CN107602692A - A kind of preparation method of collagen diaphragm material - Google Patents
A kind of preparation method of collagen diaphragm material Download PDFInfo
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- CN107602692A CN107602692A CN201710659257.1A CN201710659257A CN107602692A CN 107602692 A CN107602692 A CN 107602692A CN 201710659257 A CN201710659257 A CN 201710659257A CN 107602692 A CN107602692 A CN 107602692A
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Abstract
The invention discloses a kind of preparation method of collagen diaphragm material, it is related to collagen extractive technique field, comprises the following steps：It is skin source to take Animal Skin, depilation processing, removes attached meat, saline sook；Degreasing agent is added into Animal Skin, the degreasing agent is made up of the raw material of following parts by weight：Defatting enzyme 3 7%, neopelex 5 10%, sorbitan ester 3 7%, high-carbon fatty alcohol polyoxyethylene ether 5 10%, surplus is water；Animal Skin is sequentially placed into immersion treatment in citric acid solution, NaCl solution, the solution of Triton X 100, crushed, centrifugation, takes Animal Skin to precipitate；Precipitated to Animal Skin and add water, then added papain, Microwave-assisted firing, extract, filtering, obtain filtrate；By filtrate through micro-filtrate membrane filtration, milipore filter concentration, concentrate is freezed, produced.Preparation method of the present invention is simple, clean, obtained collagen quality better is removed for compositions such as foreign proteins, purity height, quality is flexible, adhesiveness is good, meets the requirement of clinic.
Description
Technical field
The present invention relates to collagen extractive technique field, and in particular to a kind of preparation side of collagen diaphragm material
Method.
Background technology
Collagen is that animal in-vivo content is most, is distributed most commonly used protein.It is the major structural protein of body,
It is the main constituents for forming tissue and connective tissue.The amino acid chain structure of its specific triple helices imparts it and permitted
More useful characteristic：Such as high tensile, biodegradability, low antigen active, low irritant and cytotoxicity and promotion
The characteristic of cell growth.All these features become preferable bio-medical material.
In medical science, particularly in surgical intervention, collagen is mainly used in following field：Trauma care and control
System, particularly special open sore part extremity ulcer as caused by diabetes, from over 2010, FDA ratifies two kinds of novel collagens successively
Protide sponge or colloid series of products control medicine as fully functioning wound, and special recommendation is used for ulcer caused by diabetes
Treatment and whole wounds control (Cellerate and Excellagen) in addition to 3 degree are burnt, and have and a variety of be used for wound control
The collagen protein sponge or colloid products of system and treatment are just in clinical test；In Orthopeadic Surgery, collagen protein sponge, collagen
Diaphragm, it is widely used in bone filling and cartilage damage reparation；Cranial surgery, the filling of brain dura mater wound repair are also a large amount of to use
Collagen membrane material.But at present, the mode such as preparation method generally use enzymolysis, acid extraction of collagen diaphragm prepares collagen
Protein raw materials, then by being crosslinked with other materials or being prepared by cross-linking agents such as isoamyl dialdehyde, preparation process is cumbersome,
Time is grown.
The content of the invention
It is an object of the invention to provide a kind of preparation method of collagen diaphragm material, the whole preparation process time compared with
Collagen diaphragm material quality better that is short, obtaining.
To realize object above, the present invention is achieved by the following technical programs：
A kind of preparation method of collagen diaphragm material, comprises the following steps：
S1, to take Animal Skin be skin source, cleans up, then carries out depilation processing, and the attached meat in Animal Skin is removed, and is used
0.5-1.0% saline sook 10-20min, are rinsed well with water；
S2, degreasing agent is added into the Animal Skin after step S1 processing, 40-60min is stirred at 40-50 DEG C；It is described de-
Fat agent is made up of the raw material of following parts by weight：Defatting enzyme 3-7%, neopelex 5-10%, sorbitan ester 3-
7%th, high-carbon fatty alcohol polyoxyethylene ether 5-10%, surplus are water；
S3, the Animal Skin after step S2 processing is put into immersion treatment 2-4h in 2-3% citric acid solutions and makes its swelling, taken
Go out, then be sequentially placed into immersion treatment 1-2h in 2-3%NaCl solution, immersion treatment in 0.2-1% Triton X-100 solution
20-40min, the Animal Skin after taking-up processing, is crushed using high-speed tissue mashing machine, is centrifuged, is removed supernatant, take Animal Skin
Precipitation；
S4, the Animal Skin precipitation into step S3 add water, then add papain, Microwave-assisted firing 3-
5min, 4-6h is extracted, filter-cloth filtering, obtains filtrate；
S5, by the filtrate that step S4 is obtained through micro-filtrate membrane filtration, milipore filter concentration, concentrate is freezed, produced.
