CN102040671A - Process for preparing and purifying ultra low molecular weight heparin - Google Patents
Process for preparing and purifying ultra low molecular weight heparin Download PDFInfo
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Abstract
The invention discloses a process for preparing and purifying ultra low molecular weight heparin sodium (calcium), which comprises the following steps of: reacting heparin with organic quaternary ammonium salt to generate heparin quaternary ammonium salt, performing nucleophilic substitution to generate heparin benzyl ester, and degrading under the alkaline condition to obtain a low molecular weight heparin fragment; and separating and purifying by an inorganic ceramic ultrafiltration and hollow fiber ultrafiltration combined method to obtain the ultra low molecular weight heparin sodium (calcium) of which the molecular weight distribution is 2,000 to 2,500D and the average molecular weight is 2,200D. The low molecular weight heparin fragment is obtained by controlling reaction conditions in the esterification process and the degradation time of ester hydrolysis; a ceramic membrane and a hollow fiber ultrafiltration membrane are combined to separate and purify the heparin fragment; and by selecting the pore diameter of the ceramic membrane, operating pressure, feed liquid temperature, and the molecular weight cutoff of the hollow ceramic membrane, the heparin fragment with a reasonable molecular weight distribution range is effectively separated.
Description
Technical field
The present invention relates to a kind of preparation and purifying process of ultra-low molecular weight heparin, particularly, relate to and utilize β-elimination edman degradation Edman to prepare ultra-low molecular weight heparin sodium or the calcium salt of molecular weight ranges between 2000D-2500D, utilize the production technique of ceramic super-filtering film separation and purification, belong to biomedicine field.
Background technology
Heparin (heparin) is a kind of glycosaminoglycan that extensively is present in animal organ and the tissue (as mucous membrane of small intestine, lung etc.), have multiple pharmacologically actives such as anti-freezing, anti-inflammatory, antianaphylaxis, antiviral, anticancer, regulating blood fat, but life-time service then may produce some untoward reaction, as hemorrhage and induced platelet minimizing etc.(low molecular weight heparin LMWH) is heparin fractionated or degraded and the less fragment of molecular mass that obtains to Low molecular heparin.Over nearly 10 years, people discover that to Low molecular heparin its anti thrombotic action is better than heparin, have long half time in the bioavailability height, body, bleeding tendency is little, characteristics such as oral easy absorption.Ultra-low molecular weight heparin (ultra low molecular weight heparin, ULMWH) be the antithrombotic reagent of another new variety of on the Low molecular heparin basis, developing, compare with Low molecular heparin, its anti thrombotic action is strong, subcutaneous injection easily absorbs, and has no side effect, and is mainly used in the prevention of thrombotic disease, as the prevention of formation, old phlebothrombosis and the cardiovascular and cerebrovascular diseases of nephrotic syndrome, operation posterior vein thrombus, anti-curing hyperlipemia etc.
Because the validity of low molecular weight heparin aspect treatment and prevention venous thromboembolism in a plurality of countries listings such as America and Europes, has the trend that replaces traditional heparin.Therefore, in conjunction with clinical practice, the Low molecular heparin of development desired molecular weight and molecular weight distribution and ultra-low molecular weight heparin product have very wide prospect.
China is the country that produces heparin amount maximum in the world, and the export volume of crude product heparin occupy the first place of biochemical drug always.Therefore the external low molecular weight heparin product of producing of a large amount of imports of China, can utilize the heparin raw material resources of China's abundant to produce its substitute products, improves added value of product but simultaneously.
Pharmacological research shows, reduction along with the heparin molecule amount, its anticoagulant active comparatively fast descends, but and the closely-related anti-xa activity of anti-thrombus function descends relatively slow, based on this discovery, people work out has higher antithrombotic acitivity (FXa) and low bleeding tendency---low anticoagulant active (F II heparin fragment a).Studies show that, the not anti-xa activity of the heparin fragment of molecular weight below 2000D, though molecular weight has higher anti-xa activity greater than the fragment of 8000D, anticoagulant active is also higher, does not meet in the pharmacopeia requirement about FXa/F II a.
