CN101205257A - Dextran 10 and production technique thereof - Google Patents
Dextran 10 and production technique thereof Download PDFInfo
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- CN101205257A CN101205257A CNA2006101476148A CN200610147614A CN101205257A CN 101205257 A CN101205257 A CN 101205257A CN A2006101476148 A CNA2006101476148 A CN A2006101476148A CN 200610147614 A CN200610147614 A CN 200610147614A CN 101205257 A CN101205257 A CN 101205257A
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Abstract
The invention relates to a dextran 10 and a manufacturing technique thereof. Weight average molecular weight of the dextran 10 is 9,000 to 11,000. The dextran 10 is prepared by the following manufacturing technique steps: (1) ferment, (2) first-level membrane separation, (3) second-level membrane separation, (4) molecular weight degradation, (5) decoloration and purification, (6) third-level and fourth-level membrane separation, (7) ion exchange desalination and purification, (8) decoloration and fine filtration and (9) spray drying. Compared with the prior art, the invention does not use ethanol or any organic solvent in the whole producing process, which improves product quality, reduces cost, and meanwhile solves the problem of comprehensive utilization of waste water containing fructose.
Description
Technical field
The present invention relates to a kind of Dextran 10, relate in particular to a kind of Dextran 10 and production technique thereof.
Background technology
Dextran (Dextran) is substrate with sucrose for intestinal mucosa leukonid Leuconnostoc mesenteroide NRRL, katalysis by enzyme makes sucrose decomposition, glucose is aggregated and generates dextran (dextran) simultaneously, and fructose then is not utilized, and stays in the fermentation conversion fluid.The polymer dextran that generates in the fermented liquid is handled through molecular weight degradation after extracting through alcohol precipitation again, obtains into the dextran saccharan molecule of nearly different polymerization degree of 1000 to tens0000.
The domestic early eighties of dextran begins suitability for industrialized production, because of having definite curative effects such as Q volume of blood expansion, antithrombotic and microcirculation improvement, is widely used clinically, is listed in national essential drugs." receive the kind of showing several specifications such as macrodex, Dextran 40, Dextran 20 in the Chinese pharmacopoeia, molecular weight is respectively about 70000,40000,20000, Dextran 10 is that molecular weight is about 10000 product, mainly is used as the raw material of veterinary drug and Iron Dextran at present.
Dextran 10 is as independent product, yet there are no production at present at home, entangling its reason mainly is because the production of present domestic dextran all is the grading technologies that adopt the pure precipitator method, desire to make low-molecular-weight product, must use high density, a large amount of alcohol, so not only cost sharply rises, and will make the volume increase of production unit become unrealistic, and safety problem becomes hidden danger all the time.External producer improves the quality to adopt the loaded down with trivial details big process for refining that consumes, and cost is heightened.
In view of many shortcomings of old technology, numerous producers set up novel process to improvement and have proposed requirement, all wish to develop the new production technique that overcomes these defectives, can obtain low-molecular-weight dextran product, and the while does not increase manufacturing cost or reduces cost.Reform technology a lot of researchs have been done both at home and abroad.But do not see industrial applications.At present both at home and abroad the traditional technology that adopts obtains the product of different size after by the ethanol sedimentation classification.
Existing traditional processing technology is: and fermentation (bio-transformation) → ethanol sedimentation thick acid anhydride → ethanol pinches and washes → and thick acid anhydride dissolving → acid hydrolysis → ethanol precipitation (Pyatyi division) → ethanol dehydration becomes powder → drying.The present following aspect of the defect map of traditional technology:
1. each operation all needs to use a large amount of ethanol, and alcohol has occupied more than 60% of raw materials cost, and safety problem becomes hidden danger all the time.
2. because of the limitation of technology itself, by traditional technology, the purity quality will reach officinal quality level and also must further make with extra care.
3. in the traditional technology, each operation material all is gel body or solid, all can't carry out pipe-line transportation, can only use the manpower transport, and these operations are opening operation, need use a large amount of alcohol again, and this gives and realizes that the GMP requirement causes very big difficulty.
4. the byproduct fructose that accounts for half quantity of raw material white sugar charging capacity can't reclaim in low-cost purification, not only causes the wasting of resources, and causes expensive wastewater treatment or serious environmental to pollute.
