CN115252881A - Human-like collagen composition and preparation method thereof - Google Patents
Human-like collagen composition and preparation method thereof Download PDFInfo
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- 102000008186 Collagen Human genes 0.000 title claims abstract description 62
- 108010035532 Collagen Proteins 0.000 title claims abstract description 62
- 229920001436 collagen Polymers 0.000 title claims abstract description 62
- 239000000203 mixture Substances 0.000 title claims abstract description 59
- 238000002360 preparation method Methods 0.000 title abstract description 7
- 239000000843 powder Substances 0.000 claims abstract description 60
- 108010010803 Gelatin Proteins 0.000 claims abstract description 45
- 229920000159 gelatin Polymers 0.000 claims abstract description 45
- 239000008273 gelatin Substances 0.000 claims abstract description 45
- 235000019322 gelatine Nutrition 0.000 claims abstract description 45
- 235000011852 gelatine desserts Nutrition 0.000 claims abstract description 45
- 241000251468 Actinopterygii Species 0.000 claims abstract description 40
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 21
- 238000004132 cross linking Methods 0.000 claims abstract description 16
- 229920001661 Chitosan Polymers 0.000 claims abstract description 15
- 239000003431 cross linking reagent Substances 0.000 claims abstract description 9
- 239000002994 raw material Substances 0.000 claims abstract description 8
- 229920002472 Starch Polymers 0.000 claims description 32
- 235000019698 starch Nutrition 0.000 claims description 29
- 239000008107 starch Substances 0.000 claims description 29
- 239000000243 solution Substances 0.000 claims description 25
- 102000004190 Enzymes Human genes 0.000 claims description 20
- 108090000790 Enzymes Proteins 0.000 claims description 20
- 229940088598 enzyme Drugs 0.000 claims description 20
- 238000003756 stirring Methods 0.000 claims description 16
- 239000007788 liquid Substances 0.000 claims description 15
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 12
- 108010073178 Glucan 1,4-alpha-Glucosidase Proteins 0.000 claims description 12
- 102100022624 Glucoamylase Human genes 0.000 claims description 12
- 102000004139 alpha-Amylases Human genes 0.000 claims description 12
- 108090000637 alpha-Amylases Proteins 0.000 claims description 12
- 229940024171 alpha-amylase Drugs 0.000 claims description 12
- 238000007710 freezing Methods 0.000 claims description 12
- 230000008014 freezing Effects 0.000 claims description 12
- 238000000874 microwave-assisted extraction Methods 0.000 claims description 12
- 238000005406 washing Methods 0.000 claims description 12
- 239000012153 distilled water Substances 0.000 claims description 11
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 8
- 238000006243 chemical reaction Methods 0.000 claims description 8
- 238000001035 drying Methods 0.000 claims description 8
- 239000000706 filtrate Substances 0.000 claims description 8
- 238000001914 filtration Methods 0.000 claims description 8
- 238000004108 freeze drying Methods 0.000 claims description 8
- 238000010438 heat treatment Methods 0.000 claims description 8
- 239000011259 mixed solution Substances 0.000 claims description 8
- 238000005507 spraying Methods 0.000 claims description 8
- 238000000034 method Methods 0.000 claims description 6
- 238000005520 cutting process Methods 0.000 claims description 5
- 239000000047 product Substances 0.000 claims description 5
- 239000007853 buffer solution Substances 0.000 claims description 4
- 238000007872 degassing Methods 0.000 claims description 4
- 238000005360 mashing Methods 0.000 claims description 4
- 235000013336 milk Nutrition 0.000 claims description 4
- 239000008267 milk Substances 0.000 claims description 4
- 210000004080 milk Anatomy 0.000 claims description 4
- 238000012545 processing Methods 0.000 claims description 4
- 239000007974 sodium acetate buffer Substances 0.000 claims description 4
- 239000011780 sodium chloride Substances 0.000 claims description 4
- UGTZMIPZNRIWHX-UHFFFAOYSA-K sodium trimetaphosphate Chemical compound [Na+].[Na+].[Na+].[O-]P1(=O)OP([O-])(=O)OP([O-])(=O)O1 UGTZMIPZNRIWHX-UHFFFAOYSA-K 0.000 claims description 4
- 239000006228 supernatant Substances 0.000 claims description 4
- 229920002261 Corn starch Polymers 0.000 claims description 3
- 239000008120 corn starch Substances 0.000 claims description 3
- 229920001592 potato starch Polymers 0.000 claims description 3
- 229940100445 wheat starch Drugs 0.000 claims description 3
- 230000002439 hemostatic effect Effects 0.000 abstract description 13
- 238000010521 absorption reaction Methods 0.000 abstract description 6
- 239000000463 material Substances 0.000 abstract description 4
- 239000011148 porous material Substances 0.000 abstract description 3
