CN112891609A - Porous material taking sheepskin as raw material and preparation method and application thereof - Google Patents
Porous material taking sheepskin as raw material and preparation method and application thereof Download PDFInfo
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- 239000002994 raw material Substances 0.000 title claims abstract description 45
- 239000011148 porous material Substances 0.000 title claims abstract description 43
- 238000002360 preparation method Methods 0.000 title claims abstract description 13
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims abstract description 86
- 238000005185 salting out Methods 0.000 claims abstract description 47
- 102000008186 Collagen Human genes 0.000 claims abstract description 45
- 108010035532 Collagen Proteins 0.000 claims abstract description 45
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims abstract description 45
- 229920001436 collagen Polymers 0.000 claims abstract description 45
- 239000011780 sodium chloride Substances 0.000 claims abstract description 43
- 239000000243 solution Substances 0.000 claims abstract description 39
- 238000005238 degreasing Methods 0.000 claims abstract description 37
- 239000000047 product Substances 0.000 claims abstract description 25
- 238000002791 soaking Methods 0.000 claims abstract description 22
- 238000001291 vacuum drying Methods 0.000 claims abstract description 22
- 102000057297 Pepsin A Human genes 0.000 claims abstract description 16
- 108090000284 Pepsin A Proteins 0.000 claims abstract description 16
- 229940111202 pepsin Drugs 0.000 claims abstract description 16
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 14
- 238000003756 stirring Methods 0.000 claims abstract description 11
- 238000004132 cross linking Methods 0.000 claims abstract description 10
- 239000000706 filtrate Substances 0.000 claims abstract description 10
- 239000003292 glue Substances 0.000 claims abstract description 10
- 238000004140 cleaning Methods 0.000 claims abstract description 9
- 238000007710 freezing Methods 0.000 claims abstract description 9
- 230000008014 freezing Effects 0.000 claims abstract description 9
- 239000007864 aqueous solution Substances 0.000 claims abstract description 8
- 239000011248 coating agent Substances 0.000 claims abstract description 8
- 238000000576 coating method Methods 0.000 claims abstract description 8
- 238000005520 cutting process Methods 0.000 claims abstract description 8
- 238000001914 filtration Methods 0.000 claims abstract description 8
- 239000003755 preservative agent Substances 0.000 claims abstract description 8
- 230000002335 preservative effect Effects 0.000 claims abstract description 8
- 210000004003 subcutaneous fat Anatomy 0.000 claims abstract description 8
- 238000005406 washing Methods 0.000 claims abstract description 7
- 239000003431 cross linking reagent Substances 0.000 claims abstract description 6
- 241001494479 Pecora Species 0.000 claims description 31
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 24
- 238000000034 method Methods 0.000 claims description 17
- 238000010438 heat treatment Methods 0.000 claims description 14
- 239000002244 precipitate Substances 0.000 claims description 14
- 239000000463 material Substances 0.000 claims description 12
- AZKVWQKMDGGDSV-BCMRRPTOSA-N Genipin Chemical group COC(=O)C1=CO[C@@H](O)[C@@H]2C(CO)=CC[C@H]12 AZKVWQKMDGGDSV-BCMRRPTOSA-N 0.000 claims description 10
- 238000001816 cooling Methods 0.000 claims description 10
- AZKVWQKMDGGDSV-UHFFFAOYSA-N genipin Natural products COC(=O)C1=COC(O)C2C(CO)=CCC12 AZKVWQKMDGGDSV-UHFFFAOYSA-N 0.000 claims description 10
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims description 8
- JMMWKPVZQRWMSS-UHFFFAOYSA-N isopropanol acetate Natural products CC(C)OC(C)=O JMMWKPVZQRWMSS-UHFFFAOYSA-N 0.000 claims description 8
- 238000000605 extraction Methods 0.000 claims description 7
- 238000000746 purification Methods 0.000 claims description 7
- 238000002156 mixing Methods 0.000 claims description 6
- 239000006228 supernatant Substances 0.000 claims description 4
- 230000002378 acidificating effect Effects 0.000 abstract 1
- 210000003491 skin Anatomy 0.000 description 29
- 239000007788 liquid Substances 0.000 description 18
- 238000010521 absorption reaction Methods 0.000 description 7
- 239000000203 mixture Substances 0.000 description 5
- 230000014759 maintenance of location Effects 0.000 description 4
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 238000004108 freeze drying Methods 0.000 description 2
- 229920005615 natural polymer Polymers 0.000 description 2
- 239000002861 polymer material Substances 0.000 description 2
- 239000002699 waste material Substances 0.000 description 2
- OFNXOACBUMGOPC-HZYVHMACSA-N 5'-hydroxystreptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](CO)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O OFNXOACBUMGOPC-HZYVHMACSA-N 0.000 description 1