Preferably, the Animal Skin is pigskin.
Preferably, degreasing agent and the solid-liquid ratio of Animal Skin are 3-5 in the step S2：1.
Preferably, citric acid solution and the solid-liquid ratio of Animal Skin are 4-8 in the step S3：1, NaCl solution and Animal Skin
Solid-liquid ratio be 6-10：1, Triton X-100 solution and the solid-liquid ratio of Animal Skin are 3-5：1.
Preferably, amount of water and the solid-liquid ratio of Animal Skin precipitation are 6-10 in the step S4：1.
Preferably, the addition of papain precipitates the 3-5% of quality for Animal Skin in the step S4.
Preferably, the power of microwave is 300-400W in the step S4.
Beneficial effect：Preparation method of the present invention is simple, and the whole preparation process time is shorter, and processing cost is low, for miscellaneous egg
White to wait composition to remove totally, obtained collagen diaphragm material quality better, purity is high, and quality is flexible, adhesiveness is good, meets and faces
The requirement of bed, has good Social benefit and economic benefit.
Embodiment
To make the purpose, technical scheme and advantage of the embodiment of the present invention clearer, below in conjunction with the embodiment of the present invention,
Technical scheme in the embodiment of the present invention is clearly and completely described, it is clear that described embodiment is the present invention one
Divide embodiment, rather than whole embodiments.Based on the embodiment in the present invention, those of ordinary skill in the art are not making
The every other embodiment obtained under the premise of creative work, belongs to the scope of protection of the invention.
Embodiment 1：
A kind of preparation method of collagen diaphragm material, comprises the following steps：
S1, to take Animal Skin be skin source, cleans up, then carries out depilation processing, and the attached meat in Animal Skin is removed, and is used
0.5% saline sook 10min, is rinsed well with water；
S2, add degreasing agent into the Animal Skin after step S1 processing, the solid-liquid ratio of degreasing agent and Animal Skin is 3：1,
40min is stirred at 40 DEG C；The degreasing agent is made up of the raw material of following parts by weight：Defatting enzyme 5%, neopelex
8%th, sorbitan ester 5%, high-carbon fatty alcohol polyoxyethylene ether 8%, surplus are water；
S3, by step S2 processing after Animal Skin be put into 2% citric acid solution, the feed liquid of citric acid solution and Animal Skin
Than for 4：1, immersion treatment 2h make its swelling, take out, then be sequentially placed into 2%NaCl solution, the material of NaCl solution and Animal Skin
Liquor ratio is 6：1, immersion treatment 1h, then place into 0.2% Triton X-100 solution, Triton X-100 solution is with moving
The solid-liquid ratio of thing skin is 4：1, immersion treatment 20min, the Animal Skin after taking-up processing, are broken using high-speed tissue mashing machine
It is broken, centrifugation, supernatant is removed, takes Animal Skin to precipitate；
S4, the Animal Skin precipitation into step S3 add the water of 6 times of Animal Skin precipitation quality, then add Papain
Enzyme, the addition of papain precipitate the 3% of quality for Animal Skin, Microwave-assisted firing 5min, microwave power 400W, carried
6h is taken, filter-cloth filtering, obtains filtrate；
S5, by the filtrate that step S4 is obtained through micro-filtrate membrane filtration, milipore filter concentration, concentrate is freezed, produced.