- 230000000740 bleeding effect Effects 0.000 description 4
- 239000004833 fish glue Substances 0.000 description 3
- 230000023597 hemostasis Effects 0.000 description 3
- 206010052428 Wound Diseases 0.000 description 2
- 208000027418 Wounds and injury Diseases 0.000 description 2
- 230000003796 beauty Effects 0.000 description 2
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- 241000588724 Escherichia coli Species 0.000 description 1
- AZKVWQKMDGGDSV-BCMRRPTOSA-N Genipin Chemical compound COC(=O)C1=CO[C@@H](O)[C@@H]2C(CO)=CC[C@H]12 AZKVWQKMDGGDSV-BCMRRPTOSA-N 0.000 description 1
- YQEZLKZALYSWHR-UHFFFAOYSA-N Ketamine Chemical compound C=1C=CC=C(Cl)C=1C1(NC)CCCCC1=O YQEZLKZALYSWHR-UHFFFAOYSA-N 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 239000002537 cosmetic Substances 0.000 description 1
- 238000001976 enzyme digestion Methods 0.000 description 1
- 210000002919 epithelial cell Anatomy 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- AZKVWQKMDGGDSV-UHFFFAOYSA-N genipin Natural products COC(=O)C1=COC(O)C2C(CO)=CCC12 AZKVWQKMDGGDSV-UHFFFAOYSA-N 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 239000007928 intraperitoneal injection Substances 0.000 description 1
- 229960003299 ketamine Drugs 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000011587 new zealand white rabbit Methods 0.000 description 1
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- 102000004169 proteins and genes Human genes 0.000 description 1
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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
- A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
- A61L24/04—Surgical adhesives or cements; Adhesives for colostomy devices containing macromolecular materials
- A61L24/10—Polypeptides; Proteins
- A61L24/102—Collagen
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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
- A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
- A61L24/0005—Ingredients of undetermined constitution or reaction products thereof
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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
- A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
- A61L24/001—Use of materials characterised by their function or physical properties
-
- 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
- A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
- A61L24/001—Use of materials characterised by their function or physical properties
- A61L24/0015—Medicaments; Biocides
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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
- A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
- A61L24/001—Use of materials characterised by their function or physical properties
- A61L24/0036—Porous materials, e.g. foams or sponges
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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
- A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
- A61L24/04—Surgical adhesives or cements; Adhesives for colostomy devices containing macromolecular materials
- A61L24/08—Polysaccharides
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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
- A61L24/00—Surgical adhesives or cements; Adhesives for colostomy devices
- A61L24/04—Surgical adhesives or cements; Adhesives for colostomy devices containing macromolecular materials
- A61L24/10—Polypeptides; Proteins
- A61L24/108—Specific proteins or polypeptides not covered by groups A61L24/102 - A61L24/106
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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
- A61L2400/00—Materials characterised by their function or physical properties
- A61L2400/04—Materials for stopping bleeding
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Abstract
The invention discloses a human-like collagen composition and a preparation method thereof, wherein the human-like collagen composition comprises the following raw materials: 5-15 parts by weight of human-like collagen; 10-20 parts by weight of a crosslinkable powder; 1-2 parts by weight of a cross-linking agent; 20-25 parts of fish gelatin extract and 50-80 parts of chitosan. According to the technical scheme, the human-like collagen-crosslinkable powder composition is obtained by crosslinking the human-like collagen and the crosslinkable powder, is a material with a good hemostatic effect, and then the fish gelatin extract, the human-like collagen-crosslinkable powder composition and the chitosan are prepared to obtain a composition which is spongy and has a uniform internal pore structure; the finally obtained human-like collagen composition has good structural stability, water absorption, comfort, human body compatibility and hemostatic effect.