The preparation method of low molecular weight heparin can be divided three classes substantially: i.e. physical partition method, cracking process and synthesis method.The mechanical degradation reaction is because the control difficulty can't realize industrialization production.Cracking process divides chemical cracking process and enzymatic lysis method, and the chemical cracking method mainly contains nitrous acid control depolymerization edman degradation Edman, peroxide degradation method, β-elimination edman degradation Edman, free radical catalyzed degradation method, phenmethylization and alkaline degradation method, high sulfonic acid oxidation degradation method etc.The enzymatic lysis method is the heparinase edman degradation Edman.DeR difference, the end structure of product are also different.The enzyme liberating method is lacked competitiveness owing to production cost is too high.Synthesis method is a direction of abroad in recent years heparin being studied, and utilizes different glycosyl synthetic heparin fragments, does not also carry out industrialization production.The low molecular weight heparin main product all makes with chemical degradation method on the world market at present, the bent heparin sodium of receiving that the dalteparin sodium that Pfizer produces, Ge Lansu produce has all adopted the Sodium Nitrite degradation method, the Enoxaparin Sodium of Sanofi-Aventis production has adopted the method for eliminating degraded, Opocin is with hydrogen peroxide-cuprous chloride degraded, and Novartis adopts the Isopentyl nitrite degraded to produce low molecular weight heparin sodium.The existing publication number of China is the patent of invention of CN101519459A, the production technique that a kind of employing Sodium Nitrite degraded, anion column fractionation by adsorption, ultrafiltration purification, ethanol sedimentation, lyophilize prepare ultra-low molecular weight heparin sodium (calcium) is disclosed, the ultra-low molecular weight heparin molecular-weight average of preparation is 2300-2900D, and the ratio of molecular weight between 1500-5000D reaches 80%.
With the low molecular weight heparin of chemical method degraded be molecular weight from hundreds of to the segmental mixture of nearly 10,000 different molecular weight, therefore need to adopt effective separation purification method.The fractionation precipitation separating and purifying technology that adopts is difficult to isolate the narrow fragment of range of molecular weight distributions at present; The gel permeation chromatography that adopts can be isolated rational fragment and distribute, but exists trivial operations, filler cost than problems such as height; The development of ultrafiltration membrane technique in recent years provides a kind of good molecular weight separation purification method, but because the organic membrane poor heat resistance, acid-and base-resisting and antimicrobial erosional competency difference etc., the application in actual production is subjected to certain restriction.
Summary of the invention
The objective of the invention is to improve present ultra-low molecular weight heparin preparation and separating and purifying technology, a kind of preparation and purifying process of ultra-low molecular weight heparin is provided.
The objective of the invention is to realize by following technical scheme:
Because no matter low molecular weight heparin is anticoagulating active or anti-thrombus activity, all be integrated as prerequisite with heparin and Antithrombin III (ATIII), this combination requires heparin that a pentasaccharides unit must be arranged, and this structure is active necessary by anti-FXa of heparin and anti-FIIa.In addition, anti-FIIa activity not only combines with ATIII on pentasaccharides, also must combine and could express with ATIII on ten trisaccharides, so segmental anti-FXa of low molecular weight heparin and anti-FIIa are active and in close relations by the palliating degradation degree of heparin.If only remaining pentasaccharides unit behind the depolymerized heparin, the Low molecular heparin of making so can only have the activity of anti-FXa, if pentasaccharides unit also has been degraded, the activity of so anti-FXa also will be lost, so reaction conditions needs precise control, guarantee that the Low molecular heparin relative molecular mass of gained is not less than 2000D.
