CN103724426A - Improved method for extracting insulin from animal pancreas - Google Patents
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- CN103724426A CN103724426A CN201310638708.5A CN201310638708A CN103724426A CN 103724426 A CN103724426 A CN 103724426A CN 201310638708 A CN201310638708 A CN 201310638708A CN 103724426 A CN103724426 A CN 103724426A
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- 102000004877 Insulin Human genes 0.000 title claims abstract description 33
- 108090001061 Insulin Proteins 0.000 title claims abstract description 33
- 210000000496 pancreas Anatomy 0.000 title claims abstract description 30
- 238000000034 method Methods 0.000 title claims abstract description 27
- 241001465754 Metazoa Species 0.000 title claims abstract description 15
- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Chemical compound N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 title abstract description 12
- 229940125396 insulin Drugs 0.000 title abstract description 6
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 45
- 239000002253 acid Substances 0.000 claims abstract description 11
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims abstract description 10
- 238000001556 precipitation Methods 0.000 claims abstract description 9
- 239000011780 sodium chloride Substances 0.000 claims abstract description 5
- 238000010438 heat treatment Methods 0.000 claims abstract 4
- 239000007788 liquid Substances 0.000 claims description 19
- 239000006228 supernatant Substances 0.000 claims description 14
- 239000000284 extract Substances 0.000 claims description 13
- 230000002378 acidificating effect Effects 0.000 claims description 9
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 8
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 8
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims description 8
- 239000000706 filtrate Substances 0.000 claims description 8
- 238000005406 washing Methods 0.000 claims description 8
- 238000003756 stirring Methods 0.000 claims description 7
- 239000002994 raw material Substances 0.000 claims description 5
- 101800000263 Acidic protein Proteins 0.000 claims description 4
- 101710093543 Probable non-specific lipid-transfer protein Proteins 0.000 claims description 4
- 238000001914 filtration Methods 0.000 claims description 4
- 238000004108 freeze drying Methods 0.000 claims description 4
- 238000007710 freezing Methods 0.000 claims description 4
- 230000008014 freezing Effects 0.000 claims description 4
- 238000000338 in vitro Methods 0.000 claims description 4
- 239000007787 solid Substances 0.000 claims description 4
- 238000010025 steaming Methods 0.000 claims description 4
- 238000001291 vacuum drying Methods 0.000 claims description 4
- 238000001816 cooling Methods 0.000 claims 3
- 241000282894 Sus scrofa domesticus Species 0.000 claims 1
- 238000001035 drying Methods 0.000 claims 1
- 238000002360 preparation method Methods 0.000 claims 1
- 108090000623 proteins and genes Proteins 0.000 abstract description 3
- 230000006837 decompression Effects 0.000 abstract 1
- 150000002148 esters Chemical class 0.000 abstract 1
- 239000012530 fluid Substances 0.000 abstract 1
- 102000004169 proteins and genes Human genes 0.000 abstract 1
- 238000005185 salting out Methods 0.000 abstract 1
- 235000019441 ethanol Nutrition 0.000 description 19
- 239000000243 solution Substances 0.000 description 11
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 6
- 238000005516 engineering process Methods 0.000 description 4
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 3
- 229960000583 acetic acid Drugs 0.000 description 3
- YAJCHEVQCOHZDC-QMMNLEPNSA-N actrapid Chemical compound C([C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@H]1CSSC[C@H]2C(=O)N[C@H](C(=O)N[C@@H](CO)C(=O)N[C@H](C(=O)N[C@@H](C(N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC=3C=CC(O)=CC=3)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC=3C=CC(O)=CC=3)C(=O)N[C@@H](CSSC[C@H](NC(=O)[C@H](C(C)C)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=3C=CC(O)=CC=3)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](C)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=3N=CNC=3)NC(=O)[C@H](CO)NC(=O)CNC1=O)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)NCC(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H]([C@H](C)O)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@H](C)O)C(O)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O)=O)CSSC[C@@H](C(N2)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](NC(=O)CN)[C@H](C)CC)[C@H](C)CC)[C@H](C)O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](NC(=O)[C@@H](NC(=O)[C@@H](N)CC=1C=CC=CC=1)C(C)C)C(N)=O)C1=CNC=N1 YAJCHEVQCOHZDC-QMMNLEPNSA-N 0.000 description 3
