CN104403025A - Heparin sodium decolorization method - Google Patents
Heparin sodium decolorization method Download PDFInfo
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- CN104403025A CN104403025A CN201410814688.7A CN201410814688A CN104403025A CN 104403025 A CN104403025 A CN 104403025A CN 201410814688 A CN201410814688 A CN 201410814688A CN 104403025 A CN104403025 A CN 104403025A
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Abstract
The invention discloses a heparin sodium decolorization method, belonging to the technical field of heparin sodium purification methods. The method comprises the following steps: (1) dissolving heparin sodium in water; (2) regulating the pH value of the heparin sodium solution with hydrochloric acid; (3) adding sodium sulfite, and stirring to react for 0.5 hour; (4) adding 1 wt% kieselguhr, filtering, and collecting the filtrate; (5) regulating the pH value of the filtrate to 4.0-4.5 with a sodium hydroxide solution; (6) adding 0.5-0.7 wt% EDTA (ethylene diamine tetraacetic acid) to react for 1 hour; (7) adding 2.5% sodium chloride solid; (8) keeping the solution temperature at 55-65 DEG C, adding 0.8 time of ethanol, stirring for 0.5 hour, and introducing cooling water to cool to 20-25 DEG C; and (9) standing the mixture obtained in the step (8) to precipitate, removing the supernatant waste ethanol, and dehydrating and drying the understratum precipitate to obtain the heparin sodium product. The method can effectively destroy the metal bridge between the heparin sodium and the protein impurity and iron element, does not destroy the heparin sodium structure, has small destroy actions on the activity of the heparin sodium, and has the advantages of stable quality and high yield.
Description
Technical field
The present invention relates to a kind of heparin sodium except color method, effectively can remove the color of heparin sodium, belong to heparin sodium purification method and technology field.
Background technology
Heparin sodium be use clinically at most, antithrombotics the most widely, occur with the composite form of albumen in vivo.This mixture does not have anticoagulant active, only has it to separate separately just can show anticoagulant active.Heparin sodium is widely distributed at organic sphere, is present in higher mammal, as in the lung tissue of pig, ox, sheep etc., intestinal tissue.Common heparin sodium separate mode has enzymolysis or salt solution, alcohol precipitation, oxidation etc., due to source and the extracting mode of heparin, must contain a large amount of nucleic acid and protein impurities in crude product heparin, darker color.
The raw materials for production of current domestic heparin sodium are mainly chitterlings, the color of heparin sodium produces has direct relation with the ferro element in protein impurities and blood, these impurity can form metal bridge with heparin sodium, use common removing protein mode or directly add metal chelating agent removing effects all not obvious, cause the step directly except look in treating process, as the potassium permanganate oxidation, hydrogen peroxide oxidation or the mode of both couplings that generally adopt, the usage quantity of oxygenant is larger, destruction is defined to the structure of heparin sodium, reduces quality and the yield of product.
Summary of the invention
The object of the present invention is to provide one can not affect heparin sodium activity, steady quality, the heparin sodium that yield is high removes color method.
The present invention adopts following technical scheme: a kind of heparin sodium, except color method, comprises the following steps:
(1) dissolved by heparin sodium with water, the heparin sodium aqua concentration finally obtained is 6000 ~ 8000IU/ml;
(2) pH of the heparin sodium aqua using hydrochloric acid step (1) to be obtained is adjusted to 1.0 ~ 2.5;
(3) step (2) gained solution is added S-WAT while stirring, stirring reaction 0.5h;
(4) adding mass concentration in the solution obtained to step (3) is 1% diatomite, filters and collects filtrate;
(5) use the filtrate of sodium hydroxide solution regulating step (4) gained, regulate while stirring, make pH=4.0 ~ 4.5;
(6) adding mass concentration in the solution obtained to step (5) is 0.5 ~ 0.7%EDTA, reaction 1h;
(7) add 2.5% solid sodium chloride in the solution obtained to step (6), stir and solid sodium chloride is dissolved;
(8) temperature of solution step (7) obtained keeps 55 ~ 65 DEG C, adds 0.8 times of ethanol, passes into water coolant, make gained solution temperature be cooled to 20 ~ 25 DEG C after stirring 0.5h;
(9) mixture that step (8) obtains is staticly settled, the supernatant liquid after precipitation is waste ethanol, lower sediment thing is mixture containing heparin sodium, pump upper strata waste ethanol, heparin sodium product is obtained to the dehydration of lower sediment thing, drying.
