CN103588901A - Method for extraction of heparin sodium from small intestines of pigs - Google Patents
Method for extraction of heparin sodium from small intestines of pigs Download PDFInfo
- Publication number
- CN103588901A CN103588901A CN201310532605.0A CN201310532605A CN103588901A CN 103588901 A CN103588901 A CN 103588901A CN 201310532605 A CN201310532605 A CN 201310532605A CN 103588901 A CN103588901 A CN 103588901A
- Authority
- CN
- China
- Prior art keywords
- sodium
- heparin sodium
- resin
- wash
- precipitation
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Polysaccharides And Polysaccharide Derivatives (AREA)
Abstract
The invention discloses a method for extraction of heparin sodium from small intestines of pigs. The method comprises the steps of: (1) adsorption; (2) washing; (3) elution; (4) separation; (5) protein removal; and (6) precipitation. The preparation method provided by the invention takes porcine small intestines' mucous membranes with extensive sources as raw materials, and the cost is low. The method employs a polyacrylonitrile membrane to perform separation, not only saves a lot of ethanol, reduces the production cost, but also can obtain refine heparin sodium with high purity, thus ensuring the heparin activity.
Description
Technical field
The invention belongs to chemical field, relate to a kind of extracting method of heparin sodium, be specifically related to a kind of method of extracting heparin sodium from chitterlings.
Background technology
Heparin mainly extracts from ox lung or intestinal mucosa, is a kind of mucopolysaccharide sulfuric acid ester being alternately comprised of glucosamine, L-idose aldehyde glycosides, N-Acetyl-D-glucosamine and D-Glucose aldehydic acid.Heparin makes it expose arginine reactive center by being combined with antithrombin Ⅲ, is combined and makes its deactivation, main deactivation X a and the II a factor with the thrombin of activation.Heparin is mainly used in prevention and treatment thrombotic disease, as myocardial infarction, pulmonary infarction, cerebral vessels embolism, peripheral vein thrombus etc., and formation and expansion that can anti-hemostasis suppository.Also can be used for the anti-freezing of early stage and other inside and outside of DIC.
At present, heparin sodium is mainly to extract from pig, sheep small intestine mucous membrane and ox lung, salted casings salt solution etc.What its extracting method was the most frequently used is salt solution or enzymolysis-ion-exchange-ethanol precipitation, but these two kinds of problems that the method ubiquity rate of recovery is low, purity is not high.
Summary of the invention
Technical problem to be solved by this invention is to provide a kind of reasonable in design, method and from chitterlings, extracts reliably the method for heparin sodium.
For solving above technical problem, a kind of method of extracting heparin sodium from chitterlings of the present invention, comprises the steps:
(1) absorption:
Take intestinal mucosa and put in reactor, thin up, regulates pH value to 9.0～9.5 with 30% sodium hydroxide solution, be heated to boil, keep 10～15min, be cooled to 35 ℃～45 ℃, filtrate siphon simultaneously, to reaction tank, adds 5～10% resins, insulated and stirred absorption 8～10h;
(2) washing:
After getting the standing 30min of liquid after step (1) absorption, filter, collect resin, and water is clean by resin rinsing, drains standby;
(3) wash-out:
The solid of getting step (2) adds 0.8～1.5mol/L sodium-chlor washing, 1～3h, after draining, adds 1～3 times of amount of 3～5mol/L sodium-chlor, stirs wash-out 5h, and wash-out 2 times again after draining, merges whole elutriants;
(4) separation:
In the situation that stirring, use polyacrylonitrile film to carry out separation the elutriant of step (3), filtering velocity remains on 30～40ml/min, and make part on film account for cumulative volume 10～20% time, stop suction filtration;
(5) isolating protein:
Get on film and hold back part, add DC-W, fully stir 20min, standing 5h under room temperature, filtering precipitation;
(6) precipitation:
Get the filtrate of step (5), add 95% ethanol of 1～2 times, after fully stirring, standing over night, siphon upper strata alcohol liquid, precipitation is respectively with 95% ethanol, acetone dehydration 2 times, and vacuum-drying, obtains refined heparin sodium.
As improvement of the present invention, described in step (1), resin is D204 resin.
During wash-out 2 times, add 3～5mol/L sodium chloride solution amount to reduce by half in above-mentioned steps (3), elution time is respectively 2h, 1h at every turn.