The present invention is raw material with the heparin sodium, in the aqueous solution, generate the heparin quaternary ammonium salt with the organic quaternary ammonium salt effect, the carboxyl of heparin quaternary ammonium salt and benzyl chlorine generation nucleophilic substitution reaction generate the heparin benzyl ester in dichloromethane solution, and under alkaline condition, the ester hydrolytic deterioration obtains low-molecular-weight heparin fragment.Can control the growing amount of heparin benzyl ester by the reaction conditions in the control esterification process, the high more segmental molecular weight of postheparin of then degrading of growing amount is low more, and the length of ester hydrolytic deterioration time is relevant with the structure and the physiologically active of the ultra-low molecular weight heparin that obtains.
The crude product that the degraded postprecipitation obtains, adopt ceramic membrane to carry out separation and purification in conjunction with hollow fiber ultrafiltration membrane, by selecting aperture, working pressure and the feed temperature of ceramic membrane, and the molecular weight cut-off of hollow-fibre membrane, the effective rational fragment of isolated molecule amount distribution range, simultaneously inorganic ceramic film also have high temperature resistant, corrosion-resistant, clean convenient, film is easily disinfected, the advantages such as long service life of satisfactory mechanical property, film.
The principal character that realizes production technique of the present invention may further comprise the steps:
(1) the heparin quaternary ammonium salt is synthetic
Taking heparin sodium (the anti-II a factor is tired about 140IU/mg), be dissolved in water, make the solution that bulking value concentration is 15-25%, stir and slowly add 20 times of plain oronain solution (bulking value concentration is 2-5%) of amount benzyl down, it is muddy that solution becomes gradually, filter cake after the filtration washs with purified water, and 65 ℃ of oven dry 6h get the plain oronain salt of heparin benzyl.
(2) the heparin benzyl ester is synthetic
Taking heparin benzethonium chloride salt adds the methylene dichloride stirring and dissolving of 5 times of amounts, slowly drips benzyl chloride, 30-45 ℃ of reaction 15-25h, control esterification yield is 14-15%, adds the methyl alcohol that 1.5 times of amounts are dissolved with 10% sodium-acetate when reaction finishes, treat that precipitation is separated out after, filtration.The filter cake methanol wash, drying gets the heparin benzyl ester.
(3) heparin benzyl ester alkaline condition degraded down
The taking heparin benzyl ester, soluble in water to concentration be 30-50%, be heated to 55-65 ℃, add sodium hydroxide and make its concentration be not less than 0.1mol/L, stir depolymerization 1-1.25h.Reaction finishes, cool to room temperature, and transferring to the pH value with hydrochloric acid is 6.0-7.0, adding sodium-chlor, to make its final concentration be 10%.Add 3 times of volumes methanol while stirring, separate out post precipitation and filter.The filter cake methanol wash, drying under reduced pressure, the crude product of must degrading.
In hydrochloric acid and the time, the pH of regulator solution is 6.0, adds calcium oxide and calcium chloride, the concentration that makes calcium chloride is 8.0mol/L, then prepares ultra-low molecular weight heparin calcium.
(4) ultra-low molecular weight heparin purifying crude
Get the ultra-low molecular weight heparin crude product after the degraded, be dissolved in water and make the solution of 4-8%, filter by the inorganic ceramic ultra-filtration membrane, remove the heparin fragment of macromolecule, adopt the Jiangsu zirconia ceramic ultra-filtration membrane of my High Seience Technology Co., Ltd.'s production for a long time among the present invention, it is to be supporter with highly purified aluminium sesquioxide, membrane pore size is at 10nm, the filtration of molecule screen grading can be realized,, the heparin fragment of molecular weight can be isolated greater than 3000D according to its molecular weight cut-off, the working pressure of ceramic membrane filter is selected 0.1-0.4MPa, preferred 0.2MPa, filtration temperature is selected 40-60 ℃, improves temperature and helps improving membrane flux.
By the liquid that sees through behind the inorganic ceramic membrane filtration, continuation is filtered by hollow fiber ultrafiltration membrane, selecting molecular weight cut-off among the present invention is the ultra-filtration membrane of 1000D, because the molecule of ultra-low molecular weight heparin is type in the shape of a spiral, actually transmitted molecular weight will be higher than 1000D, and ultrafiltration pressure is at 0.05～0.07MPa, and circulation velocity is controlled at about 2d/s, feed temperature 20-40 ℃, the solution of holding back is spissated ultra-low molecular weight heparin solution.The ethanol that adds 3 times of amounts in concentrated solution precipitates, and filters back filter cake washing with alcohol, adds a small amount of purified water lyophilize in the precipitation, and obtaining the white powder solid is ultra-low molecular weight heparin sodium (calcium).