- 238000013019 agitation Methods 0.000 description 3
- 206010012601 diabetes mellitus Diseases 0.000 description 3
- 238000000605 extraction Methods 0.000 description 3
- 239000012362 glacial acetic acid Substances 0.000 description 3
- 239000008103 glucose Substances 0.000 description 3
- 229950004152 insulin human Drugs 0.000 description 3
- 239000002244 precipitate Substances 0.000 description 3
- 239000012266 salt solution Substances 0.000 description 3
- 230000008901 benefit Effects 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 108090000765 processed proteins & peptides Proteins 0.000 description 2
- BDERNNFJNOPAEC-UHFFFAOYSA-N propan-1-ol Chemical compound CCCO BDERNNFJNOPAEC-UHFFFAOYSA-N 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- BWGNESOTFCXPMA-UHFFFAOYSA-N Dihydrogen disulfide Chemical compound SS BWGNESOTFCXPMA-UHFFFAOYSA-N 0.000 description 1
- 229920002527 Glycogen Polymers 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 102000035195 Peptidases Human genes 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 239000003513 alkali Substances 0.000 description 1
- 230000006538 anaerobic glycolysis Effects 0.000 description 1
- 210000000227 basophil cell of anterior lobe of hypophysis Anatomy 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000023852 carbohydrate metabolic process Effects 0.000 description 1
- 235000021256 carbohydrate metabolism Nutrition 0.000 description 1
- 239000000747 designer drug Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000012407 engineering method Methods 0.000 description 1
- 230000004110 gluconeogenesis Effects 0.000 description 1
- 229940096919 glycogen Drugs 0.000 description 1
- 230000004116 glycogenolysis Effects 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000002797 proteolythic effect Effects 0.000 description 1
- 238000010298 pulverizing process Methods 0.000 description 1
- 239000002510 pyrogen Substances 0.000 description 1
- 238000007634 remodeling Methods 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 230000001568 sexual effect Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
Classifications
-
- 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
- C07K14/575—Hormones
- C07K14/62—Insulins
- C07K14/625—Extraction from natural sources
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biochemistry (AREA)
- Genetics & Genomics (AREA)
- Gastroenterology & Hepatology (AREA)
- Toxicology (AREA)
- Endocrinology (AREA)
- Biophysics (AREA)
- General Health & Medical Sciences (AREA)
- Zoology (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Diabetes (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Peptides Or Proteins (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The invention provides an improved method for extracting insulin from animal pancreas. The technical scheme adopted is that the method comprises the following steps: adopting an acid alcohol fluid method to damage the pancreas for extracting the insulin; adjusting pH according to an isoelectric precipitation principle; performing alkalifying and acidation; removing impure protein; carrying decompression concentration for further purifying; removing ethanol; heating to remove ester; performing sodium chloride salting-out to prepare the insulin. Due to the adoption of the method, the insulin more than 49 g can be obtained from the pancreas of per kg.
Description
Technical field
The invention belongs to field of biological pharmacy, the biochemical isolation technique of Regular Insulin for designer drug.Specifically adopt acid alcohol to extract concentrating under reduced pressure method and extract the Regular Insulin in animal pancreas.
Background technology
Regular Insulin is the albumen sexual hormoue of Mammals beta Cell of islet secretion, contains two polypeptide chains that are connected with disulfide linkage in molecule.Regular Insulin dissolves in the rare alcohol below 80%, rare propyl alcohol and acid, alkali, be insoluble to more than 90% ethanol and other organic solvents, pH value be 2～4 o'clock more stable, meet proteolytic enzyme, strong acid, highly basic easily destroyed, add thermal lability, its iso-electric point is 5.3～5.4.
Regular Insulin can regulate carbohydrate metabolism, reduces gluconeogenesis, promotes glycogen synthetic, suppresses glycogenolysis, accelerates anaerobic glycolysis and the aerobic oxidation of glucose, promotes the utilization of tissue to glucose, and can promote that glucose changes fat into, thereby can reduce blood sugar.