Because the technology of preparing of film reaches its maturity, make and in dextran production, adopt membrane separation technique to become possibility.The membrane separation technique principle is: utilize the not of uniform size through molecular weight of material of all size film, by ultrafiltration, nanofiltration, realize the separation of differing molecular quantity of material with physical method.The molecular weight difference of non-polymer material and polymkeric substance dextran is bigger, can select suitable film to realize separating, and realizes extracting the purifying purpose.The range of molecular weight distributions pharmacopeia that dextran allows has defined, selects suitable film can realize the molecular weight distribution classification, can obtain the dextran product of desired molecule amount distribution range.
P.E.Barker., people such as A.Till once carried out gel chromatography and in conjunction with ultra-filtration membrane the dextran hydrolyzed solution carried out the research of dextran molecule amount fractionated, adopted gel chromatography to separate earlier and removed than the macromole part, adopted 4 group 5 * 10 then
3Molecular weight tandem ultra-filtration membrane group, wherein trapped fluid is opposite with the filtrate delivery direction, is used to remove less molecular moiety, so handles the dextran product that the back obtains single specification.But it is uneconomical that the gel chromatography separation method is implemented in commercial scale production, and the processing method that obtains before the dextran hydrolyzed solution still is a traditional technology.
People such as Guo Zhenyou once carried out the research that ultra-filtration technique replaces ethanol sedimentation crude dextran technology, adopted the pre-treatment and the ultra-filtration technique of fermented liquid to unite utilization, replace the thick anhydridization process of ethanol sedimentation in the traditional technology, and other operation were constant, comprises the alcohol precipitation classification.A step in the traditional technology has only been reformed in this research, has not considered to solve the problem of whole technologies without alcohol, and without pure water rinsing dialysis process operations, separates the dextran that obtains and be difficult to reach back operation ultrafiltration fractionated specification of quality.
Summary of the invention
Purpose of the present invention is exactly that provide in order to overcome the defective that above-mentioned technology exists a kind of improves the quality, reduces cost, solved simultaneously the Dextran 10 and the production technique thereof that contain the comprehensive utilization of fructose waste water.
Purpose of the present invention can be achieved through the following technical solutions: a kind of Dextran 10, it is characterized in that the weight-average molecular weight of this Dextran 10 is 9000～11000, and its structural formula is as follows:
Described Dextran 10 is a kind of white powder, and odorless is tasteless; Easily molten, colourless in the normal-temperature water, clarification, characteristic reaction is positive reaction, and specific optical rotation is+180 °～+ 190 °, muriate≤0.15%, weight loss on drying≤7%, heavy metal≤8ppm, residue on ignition≤0.3%.
A kind of production technique of Dextran 10 is characterized in that, this technology may further comprise the steps:
(1) fermentation is a substrate with sucrose, through dextran bacterial classification L.N.1226-a fermentation, again through suitable pretreated fermented liquid, put and carry out uf processing in the ultrafiltration storage tank, service temperature is 10～80 ℃, working pressure 0.1～1.5Mpa, add suitable water dialysis, obtain filtrate, the filtrate clarification;
(2) one-level membrane sepn: the filtrate in the above-mentioned steps (1) inserted carry out uf processing in the ultrafiltration storage tank, service temperature is 10 ℃～80 ℃, working pressure 0.1～1.5Mpa, and with an amount of pure water rinsing trapped fluid, the filtrate clarification;
(3) secondary membrane sepn: with ultra-filtration membrane or nanofiltration membrane the ultrafiltration membrane treatment filtrate in the above-mentioned steps (2) is carried out dextran and separate with list, oligosaccharides and other non-polymers such as fructose, separate dialysis simultaneously, obtain " filtrate " and " trapped fluid ", dextran is present in trapped fluid, service temperature is 10 ℃～80 ℃, working pressure 0.7～3.5Mpa;
(4) molecular weight degradation is handled: with suitable sour heating hydrolysis method the solution that contains dextran in the step (3) is carried out molecular weight degradation and handle, follow the tracks of detection molecules amount and distribution thereof, obtaining molecular-weight average is 9 * 10
3～1.1 * 10
4Molecular weight adjust treatment solution;
