylmethyldiethoxysilane used is 150wt% of water, the amount of diethyldiethoxysilane used is 200wt% of γ-aminopropylmethyldiethoxysilane, the alkaline agent is potassium hydroxide, the amount of the alkaline agent used is 0.2wt% of γ-aminopropylmethyldiethoxysilane, the amount of DMSO used is 1wt% of γ-aminopropylmethyldiethoxysilane, and the amount of hexamethyldisiloxane used is 10wt% of γ-aminopropylmethyldiethoxysilane.

[0083] Preparation of functionalized aminopolysiloxane: In an inert gas atmosphere, aminopolysiloxane and a functional agent are added to methanol and mixed, and refluxed at 80°C for 24 hours. After the reaction is completed, the methanol is removed and dried to obtain functionalized aminopolysiloxane. The inert gas is nitrogen, the amount of aminopolysiloxane used is 6wt% of methanol, and the functional agent is cinnamaldehyde, and the amount of the functional agent used is 5wt% of the aminopolysiloxane.

[0084] Preparation of crude collagen extract: Add protease to the acidic solution, add functionalized aminopolysiloxane and pretreated bovine Achilles tendon slices at a temperature of 5°C for enzymatic hydrolysis for 72 hours, centrifuge, and take the supernatant to obtain crude collagen extract. The acidic medium in the acidic solution is hydrochloric acid, and the concentration of the acidic medium in the acidic solution is 0.01 mol / L. When the concentration of the acidic medium meets the requirements, adjust the pH to 2.0; the protease is pepsin, and the usage ratio of the protease to the acidic solution is 1g:4L; the usage ratio of the pretreated bovine Achilles tendon slices to the acidic solution is 100g:4L, and the usage ratio of the functionalized aminopolysiloxane is 5wt% of the pretreated bovine Achilles tendon slices. During centrifugation, the centrifugal speed is 10000rpm / min, and the centrifugal time is 60min.

[0085] Example 5: A method for preparing solid collagen

[0086] The difference between this embodiment and embodiment 4 lies in the preparation of functionalized aminopolysiloxane.

[0087] Preparation of functionalized aminopolysiloxane: In an inert gas atmosphere, aminopolysiloxane and a functional agent are added to methanol and mixed, and refluxed at 80°C for 24 hours. After the reaction is completed, the methanol is removed and dried to obtain functionalized aminopolysiloxane. The inert gas is nitrogen, the amount of aminopolysiloxane used is 6wt% of methanol, and the functional agent is cinnamaldehyde, and the amount of the functional agent used is 8wt% of the aminopolysiloxane.

[0088] Example 6: A method for preparing solid collagen

[0089] The difference between this embodiment and embodiment 4 lies in the preparation of functionalized aminopolysiloxane.

[0090] Preparation of functionalized aminopolysiloxane: In an inert gas atmosphere, aminopolysiloxane and a functional agent are added to methanol and mixed, and refluxed at 80°C for 24 hours. After the reaction is completed, the methanol is removed and dried to obtain functionalized aminopolysiloxane. The inert gas is nitrogen, the amount of aminopolysiloxane used is 6wt% of methanol, and the functional agent is cinnamaldehyde and methacrylic acid glycidyl ether. Cinnamaldehyde and methacrylic acid glycidyl ether are used in a mass ratio of 1:2, and the amount of the functional agent used is 5wt% of the aminopolysiloxane.

[0091] Example 7: A method for preparing solid collagen

[0092] The difference between this embodiment and embodiment 4 lies in the preparation of functionalized aminopolysiloxane.

[0093] Preparation of functionalized aminopolysiloxane: In an inert gas atmosphere, aminopolysiloxane and a functional agent are added to methanol and mixed, and refluxed at 80°C for 24 hours. After the reaction is completed, the methanol is removed and dried to obtain functionalized aminopolysiloxane. The inert gas is nitrogen, the amount of aminopolysiloxane used is 6wt% of methanol, and the functional agent is cinnamaldehyde and methacrylic acid glycidyl ether. Cinnamaldehyde and methacrylic acid glycidyl ether are used in a mass ratio of 1:5, and the amount of the functional agent used is 5wt% of the aminopolysiloxane.