Description
Technical Field
The invention relates to the technical field of collagen, in particular to a human-like collagen composition and a preparation method thereof.
Background
Human-like Collagen (Human-like Collagen) is a high molecular biological protein produced by reverse transcription of mRNA of Human Collagen into cDNA, enzyme digestion, recombination of a section of gene in E.coli, high-density fermentation, separation, renaturation and purification. Due to inherent biocompatibility, biodegradability and absorbability, and the functions of promoting formation of new cells and epithelial cells, the potential application of collagen in the medical field, the beauty treatment, the cosmetics and the health care product industry is continuously widened.
In the prior art, human-like collagen has more applications and researches on beauty treatment and skin care, less applications and researches on the aspect of hemostatic efficacy, and a human-like collagen composition with excellent hemostatic effect is lacked.
Disclosure of Invention
The invention mainly aims to provide a human-like collagen composition, and aims to solve the technical problem that a human-like collagen composition with an excellent hemostatic effect is lacked in the prior art.
In order to achieve the above purpose, the human-like collagen composition provided by the invention comprises the following raw materials:
5-15 parts by weight of human-like collagen;
10-20 parts by weight of a crosslinkable powder;
1-2 parts by weight of a cross-linking agent;
20-25 parts by weight of fish gelatin extract; and
50-80 parts by weight of chitosan.
Optionally, the crosslinkable powder comprises the following starting materials: the starch treatment enzyme comprises starch and treatment enzyme, wherein the treatment enzyme accounts for 1-5% of the mass of the starch.
Optionally, the treatment enzyme comprises the following raw materials: alpha-amylase and glucoamylase; the mass ratio of the alpha-amylase to the glucoamylase is 1: (4-6).
Optionally, the starch is selected from at least one of corn starch, potato starch, or wheat starch.
Optionally, the ratio of the total weight of the human-like collagen and the fish gelatin extract to the weight of the chitosan is 1: (1.8-2.1).
The invention also provides a preparation method of the human-like collagen composition, which comprises the following steps:
preparing crosslinkable powder;
taking the crosslinkable powder with the proportion amount, adding water to form a solution, adding the human-like collagen with the proportion amount, and uniformly stirring; after centrifugal treatment, adding a cross-linking agent with a proportional amount to cross-link the human-like collagen and the cross-linkable powder, wherein the cross-linking temperature is 55-65 ℃, and the cross-linking time is 10-12 hours; after crosslinking, washing, centrifuging and freeze-drying to obtain a human-like collagen-crosslinkable powder composition;
preparing a fish gelatin extract;
dissolving chitosan in 0.6-0.8% acetic acid solution, adding the human-like collagen-crosslinkable powder composition and fish gelatin extract, stirring, and ultrasonically degassing to obtain mixed solution; uniformly spraying the mixed solution on a freezing tray, wherein the spraying thickness is 1-2mm, and pre-freezing for 40-50min at-80 ℃ to-60 ℃; and after pre-freezing, placing the mixture in a freeze dryer for freeze drying, and obtaining the human-like collagen composition after complete drying.
Alternatively, the step of preparing the crosslinkable powder comprises the steps of:
preparing 16-20% of starch milk by taking starch according to the proportion, adding sodium chloride which is 1-2% of the mass of the starch and sodium trimetaphosphate which is 1-2% of the mass of the starch, adjusting the pH value to 10.1-10.2, heating to 40-50 ℃ under the condition of stirring, reacting for 20-22 hours, adjusting the pH value to 4-5 after the reaction is finished, filtering and washing to obtain the crosslinkable powder.