The polyacrylonitrile film of above-mentioned steps (4) is the polyacrylonitrile film of holding back Mr3000～4000.
Compared with prior art, the beneficial effect that the present invention has is:
Preparation method provided by the invention be take the intestinal mucosa widely of originating and is raw material, with low cost.The present invention uses polyacrylonitrile film to carry out separation, not only can save a large amount of ethanol, reduce production costs, and the refined heparin sodium purity of preparation is high, has guaranteed the activity of heparin.
Embodiment
For ease of understanding the present invention, it is as follows that the present invention enumerates embodiment.Those skilled in the art should understand, described embodiment only, for helping to understand the present invention, should not be considered as concrete restriction of the present invention.
As no specific instructions, various raw material of the present invention all can obtain by commercially available; Or prepare according to the ordinary method of this area.Unless otherwise defined or described herein, all specialties used herein and scientific words and the art technology same meaning that skillfully person of entering is familiar.In addition any method similar or impartial to described content and material all can be applicable in the inventive method.
Unless otherwise defined or described herein, the familiar same meaning of all specialties used herein and scientific words and those skilled in the art.In addition any method similar or impartial to described content and material all can be applicable in the inventive method.
Embodiment 1
A method of extracting heparin sodium from chitterlings, comprises the steps:
(1) absorption:
Take intestinal mucosa 500kg and put in reactor, thin up, regulates pH value to 9.0～9.5 with 30% sodium hydroxide solution, be heated to boil, keep 15min, be cooled to 35 ℃, filtrate siphon simultaneously, to reaction tank, adds 10%D204 resin resin, insulated and stirred absorption 10h;
(2) washing:
After getting the standing 30min of liquid after step (1) absorption, filter, collect resin, and water is clean by resin rinsing, drains standby;
(3) wash-out:
The solid of getting step (2) adds 1mol/L sodium-chlor washing 1～3h, after draining, adds 1～3 times of amount of 4mol/L sodium-chlor, stir wash-out 5h, wash-out 2 times again after draining adds 4mol/L sodium chloride solution amount to reduce by half at every turn, elution time is respectively 2h, 1h, merges whole elutriants.
(4) separation:
In the situation that stirring, use the polyacrylonitrile film of Mr3000～4000 to carry out separation the elutriant of step (3), filtering velocity remains on 30ml/min, and make part on film account for cumulative volume 15% time, stop suction filtration;
(5) isolating protein:
Get on film and hold back part, in the ratio of DC-W:TQ=1.0:50.0～1.0:80.0, add DC-W, fully stir 20min, standing 5h under room temperature, filtering precipitation;
(6) precipitation:
Get the filtrate of step (5), add 95% ethanol of 1.5 times, after fully stirring, standing over night, siphon upper strata alcohol liquid, precipitation is respectively with 95% ethanol, acetone dehydration 2 times, and vacuum-drying, obtains refined heparin sodium.
Extracting method of the present invention is compared with conventional salt solution-ion-exchange-ethanol precipitation, saves ethanol 85% left and right, and the rate of recovery will improve more than 20%, and the rate of recovery improves more than 1% than the membrane separation process of the conventional use of the industry.
Embodiment 2
A method of extracting heparin sodium from chitterlings, comprises the steps:
(1) absorption:
Take intestinal mucosa 500kg and put in reactor, thin up, regulates pH value to 9.0 with 30% sodium hydroxide solution, be heated to boil, keep 15min, be cooled to 45 ℃, filtrate siphon simultaneously, to reaction tank, adds 10%D204 resin resin, insulated and stirred absorption 10h;
(2) washing:
After getting the standing 30min of liquid after step (1) absorption, filter, collect resin, and water is clean by resin rinsing, drains standby;
(3) wash-out:
The solid of getting step (2) adds 0.8mol/L sodium-chlor washing 3h, after draining, adds 3 times of amounts of 4mol/L sodium-chlor, stir wash-out 5h, wash-out 2 times again after draining adds 4mol/L sodium chloride solution amount to reduce by half at every turn, elution time is respectively 2h, 1h, merges whole elutriants.