Embodiment 2：
A kind of preparation method of collagen diaphragm material, comprises the following steps：
S1, to take pigskin be skin source, cleans up, then carries out depilation processing, and the attached meat in Animal Skin is removed, and is used
0.6% saline sook 15min, is rinsed well with water；
S2, add degreasing agent into the Animal Skin after step S1 processing, the solid-liquid ratio of degreasing agent and Animal Skin is 3.5：1,
45min is stirred at 45 DEG C；The degreasing agent is made up of the raw material of following parts by weight：Defatting enzyme 6%, neopelex
7%th, sorbitan ester 5%, high-carbon fatty alcohol polyoxyethylene ether 7%, surplus are water；
S3, by step S2 processing after Animal Skin be put into 2% citric acid solution, the feed liquid of citric acid solution and Animal Skin
Than for 5：1, immersion treatment 2.5h make its swelling, take out, then are sequentially placed into 3%NaCl solution, NaCl solution and Animal Skin
Solid-liquid ratio is 8：1, immersion treatment 1h, then place into 0.5% Triton X-100 solution, Triton X-100 solution with
The solid-liquid ratio of Animal Skin is 3：1, immersion treatment 30min, the Animal Skin after taking-up processing, are broken using high-speed tissue mashing machine
It is broken, centrifugation, supernatant is removed, takes Animal Skin to precipitate；
S4, the Animal Skin precipitation into step S3 add the water of 7 times of Animal Skin precipitation quality, then add Papain
Enzyme, the addition of papain precipitate the 5% of quality for Animal Skin, Microwave-assisted firing 3min, microwave power 300W, carried
4h is taken, filter-cloth filtering, obtains filtrate；
S5, by the filtrate that step S4 is obtained through micro-filtrate membrane filtration, milipore filter concentration, concentrate is freezed, produced.
Embodiment 3：
A kind of preparation method of collagen diaphragm material, comprises the following steps：
S1, to take sheepskin be skin source, cleans up, then carries out depilation processing, and the attached meat in Animal Skin is removed, and is used
0.8% saline sook 15min, is rinsed well with water；
S2, add degreasing agent into the Animal Skin after step S1 processing, the solid-liquid ratio of degreasing agent and Animal Skin is 4：1,
50min is stirred at 45 DEG C；The degreasing agent is made up of the raw material of following parts by weight：Defatting enzyme 4%, neopelex
8%th, sorbitan ester 6%, high-carbon fatty alcohol polyoxyethylene ether 7%, surplus are water；
S3, by step S2 processing after Animal Skin be put into 2.5% citric acid solution, the material of citric acid solution and Animal Skin
Liquor ratio is 6：1, immersion treatment 3h make its swelling, take out, then be sequentially placed into 2.5%NaCl solution, NaCl solution and Animal Skin
Solid-liquid ratio be 8：1, immersion treatment 1.5h, then place into 0.5% Triton X-100 solution, Triton X-100 are molten
Liquid and the solid-liquid ratio of Animal Skin are 5：1, immersion treatment 30min, the Animal Skin after taking-up processing, are entered using high-speed tissue mashing machine
Row is broken, centrifugation, removes supernatant, takes Animal Skin to precipitate；
S4, the Animal Skin precipitation into step S3 add the water of 8 times of Animal Skin precipitation quality, then add Papain
Enzyme, the addition of papain precipitate the 3% of quality for Animal Skin, Microwave-assisted firing 3min, microwave power 300W, carried
4h is taken, filter-cloth filtering, obtains filtrate；
S5, by the filtrate that step S4 is obtained through micro-filtrate membrane filtration, milipore filter concentration, concentrate is freezed, produced.