- 102000009027 Albumins Human genes 0.000 description 1
- 108010088751 Albumins Proteins 0.000 description 1
- 239000002028 Biomass Substances 0.000 description 1
- 241000282817 Bovidae Species 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- 102000016942 Elastin Human genes 0.000 description 1
- 108010014258 Elastin Proteins 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 102000006395 Globulins Human genes 0.000 description 1
- 108010044091 Globulins Proteins 0.000 description 1
- 108010076876 Keratins Proteins 0.000 description 1
- 102000011782 Keratins Human genes 0.000 description 1
- 102000015728 Mucins Human genes 0.000 description 1
- 108010063954 Mucins Proteins 0.000 description 1
- 108010081750 Reticulin Proteins 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 230000002500 effect on skin Effects 0.000 description 1
- 229920002549 elastin Polymers 0.000 description 1
- 210000002615 epidermis Anatomy 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- OFNXOACBUMGOPC-UHFFFAOYSA-N hydroxystreptomycin Natural products CNC1C(O)C(O)C(CO)OC1OC1C(C=O)(O)C(CO)OC1OC1C(N=C(N)N)C(O)C(N=C(N)N)C(O)C1O OFNXOACBUMGOPC-UHFFFAOYSA-N 0.000 description 1
- OKPOKMCPHKVCPP-UHFFFAOYSA-N isoorientaline Natural products C1=C(O)C(OC)=CC(CC2C3=CC(OC)=C(O)C=C3CCN2C)=C1 OKPOKMCPHKVCPP-UHFFFAOYSA-N 0.000 description 1
- 239000012046 mixed solvent Substances 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- JTQHYPFKHZLTSH-UHFFFAOYSA-N reticulin Natural products COC1CC(OC2C(CO)OC(OC3C(O)CC(OC4C(C)OC(CC4OC)OC5CCC6(C)C7CCC8(C)C(CCC8(O)C7CC=C6C5)C(C)O)OC3C)C(O)C2OC)OC(C)C1O JTQHYPFKHZLTSH-UHFFFAOYSA-N 0.000 description 1
- 238000010583 slow cooling Methods 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 229920001059 synthetic polymer Polymers 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
- A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
- A61L15/42—Use of materials characterised by their function or physical properties
- A61L15/425—Porous materials, e.g. foams or sponges
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
- A61L15/00—Chemical aspects of, or use of materials for, bandages, dressings or absorbent pads
- A61L15/16—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons
- A61L15/22—Bandages, dressings or absorbent pads for physiological fluids such as urine or blood, e.g. sanitary towels, tampons containing macromolecular materials
- A61L15/32—Proteins, polypeptides; Degradation products or derivatives thereof, e.g. albumin, collagen, fibrin, gelatin
- A61L15/325—Collagen
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/78—Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG]
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
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- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Animal Behavior & Ethology (AREA)
- Hematology (AREA)
- Public Health (AREA)
- Wood Science & Technology (AREA)
- Veterinary Medicine (AREA)
- Biochemistry (AREA)
- Epidemiology (AREA)
- Materials Engineering (AREA)
- Molecular Biology (AREA)
- Genetics & Genomics (AREA)
- General Chemical & Material Sciences (AREA)
- Dispersion Chemistry (AREA)
- General Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Microbiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biotechnology (AREA)
- Toxicology (AREA)
- Gastroenterology & Hepatology (AREA)
- Biophysics (AREA)
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- Proteomics, Peptides & Aminoacids (AREA)
- Materials For Medical Uses (AREA)
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Abstract
The invention discloses a porous material taking sheepskin as a raw material. The invention discloses a preparation method of a porous material taking sheepskin as a raw material, which comprises the following steps: degreasing treatment: cleaning fresh sheepskin, removing hair by a scraper, removing subcutaneous fat, cutting into blocks, soaking in a sodium chloride aqueous solution, draining, soaking in a degreasing solution, and washing to obtain a degreasing product; extracting and purifying collagen: adding water into the degreased product, adjusting to an acidic environment, adding pepsin for enzymolysis, filtering, adding powdery sodium chloride into the filtrate, stirring uniformly, and salting out to obtain a collagen extract; plasticizing: adding glycerol into the collagen extract, uniformly stirring, standing, coating with a preservative film, pre-freezing at a low temperature of-55 to-65 ℃ for 2.5 to 3.5 hours, and performing vacuum drying at a low temperature to obtain a porous sheepskin glue raw material; and (3) crosslinking: adding the porous sheepskin glue raw material into a cross-linking agent, and preserving heat for 22-26h at the temperature of 30-36 ℃ to obtain the porous material taking sheepskin as the raw material.