(4) separation:
In the situation that stirring, use the polyacrylonitrile film of Mr3000～4000 to carry out separation the elutriant of step (3), filtering velocity remains on 30ml/min, and make part on film account for cumulative volume 10% time, stop suction filtration;
(5) isolating protein:
Get on film and hold back part, in the ratio of DC-W:TQ=1.0:50.0～1.0:80.0, add DC-W, fully stir 20min, standing 5h under room temperature, filtering precipitation;
(6) precipitation:
Get the filtrate of step (5), add 95% ethanol of 1.5 times, after fully stirring, standing over night, siphon upper strata alcohol liquid, precipitation is respectively with 95% ethanol, acetone dehydration 2 times, and vacuum-drying, obtains refined heparin sodium.
Embodiment 3
A method of extracting heparin sodium from chitterlings, comprises the steps:
(1) absorption:
Take intestinal mucosa 500kg and put in reactor, thin up, regulates pH value to 9.5 with 30% sodium hydroxide solution, be heated to boil, keep 15min, be cooled to 35 ℃, filtrate siphon simultaneously, to reaction tank, adds 10%D204 resin resin, insulated and stirred absorption 10h;
(2) washing:
After getting the standing 30min of liquid after step (1) absorption, filter, collect resin, and water is clean by resin rinsing, drains standby;
(3) wash-out:
The solid of getting step (2) adds 1.5mol/L sodium-chlor washing 1～3h, after draining, adds 2 times of amounts of 4mol/L sodium-chlor, stir wash-out 5h, wash-out 2 times again after draining adds 4mol/L sodium chloride solution amount to reduce by half at every turn, elution time is respectively 2h, 1h, merges whole elutriants.
(4) separation:
In the situation that stirring, use the polyacrylonitrile film of Mr3000～4000 to carry out separation the elutriant of step (3), filtering velocity remains on 30ml/min, and make part on film account for cumulative volume 20% time, stop suction filtration;
(5) isolating protein:
Get on film and hold back part, in the ratio of DC-W:TQ=1.0:50.0～1.0:80.0, add DC-W, fully stir 20min, standing 5h under room temperature, filtering precipitation;
(6) precipitation:
Get the filtrate of step (5), add 95% ethanol of 2 times, after fully stirring, standing over night, siphon upper strata alcohol liquid, precipitation is respectively with 95% ethanol, acetone dehydration 2 times, and vacuum-drying, obtains refined heparin sodium.
Applicant's statement, person of ordinary skill in the field is on the basis of above-described embodiment, by the concrete content point value of above-described embodiment component, combined with the technical scheme of summary of the invention part, thereby the new numerical range producing, also be one of record scope of the present invention, the application, for making specification sheets simple and clear, is no longer enumerated these numerical ranges.
Claims (4)
1. from chitterlings, extract a method for heparin sodium, it is characterized in that comprising the steps:
(1) absorption:
Take intestinal mucosa and put in reactor, thin up, regulates pH value to 9.0～9.5 with 30% sodium hydroxide solution, be heated to boil, keep 10～15min, be cooled to 35 ℃～45 ℃, filtrate siphon simultaneously, to reaction tank, adds 5～10% resins, insulated and stirred absorption 8～10h;
(2) washing:
After getting the standing 30min of liquid after step (1) absorption, filter, collect resin, and water is clean by resin rinsing, drains standby;
(3) wash-out:
The solid of getting step (2) adds 0.8～1.5mol/L sodium-chlor washing, 1～3h, after draining, adds 1～3 times of amount of 3～5mol/L sodium-chlor, stirs wash-out 5h, and wash-out 2 times again after draining, merges whole elutriants;
(4) separation:
In the situation that stirring, use polyacrylonitrile film to carry out separation the elutriant of step (3), filtering velocity remains on 30～40ml/min, and make part on film account for cumulative volume 10～20% time, stop suction filtration;
(5) isolating protein:
Get on film and hold back part, add DC-W, fully stir 20min, standing 5h under room temperature, filtering precipitation;
(6) precipitation:
Get the filtrate of step (5), add 95% ethanol of 1～2 times, after fully stirring, standing over night, siphon upper strata alcohol liquid, precipitation is respectively with 95% ethanol, acetone dehydration 2 times, and vacuum-drying, obtains refined heparin sodium.
2. according to a kind of method of extracting heparin sodium from chitterlings described in claim 1, it is characterized in that: described in step (1), resin is D204 resin.