Optionally, the step of preparing the crosslinkable powder further comprises the steps of:
taking alpha-amylase and glucoamylase with a proportioning amount, and adding a buffer solution to prepare an enzyme solution;
adding the crosslinkable powder into a sodium acetate buffer solution to prepare a solution with the mass fraction of 20-25%, and heating to 50-55 ℃; adding the enzyme solution, stirring until the reaction is complete, centrifuging, washing and drying the product, and crushing the dried powder to obtain the crosslinkable powder with a microporous structure.
Optionally, the step of preparing the fish gelatin extract comprises the steps of:
providing fish gelatin;
chopping the fish gelatin, adding distilled water, and mashing the fish gelatin tissue to obtain a feed liquid;
treating the feed liquid by microwave, and filtering the feed liquid after microwave extraction is finished to obtain filtrate;
and centrifuging the filtrate, and taking supernatant to obtain the fish gelatin extract.
Optionally, in the step of adding distilled water after the fish gelatin is chopped, the mass ratio of the fish gelatin to the distilled water is 1: (40-50);
and/or in the step of treating the feed liquid by microwaves, the microwave extraction time is 3-5 minutes, and the microwave extraction power is 350-400W.
According to the technical scheme, the human-like collagen-crosslinkable powder composition is obtained by crosslinking the human-like collagen and the crosslinkable powder, is a material with a good hemostatic effect, and then the fish gelatin extract, the human-like collagen-crosslinkable powder composition and chitosan are prepared to obtain a composition which is spongy and has a uniform internal pore structure; the human-like collagen composition obtained finally has good structural stability, water absorption, comfort, human body compatibility and hemostatic effect.
Detailed Description
The technical solutions in the embodiments of the present invention will be described clearly and completely below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be obtained by a person skilled in the art without inventive step based on the embodiments of the present invention, are within the scope of protection of the present invention.
The invention provides a human-like collagen composition.
In the embodiment of the invention, the human-like collagen composition comprises 5-15 parts by weight of human-like collagen, 10-20 parts by weight of crosslinkable powder, 1-2 parts by weight of crosslinking agent, 20-25 parts by weight of fish gelatin extract and 50-80 parts by weight of chitosan.
In an embodiment of the invention, the crosslinkable powder comprises the following starting materials: starch and processing enzyme, wherein the processing enzyme accounts for 1-5% of the mass of the starch. Specifically, the treatment enzyme is 1%, 3% or 5% of the mass of the starch.
In the embodiment of the invention, the processing enzyme comprises the following raw materials: alpha-amylase and glucoamylase; the mass ratio of the alpha-amylase to the glucoamylase is 1 to (4-6). Specifically, the mass ratio may be 1: 4, 1: 5 or 1: 6, or may be any ratio of 1: 4-6.
In an embodiment of the invention, the starch is selected from at least one of corn starch, potato starch or wheat starch.
In the embodiment of the invention, the ratio of the total weight of the human-like collagen and the fish gelatin extract to the weight of the chitosan is 1 to (1.8-2.1). Specifically, the ratio may be 1: 1.8, 1: 2, or 1: 2.1, or may be any ratio of 1: (1.8-2.1).
The technical scheme of the invention is that the human-like collagen-crosslinkable powder composition is obtained by crosslinking the human-like collagen and the crosslinkable powder, and is a material with good hemostatic effect. Compared with single hemostatic powder, the human collagen-crosslinkable powder composition has better human body compatibility and has the effect of promoting the growth of human body cells.
Then taking the fish gelatin extract and the human-like collagen-crosslinkable powder composition as collagen components and synthesizing and preparing chitosan to obtain a new composition, wherein the composition is in a sponge shape and has a uniform internal pore structure; the finally obtained human-like collagen composition has good structural stability, water absorption, comfort, human body compatibility, antibacterial property, shielding effect and hemostatic effect.
