CN102952837A - Medical fish collagen polypeptide raw material and its preparation technology - Google Patents
Medical fish collagen polypeptide raw material and its preparation technology Download PDFInfo
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- CN102952837A CN102952837A CN2011102405136A CN201110240513A CN102952837A CN 102952837 A CN102952837 A CN 102952837A CN 2011102405136 A CN2011102405136 A CN 2011102405136A CN 201110240513 A CN201110240513 A CN 201110240513A CN 102952837 A CN102952837 A CN 102952837A
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Abstract
The invention relates to a preparation technology of a medical fish collagen polypeptide raw material. The technology comprises the following steps: cleaning fish scales/fish skins, pickling, washing through using water, extracting, performing coarse filtration, decoloring and deodorizing, refining, and performing dry molding. The invention also provides the medical fish collagen polypeptide raw material. The technology is simple and practical, and the fish collagen polypeptide raw material formed through the technology can reach a medical level, can be used for modifying tissue engineering support materials, medicinal sustained-release materials, coat materials and the like as a functional active factor, also can be directly used for the auxiliary treatment of diseases, avoids the virus propagation risks of animal collagens, and has a high biological safety.
Description
Technical field
The present invention relates to the animal collagen field, relate to more specifically a kind of medical Isin glue collagen polypeptide raw material and preparation technology thereof, take fish scale/fish-skin as raw material extracts medical Isin glue collagen polypeptide raw material, reach aseptic pyrogen-free level.
Background technology
Collagen protein and cell regeneration, differentiation, motion, immunity, joint lubrication, blood pressure inhibition, wound healing, aging and disease etc. have extremely close relationship.Because it is the chief component composition of many tissues in the body and is exercising a lot of very important functions, so hemostasis, reparation and the damaged property filling of Exogenous Collagen protein-based biomaterial behind tissue injury is until all bringing into play huge effect in the regenerative medicine.At present, used collagen product is mainly derived from the livestock resource on land in biological medicine and the clinical medicine, as preparing collagen protein with the ox heel string.Owing to there being the great risk of PI mad cow disease, reproductive and respiratory syndrome virus, Gu Geguo is strictly control.If extract collagen protein take processing of aquatic products waste material fish-skin or fish scale as raw material, not only security improves greatly, and takes full advantage of the fish processing waste, thereby creates huge economy and social value.
The application of Isin glue collagen polypeptide is vigorously launching aspect functional health-care food and the cosmetic, is social recognition and acceptance not only, and has obtained good economic benefit.Already confirmed, the Isin glue collagen polypeptide can increase calcium absorption, promote the effects such as collagen protein generation, stimulate cell growth.But take a broad view of domestic and international present situation, utilize collagen of fish skin to carry out biological medicine and clinical medical application, still be in bud.At biomedicine field, because unique function and the character of collagen protein, its be widely used potential and spectacular development prospect, but also do not utilize fish scale/fish-skin to prepare the research report of medical grade collagen product at present.Up to now, the Isin glue collagen polypeptide still only is used for food, makeup and field of health care products as additive, technical value added is lower, be used for biomedicine field if can prepare the medical grade Isin glue collagen, then be expected to be applied to clinical as the substitute of animal-origin collagen product, not only significantly promote its added value, and greatly improved the biological safety of collagen product.
Therefore, medical in view of Isin glue collagen polypeptide raw material, as: the modification of tissue engineering bracket material, Thermosensitive Material Used for Controlled Releasing of Medicine, coated material etc., be badly in need of meeting in the market the requirement of medical grade raw material, can be used as Isin glue collagen polypeptide raw material and preparation technology that biologically active factors is used for biomedicine field.
Summary of the invention
For the defective of above-mentioned prior art, the object of the present invention is to provide a kind of medical Isin glue collagen polypeptide raw material and preparation technology thereof.Through processing, refining, the Isin glue collagen of the present invention's preparation is white powder, and is tasteless, heavy metal and ash content equal size are lower, weight-average molecular weight is 1000～3000Dolton, and aseptic, pyrogen-free can be used as active factor or auxiliary treatment means is applied to biomedicine field.