[0094] Example 8: A method for preparing solid collagen

[0095] The difference between this embodiment and embodiment 4 lies in the preparation of functionalized aminopolysiloxane.

[0096] Preparation of functionalized aminopolysiloxane: In an inert gas atmosphere, aminopolysiloxane and a functional agent are added to methanol and mixed, and refluxed at 80°C for 24 hours. After the reaction is completed, the methanol is removed and dried to obtain functionalized aminopolysiloxane. The inert gas is nitrogen, the amount of aminopolysiloxane used is 6wt% of methanol, and the functional agent is cinnamaldehyde and methacrylic acid glycidyl ether. Cinnamaldehyde and methacrylic acid glycidyl ether are used in a mass ratio of 1:5, and the amount of the functional agent used is 8wt% of the aminopolysiloxane.

[0097] Example 9: A method for preparing liquid collagen

[0098] Preparation of collagen solution: Collagen is added to a dispersion system to prepare a collagen solution, i.e., liquid collagen. The dispersion system includes a phosphate buffer. The pH of the phosphate buffer is 6.5, and the molar concentration of the substance in the phosphate buffer is 0.01 mol / L. The concentration of the collagen solution is 35 mg / mL. The collagen is from Example 1.

[0099] The collagen in Example 9 can also be selected from collagen prepared by any method in Examples 2-8.

[0100] Comparative Example 1: A method for preparing solid collagen

[0101] The difference between this comparative example and Example 1 is that high-pressure homogenization treatment is not used in the preparation of the ultrafiltration collagen solution.

[0102] Comparative Example 2: A method for preparing solid collagen

[0103] The difference between this comparative example and Example 1 lies in the preparation of collagen.

[0104] Preparation of collagen: The pH of the ultrafiltered collagen solution was adjusted to 7.5, stirred for 6 hours, centrifuged, and the precipitate was collected to obtain collagen, i.e., solid collagen. During the centrifugation, the centrifugal speed was 10,000 rpm / min, and the centrifugal time was 30 minutes.

[0105] Comparative Example 3: A method for preparing solid collagen

[0106] The difference between this comparative example and Example 4 lies in the preparation of the crude collagen extract.

[0107] Preparation of crude collagen extract: Add protease to the acidic solution, add functionalized aminopolysiloxane and pretreated bovine Achilles tendon slices at a temperature of 5°C for enzymatic hydrolysis for 72 hours, centrifuge, and take the supernatant to obtain crude collagen extract. The acidic medium in the acidic solution is hydrochloric acid, and the concentration of the acidic medium in the acidic solution is 0.01 mol / L. When the concentration of the acidic medium meets the requirements, adjust the pH to 2.0; the protease is pepsin, and the usage ratio of the protease to the acidic solution is 1g:4L; the usage ratio of the pretreated bovine Achilles tendon slices to the acidic solution is 100g:4L, and the usage ratio of the functionalized aminopolysiloxane is 1wt% of the pretreated bovine Achilles tendon slices. During centrifugation, the centrifugal speed is 10000rpm / min, and the centrifugal time is 60min.

[0108] Comparative Example 4: A method for preparing solid collagen

[0109] The difference between this comparative example and Example 4 lies in the preparation of the crude collagen extract.

[0110] Preparation of crude collagen extract: Add protease to the acidic solution, add functionalized aminopolysiloxane and pretreated bovine Achilles tendon slices at a temperature of 5°C for enzymatic hydrolysis for 72 hours, centrifuge, and take the supernatant to obtain crude collagen extract. The acidic medium in the acidic solution is hydrochloric acid, and the concentration of the acidic medium in the acidic solution is 0.01 mol / L. When the concentration of the acidic medium meets the requirements, adjust the pH to 2.0; the protease is pepsin, and the usage ratio of the protease to the acidic solution is 1g:4L; the usage ratio of the pretreated bovine Achilles tendon slices to the acidic solution is 100g:4L, and the usage ratio of the functionalized aminopolysiloxane is 9wt% of the pretreated bovine Achilles tendon slices. During centrifugation, the centrifugal speed is 10000rpm / min, and the centrifugal time is 60min.

