CN110066845A - The preparation method of pig gall collagen - Google Patents
The preparation method of pig gall collagen Download PDFInfo
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- CN110066845A CN110066845A CN201910469883.3A CN201910469883A CN110066845A CN 110066845 A CN110066845 A CN 110066845A CN 201910469883 A CN201910469883 A CN 201910469883A CN 110066845 A CN110066845 A CN 110066845A
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- 102000008186 Collagen Human genes 0.000 title claims abstract description 110
- 108010035532 Collagen Proteins 0.000 title claims abstract description 110
- 229920001436 collagen Polymers 0.000 title claims abstract description 109
- 238000002360 preparation method Methods 0.000 title claims abstract description 49
- 210000003491 skin Anatomy 0.000 claims abstract description 39
- 210000002615 epidermis Anatomy 0.000 claims abstract description 4
- 239000000284 extract Substances 0.000 claims abstract description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 46
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 45
- 229910001868 water Inorganic materials 0.000 claims description 28
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 22
- 239000007787 solid Substances 0.000 claims description 21
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 20
- 238000003756 stirring Methods 0.000 claims description 20
- 239000008367 deionised water Substances 0.000 claims description 18
- 229910021641 deionized water Inorganic materials 0.000 claims description 18
- 238000000746 purification Methods 0.000 claims description 11
- 102000057297 Pepsin A Human genes 0.000 claims description 10
- 108090000284 Pepsin A Proteins 0.000 claims description 10
- 239000007788 liquid Substances 0.000 claims description 10
- 229940111202 pepsin Drugs 0.000 claims description 10
- 239000011780 sodium chloride Substances 0.000 claims description 10
- 229960000583 acetic acid Drugs 0.000 claims description 9
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 7
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims description 6
- 238000012545 processing Methods 0.000 claims description 6
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 5
- 238000000502 dialysis Methods 0.000 claims description 5
- 238000011010 flushing procedure Methods 0.000 claims description 5
- 239000012362 glacial acetic acid Substances 0.000 claims description 5
- 239000002245 particle Substances 0.000 claims description 5
- 239000006228 supernatant Substances 0.000 claims description 5
- 239000007844 bleaching agent Substances 0.000 claims description 4
- 239000003153 chemical reaction reagent Substances 0.000 claims description 4
- 238000002156 mixing Methods 0.000 claims description 3
- 235000019441 ethanol Nutrition 0.000 claims description 2
- 244000144992 flock Species 0.000 claims description 2
- 238000009777 vacuum freeze-drying Methods 0.000 claims description 2
- 239000013049 sediment Substances 0.000 claims 1
- 238000000034 method Methods 0.000 abstract description 9
- 210000000232 gallbladder Anatomy 0.000 abstract description 6
- 239000002994 raw material Substances 0.000 abstract description 6
- 238000005238 degreasing Methods 0.000 abstract description 3
- 238000011161 development Methods 0.000 abstract description 3
- 238000004108 freeze drying Methods 0.000 abstract description 3
- 210000001519 tissue Anatomy 0.000 abstract description 3
- 230000008901 benefit Effects 0.000 description 5
- 238000005119 centrifugation Methods 0.000 description 5
- 238000012986 modification Methods 0.000 description 5
- 230000004048 modification Effects 0.000 description 5
- 239000000126 substance Substances 0.000 description 5
- 210000000941 bile Anatomy 0.000 description 4
- BHQCQFFYRZLCQQ-OELDTZBJSA-N cholic acid Chemical compound C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC(O)=O)C)[C@@]2(C)[C@@H](O)C1 BHQCQFFYRZLCQQ-OELDTZBJSA-N 0.000 description 4
- 239000003814 drug Substances 0.000 description 4
- 229940088598 enzyme Drugs 0.000 description 4
- 238000001914 filtration Methods 0.000 description 4
- 239000000843 powder Substances 0.000 description 4
- 239000002244 precipitate Substances 0.000 description 4
- BHQCQFFYRZLCQQ-UHFFFAOYSA-N (3alpha,5alpha,7alpha,12alpha)-3,7,12-trihydroxy-cholan-24-oic acid Natural products OC1CC2CC(O)CCC2(C)C2C1C1CCC(C(CCC(O)=O)C)C1(C)C(O)C2 BHQCQFFYRZLCQQ-UHFFFAOYSA-N 0.000 description 3
- 239000004380 Cholic acid Substances 0.000 description 3
- 238000013475 authorization Methods 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 229960002471 cholic acid Drugs 0.000 description 3
- 235000019416 cholic acid Nutrition 0.000 description 3