For achieving the above object, the technical solution used in the present invention is: a kind of Isin glue collagen polypeptide raw material and the side's of preparation technique thereof that satisfies the medical grade requirement, and decoloration and deodorization and degerming, depyrogenation, its method is:
A: clean: fish scale/fish-skin first through slightly washing, is repeatedly washed with deionized water again, drain;
B: pickling: described fish scale/fish-skin soaks with pH3～5 hydrochloric acid solns, stirring and evenly mixing;
C: washing: the fish scale/fish-skin of described acid treatment washes with water regulates pH to neutral, and then a small amount of water repeatedly washs, and drains;
D: extract: in 2～10: 1 ratio adds entry in fish scale/fish-skin, regulates enzymolysis 4～12h under ℃ condition of pH to 8～9,30～50; Be warming up to 70～100 ℃, keep 5～20min;
E: coarse filtration: the solution that extracts is removed precipitation by filter type;
F: decoloration and deodorization: described step e filtrate is crossed post process decoloration and deodorization;
G: refining: described step f filtrate is passed through the membrane filtration system;
H: drying and moulding.
Described fish-skin/fish scale should be fresh, and pollution-free and source can be reviewed, and meets the inspection and quarantine requirement;
Another aspect of the present invention is the deionized water preparation at hydrochloric acid soln described in the step b, and the volume ratio of described fish scale/fish-skin and hydrochloric acid soln is 4～8: 1, and described stir process condition is: 30～50 ℃ of stir process 4～8h;
Another aspect of the present invention is deionized water at the water of the adjusting pH described in the step c, and washing used water is injection water;
Another aspect of the present invention, water used in steps d～g is water for injection;
Another aspect of the present invention all carries out in gnotobasis in steps d～g;
Another aspect of the present invention is in diatomite, kaolin, Plate Filtration, the membrane filtration one or more at filter type described in the step e;
Another aspect of the present invention is processed and is comprised in macropore ion exchange resin, gac, Q-sepharose, the C-18 post one or more crossing post described in the step f;
Another aspect of the present invention, the system of membrane filtration described in the step g can desalination, removing impurities albumen, degerming, depyrogenation and concentrated.
Another aspect of the present invention is a kind of in lyophilize, spraying drying, the vacuum concentration drying at drying technology described in the step h.
Another embodiment of the present invention, a kind of medical Isin glue collagen polypeptide raw material.
Another aspect of the present invention, the weight-average molecular weight of described medical Isin glue collagen polypeptide raw material is 1000～3000Dolton.
Another aspect of the present invention, described medical Isin glue collagen polypeptide raw material is aseptic, pyrogen-free.
The invention solves the medical technical difficult problem that will solve, major embodiment in the following areas:
(1) aseptic controlling in the medical Isin glue collagen polypeptide preparation process;
(2) decoloration and deodorization in the medical Isin glue collagen polypeptide preparation process is processed;
(3) processing such as degerming, depyrogenation and desalination in the medical Isin glue collagen polypeptide preparation process;
Beneficial effect of the present invention is: preparation technology of the present invention has effectively solved the technical difficult problem of above-mentioned three aspects, makes the Isin glue collagen polypeptide for preparing satisfy preferably medical application.The medical Isin glue collagen polypeptide of the present invention preparation is effective decoloration and deodorization not only, range of molecular weight distributions is 1000～3000Dolton, especially concentrate on 1000～2000Dolton, heavy metal content＜0.05mg/kg, ash oontent＜0.5%, the aseptic controlling level is not less than 10-6, endotoxin content＜0.25Eu/mg, meet the requirement of medical grade raw material, can be used as biologically active factors and be used for biomedicine field.