The invention also provides a preparation method of the human-like collagen composition, which comprises the following steps:
preparing crosslinkable powder;
taking the crosslinkable powder with the proportion amount, adding water to form a solution, adding the human-like collagen with the proportion amount, and uniformly stirring; after centrifugal treatment, cross-linking agent with a certain proportion is added to cross-link human collagen and cross-linkable powder, the cross-linking temperature is 55-65 ℃, and the cross-linking time is 10-12 hours; after crosslinking, washing, centrifuging and freeze-drying to obtain a human-like collagen-crosslinkable powder composition;
preparing a fish gelatin extract;
dissolving chitosan in 0.6-0.8% acetic acid solution, adding human-like collagen-crosslinkable powder composition and fish gelatin extract, stirring, and ultrasonically degassing to obtain mixed solution; uniformly spraying the mixed solution on a freezing plate with the spraying thickness of 1-2mm, and pre-freezing for 40-50min at-80 ℃ to-60 ℃; and after pre-freezing, placing the mixture in a freeze dryer for freeze drying, and obtaining the human-like collagen composition after complete drying.
In an embodiment of the invention, the step of preparing the crosslinkable powder comprises the steps of:
preparing 16-20% of starch milk by taking starch in proportion, adding sodium chloride which is 1-2% of the mass of the starch and sodium trimetaphosphate which is 1-2% of the mass of the starch, adjusting the pH value to 10.1-10.2, heating to 40-50 ℃ under the condition of stirring, reacting for 20-22 hours, adjusting the pH value to 4-5 after the reaction is finished, filtering and washing to obtain the crosslinkable powder.
In an embodiment of the invention, the step of preparing the crosslinkable powder further comprises the steps of:
taking alpha-amylase and glucoamylase with a proportioning amount, and adding a buffer solution to prepare an enzyme solution;
adding the crosslinkable powder into a sodium acetate buffer solution to prepare a solution with the mass fraction of 20-25%, and heating to 50-55 ℃; adding enzyme solution, stirring until the reaction is complete, centrifuging, washing and drying the product, and crushing the dried powder to obtain the crosslinkable powder with the microporous structure.
In the embodiment of the invention, the step of preparing the fish gelatin extract comprises the following steps:
providing fish gelatin;
cutting fish gelatin, adding distilled water, and mashing fish gelatin tissue to obtain feed liquid;
treating the feed liquid by microwave, and filtering the feed liquid after microwave extraction to obtain filtrate;
centrifuging the filtrate, and collecting supernatant to obtain fish gelatin extract.
In the embodiment of the invention, in the step of adding distilled water after cutting fish glue, the mass ratio of the fish glue to the distilled water is 1: (40-50).
In the embodiment of the invention, in the step of treating the feed liquid by microwaves, the microwave extraction time is 3-5 minutes, and the microwave extraction power is 350-400W.
For a further understanding of the present invention, reference will now be made to the following preferred embodiments of the invention in conjunction with the examples, but it is to be understood that the description is intended to further illustrate the features and advantages of the invention and is not intended to limit the scope of the claims which follow.
The raw material ratios of examples 1 to 3 are shown in Table 1.
TABLE 1
| Example 1 | Example 2 | Example 3 | |
| Human-like collagen (parts by weight) | 5 | 10 | 15 |
| Crosslinkable powder (parts by weight) | 10 | 15 | 20 |
| Genipin cross-linking agent (parts by weight) | 1 | 1.5 | 2 |
| Fish gelatin extract (parts by weight) | 20 | 25 | 25 |
| Chitosan (parts by weight) | 50 | 70 | 80 |
Examples 1 to 3
The preparation method of the human-like collagen composition comprises the following steps:
preparing 18% of starch milk by taking starch in proportion, adding sodium chloride which is 1.5% of the mass of the starch and sodium trimetaphosphate which is 1.5% of the mass of the starch, adjusting the pH value to 10.1, heating to 50 ℃ under the stirring condition, reacting for 21 hours, adjusting the pH value to 5 after the reaction is finished, filtering and washing to obtain the crosslinkable powder.