In step (3), the mass concentration of the S-WAT added is 0.2 ~ 1.0%.
In step (4), after collecting by filtration filtrate, with water, to rinse in diatomite residual tires, and residual tiring to be collected and in the filtrate be mixed into collected by step (4).The object economized on resources can be reached like this.
In step (6), while adding 0.5 ~ 0.7%EDTA, solution is heated to 55 ~ 65 DEG C, makes solution keep 55 ~ 65 DEG C of reaction 1h.
The invention has the beneficial effects as follows: the present invention uses hydrochloric acid by the pH regulator of heparin sodium aqua to acid, and can either make partial impurities sex change, playing the effect removing albumen, can be again follow-up removal ferro element create environment condition.Through the heparin solution of pre-treatment, can be good at removing ferro element under the coupling effect of S-WAT and metal chelating agent EDTA, serve the single complexing agent that adds and to be beyond one's reach effect.Meanwhile, the good separate part protein impurities of mode of subsequent thermal precipitation and by the ferro element of complexing, these impurity are separated totally with heparin sodium, and follow precipitation supernatant waste liquid and be separated, removal color effects is good.By the heparin sodium of method process of the present invention, except directly color can be reduced, decrease the impact of wherein iron ion simultaneously, decrease the consumption of potassium permanganate or hydrogen peroxide in follow-up treating process, reduce oxygenant to heparin sodium structural damage, improve product quality, improve product yield.The present invention is by regulating reaction environment, promote that metal chelating agent plays a role, metal bridge between effective destruction heparin sodium and protein impurities and ferro element, do not destroy heparin sodium structure again, then, while removal protein impurities with removal ferro element, effectively reduce the consumption of oxygenant in follow-up treating process, reduce the destruction of heparin sodium activity, stabilised quality, improves yield.
Embodiment
Below in conjunction with specific embodiment, the present invention is described in detail.
Embodiment 1:
The heparin sodium of the present embodiment, except color method, comprises the following steps:
(1) dissolved by heparin sodium with water, the heparin sodium aqua concentration finally obtained is 6000IU/ml;
(2) pH of the heparin sodium aqua using hydrochloric acid step (1) to be obtained is adjusted to 2.5;
(3) step (2) gained solution being added mass concentration is while stirring 1.0% S-WAT, stirring reaction 0.5h;
(4) adding mass concentration in the solution obtained to step (3) is 1% diatomite, filters and collects filtrate, and with water, to rinse in diatomite residual tires, and residual tiring collected and be mixed in collected filtrate;
(5) use the filtrate of sodium hydroxide solution regulating step (4) gained, regulate while stirring, make pH=4.0;
(6) adding mass concentration in solution step (5) obtained is 0.5%EDTA, and reaction 1h, while adding EDTA, is heated to 55 DEG C by solution, makes solution keep 55 DEG C to react 1h;
(7) add 2.5% solid sodium chloride in the solution obtained to step (6), stir and solid sodium chloride is dissolved;
(8) temperature of the solution obtained to step (7) keeps 55 DEG C, adds 0.8 times of ethanol, passes into water coolant, make gained solution temperature be cooled to 20 DEG C after stirring 0.5h;
(9) mixture that step (8) obtains is staticly settled, the supernatant liquid after precipitation is waste ethanol, lower sediment thing is mixture containing heparin sodium, pump upper strata waste ethanol, heparin sodium product is obtained to the dehydration of lower sediment thing, drying.
Result the present embodiment being obtained to heparin sodium product is analyzed: in the per-cent reduce heparin sodium color and standing process, the minimizing ratio of potassium permanganate usage quantity is tested.
Through the numerical value of UV spectrophotometer measuring OD400nm, by this index Show Color clearance, detect the method colour removal rate of this embodiment is 30%.