Embodiment
Embodiment 1
The preparation technology of a kind of medical Isin glue collagen polypeptide raw material of the present invention is:
A. select the source can review, meet the fresh fish scale of inspection and quarantine requirement, with the tap water cleaning of 8 times of volumes, behind 45 ℃ of stirring 1h, draining repeats above-mentioned steps 3 times; After thorough emptying, with the washed with de-ionized water of 8 times of volumes, behind 45 ℃ of stirring 1h, draining repeats above-mentioned steps 3 times, drains again;
B. the hydrochloric acid soln (pH=4.5) with 8 times of volumes soaks 45 ℃ of stir process 8h.The configuration water of hydrochloric acid soln is deionized water, drains;
C. deionized water rinsing drains to neutral; Repeatedly wash with injection water again, remove remaining deionized water;
D. in aseptic clean room, add injection water in 8: 1 ratios in fish scale/fish-skin raw material, aseptic NaOH regulates pH to 8, enzymolysis 12h under 50 ℃ of conditions; Be warming up to 75 ℃, keep 15min, the deactivation biological enzyme; Used vessel are all processed through aseptic, pyrogen-free; Preparation institute water is water for injection;
E. in the aseptic clean room, adopt aseptic Plate Filtration device that mentioned solution is filtered, remove foreign material;
F. in the aseptic clean room, above-mentioned filtrate is crossed resinene post and middle granulated active carbon post; Used vessel and chromatographic column are all processed through aseptic, pyrogen-free;
G. in the aseptic clean room, above-mentioned filtrate is crossed film processing system, carry out desalination, remove foreign protein, degerming, depyrogenation, concentration; Film processing system is aseptic pyrogen-free state; Used vessel are all processed through aseptic, pyrogen-free;
H. in the aseptic clean room, lyophilize prepares medical Isin glue collagen polypeptide powder; Freeze drier is sterile state; Used vessel are all processed through aseptic, pyrogen-free;
Embodiment 2
The preparation technology of a kind of medical Isin glue collagen polypeptide raw material of the present invention is:
A. with embodiment 1;
B. the hydrochloric acid soln (pH=4) with 8 times of volumes soaks 50 ℃ of stir process 4h.The configuration water of hydrochloric acid soln is deionized water, drains;
C. with embodiment 1;
D. in aseptic clean room, add injection water in 10: 1 ratios in fish scale/fish-skin raw material, aseptic NaOH regulates pH to 8.5, enzymolysis 12h under 40 ℃ of conditions; Be warming up to 70 ℃, keep 20min, the deactivation biological enzyme; Used vessel are all processed through aseptic, pyrogen-free; Preparation institute water is water for injection;
E. in the aseptic clean room, adopt aseptic diatomaceous earth filter that mentioned solution is filtered, remove foreign material;
F. in the aseptic clean room, above-mentioned filtrate is crossed middle granulated active carbon post; Used vessel and chromatographic column are all processed through aseptic, pyrogen-free;
G. with embodiment 1;
H. aseptic spraying drying prepares medical Isin glue collagen polypeptide powder; Used vessel are all processed through aseptic, pyrogen-free.