Description
Technical Field
The invention relates to the technical field of biological medicines, in particular to a porous material taking sheep skin as a raw material, and a preparation method and application thereof.
Background
With the continuous development of society, people increasingly demand environment-friendly materials. In recent years, natural polymer materials derived from renewable and degradable have received much attention. Every year, about 100 ten thousand tons of waste sheepskins are produced in China, and most of the waste sheepskins are buried but not fully utilized.
The sheepskin is skin of goat or sheep of the family Bovidae, the sheepskin contains water, protein, fat and inorganic substances, the protein forming epidermal layer is mainly keratin, the protein forming dermal layer is mainly collagen and reticulin, and in addition, the skin contains elastin, albumin, globulin and mucin.
In natural and synthetic polymer materials, the sheep skin has less fat content in the skin layer, full fiber tissue, clear pores, elastic cortex, firmness and durability, and at present, the research on preparing porous materials by using sheep skin is few.
Disclosure of Invention
Based on the technical problems in the background art, the invention provides a porous material taking sheepskin as a raw material, and a preparation method and application thereof.
A preparation method of a porous material taking sheepskin as a raw material comprises the following steps:
s1, degreasing treatment: cleaning fresh sheepskin, removing hair by a scraper, removing subcutaneous fat, cutting into blocks, soaking in a sodium chloride aqueous solution, draining, soaking in a degreasing solution, and washing to obtain a degreasing product;
s2, collagen extraction and purification: adding water into the defatted product, adjusting pH to 2.0-3.0, adding pepsin for enzymolysis, filtering, adding powdered sodium chloride into the filtrate, stirring, and salting out to obtain collagen extract;
s3, plasticizing: adding glycerol into the collagen extract, stirring uniformly, standing for 6-10h for defoaming, coating with a preservative film until no bubbles exist on the surface, pre-freezing at the low temperature of-55 to-65 ℃ for 2.5-3.5h, and drying in vacuum at low temperature to obtain a porous sheepskin gelatin raw material;
s4, crosslinking: adding the porous sheepskin glue raw material into a cross-linking agent, and preserving heat for 22-26h at the temperature of 30-36 ℃ to obtain the porous material taking sheepskin as the raw material.
Preferably, in S1, the mass fraction of the sodium chloride aqueous solution is 0.8-1.2%, and the soaking time in the sodium chloride aqueous solution is 10-14 h.
Preferably, in S1, the degreasing solution is prepared from isopropanol and ethyl acetate in a volume ratio of 1.5-2.5: 1, the volume ratio of the drained sheep skin blocks to the degreasing solution is 1: 3.5-4.5, the soaking time in the degreasing solution is 4.5-5.5h, and the soaking temperature in the degreasing solution is 28-32 ℃.
Preferably, in S2, the mass ratio of pepsin to defatted product is 0.5-1.5: 100.
preferably, in S2, the enzymolysis time is 70-74h, and the enzymolysis temperature is 28-32 ℃.
Preferably, in S2, the specific operation of salting out is as follows: adding powdered sodium chloride into the filtrate until the concentration of the sodium chloride is 43-45mol/L for primary salting out, then taking the upper layer of flocculent precipitate, centrifuging, discarding the supernatant, re-dissolving, then adding powdered sodium chloride again until the concentration of the sodium chloride is 1.5-2.0mol/L for secondary salting out, taking the upper layer of flocculent precipitate, centrifuging, discarding the supernatant, and thus obtaining a collagen extract; the temperature of the primary salting-out and the secondary salting-out is 33-37 ℃, and the time of the primary salting-out and the secondary salting-out is 22-26 h.
Preferably, in S3, the volume ratio of the collagen extract to the glycerol is 100: 0.1-0.3.
Preferably, in S3, the low-temperature vacuum drying specifically operates as follows: the vacuum degree is always maintained at 150-200Pa in the low-temperature vacuum drying process; heating the pre-frozen material to-10-20 ℃ at the speed of 6-10 ℃/h, and preserving heat for 1-2 h; then cooling to-30-40 ℃ at the speed of 1-2 ℃/h, and preserving heat for 1-2 h; then heating to-5-8 ℃ at the speed of 6-10 ℃/h, and preserving heat for 1-2 h; then the temperature is reduced to-20 to-30 ℃ at the speed of 1-2 ℃/h, and the temperature is preserved for 10-20 h.
Preferably, in S4, the cross-linking agent is genipin, and preferably a genipin solution with the mass fraction of 0.05-0.75%.