Adding a buffer solution into alpha-amylase and glucoamylase in a proportioning amount to prepare an enzyme solution, wherein the total mass of the alpha-amylase and the glucoamylase is 3% of the mass of the starch, and the mass ratio of the alpha-amylase to the glucoamylase is 1: 5;
adding the crosslinkable powder into a sodium acetate buffer solution to prepare a solution with the mass fraction of 24%, and heating to 50 ℃; adding the enzyme solution, stirring until the reaction is complete, centrifuging, washing and drying the product, and crushing the dried powder to obtain the crosslinkable powder with a microporous structure;
taking the crosslinkable powder with the proportion amount, adding water to form a solution, adding the human-like collagen with the proportion amount, and uniformly stirring; after centrifugal treatment, adding a cross-linking agent in a proportion for cross-linking the human-like collagen and the cross-linkable powder, wherein the cross-linking temperature is 55 ℃, and the cross-linking time is 11 hours; after crosslinking, washing, centrifuging and freeze-drying to obtain a human-like collagen-crosslinkable powder composition;
providing fish gelatin; cutting fish gelatin, adding distilled water, and mashing fish gelatin tissue to obtain feed liquid; the mass ratio of the fish glue to the distilled water is 1: 45;
treating the feed liquid by microwave, wherein the microwave extraction time is 4 minutes, the microwave extraction power is 370W, and filtering the feed liquid after the microwave extraction is finished to obtain filtrate;
centrifuging the filtrate, and collecting supernatant to obtain fish gelatin extract;
dissolving chitosan in 0.7% acetic acid solution, adding human-like collagen-crosslinkable powder composition and fish gelatin extract, stirring, and ultrasonically degassing to obtain mixed solution; uniformly spraying the mixed solution on a freezing plate, wherein the spraying thickness is 1mm, and pre-freezing for 45min at-70 ℃; and after pre-freezing, placing the mixture in a freeze dryer for freeze drying, and obtaining the human-like collagen composition after complete drying.
The hemostatic effect of the human-like collagen compositions prepared in examples 1-3 was tested, and compared with commercially available gelatin sponge (Nanjing Jinling pharmaceutical Co., ltd.), by the following method:
experimental rabbits (new zealand white rabbits, weighing 2.5 ± 0.2 kg) were anesthetized by intraperitoneal injection with ketamine, opened layer by layer, and subjected to bleeding wound of 1cm × 1cm on the left lobe of the liver by local clamping, hemostasis was performed using commercially available gelatin sponges and the human-like collagen compositions of examples 1-3, respectively, and local hemostasis time was observed: the bleeding wounds were laid down with hemostatic material, covered with gauze, and the bleeding was observed at time points 65, 85 and 150s, respectively, and the bleeding stop time was recorded. The test results are shown in Table 2.
TABLE 2
| Example 1 | Example 2 | Example 3 | Gelatin sponge | |
| Hemostasis time (second) | 65 | 70 | 68 | 120 |
Compared with the gelatin sponge in the prior art, the human-like collagen composition has better hemostatic effect.
The human-like collagen compositions prepared in examples 1-3 were tested for porosity and water absorption. The test results are shown in Table 3.
TABLE 3
| Example 1 | Example 2 | Example 3 | |
| Porosity (%) | 61 | 66 | 63 |
| Water absorption (%) | 195 | 210 | 205 |
It can be seen that the human-like collagen composition of the present invention has good water absorption, and the porosity reflects the air permeability.
The above description is only a preferred embodiment of the present invention, and is not intended to limit the scope of the present invention, and all equivalent modifications made by the present invention in the light of the present specification, or directly/indirectly applied to other related technical fields are included in the scope of the present invention.