(5) decolouring purifying: the gac that in the decolouring container, adds 0.5～2% (W/V), heating, after molecular weight adjustment treatment solution is regulated pH to 6～7 in the step (4), decolour, filter then, remove nitrogenous class impurity such as free protein and can be obtained clarifying colourless " decolouring filtrate " by other impurity of charcoal absorption;
(6) three, level Four membrane sepn classification: with 100
～10
3 
The molecular weight of three grades of films of ultrafiltration after to decolouring adjust treatment solution and carry out molecular-weight gradation, make wherein relatively that the macromole part is trapped in three grades of membrane sepn, small molecules partly filters relatively, to remove macromole part, filtrate M
W8.8 * 10
3～10.5 * 10
3Between relative small molecules partly use ultra-filtration membrane molecular weight specification 20
～50
The level Four film carry out classification, to remove small molecules, hold back M
W9.5 * 10
3～11 * 10
3Between trapped fluid; Working pressure is 0.5～3.5Mpa, and service temperature is 10 ℃～80 ℃;
(7) from handing over desalting and purifying: the filtrate in the above-mentioned steps (6) is concentrated with nanofiltration membrane, perhaps with evaporation concentration to 10%～30% concentration, then by the yin, yang ion exchange column, remove remaining salt and charged all kinds of impurity, obtain " from handing over solution ", " from handing over solution " after the ion-exchange requires chlorine ion concentration below 0.15%; PH=4～7; Below the heavy metal 8ppm;
(8) the smart filter of decolouring: " from handing over solution " that obtain in the step (7) added 0.5%～2% (W/W) gac heating decolouring once more in suitable container, be further purified refining purpose, by the microporous membrane fine filter sterilizing and remove particulate matter to reach;
(9) spraying drying: by spraying drying powder-forming, and control moisture content below 7% promptly.
The membrane sepn of uf processing is the tangential flow filtration that adopts in described step (1), (2), and promptly the feed liquid flow direction becomes tangential direction with the filter membrane face, leaches direction and becomes vertical direction with film, and the specification of film is 300
10
4 , the form of film can be tubular fibre, tubular type or rolling, material comprises polysulfones, improvement polysulfones, polyethersulfone.
Tubular fibre working pressure in described step (1), (2) working pressure is selected 0.1～0.3Mpa for use; The working pressure of rolling and tubular membrane is selected 0.7～1.5Mpa for use; Service temperature is 40～60 ℃.
The membrane sepn of uf processing is the tangential flow filtration that adopts in the described step (3), be that the feed liquid flow direction becomes tangential direction with the filter membrane face, leach direction and become vertical direction with film, the specification of film is selected according to the molecular-weight average of dextran in the fermented liquid, and wherein the film specification of ultra-filtration membrane is a molecular weight 20
～10
3 
, the molecular weight 100～1000 of nanofiltration membrane.
The form of the film of membrane sepn comprises tubular type or rolling in the described step (3), and material comprises polysulfones, improvement polysulfones, polyethersulfone etc., and service temperature is 40～60 ℃.
Described step (2) (3) two step operation also can be undertaken by reversed sequence, but the ultrafiltration filtrate must just can be for further processing after concentrating.
Acid in the described step (4) comprises hydrochloric acid, sulfuric acid, nitric acid, citric acid.
Described step (6) membrane sepn is to adopt tangential flow filtration, and the form of film comprises tubular fibre, flat board, tubular type or rolling, and material comprises polysulfones, improvement polysulfones, polyethersulfone etc., and service temperature is 40～60 ℃.
Compared with prior art, the present invention adopts film and membrane technique that tunning is carried out effective separation and Extraction and the dextran of different molecular weight is carried out fractional separation, can effectively obtain scintilla dextran products such as Dextran 10; Whole process of production is without ethanol or any organic solvent; Realized improving the quality, reduced cost, solved simultaneously and contain fructose waste water comprehensive utilization, membrane separating method of the present invention is not only applicable to the dextran-fermentation liquid that traditional technology obtains, and the dextran-fermentation liquid that obtains such as fermentation method, enzyme engineering method of being applicable to all kinds of control molecular weight.Its comprehensive superiority is:
1. all technology improves yield simultaneously and can significantly improve without alcohol or other organic solvent, has exempted the alcohol expense in the material cost.Measures for the prevention of explosion need not be taked in the workshop, has saved the cost in workshop, has eliminated potential safety hazard.