Effect of the present invention is:
(1) temperature and time that reacts by the control depolymerized heparin, the control degradation product reaches lower molecular weight and narrower range of molecular weight distributions, obtain range of molecular weight distributions at 2000D-2500D, molecular-weight average is the ultra-low molecular weight heparin sodium of 2200D, its anti-Xa factor is tired at 100-125IU/mg, the anti-II a factor is tired at 10-25IU/mg, the ratio range of FXa/FII a is between 5.0-10.0, solubleness and clarity reach the requirement of international superior product, the absorbancy at wavelength 280nm place is 0.06-0.10, and the absorbancy at wavelength 260nm place is 0.4-0.6.
Add sodium-chlor or calcium chloride during (2) by depolymerization under alkaline condition, can correspondingly obtain ultra-low molecular weight heparin sodium or ultra-low molecular weight heparin calcium.
(3) the present invention is applied to the membrane ultrafiltration technology effectively solve the separation problem of ultra-low molecular weight heparin in the separation of different molecular weight section cleverly, has avoided using in purifying the gel column adsorption method of separation of loaded down with trivial details costliness.The separation of ultra-low molecular weight heparin sodium adopts inorganic ceramic film and hollow fiber ultrafiltration membrane bonded method to realize, simultaneously at first carry out the inorganic ceramic membrane filtration and also effectively reduced the membrane pollution problem that occurs in the membrane filtration processes, satisfied the industrial continuous production condition.The two-step approach ultra-filtration technique that adopts has solved the stability of quality product, production continuity.Production process does not produce environmental pollution, and production cost of products is lower, corresponding yield height.
Embodiment
Embodiment 1
Get unfraction heparin sodium (the anti-II a factor tire 140IU/mg) 100g, add water 500ml dissolving, stir and slowly add 25% the plain oronain solution of benzyl 10000ml down, it is muddy that solution becomes gradually, filter cake after the filtration washs with purified water, and 65 ℃ of oven dry 6h get the plain oronain salt of heparin benzyl 325g.
Taking heparin benzethonium chloride salt 200g adds methylene dichloride 1000ml stirring and dissolving, slowly drips benzyl chloride 200ml, 40 ℃ of reaction 20h, the control esterification yield is 14.5%, adds the methyl alcohol 1800ml that is dissolved with 10% sodium-acetate when reaction finishes, treat that precipitation is separated out after, filter.The filter cake methanol wash, drying gets heparin benzyl ester 71.50g.
Taking heparin benzyl ester 50g is dissolved in the 1500ml water, is heated to 65 ℃, adds sodium hydroxide 6.4g, stirs depolymerization 1.25h.Reaction finishes, cool to room temperature, and transferring to the pH value with hydrochloric acid is 6.5, adds analytical pure sodium-chlor 150g stirring and dissolving.Add 3 times of volumes methanol while stirring, separate out post precipitation and filter.The filter cake methanol wash, drying under reduced pressure, crude product 31.8g must degrade.
Get the ultra-low molecular weight heparin crude product 20g after the degraded, add the 400ml water dissolution and make about 5% solution, filter by the inorganic ceramic ultra-filtration membrane, adopt the Jiangsu zirconia ceramic ultra-filtration membrane of my High Seience Technology Co., Ltd.'s production for a long time, membrane pore size is at 10nm, filtering working pressure is 0.2MPa, and 60 ℃ of filtration temperatures obtain to filter through liquid.By the liquid that sees through behind the inorganic ceramic membrane filtration, continuation is that the hollow fiber ultrafiltration membrane of 1000D is filtered by molecular weight cut-off, ultrafiltration pressure is at 0.06MPa, circulation velocity is controlled at about 2d/s, 30 ℃ of feed temperatures, the solution of holding back is spissated ultra-low molecular weight heparin solution, and volume is 68ml.