Diabetes are one of modal metabolic troubles in the world, and the latest data of announcing for 2011 according to IDF shows, global diabetes number of patients has reached 3.66 hundred million.Clinical diabetes patient uses insulin treatment to maintain the normal level of blood sugar more.Insulin human is also widely used clinically in recent years.At present, the peptide method that turns of using gene engineering and proteolytic ferment is successfully produced insulin human.Gene engineering method is produced insulin human and is invested greatly, complex process.
Traditional processing technology, from from animal (pig) purification Regular Insulin, adopts sour-ol liquid by its extracting and separating, after being separated out, carries out separation and purification by saltouing.Result: remove impurity albumen, zwitterion, pyrogen material etc., can obtain Regular Insulin.
Summary of the invention
Object of the present invention is exactly the yield in order to improve Regular Insulin, and traditional technology is transformed, and controls alkalization, the acidizing PH value of sour-ol extraction liquid well, has obtained a kind of method of extracting Regular Insulin from animal pancreas of improvement, and the method comprises the steps:
1, raw materials pretreatment: after pancreas is in vitro, deeply freeze immediately, first proceed to-15～28 ℃ ℃ after anxious freezing at-20～-40 ℃ and save backup.
2, acid alcohol liquid extracts: pancreas freeze-drying adds the acidic ethanol glacial acetic acid of 2.0～3.0 times of amounts to adjust pH2.0～3.2 after pulverizing, and ethanol content is 60%～75%, at 8～15 ℃, stirs and extracts 3h, centrifugal, gets supernatant liquor.Filter residue extracts with aforesaid method with 1.0～2.0 times of amount acidic ethanols again.Merge extracted twice liquid.
3, alkalization, acidifying: extracting solution adds strong aqua to adjust pH7.8～8.5 (10～15 ℃ of liquid temperatures) under constantly stirring, carry out immediately press filtration, remove basic protein, filtrate should be clarified, and in time with sulfuric acid acidation to pH3.5～3.8, be cooled to 4～5 ℃, standing 4～8h, fully precipitates acidic protein.
4, concentrating under reduced pressure: inhale supernatant liquid to concentrating under reduced pressure pot, lower floor filters, and throw out discards.Filtrate is incorporated to supernatant liquor, at 30 ℃ of following pressure reducing and steaming ethanol, is concentrated into concentrated solution proportion and is 1.04～1.06 (be about original volume 1/10～1/9 till).
5, degrease, saltout: concentrated solution proceeds in degrease pot and in 5min, is heated to after 48～55 ℃, is cooled to 4～5 ℃ immediately with icy salt solution, and standing 3～4h, isolates subnatant.Adjust pH2.3～2.5 with hydrochloric acid, under agitation add 0.25～0.3 times of amount solid sodium chloride in 20～25 ℃, be incubated standing 3～8h, separate out precipitation.
6, washing, dry: filter, precipitation washing with acetone three times, 10～30 ℃ of vacuum-dryings, obtain Regular Insulin, weigh.
Compared with prior art, advantage of the present invention and positively effect are:
Adopt acid alcohol to extract concentrating under reduced pressure method and extract the Regular Insulin in animal pancreas, can improve the efficiency of extracting Regular Insulin, and make its physico-chemical property and biological activity keep stable, thereby be conducive to next step, Regular Insulin is refined, finally made the indices of Regular Insulin all meet Chinese Pharmacopoeia standard.The more important thing is, by technique update and perfect, make Regular Insulin yield bring up to 0.4%～0.6%, cost-saving, improved economic benefit.
Embodiment
Below in conjunction with specific embodiment, the present invention will be further described.
Embodiment 1
(1) raw materials pretreatment: after pancreas is in vitro, deeply freeze immediately, first proceed to-20 ℃ after-30 ℃ of following anxious freezing and save backup.
(2) acid alcohol liquid extracts: get freeze-drying pancreas piece 4kg, add the acidic ethanol (acidic ethanol control content is 68%, and adjusts pH2.5 with glacial acetic acid) of 11.2L after broken with plane pancreas power blader,, at 12 ℃, stir extraction 3h, centrifugal, get supernatant liquor 10.1L.Filter residue extracts 1h with 10.1L acidic ethanol with aforesaid method again, centrifugal, gets supernatant liquor.Merge extracted twice liquid.