A porous material taking sheepskin as a raw material is prepared by adopting the preparation method of the porous material taking sheepskin as the raw material.
The porous material taking the sheepskin as the raw material is applied as the wound dressing.
The method takes the sheepskin as a raw material, firstly, the sheepskin is degreased, and according to the similar compatibility principle, mixed solvent of isopropanol and ethyl acetate is adopted to degrease sheepskin particles; then, performing enzymolysis on the sheepskin by adopting a method combining acetic acid and pepsin to obtain a collagen extract with a natural triple helical structure and biological activity; then pre-freezing, then freeze-drying the extract under the conditions of low temperature and vacuum, and performing low-temperature vacuum drying on the pre-frozen collagen product, particularly controlling the rapid heating speed to be matched with the slow cooling speed, so that the structure of the collagen can be uniformly destroyed, the solvent molecules are separated, the product is in a uniform flocculent structure, compared with the common freeze-drying, the structure surface is smooth and flat, and the porous material with a three-dimensional porous structure is prepared; and finally, crosslinking the obtained sheepskin porous glue raw material, wherein the pore size formed by crosslinking is uniform in distribution and extremely good in moisture absorption state.
The collagen porous material prepared by taking sheepskin as a raw material and genipin as a cross-linking agent is a biomass material which is used as a natural high molecular material, and the macromolecular chain of the collagen porous material is rich in-OH and-NH2And hydrophilic groups such as-COOH and the like, has good moisture absorption performance, and has good prospect when being applied to medical wound dressings.
Detailed Description
The technical solution of the present invention will be described in detail below with reference to specific examples.
Example 1
A preparation method of a porous material taking sheepskin as a raw material comprises the following steps:
s1, degreasing treatment: cleaning fresh sheep skin, removing hair by a scraper, removing subcutaneous fat, cutting into blocks of 2mm × 2mm, soaking in 0.8% sodium chloride water solution for 14h, taking out, draining, soaking in degreasing solution for 4.5h at 32 deg.C, and washing to obtain degreased product;
the volume ratio of the drained sheep skin blocks to the degreasing solution is 1: 3.5, the degreasing solution is prepared by mixing isopropanol and ethyl acetate in a volume ratio of 2.5: 1, preparing a composition;
s2, collagen extraction and purification: adding water into the degreased product, adjusting the pH value of the system to 2.0, adding pepsin for enzymolysis for 70 hours, wherein the enzymolysis temperature is 32 ℃, and the mass ratio of the pepsin to the degreased product is 0.7: 100, filtering, adding powdered sodium chloride into the filtrate until the concentration of the sodium chloride is 45mol/L for primary salting out, then taking the upper layer of flocculent precipitate, centrifuging, discarding the upper layer of clear liquid, re-dissolving, then adding the powdered sodium chloride again until the concentration of the sodium chloride is 1.5mol/L for secondary salting out, taking the upper layer of flocculent precipitate, centrifuging, discarding the upper layer of clear liquid, and obtaining a collagen extract;
the temperature of the primary salting-out and the secondary salting-out is 37 ℃, and the time of the primary salting-out and the secondary salting-out is 22 hours;
s3, plasticizing: adding glycerol into the collagen extract, and uniformly stirring, wherein the volume ratio of the collagen extract to the glycerol is 100: 0.25, standing for 6h for defoaming, coating the surface of the sheep skin by using a preservative film until no bubbles exist on the surface, pre-freezing the sheep skin at the low temperature of-65 ℃ for 2.5h, and performing vacuum drying at the low temperature to obtain a porous sheep skin glue raw material;
the low-temperature vacuum drying operation is as follows: the vacuum degree is always maintained to be 200Pa in the low-temperature vacuum drying process; heating the pre-frozen material to-20 ℃ at the speed of 6 ℃/h, and preserving heat for 1 h; then cooling to-30 ℃ at the speed of 2 ℃/h, and preserving heat for 2 h; then heating to-8 ℃ at the speed of 6 ℃/h, and preserving heat for 1 h; then cooling to-20 ℃ at the speed of 2 ℃/h, and preserving heat for 20 h;
s4, crosslinking: adding the porous collagen material of the sheepskin into 0.015 percent genipin solution by mass fraction, and keeping the temperature at 36 ℃ for 22 hours to obtain the porous material taking the sheepskin as the raw material.