The ethanol that adds 200ml 95% in concentrated solution precipitates, and filters back filter cake absolute ethanol washing.Add about 15ml purified water dissolving in the precipitation of collecting, the precooling postlyophilization obtains white powder solid 13.6g, and purification yield is 68%.
Through check, the ultra-low molecular weight heparin sodium molecule amount distribution range of preparation accounts for 92% in the ratio of 2000D-2500D, and molecular-weight average is 2200D.Its anti-Xa factor is tired and is 124IU/mg, and the anti-II a factor is tired and is 16IU/mg, and the ratio of FXa/F II a is 7.75, and solubleness and clarity detect according to official method, and the absorbancy at wavelength 280nm place is 0.08, and the absorbancy at wavelength 260nm place is 0.52.
Embodiment 2
Get unfraction heparin sodium (the anti-II a factor tire 140IU/mg) 150g, add water 750ml dissolving, stir and slowly add 25% the plain oronain solution of benzyl 15000ml down, it is muddy that solution becomes gradually, filter cake after the filtration washs with purified water, and 65 ℃ of oven dry 6h get the plain oronain salt of heparin benzyl 477g.
Taking heparin benzethonium chloride salt 400g adds methylene dichloride 2000ml stirring and dissolving, slowly drips benzyl chloride 400ml, 35 ℃ of reaction 25h, the control esterification yield is 13.8%, adds the methyl alcohol 3500ml that is dissolved with 10% sodium-acetate when reaction finishes, treat that precipitation is separated out after, filter.The filter cake methanol wash, drying gets heparin benzyl ester 142g.
Taking heparin benzyl ester 100g is dissolved in the 3000ml water, is heated to 62 ℃, adds sodium hydroxide 12g, stirs depolymerization 1h.Reaction finishes, cool to room temperature, and transferring to the pH value with hydrochloric acid is 6.0, adds analytical pure calcium chloride 1800g, stirring and dissolving.Add 3 times of volumes methanol while stirring, separate out post precipitation and filter.The filter cake methanol wash, drying under reduced pressure, crude product 75g must degrade.
Get the ultra-low molecular weight heparin calcium crude product 60g after the degraded, add the 1200ml water dissolution and make about 5% solution, filter by the inorganic ceramic ultra-filtration membrane, adopt the Jiangsu zirconia ceramic ultra-filtration membrane of my High Seience Technology Co., Ltd.'s production for a long time, membrane pore size is at 10nm, filtering working pressure is 0.3MPa, and 60 ℃ of filtration temperatures obtain to filter through liquid.By the liquid that sees through behind the inorganic ceramic membrane filtration, continuation is that the hollow fiber ultrafiltration membrane of 1000D is filtered by molecular weight cut-off, ultrafiltration pressure is at 0.08MPa, circulation velocity is controlled at about 2d/s, 30 ℃ of feed temperatures, the solution of holding back is spissated ultra-low molecular weight heparin solution, and volume is 145ml.
The ethanol that adds 450ml 95% in concentrated solution precipitates, and filters back filter cake absolute ethanol washing.Add about 100ml purified water dissolving in the precipitation of collecting, the precooling postlyophilization obtains white powder solid 39.6g, and purification yield is 66%.
Through check, the ultra-low molecular weight heparin calcium range of molecular weight distributions of preparation accounts for 90% in the ratio of 2000D-2500D, and molecular-weight average is 2200D.Its anti-Xa factor is tired and is 105IU/mg, and the anti-II a factor is tired and is that 12.8IU/mg, the ratio of FXa/FII a are 8.20, and solubleness and clarity detect according to official method, and the absorbancy at wavelength 280nm place is 0.082, and the absorbancy at wavelength 260nm place is 0.55.Calcium contents 10.80%.