Claims (10)
1. A human-like collagen composition, comprising the following raw materials:
5-15 parts by weight of human-like collagen;
10-20 parts by weight of a crosslinkable powder;
1-2 parts by weight of a cross-linking agent;
20-25 parts by weight of fish gelatin extract; and
50-80 parts by weight of chitosan.
2. The human-like collagen composition according to claim 1, wherein said crosslinkable powder comprises the following raw materials: the starch treatment enzyme comprises starch and treatment enzyme, wherein the treatment enzyme accounts for 1-5% of the mass of the starch.
3. The human-like collagen composition according to claim 2, wherein said processing enzyme comprises the following raw materials: alpha-amylase and glucoamylase; the mass ratio of the alpha-amylase to the glucoamylase is 1: (4-6).
4. The human-like collagen composition according to claim 2, wherein said starch is selected from at least one of corn starch, potato starch or wheat starch.
5. The human-like collagen composition according to claim 1, wherein the ratio of the total weight of said human-like collagen and said fish gelatin extract to the weight of said chitosan is 1: (1.8-2.1).
6. A method for preparing a human-like collagen composition, comprising the steps of:
preparing crosslinkable powder;
taking the crosslinkable powder with the proportion amount, adding water to form a solution, adding the human-like collagen with the proportion amount, and uniformly stirring; after centrifugal treatment, adding a cross-linking agent with a proportional amount to cross-link the human-like collagen and the cross-linkable powder, wherein the cross-linking temperature is 55-65 ℃, and the cross-linking time is 10-12 hours; after crosslinking, washing, centrifuging and freeze-drying to obtain a human-like collagen-crosslinkable powder composition;
preparing a fish gelatin extract;
dissolving chitosan in 0.6-0.8% acetic acid solution, adding the human-like collagen-crosslinkable powder composition and fish gelatin extract, stirring, and ultrasonically degassing to obtain mixed solution; uniformly spraying the mixed solution on a freezing tray, wherein the spraying thickness is 1-2mm, and pre-freezing for 40-50min at-80 ℃ to-60 ℃; and after pre-freezing, placing the mixture in a freeze dryer for freeze drying, and obtaining the human-like collagen composition after complete drying.
7. The method of preparing a human-like collagen composition according to claim 6, wherein the step of preparing a cross-linkable powder comprises the steps of:
preparing 16-20% of starch milk by taking starch in proportion, adding sodium chloride which is 1-2% of the mass of the starch and sodium trimetaphosphate which is 1-2% of the mass of the starch, adjusting the pH value to 10.1-10.2, heating to 40-50 ℃ under the condition of stirring, reacting for 20-22 hours, adjusting the pH value to 4-5 after the reaction is finished, filtering and washing to obtain the crosslinkable powder.
8. The method of preparing a human-like collagen composition according to claim 7, wherein the step of preparing the cross-linkable powder further comprises the steps of:
taking alpha-amylase and glucoamylase in a proportion amount, and adding a buffer solution to prepare an enzyme solution;
adding the crosslinkable powder into a sodium acetate buffer solution to prepare a solution with the mass fraction of 20-25%, and heating to 50-55 ℃; and adding the enzyme solution, stirring until the reaction is complete, centrifuging, washing and drying the product, and crushing the dried powder to obtain the crosslinkable powder with the microporous structure.
9. The method for preparing a human-like collagen composition according to claim 6, wherein the step of preparing a fish gelatin extract comprises the steps of:
providing fish gelatin;
cutting the fish gelatin, adding distilled water, and mashing fish gelatin tissue to obtain feed liquid;
treating the feed liquid by microwave, and filtering the feed liquid after the microwave extraction is finished to obtain filtrate;
and centrifuging the filtrate, and taking supernatant to obtain the fish gelatin extract.
10. The method for preparing a human-like collagen composition according to claim 9, wherein in the step of adding distilled water after the cutting of said fish gelatin, the mass ratio of said fish gelatin to said distilled water is 1: (40-50);
and/or in the step of treating the feed liquid by microwaves, the microwave extraction time is 3-5 minutes, and the microwave extraction power is 350-400W.
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