Example 2
A preparation method of a porous material taking sheepskin as a raw material comprises the following steps:
s1, degreasing treatment: cleaning fresh sheep skin, removing hair by a scraper, removing subcutaneous fat, cutting into blocks of 2mm × 2mm, soaking in 1.2 wt% sodium chloride water solution for 10h, taking out, draining, soaking in degreasing solution for 5.5h at 28 deg.C, and washing to obtain degreased product;
the volume ratio of the drained sheep skin blocks to the degreasing solution is 1: 4.5, the degreasing solution is prepared by mixing isopropanol and ethyl acetate according to a volume ratio of 1.5: 1, preparing a composition;
s2, collagen extraction and purification: adding water into the degreased product, adjusting the pH value of the system to 3.0, adding pepsin for enzymolysis for 74 hours, wherein the enzymolysis temperature is 28 ℃, and the mass ratio of the pepsin to the degreased product is 1.3: 100, filtering, adding powdered sodium chloride into the filtrate until the concentration of the sodium chloride is 43mol/L, performing primary salting-out, then taking the upper-layer flocculent precipitate, centrifuging, discarding the upper-layer clear liquid, re-dissolving, then adding powdered sodium chloride again until the concentration of the sodium chloride is 2.0mol/L, performing secondary salting-out, taking the upper-layer flocculent precipitate, centrifuging, discarding the upper-layer clear liquid, and obtaining a collagen extract;
the temperature of the primary salting-out and the secondary salting-out is 33 ℃, and the time of the primary salting-out and the secondary salting-out is 26 hours;
s3, plasticizing: adding glycerol into the collagen extract, and uniformly stirring, wherein the volume ratio of the collagen extract to the glycerol is 100: 0.15, standing for 10h for defoaming treatment, coating the surface of the sheep skin with a preservative film until no bubbles exist, pre-freezing at the low temperature of-55 ℃ for 3.5h, and performing vacuum drying at the low temperature to obtain a porous sheep skin glue raw material;
the low-temperature vacuum drying operation is as follows: the vacuum degree is always maintained to be 150Pa in the low-temperature vacuum drying process; heating the pre-frozen material to-10 ℃ at the speed of 10 ℃/h, and preserving heat for 2 h; then cooling to-40 ℃ at the speed of 1 ℃/h, and preserving heat for 1 h; then heating to-5 ℃ at the speed of 10 ℃/h, and preserving heat for 2 h; then cooling to-30 ℃ at the speed of 1 ℃/h, and preserving heat for 10 h;
s4, crosslinking: adding the porous sheepskin collagen into 0.50 mass percent genipin solution, and keeping the temperature at 33 ℃ for 26h to obtain the porous material taking sheepskin as a raw material.
Example 3
A preparation method of a porous material taking sheepskin as a raw material comprises the following steps:
s1, degreasing treatment: cleaning fresh sheep skin, removing hair by a scraper, removing subcutaneous fat, cutting into blocks of 2mm × 2mm, soaking in 1% sodium chloride aqueous solution for 12h, taking out, draining, soaking in degreasing solution for 5h at 30 deg.C, and repeatedly cleaning with distilled water for 4 times to obtain degreased product;
the volume ratio of the drained sheep skin blocks to the degreasing solution is 1: 4, degreasing solution is prepared by mixing isopropanol and ethyl acetate according to a volume ratio of 2: 1, preparing a composition;
s2, collagen extraction and purification: adding water into the degreased product, adjusting the pH value of the system to 2.5, adding pepsin for enzymolysis for 72 hours, wherein the enzymolysis temperature is 30 ℃, and the mass ratio of the pepsin to the degreased product is 1.5: 100, filtering, adding powdered sodium chloride into the filtrate until the concentration of the sodium chloride is 44mol/L, performing primary salting-out, then taking the upper layer of flocculent precipitate, centrifuging, discarding the upper layer of clear liquid, re-dissolving, then adding powdered sodium chloride again until the concentration of the sodium chloride is 1.7mol/L, performing secondary salting-out, taking the upper layer of flocculent precipitate, centrifuging, discarding the upper layer of clear liquid, and obtaining a collagen extract;
the temperature of the primary salting-out and the secondary salting-out is 35 ℃, and the time of the primary salting-out and the secondary salting-out is 24 hours;
s3, plasticizing: adding glycerol into the collagen extract, and uniformly stirring, wherein the volume ratio of the collagen extract to the glycerol is 100: 0.2, standing for 8h for defoaming treatment, coating the surface of the sheep skin with a preservative film until no bubbles exist, pre-freezing at the low temperature of-60 ℃ for 3h, and performing vacuum drying at the low temperature to obtain a porous sheep skin glue raw material;
the low-temperature vacuum drying operation is as follows: the vacuum degree is always maintained to be 160Pa in the low-temperature vacuum drying process; heating the pre-frozen material to-12 ℃ at the speed of 9 ℃/h, and preserving heat for 1.8 h; then cooling to-37 ℃ at the speed of 1.3 ℃/h, and preserving heat for 1.2 h; then the temperature is raised to-6 ℃ at the speed of 9 ℃/h, and the temperature is kept for 1.7 h; then, cooling to-28 ℃ at the speed of 1.3 ℃/h, and preserving heat for 13 h;
s4, crosslinking: adding the porous sheepskin collagen into 0.25 mass percent genipin solution, and keeping the temperature at 30 ℃ for 24 hours to obtain the porous material taking sheepskin as a raw material.