Embodiment 3
The preparation technology of a kind of medical Isin glue collagen polypeptide raw material of the present invention is:
A. select the source can review, meet the fresh fish scale of inspection and quarantine requirement, with the tap water cleaning of 5 times of volumes, behind 40 ℃ of stirring 2.5h, draining repeats above-mentioned steps 3 times; After thorough emptying, with the washed with de-ionized water of 5 times of volumes, behind 40 ℃ of stirring 2.5h, draining repeats above-mentioned steps 3 times, drains again;
B. the hydrochloric acid soln (pH=4.0) with 10 times of volumes soaks 45 ℃ of stir process 5h.The configuration water of hydrochloric acid soln is deionized water, drains;
C. with embodiment 1;
D. in aseptic clean room, add injection water in 9: 1 ratios in fish scale/fish-skin raw material, aseptic NaOH regulates pH to 9.0, enzymolysis 10h under 50 ℃ of conditions; Be warming up to 80 ℃, keep 15min, the deactivation biological enzyme; Used vessel are all processed through aseptic, pyrogen-free; Preparation institute water is water for injection;
E～h is with embodiment 1;
Embodiment 4
The preparation technology of a kind of medical Isin glue collagen polypeptide raw material of the present invention is:
A. with embodiment 3;
B. the hydrochloric acid soln (pH=3.5) with 8 times of volumes soaks 45 ℃ of stir process 5h.The configuration water of hydrochloric acid soln is deionized water, drains;
C. with embodiment 1;
D. in aseptic clean room, add injection water in 8: 1 ratios in fish scale/fish-skin raw material, aseptic NaOH regulates pH to 8.5, enzymolysis 10h under 45 ℃ of conditions; Be warming up to 75 ℃, keep 20min, the deactivation biological enzyme; Used vessel are all processed through aseptic, pyrogen-free; Preparation institute water is water for injection;
E. with embodiment 2;
F. with embodiment 2;
G. with embodiment 1;
H. aseptic vacuum-drying prepares medical Isin glue collagen polypeptide powder; Used vessel are all processed through aseptic, pyrogen-free.
Get respectively the medical Isin glue collagen polypeptide portion of said process gained sample, adopt high performance liquid chromatography (HPLC) method detection molecules amount, limulus reagent test bacterial detection endotoxin content.Its ash content, heavy metal and sterilized water equality index all detect by method under Pharmacopoeia of People's Republic of China (2007 editions), the GB/T 16886 series standard corresponding entry, the results are shown in Table 1.
Each embodiment sample detection result of table 1.
From table 1, the medical Isin glue collagen polypeptide of the present invention preparation is effective decoloration and deodorization not only, and range of molecular weight distributions is 1000～3000Dolton, especially concentrate on 1000～2000Dolton, heavy metal content＜0.05mg/kg, ash oontent＜0.5%, the aseptic controlling level is not less than 10
-6, endotoxin content＜0.25Eu/mg meets the requirement of medical grade raw material, can be used as biologically active factors and is used for biomedicine field.
So far invention has been described in conjunction with the embodiments.Those skilled in the art should be appreciated that can easily make various other modifications to described embodiment in the situation that do not depart from the scope of the present invention and spirit.Therefore, the scope of appended claims is not limited to above-mentioned explanation, but will broadly explain claim.
Claims (13)
1. the preparation technology of a medical Isin glue collagen polypeptide raw material, it is characterized in that: it may further comprise the steps:
A: clean: fish scale/fish-skin first through slightly washing, is repeatedly washed with deionized water again, drain;
B: pickling: described fish scale/fish-skin soaks with pH3～5 hydrochloric acid solns, stirring and evenly mixing;
C: washing: the fish scale/fish-skin of described acid treatment washes with water regulates pH to neutral, and then a small amount of water repeatedly washs, and drains;
D: extract: in 2～10: 1 ratio adds entry in fish scale/fish-skin, regulates enzymolysis 4～12h under ℃ condition of pH to 8～9,30～50; Be warming up to 70～100 ℃, keep 5～20min;
E: coarse filtration: the solution that extracts is removed precipitation by filter type;
F: decoloration and deodorization: described step e filtrate is crossed post process decoloration and deodorization;
G: refining: described step f filtrate is passed through the membrane filtration system;
H: drying and moulding.
2. technique according to claim 1 is characterized in that: be fresh at fish scale/fish-skin described in the step a, and free of contamination fish scale/fish-skin.
3. technique according to claim 1 is characterized in that: be the deionized water preparation at hydrochloric acid soln described in the step b, the volume ratio of described fish scale/fish-skin and hydrochloric acid soln is 4～8: 1, and described stir process condition is: 30～50 ℃ of stir process 4～8h.
4. technique according to claim 1, it is characterized in that: the water at the adjusting pH described in the step c is deionized water, washing used water is injection water.
5. technique according to claim 1, it is characterized in that: water used in steps d～g is water for injection.