- KXGVEGMKQFWNSR-UHFFFAOYSA-N deoxycholic acid Natural products C1CC2CC(O)CCC2(C)C2C1C1CCC(C(CCC(O)=O)C)C1(C)C(O)C2 KXGVEGMKQFWNSR-UHFFFAOYSA-N 0.000 description 3
- 238000004090 dissolution Methods 0.000 description 3
- 239000004744 fabric Substances 0.000 description 3
- 239000010419 fine particle Substances 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 150000003384 small molecules Chemical class 0.000 description 3
- 241000282994 Cervidae Species 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 230000007613 environmental effect Effects 0.000 description 2
- 238000009472 formulation Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 239000002699 waste material Substances 0.000 description 2
- 241000251468 Actinopterygii Species 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000010834 Extracellular Matrix Proteins Human genes 0.000 description 1
- 108010037362 Extracellular Matrix Proteins Proteins 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241000237860 Sipunculidae Species 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 210000002808 connective tissue Anatomy 0.000 description 1
- 239000002537 cosmetic Substances 0.000 description 1
- 229960000935 dehydrated alcohol Drugs 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 229960004756 ethanol Drugs 0.000 description 1
- MJEMIOXXNCZZFK-UHFFFAOYSA-N ethylone Chemical compound CCNC(C)C(=O)C1=CC=C2OCOC2=C1 MJEMIOXXNCZZFK-UHFFFAOYSA-N 0.000 description 1
- 210000002744 extracellular matrix Anatomy 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 239000000052 vinegar Substances 0.000 description 1
- 235000021419 vinegar Nutrition 0.000 description 1
Classifications
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- 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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- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Wood Science & Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Microbiology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- Toxicology (AREA)
- Gastroenterology & Hepatology (AREA)
- Biophysics (AREA)
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- Proteomics, Peptides & Aminoacids (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Cosmetics (AREA)
Abstract
The present invention relates to collagen preparation technical fields, and in particular to the preparation method of pig gall collagen.This method is using pig gall skin as raw material, pig gall skin first passes through attachment, the decoloration, degreasing that gallbladder epidermis face is removed by mechanical means, it is extracted again by acid-enzyme binding-method, further by saltouing and dialysing, obtains spongiform pure white collagen finally by freeze-drying.The present invention extracts high-quality collagen from the pig gall skin tissue of abandonment, and deep development high value added product, preparation method is simple, improves the value of pig gall skin.
Description
Technical field
The present invention relates to collagen preparation technical fields, and in particular to the preparation method of pig gall collagen.
Background technique
Collagen is boiomacromolecule, the main component in animal connective tissue, and content is most in the mammalian body
Functional protein more, distribution is most wide, accounts for the 25%~30% of total protein；Collagen is extracellular matrix ingredient,
In vivo with the presence of insoluble macromolecular structure, the unique triple helices structure of collagen keeps molecular structure highly stable；Collagen
Albumen has good biocompatibility, biodegradable and bioactivity, therefore in food, medicine, organizational project, change
The fields such as cosmetic are widely applied.
Pig gall is the gall bladder of animal pig, and main composition is its pig's bile, contains a large amount of cholic acid substances in bile；
Pig gall can directly be used as medicine by Traditional Chinese Medicine, and modern medicine mainly takes its bile, isolate and purify the cholic acid substance for obtaining high-purity,
Further cholic acid analog is obtained by chemical synthesis means.Pig gall skin removes the crust after its bile for the gall bladder of animal pig,
Usually abandoned or simple process is added in feed during extracting cholic acid, causes a large amount of wasting of resources.From the pig of abandonment
High-quality collagen is extracted in gallbladder skin tissue, deep development high value added product not only can solve the pollution problem of waste,
And by comprehensive exploitation and high-value-use, huge environmental benefit and social benefit will be generated.