The porous material sample obtained in this example was tested, and the liquid absorption rate was 18% and the liquid retention rate was 13%.
Example 4
A preparation method of a porous material taking sheepskin as a raw material comprises the following steps:
s1, degreasing treatment: cleaning fresh sheep skin, removing hair by a scraper, removing subcutaneous fat, cutting into blocks of 2mm × 2mm, soaking in 1% sodium chloride water solution for 12h, taking out, draining, soaking in degreasing solution for 5h at 30 deg.C, and washing to obtain degreased product;
the volume ratio of the drained sheep skin blocks to the degreasing solution is 1: 4, degreasing solution is prepared by mixing isopropanol and ethyl acetate according to a volume ratio of 2: 1, preparing a composition;
s2, collagen extraction and purification: adding water into the degreased product, adjusting the pH value of the system to 2.5, adding pepsin for enzymolysis for 72 hours, wherein the enzymolysis temperature is 30 ℃, and the mass ratio of the pepsin to the degreased product is 1: 100, filtering, adding powdered sodium chloride into the filtrate until the concentration of the sodium chloride is 44mol/L, performing primary salting-out, then taking the upper layer of flocculent precipitate, centrifuging, discarding the upper layer of clear liquid, re-dissolving, then adding powdered sodium chloride again until the concentration of the sodium chloride is 1.7mol/L, performing secondary salting-out, taking the upper layer of flocculent precipitate, centrifuging, discarding the upper layer of clear liquid, and obtaining a collagen extract;
the temperature of the primary salting-out and the secondary salting-out is 35 ℃, and the time of the primary salting-out and the secondary salting-out is 24 hours;
s3, plasticizing: adding glycerol into the collagen extract, and uniformly stirring, wherein the volume ratio of the collagen extract to the glycerol is 100: 0.1, standing for 8h for defoaming treatment, coating the surface of the sheep skin with a preservative film until no bubbles exist, pre-freezing at the low temperature of-60 ℃ for 3h, and performing vacuum drying at the low temperature to obtain a porous sheep skin glue raw material;
the low-temperature vacuum drying operation is as follows: the vacuum degree is always maintained to be 180Pa in the low-temperature vacuum drying process; heating the pre-frozen material to-18 ℃ at the speed of 7 ℃/h, and preserving heat for 1.2 h; then, the temperature is reduced to minus 33 ℃ at the speed of 1.7 ℃/h, and the temperature is preserved for 1.8 h; then heating to-7 ℃ at the speed of 7 ℃/h, and preserving heat for 1.3 h; then, the temperature is reduced to-22 ℃ at the speed of 1.7 ℃/h, and the temperature is kept for 17 h;
s4, crosslinking: adding the porous sheepskin collagen into 0.05 percent genipin solution by mass fraction, and keeping the temperature at 30 ℃ for 24 hours to obtain the porous material taking sheepskin as a raw material.
The porous material sample obtained in this example was tested, and the liquid absorption rate was 14% and the liquid retention rate was 7%.