(3) alkalization, acidifying: extracting solution adds strong aqua to adjust pH8.0 (12 ℃ of liquid temperatures) under constantly stirring, and carries out immediately press filtration, removes basic protein, filtrate should be clarified, and in time with sulfuric acid acidation to pH3.6, be cooled to 5 ℃, standing 4h, fully precipitates acidic protein.
(4) concentrating under reduced pressure: inhale supernatant liquid to concentrating under reduced pressure pot, lower floor filters with canvas, and throw out discards.Filtrate is incorporated to supernatant liquor, and pressure reducing and steaming ethanol at 20 ℃ is concentrated into concentrated solution proportion and is 1.04 (be about original volume 1/10).
(5) degrease, saltout: concentrated solution proceeds in degrease pot with in 5min and is heated to after 50 ℃, is cooled to 5 ℃ immediately with icy salt solution, and standing 4h, isolates subnatant.Adjust pH2.3 with hydrochloric acid, under agitation add 0.25 times of amount solid sodium chloride in 20 ℃, be incubated standing 4h, separate out precipitation.
(6) washing, dry: filter, precipitation washing with acetone three times, 20 ℃ of vacuum-dryings, obtain Regular Insulin, and 20.4g weighs.
Embodiment 2
(1) raw materials pretreatment: after pancreas is in vitro, deeply freeze immediately, first proceed to-15 ℃ after-35 ℃ of following anxious freezing and save backup.
(2) acid alcohol liquid extracts: get freeze-drying pancreas piece 4kg, add the acidic ethanol (acidic ethanol control content is 75%, and adjusts pH2.9 with glacial acetic acid) of 9.2L after broken with plane pancreas power blader,, at 10 ℃, stir extraction 3h, centrifugal, get supernatant liquor 7.95L.Filter residue extracts 1h with 7.95L acidic ethanol with aforesaid method again, centrifugal, gets supernatant liquor.Merge extracted twice liquid.
(3) alkalization, acidifying: extracting solution adds strong aqua to adjust pH8.3 (10 ℃ of liquid temperatures) under constantly stirring, and carries out immediately press filtration, removes basic protein, filtrate should be clarified, and in time with sulfuric acid acidation to pH3.4, be cooled to 4 ℃, standing 6h, fully precipitates acidic protein.
(4) concentrating under reduced pressure: inhale supernatant liquid to concentrating under reduced pressure pot, lower floor filters with canvas, and throw out discards.Filtrate is incorporated to supernatant liquor, and pressure reducing and steaming ethanol at 20 ℃ is concentrated into concentrated solution proportion and is 1.06 (be about original volume 1/9).
(5) degrease, saltout: concentrated solution proceeds in degrease pot with in 5min and is heated to after 55 ℃, is cooled to 4 ℃ immediately with icy salt solution, and standing 6h, isolates subnatant.Adjust pH2.5 with hydrochloric acid, under agitation add 0.28 times of amount solid sodium chloride in 20 ℃, be incubated standing 6h, separate out precipitation.
(6) washing, dry: filter, precipitation washing with acetone three times, 20 ℃ of vacuum-dryings, obtain Regular Insulin, and 19.2g weighs.
The above, be only preferred embodiment of the present invention, is not the restriction of the present invention being made to other form, and any those skilled in the art may utilize the technology contents of above-mentioned announcement to be changed or be modified as the equivalent embodiment of equivalent variations.But every technical solution of the present invention content that do not depart from, any simple modification, equivalent variations and the remodeling above embodiment done according to technical spirit of the present invention, still belong to the protection domain of technical solution of the present invention.
Claims (9)
1. a method of extracting Regular Insulin from animal pancreas for improvement, is characterized in that, comprises the following steps: (1) raw materials pretreatment; (2) acid alcohol liquid extracts; (3) acidifying, alkalization; (4) concentrating under reduced pressure; (5) degrease, saltout.