Embodiment 4：
A kind of preparation method of collagen diaphragm material, comprises the following steps：
S1, to take Animal Skin be skin source, cleans up, then carries out depilation processing, and the attached meat in Animal Skin is removed, and is used
0.8% saline sook 20min, is rinsed well with water；
S2, add degreasing agent into the Animal Skin after step S1 processing, the solid-liquid ratio of degreasing agent and Animal Skin is 4.5：1,
55min is stirred at 50 DEG C；The degreasing agent is made up of the raw material of following parts by weight：Defatting enzyme 3%, neopelex
10%th, sorbitan ester 3%, high-carbon fatty alcohol polyoxyethylene ether 10%, surplus are water；
S3, by step S2 processing after Animal Skin be put into 2.5% citric acid solution, the material of citric acid solution and Animal Skin
Liquor ratio is 7：1, immersion treatment 3.5h make its swelling, take out, then be sequentially placed into 3%NaCl solution, NaCl solution and Animal Skin
Solid-liquid ratio be 8：1, immersion treatment 2h, then place into 0.8% Triton X-100 solution, Triton X-100 solution
Solid-liquid ratio with Animal Skin is 4：1, immersion treatment 30min, the Animal Skin after taking-up processing, are carried out using high-speed tissue mashing machine
It is broken, centrifugation, supernatant is removed, takes Animal Skin to precipitate；
S4, the Animal Skin precipitation into step S3 add the water of 9 times of Animal Skin precipitation quality, then add Papain
Enzyme, the addition of papain precipitate the 3% of quality for Animal Skin, Microwave-assisted firing 5min, microwave power 350W, carried
5h is taken, filter-cloth filtering, obtains filtrate；
S5, by the filtrate that step S4 is obtained through micro-filtrate membrane filtration, milipore filter concentration, concentrate is freezed, produced.
Embodiment 5：
A kind of preparation method of collagen diaphragm material, comprises the following steps：
S1, to take Animal Skin be skin source, cleans up, then carries out depilation processing, and the attached meat in Animal Skin is removed, and is used
1.0% saline sook 20min, is rinsed well with water；
S2, add degreasing agent into the Animal Skin after step S1 processing, the solid-liquid ratio of degreasing agent and Animal Skin is 5：1,
60min is stirred at 50 DEG C；The degreasing agent is made up of the raw material of following parts by weight：Defatting enzyme 7%, neopelex
5%th, sorbitan ester 7%, high-carbon fatty alcohol polyoxyethylene ether 5%, surplus are water；
S3, by step S2 processing after Animal Skin be put into 3% citric acid solution, the feed liquid of citric acid solution and Animal Skin
Than for 8：1, immersion treatment 4h make its swelling, take out, then be sequentially placed into 2%NaCl solution, the material of NaCl solution and Animal Skin
Liquor ratio is 10：1, immersion treatment 2h, then place into 1% Triton X-100 solution, Triton X-100 solution is with moving
The solid-liquid ratio of thing skin is 5：1, immersion treatment 40min, the Animal Skin after taking-up processing, are broken using high-speed tissue mashing machine
It is broken, centrifugation, supernatant is removed, takes Animal Skin to precipitate；
S4, the Animal Skin precipitation into step S3 add the water of 10 times of Animal Skin precipitation quality, then add Papain
Enzyme, the addition of papain precipitate the 4% of quality for Animal Skin, Microwave-assisted firing 5min, microwave power 400W, carried
6h is taken, filter-cloth filtering, obtains filtrate；
S5, by the filtrate that step S4 is obtained through micro-filtrate membrane filtration, milipore filter concentration, concentrate is freezed, produced.
The above embodiments are merely illustrative of the technical solutions of the present invention, rather than its limitations；Although with reference to the foregoing embodiments
The present invention is described in detail, it will be understood by those within the art that：It still can be to foregoing each implementation
Technical scheme described in example is modified, or carries out equivalent substitution to which part technical characteristic；And these modification or
Replace, the essence of appropriate technical solution is departed from the spirit and scope of various embodiments of the present invention technical scheme.
Claims (7)
1. a kind of preparation method of collagen diaphragm material, it is characterised in that comprise the following steps：
S1, to take Animal Skin be skin source, cleans up, then carries out depilation processing, and the attached meat in Animal Skin is removed, uses 0.5-
1.0% saline sook 10-20min, is rinsed well with water；
S2, degreasing agent is added into the Animal Skin after step S1 processing, 40-60min is stirred at 40-50 DEG C；The degreasing agent
It is made up of the raw material of following parts by weight：Defatting enzyme 3-7%, neopelex 5-10%, sorbitan ester 3-7%,
High-carbon fatty alcohol polyoxyethylene ether 5-10%, surplus are water；
S3, the Animal Skin after step S2 processing is put into immersion treatment 2-4h in 2-3% citric acid solutions and makes its swelling, taken out,
It is sequentially placed into immersion treatment 1-2h in 2-3%NaCl solution again, immersion treatment 20- in 0.2-1% Triton X-100 solution
40min, the Animal Skin after taking-up processing, is crushed using high-speed tissue mashing machine, is centrifuged, is removed supernatant, take Animal Skin to sink
Form sediment；
S4, the Animal Skin precipitation into step S3 add water, then add papain, Microwave-assisted firing 3-5min, carry
4-6h is taken, filter-cloth filtering, obtains filtrate；
S5, by the filtrate that step S4 is obtained through micro-filtrate membrane filtration, milipore filter concentration, concentrate is freezed, produced.