3. according to a kind of method of extracting heparin sodium from chitterlings described in claim 1, it is characterized in that: during wash-out 2 times, add 3～5mol/L sodium chloride solution amount to reduce by half in described step (3), elution time is respectively 2h, 1h at every turn.
4. according to a kind of method of extracting heparin sodium from chitterlings described in claim 1, it is characterized in that: the polyacrylonitrile film of described step (4) is the polyacrylonitrile film of holding back Mr3000～4000.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310532605.0A CN103588901A (en) | 2013-10-31 | 2013-10-31 | Method for extraction of heparin sodium from small intestines of pigs |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310532605.0A CN103588901A (en) | 2013-10-31 | 2013-10-31 | Method for extraction of heparin sodium from small intestines of pigs |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN103588901A true CN103588901A (en) | 2014-02-19 |
Family
ID=50079248
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310532605.0A Pending CN103588901A (en) | 2013-10-31 | 2013-10-31 | Method for extraction of heparin sodium from small intestines of pigs |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103588901A (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4409103A (en) * | 1980-02-29 | 1983-10-11 | Italfarmaco S.P.A. | Method for the preparation of calcium heparinate |
| CN101597344A (en) * | 2009-05-07 | 2009-12-09 | 张丽萍 | A kind of extraction of heparin, separation, purification process |
-
2013
- 2013-10-31 CN CN201310532605.0A patent/CN103588901A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4409103A (en) * | 1980-02-29 | 1983-10-11 | Italfarmaco S.P.A. | Method for the preparation of calcium heparinate |
| CN101597344A (en) * | 2009-05-07 | 2009-12-09 | 张丽萍 | A kind of extraction of heparin, separation, purification process |
Non-Patent Citations (1)
| Title |
|---|
| 夏华等: "膜分离与沉淀结合法提取猪小肠黏膜中肝素钠的工艺研究", 《卫生职业教育》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103183747B (en) | Trypsin method extracts the technique of high-purity heparin sodium from intestinal mucosa | |
| CN102229681A (en) | Preparation method for producing heparin sodium by using porcine small intestines | |
| CN106589167B (en) | The preparation method and its process units of high-titer crude heparin sodium | |
| CN103951768A (en) | Method for extraction of heparin sodium and co-production of polypeptide protein powder and amino acid protein powder by utilizing porcine and bovine lungs | |
| CN102952204A (en) | Novel production technique of heparin sodium | |
| CN1978467B (en) | Method for separating and purifying lentinan | |
| CN103183746B (en) | Process for extracting heparin sodium comprehensively by small intestines of pigs | |
| CN103183748B (en) | Technology for extracting heparin sodium from intestinal mucosa with trypsin method | |
| CN106046188B (en) | A kind of preparation method of fucoidin | |
| CN103275206B (en) | Pregnant mare serum gonadotropin (PMSG) purifying method | |
| CN104072639A (en) | Production method for nadroparin calcium | |
| CN103183744B (en) | Using alkali proteinase method extracts the technique of high-purity heparin sodium from intestinal mucosa | |
| CN105621478A (en) | Washing impurity removal method for metatitanic acid in production of titanium dioxide by sulfuric acid method | |
| CN103848929A (en) | Process for high-efficiently extracting sodium heparin | |
| CN104387504A (en) | Method for preparing heparin sodium by using small intestines of pigs | |
| CN106967186B (en) | A method of extracting heparin-like compound from air bladder | |
| CN103588902A (en) | Separation purification method of refined heparin sodium | |
| CN104829750A (en) | Refining process of heparin lithium | |
| CN104530260A (en) | Method for co-producing high-purity heparin sodium and dermatan sulfate from pig lungs | |
| CN104119457A (en) | Heparin sodium and dermatan sulfate combined production process | |
| CN103588901A (en) | Method for extraction of heparin sodium from small intestines of pigs | |
| CN103601820A (en) | Preparation method of heparin lithium | |
| CN104945539A (en) | Energy-saving low-temperature heparin sodium extraction technology | |
| CN105440164A (en) | Method for improving purity of heparin sodium | |
| CN107641161A (en) | A kind of optimal reparation technology of casing accessory substance liquaemin |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20140219 |
|
| WD01 | Invention patent application deemed withdrawn after publication |