[0111] Comparative Example 5: A method for preparing solid collagen

[0112] The difference between this comparative example and Example 4 lies in the preparation of the functionalized aminopolysiloxane.

[0113] Preparation of functionalized aminopolysiloxane: In an inert gas atmosphere, aminopolysiloxane and a functional agent are added to methanol and mixed, and refluxed at 80°C for 24 hours. After the reaction is completed, the methanol is removed and dried to obtain functionalized aminopolysiloxane. The inert gas is nitrogen, the amount of aminopolysiloxane used is 6wt% of methanol, and the functional agent is cinnamaldehyde, and the amount of the functional agent used is 1wt% of the aminopolysiloxane.

[0114] Comparative Example 6: A method for preparing solid collagen

[0115] The difference between this comparative example and Example 4 lies in the preparation of the functionalized aminopolysiloxane.

[0116] Preparation of functionalized aminopolysiloxane: In an inert gas atmosphere, aminopolysiloxane and a functional agent are added to methanol and mixed, and refluxed at 80°C for 24 hours. After the reaction is completed, the methanol is removed and dried to obtain functionalized aminopolysiloxane. The inert gas is nitrogen, the amount of aminopolysiloxane used is 6wt% of methanol, and the functional agent is cinnamaldehyde, and the amount of the functional agent used is 11wt% of the aminopolysiloxane.

[0117] Comparative Example 7: A method for preparing solid collagen

[0118] The difference between this comparative example and Example 4 lies in the preparation of the functionalized aminopolysiloxane.

[0119] Preparation of functionalized aminopolysiloxane: In an inert gas atmosphere, aminopolysiloxane and a functional agent are added to methanol and mixed, and refluxed at 80°C for 24 hours. After the reaction is completed, the methanol is removed and dried to obtain functionalized aminopolysiloxane. The inert gas is nitrogen, the amount of aminopolysiloxane used is 6wt% of methanol, and the functional agent is glycidyl methacrylate, and the amount of the functional agent used is 5wt% of the aminopolysiloxane.

[0120] Comparative Example 8: A method for preparing solid collagen

[0121] The difference between this comparative example and Example 4 lies in the preparation of the functionalized aminopolysiloxane.

[0122] Preparation of functionalized aminopolysiloxane: In an inert gas atmosphere, aminopolysiloxane and a functional agent are added to methanol and mixed, and refluxed at 80°C for 24 hours. After the reaction is completed, the methanol is removed and dried to obtain functionalized aminopolysiloxane. The inert gas is nitrogen, the amount of aminopolysiloxane used is 6wt% of methanol, and the functional agent is glycidyl methacrylate, and the amount of the functional agent used is 8wt% of the aminopolysiloxane.

[0123] Test example:

[0124] 1. Infrared spectroscopy characterization

[0125] The present invention performs infrared characterization on the functionalized aminopolysiloxane prepared in Example 4, and the results are as follows: Figure 1 As shown, at 1648cm -1 The absorption peak of carbon-nitrogen double bond is at 1591cm -1 and 1447cm -1 The absorption peak of the benzene ring is at 1264 cm -1 The absorption peak of silicon-carbon bond is at 996 cm -1 The absorption peak at is the silicon-oxygen-silicon bond, indicating that functionalized aminopolysiloxane is obtained.

[0126] 2. Collagen purity

[0127] The present invention tested the purity of the solid collagen prepared in each embodiment and comparative example. The test results are as follows: Figure 2 As shown, S1 is Example 1, S2 is Example 2, S3 is Example 3, S4 is Example 4, S5 is Example 5, S6 is Example 6, S7 is Example 7, S8 is Example 8, D1 is Comparative Example 1, D2 is Comparative Example 2, D3 is Comparative Example 3, D4 is Comparative Example 4, D5 is Comparative Example 5, D6 is Comparative Example 6, D7 is Comparative Example 7, and D8 is Comparative Example 8. By comparing the results, it is found that the purity of the solid collagen prepared by the method of the present invention is relatively high, indicating that the method of the present invention can effectively retain the components of collagen and avoid the retention of non-collagen components. Functionalized aminopolysiloxane is used in the present invention, and the purity of the obtained collagen is also relatively high, indicating that less non-collagen components are retained, and the use of functionalized aminopolysiloxane will not increase the non-collagen components in the collagen product.