The material source for preparing of collagen has very much, can pass through the preparations sides such as sour formulation, enzyme formulation, acid-enzyme binding-method
Method obtains.Such as the patent of invention that Authorization Notice No. is CN100345867C, a kind of entitled " system of acid soluble fish skin collagen
Preparation Method " discloses one kind using fish-skin as raw material, obtains collagen by serial extraction purification means；Authorization is public
Number patent of invention for being CN101805775B is accused, it is entitled " a kind of preparation method of collagen from deer sinew ", be exactly with deer's sinew
Collagen is prepared in raw material；The patent of invention of Authorization Notice No. CN101381411B, it is entitled " Sipunculid collagen protein and its
Preparation method " is exactly that collagen is prepared in raw material with siphon-worm body wall；And currently, not yet discovery is using pig gall skin as raw material
Prepare the technical study of collagen.
Summary of the invention
The object of the present invention is to provide a kind of preparation methods of pig gall collagen, using pig gall skin as raw material, pig gall skin
Attachment, the decoloration, degreasing for removing gallbladder epidermis face by mechanical means are first passed through, then is extracted by acid-enzyme binding-method, further
By saltouing and dialysing, spongiform pure white collagen is obtained finally by freeze-drying.
The preparation method of pig gall collagen provided by the invention, comprising the following steps:
S1, the processing of pig gall skin
Fresh pig gall skin removal is attached to the fat and other attachments on surface, with clear water repeated flushing to filtering out
It until water is without obvious color, drains, shreds, be that bleaching agent is added in 1kg:10-30L according to solid-liquid ratio, after impregnating 24-48h, take out
Then pig gall skin is after organic reagent stirring is added in 1kg:5-20L, to take out pig gall skin and cleaned 3-5 times with clear water according to solid-liquid ratio,
It is drained and standby；
S2, the preparation of collagen solution
Deionized water is added according to solid-liquid ratio 1kg:5-10L in the pig gall skin that S1 is handled well, is beaten with refiner homogenate
Broken, continuously adding deionized water to solid-liquid ratio is 1kg:30-50L, and glacial acetic acid, which is added, makes the concentration 0.2- of acetum
1.0mol/L after stirring and dissolving, is added pepsin and the mass concentration of pepsin is made to be 0.1-2.0%, then extract 24-
72h, high speed refrigerated centrifuge, takes supernatant to obtain collagen solution；
S3, the preparation of solid collagen
The collagen solution that S2 is obtained is added solid NaCl and the mass concentration of NaCl is made to be 1.0-5.0%, Bian Jia
Side stirring, obtains white flock precipitate, is cleaned 3-5 times with deionized water, is dehydrated, obtains the solid collagen of water content 80%-95%
Albumen；
S4, the purifying of collagen solution
The solid collagen that S3 is obtained, the acetum that 0.1-2.0mol/L is added are redissolved, after stirring and dissolving, are spent
Ionized water is dialyzate dialysis 2-3d, changes water 4-6 times halfway, obtains collagen solution after purification；
S5, the preparation of pig gall collagen
By the collagen solution after purification of S4, vacuum freeze drying obtains that purity is high, color be white, spongiform pig gall
Collagen.
Preferably, the preparation method of above-mentioned pig gall collagen, in S1, the chopping of pig gall skin refers to being cut into≤0.5cm3
Particle.
Preferably, the preparation method of above-mentioned pig gall collagen, in S1, the bleaching agent is to be containing volume fraction
0.5-5%H2O20.01-0.2mol/L NaOH solution.
Preferably, the preparation method of above-mentioned pig gall collagen is 0.5-5%H2O containing volume fraction20.01-
0.2mol/L NaOH solution is the H for being 0.5-5% by volume fraction2O20.01-0.2mol/L NaOH is added under conditions of stirring
In solution, whipping temp is 15-25 DEG C.
Preferably, the preparation method of above-mentioned pig gall collagen, in S1, the organic reagent be ethyl alcohol, acetone or
One of isopropanol.
Preferably, the preparation method of above-mentioned pig gall collagen, in S1, mixing time 8-24h.
Preferably, the preparation method of above-mentioned pig gall collagen, in S2, it is 8000- that high speed, which freezes centrifugal rotational speed,
12000r/min, time 8-12min.
Preferably, the preparation method of above-mentioned pig gall collagen, in S4, the amount that acetum is added there was not solid collagen
Albumen.
Compared with prior art, the preparation method of pig gall collagen provided by the invention has the advantages that
(1) present invention extracts high-quality collagen from the pig gall skin tissue of abandonment, deep development high value added product, no
Only can solve the pollution problem of waste, and by comprehensive exploitation and high-value-use, will generate huge environmental benefit and
Social benefit；
(2) pig gall skin is first passed through attachment, the decoloration, degreasing that gallbladder epidermis face is removed by mechanical means by the present invention, then
It is extracted by acid-enzyme binding-method, further by saltouing and dialysing, obtains spongiform pure white coloring agent finally by freeze-drying
Former albumen, preparation method is simple, improves the value of pig gall skin.