2. preparation method described in claim 1, is characterized in that, each step is specially:
(1) raw materials pretreatment: after pancreas is in vitro, deeply freeze immediately.
(2) acid alcohol liquid extracts: after pancreas freeze-drying shatters, add acidic ethanol to stir and extract, and centrifugal, get supernatant liquor.Filter residue extracts with aforesaid method again.Merge extracted twice liquid.
(3) alkalization, acidifying: extracting solution adds strong aqua to adjust pH, and press filtration, removes basic protein, filtrate sulfuric acid acidation, cooling, standing, acidic protein is fully precipitated.
(4) concentrating under reduced pressure: inhale supernatant liquid to concentrating under reduced pressure pot, lower floor filters, and throw out discards.Filtrate is incorporated to supernatant liquor, and pressure reducing and steaming ethanol is concentrated.
(5) degrease, saltout: concentrated solution heating cooling degrease.With hydrochloric acid, adjust pH, add 0.25～0.3 times of amount solid sodium chloride, be incubated standingly, separate out precipitation.
(6) washing, dry: filter, precipitation washing with acetone, vacuum-drying, obtains Regular Insulin.
3. the method for extracting Regular Insulin from animal pancreas of a kind of improvement according to claim 2, is characterized in that, described pancreas is Pancreas Bovis seu Bubali or Pancreas Sus domestica.
4. the method for extracting Regular Insulin from animal pancreas of a kind of improvement according to claim 2, is characterized in that, in described step (1), pancreas adopts-20～-40 ℃ of quick freezings, and storage temperature is-15～28 ℃.
5. the method for extracting Regular Insulin from animal pancreas of a kind of improvement according to claim 2, is characterized in that, in described step (2), acid alcohol liquid pH is 2.0～2.9, and ethanol content is 60%～75%, and whipping temp is 8～15 ℃.
6. the method for extracting Regular Insulin from animal pancreas of a kind of improvement according to claim 2, is characterized in that, in described step (3), alkalization pH is 7.8～8.5, and acidifying pH is 3.4～3.8.
7. the method for extracting Regular Insulin from animal pancreas of a kind of improvement according to claim 2, is characterized in that, adopts concentrating under reduced pressure method to boil off ethanol in described step (4), and temperature is 25～35 ℃.
8. the method for extracting Regular Insulin from animal pancreas of a kind of improvement according to claim 2, is characterized in that, in described step (5), concentrated solution adds rapid heating cooling degrease in degrease pot, and the temperature of heating degrease is: 45～52 ℃.
9. the method for extracting Regular Insulin from animal pancreas of a kind of improvement according to claim 2, is characterized in that, adopts electric vacunm drying case to be dried in described step (6).
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103739702A (en) * | 2013-11-30 | 2014-04-23 | 青岛康原药业有限公司 | Method for extraction of insulin |
| CN105384808A (en) * | 2015-11-25 | 2016-03-09 | 青岛康原药业有限公司 | Method for extracting insulin from hog pancreas |
| CN116874586A (en) * | 2023-07-24 | 2023-10-13 | 盐城凯利药业有限公司 | Insulin extraction process |
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| CN1076931A (en) * | 1992-03-31 | 1993-10-06 | 中国科学院上海生物化学研究所 | Insulin antagonistic peptide and its separation purification method |
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2013
- 2013-11-29 CN CN201310638708.5A patent/CN103724426A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1076931A (en) * | 1992-03-31 | 1993-10-06 | 中国科学院上海生物化学研究所 | Insulin antagonistic peptide and its separation purification method |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103739702A (en) * | 2013-11-30 | 2014-04-23 | 青岛康原药业有限公司 | Method for extraction of insulin |
| CN105384808A (en) * | 2015-11-25 | 2016-03-09 | 青岛康原药业有限公司 | Method for extracting insulin from hog pancreas |
| CN116874586A (en) * | 2023-07-24 | 2023-10-13 | 盐城凯利药业有限公司 | Insulin extraction process |
| CN116874586B (en) * | 2023-07-24 | 2025-03-18 | 盐城凯利药业有限公司 | A kind of insulin extraction process |
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Application publication date: 20140416 |