[0128] 3. Collagen concentration

[0129] The present invention tests the concentration of collagen in the ultrafiltered collagen solution in each embodiment and comparative example, and calculates the collagen concentration by the ratio of the content of the most solid collagen to the volume of the initial ultrafiltered collagen solution. The collagen concentration in this experiment represents the extraction effect of the method of the present invention on collagen in bovine Achilles tendon. The volume of the initial ultrafiltered collagen solution in the present invention is basically the same. The test results are as follows Figure 3As shown, S1 is Example 1, S2 is Example 2, S3 is Example 3, S4 is Example 4, S5 is Example 5, S6 is Example 6, S7 is Example 7, S8 is Example 8, D1 is Comparative Example 1, D2 is Comparative Example 2, D3 is Comparative Example 3, D4 is Comparative Example 4, D5 is Comparative Example 5, D6 is Comparative Example 6, D7 is Comparative Example 7, and D8 is Comparative Example 8. The present invention is to extract collagen from bovine Achilles tendon, mainly by using enzymatic treatment, high-pressure homogenization and ultrafiltration treatment, and finally by adjusting the ionic strength of the solution to separate the collagen, wherein the enzymatic treatment is the key step in extracting collagen from the bovine Achilles tendon into the solution, and only by separating more collagen from the bovine Achilles tendon and entering the solution can the content of the collagen finally prepared be improved. amount, and also ensure the structure of collagen so that collagen has high activity retention. After exploration and research, the present invention finds that by adjusting the high-pressure homogenization treatment or adjusting the ionic strength, more collagen extracted to a certain amount after enzymatic hydrolysis can be precipitated. However, the high-pressure homogenization treatment or the adjustment of the ionic strength are both after the enzymatic hydrolysis step. Therefore, the high-pressure homogenization treatment or the adjustment of the ionic strength can precipitate the collagen that cannot be precipitated under ordinary treatment, and obtain a high collagen yield, which is manifested as a high collagen concentration. Further, the present invention finds that functionalized aminopolysiloxane can be added in the enzymatic hydrolysis step. After the enzymatic hydrolysis treatment, the content of collagen in the solution after the enzymatic hydrolysis treatment can be increased, and then in the subsequent separation treatment step, a higher collagen concentration can be prepared, that is, a higher yield. In the present invention, aminopolysiloxane can be functionalized by using cinnamaldehyde and / or glycidyl methacrylate. It is found that the concentration of collagen prepared by aminopolysiloxane functionalized by cinnamaldehyde is higher. When the amount of the functionalized aminopolysiloxane is within a certain range, it has the effect of increasing the yield of collagen when applied to enzymatic hydrolysis. If the amount is too high or too low, the effect is not good. Furthermore, in the preparation of functionalized aminopolysiloxane, if the amount of cinnamaldehyde is too high or too low, the use effect of the obtained functionalized aminopolysiloxane will also be poor. When cinnamaldehyde is used, glycidyl methacrylate can be further used to functionalize the aminopolysiloxane. The aminopolysiloxane functionalized by cinnamaldehyde and glycidyl methacrylate can better increase the yield of collagen.

[0130] The present invention selects representative collagen prepared in Example 1, Example 2, Example 3, Example 4 and Example 6 for electrophoresis test. The electrophoresis diagram is as follows: Figure 4As shown, M is a protein molecular weight standard, 1 is a collagen reference, 2 is the collagen prepared in Example 1, 3 is the collagen prepared in Example 2, 4 is the collagen prepared in Example 3, 5 is the collagen prepared in Example 4, and 6 is the collagen prepared in Example 1. Figure 1 That is, the collagen extracted in the present invention maintains the original triple helical active structure.