To the product of process of the present invention be used, and use potassium permanganate oxidation except look, use the numerical value of UV spectrophotometer measuring OD400nm, by this index Show Color index, result be presented at make color up to standard basis on, potassium permanganate consumption reduces 55%.
Embodiment 2:
The heparin sodium of the present embodiment, except color method, comprises the following steps:
(1) dissolved by heparin sodium with water, the heparin sodium aqua concentration finally obtained is 8000IU/ml;
(2) pH of the heparin sodium aqua using hydrochloric acid step (1) to be obtained is adjusted to 1.0;
(3) step (2) gained solution being added mass concentration is while stirring 0.2% S-WAT, stirring reaction 0.5h;
(4) adding mass concentration in the solution obtained to step (3) is 1% diatomite, filters and collects filtrate, and with water, to rinse in diatomite residual tires, and residual tiring collected and be mixed in collected filtrate;
(5) use the filtrate of sodium hydroxide solution regulating step (4) gained, regulate while stirring, make pH=4.5;
(6) adding mass concentration in solution step (5) obtained is 0.7%EDTA, and reaction 1h, while adding EDTA, is heated to 65 DEG C by solution, makes solution keep 65 DEG C to react 1h;
(7) add 2.5% solid sodium chloride in solution step (6) obtained, stir and solid sodium chloride is dissolved;
(8) temperature of solution step (7) obtained keeps 65 DEG C, adds 0.8 times of ethanol, passes into water coolant, make gained solution temperature be cooled to 25 DEG C after stirring 0.5h;
(9) mixture that step (8) obtains is staticly settled, the supernatant liquid after precipitation is waste ethanol, lower sediment thing is mixture containing heparin sodium, pump upper strata waste ethanol, heparin sodium product is obtained to the dehydration of lower sediment thing, drying.
Result the present embodiment being obtained to heparin sodium product is analyzed: in the per-cent reduce heparin sodium color and standing process, the minimizing ratio of potassium permanganate usage quantity is tested.Through the numerical value of UV spectrophotometer measuring OD400nm, by this index Show Color clearance, detect this test colour removal rate is 38%.
To the product of process of the present invention be used, and use potassium permanganate oxidation except look, use the numerical value of UV spectrophotometer measuring OD400nm, by this index Show Color index, result be presented at make color up to standard basis on, potassium permanganate consumption reduces 61%.
The present invention uses hydrochloric acid by the pH regulator of heparin sodium aqua to acid, and can either make partial impurities sex change, playing the effect removing albumen, can be again follow-up removal ferro element create environment condition.Through the heparin solution of pre-treatment, can be good at removing ferro element under the coupling effect of S-WAT and metal chelating agent EDTA, serve the single complexing agent that adds and to be beyond one's reach effect.Meanwhile, the good separate part protein impurities of mode of subsequent thermal precipitation and by the ferro element of complexing, these impurity are separated totally with heparin sodium, and follow precipitation supernatant waste liquid and be separated, removal color effects is good.By the heparin sodium of method process of the present invention, except directly color can be reduced, decrease the impact of wherein iron ion simultaneously, decrease the consumption of potassium permanganate or hydrogen peroxide in follow-up treating process, reduce oxygenant to heparin sodium structural damage, improve product quality, improve product yield.The present invention is by regulating reaction environment, promote that metal chelating agent plays a role, metal bridge between effective destruction heparin sodium and protein impurities and ferro element, do not destroy heparin sodium structure again, then, while removal protein impurities with removal ferro element, effectively reduce the consumption of oxygenant in follow-up treating process, reduce the destruction of heparin sodium activity, stabilised quality, improves yield.
In other embodiments of the present invention, while adding EDTA, solution can be heated to the arbitrary temp between 55 ~ 65 DEG C.
In other embodiments of the present invention, the mass concentration of the S-WAT added can be the arbitrary value between 0.2 ~ 1.0%.
In other embodiments of the present invention, after collecting by filtration filtrate, can save with water that to rinse in diatomite residual tires, step.