2. the preparation method of collagen diaphragm material as claimed in claim 1, it is characterised in that the Animal Skin is pig
Skin.
3. the preparation method of collagen diaphragm material as claimed in claim 1, it is characterised in that degreasing in the step S2
The solid-liquid ratio of agent and Animal Skin is 3-5：1.
4. the preparation method of collagen diaphragm material as claimed in claim 1, it is characterised in that lemon in the step S3
Acid solution and the solid-liquid ratio of Animal Skin are 4-8：1, the solid-liquid ratio of NaCl solution and Animal Skin is 6-10：1, Triton X-100 is molten
Liquid and the solid-liquid ratio of Animal Skin are 3-5：1.
5. the preparation method of collagen diaphragm material as claimed in claim 1, it is characterised in that add water in the step S4
The solid-liquid ratio of amount and Animal Skin precipitation is 6-10：1.
6. the preparation method of collagen diaphragm material as claimed in claim 1, it is characterised in that pawpaw in the step S4
The addition of protease is the 3-5% that Animal Skin precipitates quality.
7. the preparation method of collagen diaphragm material as claimed in claim 1, it is characterised in that microwave in the step S4
Power be 300-400W.
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| CN1903918A (en) * | 2006-07-31 | 2007-01-31 | 华南理工大学 | Method of extracting collagen and method of using collagen to prepare collagen protein |
| CN102559826A (en) * | 2012-02-15 | 2012-07-11 | 胡如桂 | Method for preparing collagen oligopeptide |
| CN102921034A (en) * | 2012-12-04 | 2013-02-13 | 广州创尔生物技术有限公司 | Composite dressing of pig accellular dermal matrix and preparation method of composite dressing |
| CN103966294A (en) * | 2013-02-02 | 2014-08-06 | 深圳兰度生物材料有限公司 | Extraction method for bioactive collagen |
| CN104098783A (en) * | 2014-06-18 | 2014-10-15 | 常州药物研究所有限公司 | Preparation method of collagen diaphragm material |
| CN105985430A (en) * | 2016-01-27 | 2016-10-05 | 成都市科乐生物技术有限公司 | Low-temperature macromolecular collagen extracting method |
| CN107164442A (en) * | 2017-06-05 | 2017-09-15 | 深圳知本康业有限公司 | A kind of collagen polypeptide and application |
-
2017
- 2017-08-04 CN CN201710659257.1A patent/CN107602692A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1903918A (en) * | 2006-07-31 | 2007-01-31 | 华南理工大学 | Method of extracting collagen and method of using collagen to prepare collagen protein |
| CN102559826A (en) * | 2012-02-15 | 2012-07-11 | 胡如桂 | Method for preparing collagen oligopeptide |
| CN102921034A (en) * | 2012-12-04 | 2013-02-13 | 广州创尔生物技术有限公司 | Composite dressing of pig accellular dermal matrix and preparation method of composite dressing |
| CN103966294A (en) * | 2013-02-02 | 2014-08-06 | 深圳兰度生物材料有限公司 | Extraction method for bioactive collagen |
| CN104098783A (en) * | 2014-06-18 | 2014-10-15 | 常州药物研究所有限公司 | Preparation method of collagen diaphragm material |
| CN105985430A (en) * | 2016-01-27 | 2016-10-05 | 成都市科乐生物技术有限公司 | Low-temperature macromolecular collagen extracting method |
| CN107164442A (en) * | 2017-06-05 | 2017-09-15 | 深圳知本康业有限公司 | A kind of collagen polypeptide and application |
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