[0131] The embodiments and / or implementation methods described above are only used to illustrate the preferred embodiments and / or implementation methods for realizing the technology of the present invention, and are not intended to limit the implementation methods of the technology of the present invention in any form. Any technical personnel in this field may make slight changes or modifications to other equivalent embodiments without departing from the scope of the technical means disclosed in the content of the present invention, but they should still be regarded as technologies or embodiments that are essentially the same as the present invention.

[0132] This article uses specific examples to illustrate the principles and implementation methods of this application. The description of the above embodiments is only used to help understand the method and its core ideas of this application. The above is only the preferred implementation method of this application. It should be pointed out that due to the limitations of textual expression and the objective existence of infinite specific structures, ordinary technicians in this technical field can make several improvements, modifications or changes without departing from the principles of this application, and can also combine the above technical features in an appropriate manner; these improvements, modifications, changes or combinations, or the direct application of the concept and technical solution of the invention to other occasions without improvement, should be regarded as the scope of protection of this application.

Claims

1. A method for preparing a collagen product, comprising preparing a collagen product from a biomass raw material through an extraction process, wherein the biomass raw material is firstly used to prepare a collagen crude extract, and then the collagen crude extract is subjected to high-pressure homogenization and ultrafiltration to prepare an ultrafiltered collagen solution, and the ultrafiltered collagen solution is subjected to ionic strength adjustment and separation to obtain a collagen product; The preparation of the crude collagen extract is as follows: enzymatically hydrolyzing the biomass raw material with a protease under acidic conditions; the protease is pepsin; an extraction aid is added during the enzymatic hydrolysis, and the extraction aid is a functionalized aminopolysiloxane; the amount of the extraction aid used is 5-8wt% of the biomass raw material; The functionalized aminopolysiloxane is prepared by adding the aminopolysiloxane and the functional agent into methanol at 80° C. and in an inert gas atmosphere, mixing, removing the methanol, and drying to obtain the functionalized aminopolysiloxane; the amount of the aminopolysiloxane used is 6wt% of the methanol; the functional agent is cinnamaldehyde, or a functional agent composited with cinnamaldehyde and glycidyl methacrylate in a mass ratio of 1:2-5, and the amount of the functional agent used is 5wt% of the aminopolysiloxane; The preparation of the aminopolysiloxane comprises the following steps: mixing γ-aminopropylmethyldiethoxysilane and diethyldiethoxysilane with water, then adding an alkaline agent and DMSO, stirring and mixing, reacting and polymerizing at 50° C., performing reduced pressure distillation, then adding a capping agent hexamethyldisiloxane for reaction, and adjusting the pH to neutral to obtain the aminopolysiloxane; the amount of γ-aminopropylmethyldiethoxysilane used is 150wt% of water; based on γ-aminopropylmethyldiethoxysilane, the amount of diethyldiethoxysilane used is 200wt%, the amount of the alkaline agent used is 0.2wt%; the amount of DMSO used is 1wt%; and the amount of hexamethyldisiloxane used is 10wt%.

2. The method for preparing a collagen product according to claim 1, characterized in that: In the high-pressure homogenization, the homogenization pressure is 100-400 bar, and the feed rate is 10-150 L / h; or an acidic reagent and an alkaline solution are used in adjusting the ionic strength; or the biomass raw material is bovine Achilles tendon.

3. The method for preparing a collagen product according to claim 2, characterized in that: The acidic reagent is at least one of acetic acid, hydrochloric acid, citric acid and phosphate; or the usage ratio of the acidic reagent to the ultrafiltration collagen solution is 0.1-0.5 mol / L; or the alkaline medium in the alkaline solution is at least one of sodium hydroxide, potassium hydroxide and sodium bicarbonate; Or the alkaline solution contains 5-20wt% alkaline medium.

4. The method for preparing a collagen product according to claim 1, characterized in that: The collagen product is solid collagen or liquid collagen.

5. The method for preparing a collagen product according to claim 4, characterized in that: The liquid collagen is solid collagen dispersed in a dispersion system, and the dispersion system includes at least one of a phosphate buffer, a sodium hyaluronate system, a hydroxyapatite system and a silicone gel system.

6. The method for preparing a collagen product according to claim 5, characterized in that: The solvent of the dispersion system is a medical solvent, and the medical solvent is water or alcohol.

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