Specific embodiment
The present invention is described in detail With reference to embodiment, it is to be understood that protection scope of the present invention is simultaneously
It is not restricted by specific implementation.The test method of actual conditions is not specified in the following example, usually according to normal condition
Operation, the test material source being not specified be it is commercially available, due to not being related to inventive point, therefore its step is not described in detail.
Embodiment 1
The preparation method of pig gall collagen, comprising the following steps:
S1, the processing of pig gall skin
Fresh pig gall skin 5kg is weighed, fresh pig gall skin removal is attached to the fat and other attachments on surface, with clear
Water repeated flushing drains until the water filtered out is without obvious color, is chopped into 0.5cm3Fine particle；Containing for 50L is added
The H that volume fraction is 0.5%2O20.01mol/L NaOH solution, impregnate for 24 hours, take out pig gall skin；It is added at 25L dehydrated alcohol
12h is managed, is at the uniform velocity stirred, pig gall Pi Houyong clear water is taken out and cleans 5 times, it is drained and standby；
S2, the preparation of collagen solution
The pig gall skin powder that S1 is handled well is taken 1000g (moisture content 90%), 5L deionized water is added, it will be broken with homogenizer
End is further smashed in fine-fibrous, is continuously added 25L deionized water, is then put into reaction kettle, and glacial acetic acid, which is added, to be made
The concentration of acetum is 0.5mol/L, and after continuing stirring dissolution in 15 minutes, the quality that pepsin makes pepsin is added
Concentration is 1%, after then extracting 24 hours, releases completely reacted collagen, refrigerated centrifuge centrifugation, 10000r/min centrifugation
It 10 minutes, takes out supernatant and obtains collagen solution, reject precipitates particle；
S3, the preparation of solid collagen
The collagen solution that S2 is obtained, which pours into agitator, saltouts, and is slowly added to sodium chloride and makes the quality of NaCl dense
Degree is 1.0%, and stirring while adding, flocculent deposit is constantly precipitated；It is used after the collagen saltoutd is cleaned 3 times with deionized water
Then filter-cloth filtering is installed with filter bag and is tightened, be then dehydrated with dewaterer, obtains the solid collagen that water content is 80%；
S4, the purifying of collagen solution
The solid collagen that S3 is obtained is placed in agitator, the uninterrupted 0.5mol/L aqueous acetic acid that is added is not to having
Solid collagen is crossed, stirring and dissolving is that dialyzate dialysis 3d removes small-molecule substance with deionized water up to being completely dissolved, in
Way is changed water 5 times, and collagen solution after purification is obtained；
S5, the preparation of pig gall collagen
It collagen solution will be freeze-dried to obtain spongy collagen 62.5g after purification.
The H for being 0.5% containing volume fraction in the present embodiment2O20.01mol/L NaOH solution be to be by volume fraction
0.5% H2O2It is added in 0.01mol/L NaOH solution under conditions of stirring, whipping temp is 20 DEG C.
Embodiment 2
The preparation method of pig gall collagen, comprising the following steps:
S1, the processing of pig gall skin
Fresh pig gall skin 5kg is weighed, fresh pig gall skin removal is attached to the fat and other attachments on surface, with clear
Water repeated flushing drains until the water filtered out is without obvious color, is chopped into 0.5cm3Fine particle；Containing for 100L is added
The H that volume fraction is 2%2O20.1mol/L NaOH solution, impregnate 48h, take out pig gall skin；100L acetone treatment 8h is added,
It at the uniform velocity stirs, takes out pig gall Pi Houyong clear water and clean 3 times, it is drained and standby；
S2, the preparation of collagen solution
The pig gall skin powder that S1 is handled well is taken 1000g (moisture content 90%), 10L deionized water is added, it will with homogenizer
Powder is further smashed in fine-fibrous, is continued to add 40L deionized water, then be put into reaction kettle, and glacial acetic acid, which is added, to be made
The concentration of acetum is 0.2mol/L, and after continuing stirring dissolution in 15 minutes, the quality that pepsin makes pepsin is added
Concentration is 0.1%, after then extracting 72 hours, releases completely reacted collagen, refrigerated centrifuge centrifugation, 12000r/min from
It the heart 8 minutes, takes out supernatant and obtains collagen solution, reject precipitates particle；
S3, the preparation of solid collagen
The collagen solution that S2 is obtained, which pours into agitator, saltouts, and is slowly added to sodium chloride and makes the quality of NaCl dense
Degree is 5.0%, and stirring while adding, flocculent deposit is constantly precipitated；It is used after the collagen saltoutd is cleaned 5 times with deionized water
Then filter-cloth filtering is installed with filter bag and is tightened, be then dehydrated with dewaterer, obtains the solid collagen that water content is 95%；
S4, the purifying of collagen solution
The solid collagen that S3 is obtained is placed in agitator, the uninterrupted 0.1mol/L aqueous acetic acid that is added is not to having
Solid collagen is crossed, stirring and dissolving is that dialyzate dialysis 2d removes small-molecule substance with deionized water up to being completely dissolved, in
Way is changed water 4 times, and collagen solution after purification is obtained；
S5, the preparation of pig gall collagen
It collagen solution will be freeze-dried to obtain spongy collagen 72.36g after purification.