Example 5
A preparation method of a porous material taking sheepskin as a raw material comprises the following steps:
s1, degreasing treatment: cleaning fresh sheep skin, removing hair by a scraper, removing subcutaneous fat, cutting into blocks of 2mm × 2mm, soaking in 1% sodium chloride water solution for 12h, taking out, draining, soaking in degreasing solution for 5h at 30 deg.C, and washing to obtain degreased product;
the volume ratio of the drained sheep skin blocks to the degreasing solution is 1: 4, degreasing solution is prepared by mixing isopropanol and ethyl acetate according to a volume ratio of 2: 1, preparing a composition;
s2, collagen extraction and purification: adding water into the degreased product, adjusting the pH value of the system to 2.5, adding pepsin for enzymolysis for 72 hours, wherein the enzymolysis temperature is 30 ℃, and the mass ratio of the pepsin to the degreased product is 0.5: 100, filtering, adding powdered sodium chloride into the filtrate until the concentration of the sodium chloride is 44mol/L, performing primary salting-out, then taking the upper layer of flocculent precipitate, centrifuging, discarding the upper layer of clear liquid, re-dissolving, then adding powdered sodium chloride again until the concentration of the sodium chloride is 1.7mol/L, performing secondary salting-out, taking the upper layer of flocculent precipitate, centrifuging, discarding the upper layer of clear liquid, and obtaining a collagen extract;
the temperature of the primary salting-out and the secondary salting-out is 35 ℃, and the time of the primary salting-out and the secondary salting-out is 24 hours;
s3, plasticizing: adding glycerol into the collagen extract, and uniformly stirring, wherein the volume ratio of the collagen extract to the glycerol is 100: 0.3, standing for 8h for defoaming treatment, coating the surface of the sheep skin with a preservative film until no bubbles exist on the surface, pre-freezing the sheep skin at the low temperature of-60 ℃ for 3h, and performing vacuum drying at the low temperature to obtain a porous sheep skin glue raw material;
the low-temperature vacuum drying operation is as follows: the vacuum degree is always maintained to be 170Pa in the low-temperature vacuum drying process; heating the pre-frozen material to-15 ℃ at the speed of 8 ℃/h, and preserving heat for 1.5 h; then cooling to-35 ℃ at the speed of 1.5 ℃/h, and preserving heat for 1.5 h; then heating to-6.5 ℃ at the speed of 8 ℃/h, and preserving the heat for 1.5 h; then, cooling to-25 ℃ at the speed of 1.5 ℃/h, and preserving heat for 15 h;
s4, crosslinking: adding the porous sheepskin collagen into 0.75% by mass of genipin solution, and keeping the temperature at 30 ℃ for 24h to obtain the porous material taking sheepskin as a raw material.
The porous material sample obtained in this example was tested, and the liquid absorption rate was 17% and the liquid retention rate was 15%.
The test results of the porous material obtained in the above embodiment prove that the liquid absorption rate of the invention is 14-18%, the liquid retention rate is 7-15%, the moisture absorption performance is good, the requirements of the field of wound dressing are met, and the application range of the sheepskin is expanded.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered to be within the technical scope of the present invention, and the technical solutions and the inventive concepts thereof according to the present invention should be equivalent or changed within the scope of the present invention.
Claims (10)
1. A preparation method of a porous material taking sheepskin as a raw material is characterized by comprising the following steps:
s1, degreasing treatment: cleaning fresh sheepskin, removing hair by a scraper, removing subcutaneous fat, cutting into blocks, soaking in a sodium chloride aqueous solution, draining, soaking in a degreasing solution, and washing to obtain a degreasing product;
s2, collagen extraction and purification: adding water into the defatted product, adjusting pH to 2.0-3.0, adding pepsin for enzymolysis, filtering, adding powdered sodium chloride into the filtrate, stirring, and salting out to obtain collagen extract;
s3, plasticizing: adding glycerol into the collagen extract, uniformly stirring, standing for 6-10h for defoaming, coating with a preservative film, pre-freezing at the low temperature of-55 to-65 ℃ for 2.5-3.5h, and performing vacuum drying at the low temperature to obtain a porous sheepskin collagen raw material;
s4, crosslinking: adding the porous sheepskin glue raw material into a cross-linking agent, and preserving heat for 22-26h at the temperature of 30-36 ℃ to obtain the porous material taking sheepskin as the raw material.
2. The method for preparing a porous material by using sheepskin as a raw material according to claim 1, wherein in S1, the mass fraction of the sodium chloride aqueous solution is 0.8-1.2%, and the soaking time in the sodium chloride aqueous solution is 10-14 h.
3. The method for preparing a porous material by using sheepskin as a raw material according to claim 1, wherein in S1, the degreasing solution is prepared by mixing isopropanol and ethyl acetate in a volume ratio of 1.5-2.5: 1, the volume ratio of the drained sheep skin blocks to the degreasing solution is 1: 3.5-4.5, the soaking time in the degreasing solution is 4.5-5.5h, and the soaking temperature in the degreasing solution is 28-32 ℃.
4. The method for preparing a porous material by using sheep skin as a raw material according to claim 1, wherein in S2, the mass ratio of pepsin to defatted product is 0.5-1.5: 100, respectively; preferably, the enzymolysis time is 70-74h, and the enzymolysis temperature is 28-32 ℃.
5. The method for preparing a porous material by using sheepskin as a raw material according to claim 1, wherein the salting out is performed in S2 by the following steps: adding powdered sodium chloride into the filtrate until the concentration of the sodium chloride is 43-45mol/L for primary salting out, then taking the upper layer of flocculent precipitate, centrifuging, discarding the supernatant, re-dissolving, then adding powdered sodium chloride again until the concentration of the sodium chloride is 1.5-2.0mol/L for secondary salting out, taking the upper layer of flocculent precipitate, centrifuging, discarding the supernatant, and thus obtaining a collagen extract; the temperature of the primary salting-out and the secondary salting-out is 33-37 ℃, and the time of the primary salting-out and the secondary salting-out is 22-26 h.