6. technique according to claim 1 is characterized in that: all carry out in gnotobasis in steps d～g.
7. technique according to claim 1 is characterized in that: be in diatomite, kaolin, Plate Filtration, the membrane filtration one or more at filter type described in the step e.
8. technique according to claim 1 is characterized in that: process and comprise in macropore ion exchange resin, gac, Q-sepharose, the C-18 post one or more crossing post described in the step f.
9. technique according to claim 1 is characterized in that: the system of membrane filtration described in the step g can desalination, removing impurities albumen, degerming, depyrogenation and concentrated.
10. technique according to claim 1 is characterized in that: be a kind of in lyophilize, spraying drying, the vacuum concentration drying at the drying technology described in the step h.
11. medical Isin glue collagen polypeptide raw material such as the arbitrary described technique preparation of claim 1 to 10.
12. medical Isin glue collagen polypeptide raw material according to claim 11, it is characterized in that: the weight-average molecular weight of described medical Isin glue collagen polypeptide raw material is 1000～3000Dolton.
13. medical Isin glue collagen polypeptide raw material according to claim 11 is characterized in that: described medical Isin glue collagen polypeptide raw material is aseptic, pyrogen-free.
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| CN2011102405136A CN102952837A (en) | 2011-08-19 | 2011-08-19 | Medical fish collagen polypeptide raw material and its preparation technology |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104498569A (en) * | 2014-11-24 | 2015-04-08 | 威海市桢昊生物技术有限公司 | Method for preparing fish-skin collagen powder by use of dry fish skin |
| CN105755078A (en) * | 2014-12-19 | 2016-07-13 | 山东国际生物科技园发展有限公司 | Preparation method and application of medical grade fish skin collagens |
| CN106615597A (en) * | 2016-11-15 | 2017-05-10 | 江南大学 | Method for preparing red snapper fish scale collagen protein |
| CN106967169A (en) * | 2017-05-05 | 2017-07-21 | 张军志 | A kind of extracting method of Isin glue collagen |
| CN115605567A (en) * | 2020-05-07 | 2023-01-13 | 诺维信公司（Dk） | Medical cleaning composition, use and cleaning method |
| CN115754092A (en) * | 2022-12-16 | 2023-03-07 | 江西师范大学 | Identification method of gelatin of cyprinus carpio |
| CN117264761A (en) * | 2023-09-25 | 2023-12-22 | 海南华研胶原科技股份有限公司 | Preparation process and system of medical fish collagen polypeptide raw material |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104498569A (en) * | 2014-11-24 | 2015-04-08 | 威海市桢昊生物技术有限公司 | Method for preparing fish-skin collagen powder by use of dry fish skin |
| CN105755078A (en) * | 2014-12-19 | 2016-07-13 | 山东国际生物科技园发展有限公司 | Preparation method and application of medical grade fish skin collagens |
| CN105755078B (en) * | 2014-12-19 | 2020-07-17 | 山东国际生物科技园发展有限公司 | Preparation method and application of medical-grade fish skin collagen |
| CN106615597A (en) * | 2016-11-15 | 2017-05-10 | 江南大学 | Method for preparing red snapper fish scale collagen protein |
| CN106967169A (en) * | 2017-05-05 | 2017-07-21 | 张军志 | A kind of extracting method of Isin glue collagen |
| CN106967169B (en) * | 2017-05-05 | 2020-01-10 | 广东时代食品与生命健康研究有限公司 | Extraction method of fish collagen |
| CN115605567A (en) * | 2020-05-07 | 2023-01-13 | 诺维信公司（Dk） | Medical cleaning composition, use and cleaning method |
| CN115754092A (en) * | 2022-12-16 | 2023-03-07 | 江西师范大学 | Identification method of gelatin of cyprinus carpio |
| CN117264761A (en) * | 2023-09-25 | 2023-12-22 | 海南华研胶原科技股份有限公司 | Preparation process and system of medical fish collagen polypeptide raw material |
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Application publication date: 20130306 |