The H for being 2% containing volume fraction in the present embodiment2O20.1mol/L NaOH solution be by volume fraction be 2%
H2O2It is added in 0.1mol/L NaOH solution under conditions of stirring, whipping temp is 15 DEG C.
Embodiment 3
The preparation method of pig gall collagen, comprising the following steps:
S1, the processing of pig gall skin
Fresh pig gall skin 5kg is weighed, fresh pig gall skin removal is attached to the fat and other attachments on surface, with clear
Water repeated flushing drains until the water filtered out is without obvious color, is chopped into 0.5cm3Fine particle；Containing for 150L is added
The H that volume fraction is 5%2O20.2mol/L NaOH solution, impregnate 36h, take out pig gall skin；50L isopropanol is added and handles 8h,
It at the uniform velocity stirs, takes out pig gall Pi Houyong clear water and clean 4 times, it is drained and standby；
S2, the preparation of collagen solution
The pig gall skin powder that S1 is handled well is taken 1000g (moisture content 90%), 8L deionized water is added, it will be broken with homogenizer
End is further smashed in fine-fibrous, is continued to add 32L deionized water, then be put into reaction kettle, and glacial acetic acid, which is added, makes vinegar
The concentration of acid solution is 1.0mol/L, after continuing stirring dissolution in 15 minutes, pepsin is added and makes the quality of pepsin dense
Degree is 2.0%, after then extracting 48 hours, releases completely reacted collagen, refrigerated centrifuge centrifugation, 8000r/min centrifugation
It 12 minutes, takes out supernatant and obtains collagen solution, reject precipitates particle；
S3, the preparation of solid collagen
The collagen solution that S2 is obtained, which pours into agitator, saltouts, and is slowly added to sodium chloride and makes the quality of NaCl dense
Degree is 2.0%, and stirring while adding, flocculent deposit is constantly precipitated；It is used after the collagen saltoutd is cleaned 4 times with deionized water
Then filter-cloth filtering is installed with filter bag and is tightened, be then dehydrated with dewaterer, obtains the solid collagen that water content is 90%；
S4, the purifying of collagen solution
The solid collagen that S3 is obtained is placed in agitator, the uninterrupted 2.0mol/L aqueous acetic acid that is added is not to having
Solid collagen is crossed, stirring and dissolving is that dialyzate dialysis 2d removes small-molecule substance with deionized water up to being completely dissolved, in
Way is changed water 6 times, and collagen solution after purification is obtained；
S5, the preparation of pig gall collagen
It collagen solution will be freeze-dried to obtain spongy collagen 81.42g after purification.
The H for being 5% containing volume fraction in the present embodiment2O20.2mol/L NaOH solution be by volume fraction be 5%
H2O2It is added in 0.2mol/L NaOH solution under conditions of stirring, whipping temp is 25 DEG C.
It should be noted that involved in the present invention when numberical range, it is thus understood that two endpoints of each numberical range with
And any one numerical value can be selected between two endpoints, since the step method of use is identical as embodiment, go to live in the household of one's in-laws on getting married in order to prevent
It states, the present invention describes preferred embodiment., but technology people in the art although preferred embodiments of the present invention have been described
Member is once know basic creative concept, then additional changes and modifications can be made to these embodiments.So appended right
It is required that being intended to be construed to includes preferred embodiment and all change and modification for falling into the scope of the invention.