Embodiment 3
Get classification heparin sodium (the anti-II a factor 150IU/mg that tires, dermatan sulfate content is lower than 2%) 100g, add water 500ml dissolving, stir the plain oronain solution of the benzyl 10000ml of slow adding 25% down, it is muddy that solution becomes gradually, filter cake after the filtration washs with purified water, and 65 ℃ of oven dry 6h get the plain oronain salt of heparin benzyl 332g.
Taking heparin benzethonium chloride salt 300g adds methylene dichloride 1500ml stirring and dissolving, slowly drips benzyl chloride 300ml, 40 ℃ of reaction 20h, the control esterification yield is 14.0-14.5%, adds the methyl alcohol 2700ml that is dissolved with 10% sodium-acetate when reaction finishes, after treating that precipitation is separated out, filter.The filter cake methanol wash, drying gets heparin benzyl ester 115g.
Taking heparin benzyl ester 100g is dissolved in the 3000ml water, is heated to 65 ℃, adds sodium hydroxide 9g, stirs depolymerization 1h.Reaction finishes, cool to room temperature, and transferring to the pH value with hydrochloric acid is 6.5, adds analytical pure sodium-chlor 300g stirring and dissolving.Add 3 times of volumes methanol while stirring, separate out post precipitation and filter.The filter cake methanol wash, drying under reduced pressure, crude product 68.60g must degrade.
Get the ultra-low molecular weight heparin crude product 50g after the degraded, add the 1000ml water dissolution and make about 5% solution, filter by the inorganic ceramic ultra-filtration membrane, adopt the Jiangsu zirconia ceramic ultra-filtration membrane of my High Seience Technology Co., Ltd.'s production for a long time, membrane pore size is at 10nm, filtering working pressure is 0.3MPa, and 60 ℃ of filtration temperatures obtain to filter through liquid.By the liquid that sees through behind the inorganic ceramic membrane filtration, continuation is that the hollow fiber ultrafiltration membrane of 1000D is filtered by molecular weight cut-off, ultrafiltration pressure is at 0.06MPa, circulation velocity is controlled at about 2d/s, 30 ℃ of feed temperatures, the solution of holding back is spissated ultra-low molecular weight heparin solution, and volume is 158ml.
The ethanol that adds 450ml 95% in concentrated solution precipitates, and filters back filter cake absolute ethanol washing.Add about 15ml purified water dissolving in the precipitation of collecting, the precooling postlyophilization obtains white powder solid 32.8g, and purification yield is 65.6%.
Through check, the ultra-low molecular weight heparin sodium molecule amount distribution range of preparation accounts for 93% in the ratio of 2000D-2500D, and molecular-weight average is 2200D.Its anti-Xa factor is tired and is 120IU/mg, and the anti-II a factor is tired and is 15IU/mg, and the ratio of FXa/F II a is 8, and solubleness and clarity detect according to official method, and the absorbancy at wavelength 280nm place is 0.068, and the absorbancy at wavelength 260nm place is 0.50.
Claims (8)
1. the preparation and the purifying process of a ultra-low molecular weight heparin sodium (calcium) is characterized in that, may further comprise the steps:
(1) the heparin quaternary ammonium salt is synthetic;
(2) the heparin benzyl ester is synthetic;
(3) heparin benzyl ester alkaline condition degraded down;
(4) ultra-low molecular weight heparin purifying crude;
(5) vacuum lyophilization of ultra-low molecular weight heparin and finished product.
Classification or classification heparin raw material do not generate the heparin quaternary ammonium salt with the organic quaternary ammonium salt effect in the aqueous solution, the carboxyl of heparin quaternary ammonium salt and benzyl chlorine generation nucleophilic substitution reaction generate the heparin benzyl ester in dichloromethane solution, under alkaline condition, the ester hydrolytic deterioration obtains low-molecular-weight heparin fragment.Carry out separation and purification by inorganic ceramic ultrafiltration and Hollow Fiber Ultrafiltration bonded method, obtain the ultra-low molecular weight heparin sodium (calcium) of reasonable range of molecular weight distributions, make finished product after the vacuum-drying.