6. The method for preparing a porous material using sheepskin as a raw material according to claim 1, wherein in S3, the volume ratio of collagen extract to glycerol is 100: 0.1-0.3.
7. The method for preparing a porous material by using sheepskin as a raw material according to claim 1, wherein in S3, the low-temperature vacuum drying is specifically performed as follows: the vacuum degree is always maintained at 150-200Pa in the low-temperature vacuum drying process; heating the pre-frozen material to-10-20 ℃ at the speed of 6-10 ℃/h, and preserving heat for 1-2 h; then cooling to-30-40 ℃ at the speed of 1-2 ℃/h, and preserving heat for 1-2 h; then heating to-5-8 ℃ at the speed of 6-10 ℃/h, and preserving heat for 1-2 h; then the temperature is reduced to-20 to-30 ℃ at the speed of 1-2 ℃/h, and the temperature is preserved for 10-20 h.
8. The method for preparing the porous material by taking the sheepskin as the raw material according to claim 1, wherein in S4, the cross-linking agent is genipin, and is preferably genipin solution with the mass fraction of 0.05-0.75%.
9. A porous material made of sheep skin, which is characterized by being prepared by the method for preparing the porous material made of sheep skin according to any one of claims 1 to 9.
10. Use of the porous sheepskin-based material of claim 9 as a wound dressing.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113502355A (en) * | 2021-07-21 | 2021-10-15 | 四川大学 | Ultrathin multifunctional microfiltration membrane based on animal hides and preparation method and application |
| CN114225112A (en) * | 2021-11-22 | 2022-03-25 | 江苏苏伯纳生物科技有限公司 | Preparation method of cerebrospinal fluid isolation repairing film |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101695581A (en) * | 2009-10-26 | 2010-04-21 | 西北大学 | Method for preparing human-like collagen haemostatic sponge in scale |
| WO2011014154A1 (en) * | 2009-07-27 | 2011-02-03 | National Cheng Kung University | Method for preparing porous collagen matrices |
| CN106866816A (en) * | 2017-04-14 | 2017-06-20 | 桂林融通科技有限公司 | The acid-enzyme binding-method of collagen is extracted from pigskin |
| CN107475337A (en) * | 2017-08-18 | 2017-12-15 | 合肥丰洁生物科技有限公司 | A kind of method that collagen is extracted in the sebum from animal |
| WO2018143548A1 (en) * | 2017-01-31 | 2018-08-09 | 주식회사 마린테크노 | Method for extracting marine collagen from fish skin |
| CN110016232A (en) * | 2019-04-12 | 2019-07-16 | 江南大学 | A kind of fish scale collagen composite sponge preparation method |
-
2021
- 2021-01-15 CN CN202110056039.5A patent/CN112891609A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2011014154A1 (en) * | 2009-07-27 | 2011-02-03 | National Cheng Kung University | Method for preparing porous collagen matrices |
| CN101695581A (en) * | 2009-10-26 | 2010-04-21 | 西北大学 | Method for preparing human-like collagen haemostatic sponge in scale |
| WO2018143548A1 (en) * | 2017-01-31 | 2018-08-09 | 주식회사 마린테크노 | Method for extracting marine collagen from fish skin |
| CN106866816A (en) * | 2017-04-14 | 2017-06-20 | 桂林融通科技有限公司 | The acid-enzyme binding-method of collagen is extracted from pigskin |
| CN107475337A (en) * | 2017-08-18 | 2017-12-15 | 合肥丰洁生物科技有限公司 | A kind of method that collagen is extracted in the sebum from animal |
| CN110016232A (en) * | 2019-04-12 | 2019-07-16 | 江南大学 | A kind of fish scale collagen composite sponge preparation method |
Non-Patent Citations (1)
| Title |
|---|
| 千恩楠: "胶原蛋白的提取及其多孔材料的制备与性能研究", 《中国优秀硕士学位论文全文数据库 工程科技I辑》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113502355A (en) * | 2021-07-21 | 2021-10-15 | 四川大学 | Ultrathin multifunctional microfiltration membrane based on animal hides and preparation method and application |
| CN113502355B (en) * | 2021-07-21 | 2022-04-01 | 四川大学 | Ultrathin multifunctional microfiltration membrane based on animal hides and preparation method and application |
| CN114225112A (en) * | 2021-11-22 | 2022-03-25 | 江苏苏伯纳生物科技有限公司 | Preparation method of cerebrospinal fluid isolation repairing film |
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