Obviously, various changes and modifications can be made to the invention without departing from essence of the invention by those skilled in the art
Mind and range.In this way, if these modifications and changes of the present invention belongs to the range of the claims in the present invention and its equivalent technologies
Within, then the present invention is also intended to include these modifications and variations.
Claims (8)
1. the preparation method of pig gall collagen, which comprises the following steps:
S1, the processing of pig gall skin
Pig gall epidermis face is removed except the fat and other attachments of attachment, with clear water repeated flushing to the water filtered out without obvious color
Until, it drains, shreds, be that bleaching agent is added in 1kg:10-30L according to solid-liquid ratio, after impregnating 24-48h, take out pig gall skin, then
It is that organic reagent processing is added in 1kg:5-20L according to solid-liquid ratio, takes out pig gall skin and cleaned with clear water, it is drained and standby；
S2, the preparation of collagen solution
Deionized water homogenate is added according to solid-liquid ratio 1kg:5-10L in pig gall skin that S1 is handled well to smash, continuously add from
Sub- water to solid-liquid ratio is 1kg:30-50L, and glacial acetic acid, which is added, makes the concentration 0.2-1.0mol/L of acetum, after stirring and dissolving,
Pepsin, which is added, makes the mass concentration of pepsin be 0.1-2.0%, then extracts 24-72h, refrigerated centrifuge takes supernatant
Liquid obtains collagen solution；
S3, the preparation of solid collagen
NaCl is added in the collagen solution of S2, so that the mass concentration of NaCl is 1.0-5.0%, it is heavy to obtain white flock
It forms sediment, is washed with deionized water, be dehydrated, obtain the solid collagen of water content 80%-95%；
S4, the purifying of collagen solution
The acetum that 0.1-2.0mol/L is added in the solid collagen that S3 is obtained redissolves, and then uses deionized water dialysis 2-
3d changes water 4-6 times halfway, obtains collagen solution after purification；
S5, the preparation of pig gall collagen
By the collagen solution vacuum freeze drying after purification of S4, pig gall collagen is obtained.
2. the preparation method of pig gall collagen according to claim 1, which is characterized in that in S1, the chopping of pig gall skin
It refers to being cut into≤0.5cm3Particle.
3. the preparation method of pig gall collagen according to claim 1, which is characterized in that in S1, the bleaching agent
To be 0.5-5%H containing volume fraction2O20.01-0.2mol/L NaOH solution.
4. the preparation method of pig gall collagen according to claim 3 is 0.5-5%H containing volume fraction2O2's
The preparation method of 0.01-0.2mol/L NaOH solution is by H2O2The NaOH solution of 0.01-0.2mol/L is added at 15-25 DEG C
In, it obtains after mixing evenly.
5. the preparation method of pig gall collagen according to claim 1, which is characterized in that in S1, described is organic
Reagent is one of ethyl alcohol, acetone or isopropanol.
6. the preparation method of pig gall collagen according to claim 1, which is characterized in that in S1, mixing time is
8-24h。
7. the preparation method of pig gall collagen according to claim 1, which is characterized in that in S2, freeze at a high speed from
Heart revolving speed is 8000-12000r/min, time 8-12min.
8. the preparation method of pig gall collagen according to claim 1, which is characterized in that in S4, it is molten that acetic acid is added
The amount of liquid there was not solid collagen.
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|---|---|---|---|---|
| CN111500522A (en) * | 2020-04-12 | 2020-08-07 | 江苏安泰康健康科技有限公司 | Culture system based on pigskin collagen and three-dimensional tissue culture method |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101092449A (en) * | 2007-06-18 | 2007-12-26 | 南京农业大学 | Technique for preparing collagen of freshwater 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 |
-
2019
- 2019-05-31 CN CN201910469883.3A patent/CN110066845A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101092449A (en) * | 2007-06-18 | 2007-12-26 | 南京农业大学 | Technique for preparing collagen of freshwater 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 |
Non-Patent Citations (2)
| Title |
|---|
| 郭振武: "《实用胆囊外科》", 30 June 1996 * |
| 马克昌等: "《骨生理学》", 31 March 2000, 河南医科大学出版社 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111500522A (en) * | 2020-04-12 | 2020-08-07 | 江苏安泰康健康科技有限公司 | Culture system based on pigskin collagen and three-dimensional tissue culture method |
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