2. the reaction conditions control of method according to claim 1 in the building-up process of heparin benzyl ester, it is characterized in that: taking heparin benzethonium chloride salt, the methylene dichloride stirring and dissolving that adds 5 times of amounts, slowly drip benzyl chloride, the condition of esterification is 30-45 ℃ of reaction 15-25h, the control esterification yield is 14-15%, 35 ℃ of reactions of preferable reaction temperature 20h.
3. method according to claim 1 is characterized in that in the control of heparin benzyl ester degradation process neutral and alkali condition: the adding concentration of sodium hydroxide is heated to 55-65 ℃ between 0.1-0.15mol/L, stirs depolymerization 1-1.25h.
4. the ultra-low molecular weight heparin purifying crude method that method according to claim 1 adopts, it is characterized in that: adopt the inorganic ceramic ultra-filtration membrane to carry out purifying, obtain the rational ultra-low molecular weight heparin of range of molecular weight distributions in conjunction with the method for hollow fiber ultrafiltration membrane.
5. the inorganic ceramic ultra-filtration membrane in the described purification process of claim 4, it is characterized in that: described inorganic ceramic film is the zirconia ceramic ultra-filtration membrane, with highly purified aluminium sesquioxide is supporter, and membrane pore size can be isolated the heparin fragment of molecular weight greater than 3000D at 10nm.
6. the hollow fiber ultrafiltration film in the described purification process of claim 4, it is characterized in that: the molecular weight cut-off of described hollow fiber ultrafiltration membrane is 1000D.
7. the operational condition of the described inorganic ceramic membrane ultrafiltration of claim 4 is characterized in that: working pressure selection 0.1-0.4MPa, preferred 0.2Mpa; Filtration temperature is selected 40-60 ℃, improves temperature and helps improving membrane flux.
8. ultra-low molecular weight heparin sodium (calcium) that requires preparation and purification process acquisition by aforesaid right, it is characterized in that: range of molecular weight distributions accounts for more than 85% at the ultra-low molecular weight heparin of 2000-2500D, molecular-weight average is 2200D, its anti-Xa factor is tired at 100-125IU/mg, the anti-II a factor is tired at 10-25IU/mg, the ratio range of FXa/F II a is between 5.0-10.0, solubleness and clarity reach the standard of international superior product, the absorbancy of 280nm place wavelength is 0.06-0.10, and the absorbancy at wavelength 260nm place is 0.4-0.6.
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| CN102633908A (en) * | 2012-05-02 | 2012-08-15 | 雷晓刚 | Method for preparing high-quality LMW (low molecular weight) heparins |
| CN102731683A (en) * | 2012-07-17 | 2012-10-17 | 湖北亿诺瑞生物制药有限公司 | Method of separating natural low molecular heparin from heparin waste liquor |
| CN102993336A (en) * | 2011-09-14 | 2013-03-27 | 浦江亚太肠衣有限公司 | Crude heparin sodium purification technology |
| CN103012620A (en) * | 2011-09-26 | 2013-04-03 | 徐美英 | High-purity heparin benzyl ester salt and preparation method and application thereof |
| CN103012621A (en) * | 2011-09-26 | 2013-04-03 | 徐美英 | High-purity heparin benzyl ester salt and preparation method and application thereof |
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| CN103012621A (en) * | 2011-09-26 | 2013-04-03 | 徐美英 | High-purity heparin benzyl ester salt and preparation method and application thereof |
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| CN103145880B (en) * | 2013-03-19 | 2015-04-22 | 山东辰中生物制药有限公司 | Method for removing free sulphur in enoxaparin sodium raw material |
| CN104193850A (en) * | 2014-08-16 | 2014-12-10 | 厦门世达膜科技有限公司 | Method for producing crude sodium heparin |
| RU2749424C1 (en) * | 2020-06-10 | 2021-06-10 | Федеральное государственное унитарное предприятие "Московский эндокринный завод" | Method for obtaining drug with anti-coagulant activity |
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