Claims (4)
1. heparin sodium is except a color method, and it is characterized in that, it comprises the following steps:
(1) dissolved by heparin sodium with water, the heparin sodium aqua concentration finally obtained is 6000 ~ 8000IU/ml;
(2) pH of the heparin sodium aqua using hydrochloric acid step (1) to be obtained is adjusted to 1.0 ~ 2.5;
(3) step (2) gained solution is added S-WAT while stirring, stirring reaction 0.5h;
(4) adding mass concentration in the solution obtained to step (3) is 1% diatomite, filters and collects filtrate;
(5) use the filtrate of sodium hydroxide solution regulating step (4) gained, regulate while stirring, make pH=4.0 ~ 4.5;
(6) adding mass concentration in the solution obtained to step (5) is 0.5 ~ 0.7%EDTA, reaction 1h;
(7) add 2.5% solid sodium chloride in the solution obtained to step (6), stir and solid sodium chloride is dissolved;
(8) temperature of solution step (7) obtained keeps 55 ~ 65 DEG C, adds 0.8 times of ethanol, passes into water coolant, make gained solution temperature be cooled to 20 ~ 25 DEG C after stirring 0.5h;
(9) mixture that step (8) obtains is staticly settled, the supernatant liquid after precipitation is waste ethanol, lower sediment thing is mixture containing heparin sodium, pump upper strata waste ethanol, heparin sodium product is obtained to the dehydration of lower sediment thing, drying.
2. heparin sodium according to claim 1 is except color method, and it is characterized in that: in step (3), the mass concentration of the S-WAT added is 0.2 ~ 1.0%.
3. heparin sodium according to claim 1 is except color method, it is characterized in that: in step (4), after collecting by filtration filtrate, and with water, to rinse in diatomite residual tires, and residual tiring to be collected and in the filtrate be mixed into collected by step (4).
4. heparin sodium according to claim 1 is except color method, it is characterized in that: in step (6), while adding 0.5 ~ 0.7%EDTA, solution is heated to 55 ~ 65 DEG C, makes solution keep 55 ~ 65 DEG C of reaction 1h.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106967188A (en) * | 2017-05-31 | 2017-07-21 | 广元市海鹏生物科技有限公司 | A kind of liquaemin removes color method |
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| CN101942039A (en) * | 2010-09-16 | 2011-01-12 | 山东海科化工集团有限公司 | Parnaparin production method |
| CN102020724A (en) * | 2010-12-23 | 2011-04-20 | 安徽丰原发酵技术工程研究有限公司 | Method for extracting sodium hyaluronate from fermentation liquor containing hyaluronic acid |
| CN102757516A (en) * | 2012-08-03 | 2012-10-31 | 常州千红生化制药股份有限公司 | Decoloration method of enoxaparin sodium |
| RU2512768C1 (en) * | 2012-12-18 | 2014-04-10 | Федеральное бюджетное учреждение "Государственный институт кровезаменителей и медицинских препаратов (ФБУ "ГИКиМП") | Method of obtaining low-molecular heparin |
| CN104098716A (en) * | 2014-07-16 | 2014-10-15 | 南京健友生化制药股份有限公司 | Production method of dalteparin sodium fine product |
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Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101942039A (en) * | 2010-09-16 | 2011-01-12 | 山东海科化工集团有限公司 | Parnaparin production method |
| CN102020724A (en) * | 2010-12-23 | 2011-04-20 | 安徽丰原发酵技术工程研究有限公司 | Method for extracting sodium hyaluronate from fermentation liquor containing hyaluronic acid |
| CN102757516A (en) * | 2012-08-03 | 2012-10-31 | 常州千红生化制药股份有限公司 | Decoloration method of enoxaparin sodium |
| RU2512768C1 (en) * | 2012-12-18 | 2014-04-10 | Федеральное бюджетное учреждение "Государственный институт кровезаменителей и медицинских препаратов (ФБУ "ГИКиМП") | Method of obtaining low-molecular heparin |
| CN104098716A (en) * | 2014-07-16 | 2014-10-15 | 南京健友生化制药股份有限公司 | Production method of dalteparin sodium fine product |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106967188A (en) * | 2017-05-31 | 2017-07-21 | 广元市海鹏生物科技有限公司 | A kind of liquaemin removes color method |
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