2. because grading technology adopts membrane separation technique, and all processes is operated, purification refine means such as decolouring, ion-exchange, pure water dialysis rinsing can be easier to implement in the aqueous solution, standards of pharmacopoeia can be reached.
3. the present invention not only can prepare the various dextran bulk drug specification products of pharmacopeia regulation, also can be equipped with molecular-weight average 3 * 10 with legal system
3～30 * 10
4The dextran product of other specifications.
4. the present invention can more easily extract byproduct fructose, not only prevents to produce the trade effluent of a large amount of high nutrition, and makes the fructose syrups of food grade, has both solved environmental protection problem, has realized comprehensive utilization again.
5. the whole operations of the present invention all can realize the canalization operation, and container all can be airtight, and smart filter, spraying drying are carried out in the cleaning shop, and it is reasonable, feasible to realize on the hardware that GMP requires.
Embodiment
Embodiment 1
1. with sucrose substrate, through dextran bacterial classification L.N.1226-a fermentation, again through suitable pretreated fermented liquid 53L, wherein dextran content 5.57%, and monose and oligosaccharide content 6.83% such as fructose contain protein 0.19%, inorganic salt 0.17% are put and are carried out uf processing in the ultrafiltration storage tank.Filter membrane specification 10
3 
The form of film is a rolling, and material can be the improvement polysulfones, and service temperature is 50 ℃, and working pressure 0.8Mpa adds suitable water dialysis, obtains filtrate 95L.The filtrate clarification.
2. above-mentioned filtrate is inserted and is carried out uf processing in the ultrafiltration storage tank, filter membrane molecular weight specification 50
The form of film is a rolling, and material is the improvement polysulfones, and service temperature is 40 ℃, and working pressure 0.7Mpa with an amount of pure water rinsing trapped fluid, obtains trapped fluid 17.1L, and right acid anhydride concentration is 16.3%; Obtain filtrate 108L.Trapped fluid HPLC detects collection of illustrative plates and shows no fructose and oligosaccharides peak.Single, oligosaccharide concentration 3.2% such as fructose in the filtrate.
3. above-mentioned 2 trapped fluids heating adds acid and relaxes degraded, and the dextran molecule amount is adjusted in degraded, detects with HPLC and follows the tracks of detection molecules amount and distribution thereof, and the dextran molecule amount is adjusted in degraded, regulates PH to 6.0 with NaOH during end, obtains molecular weight M
W=10500 adjust treatment solution, add 2% (W/V) gac, filter, and obtain clarifying colourless filtrate 23L, right acid anhydride concentration 12%.N content＜50ppm.
4. the molecular weight after above-mentioned 3 the decolouring is adjusted treatment solution and is carried out the ultrafiltration stage treatment, ultra-filtration membrane molecular weight specification 200
, the dextran dry-matter of ultrafiltration dialysis to 90% leaches, and obtains trapped fluid 4.5L, dextran concentration 6.2%; Obtain filtrate 63.5L, dextran concentration 3.9%; HPLC detects and the GPC software analysis shows: filtrate M
W=10100.
5. above-mentioned 4 filtrate is carried out the ultrafiltration stage treatment, ultra-filtration membrane molecular weight specification 30
, ultrafiltration dialysis has 15% dextran dry-matter to leach, and obtains trapped fluid 10.2L, dextran concentration 20.5%, and HPLC detects and the GPC software analysis shows: trapped fluid M
W=9900.
6. above-mentioned 5 trapped fluid carries out the ion exchange column desalination, and activated carbon decolorizing is handled, and carries out spraying drying after concentrating, and obtains dextran powder 1805kg.Mass analysis meets the requirements.
Embodiment 2
1. with sucrose substrate, through dextran bacterial classification L.N.1226-a fermentation, again through suitable pretreated fermented liquid 53L, wherein dextran content 5.57%, and monose and oligosaccharide content 6.83% such as fructose contain protein 0.19%, inorganic salt 0.17% are put and are carried out uf processing in the ultrafiltration storage tank.Filter membrane specification 300
The form of film is a tubular fibre, and material can be a polysulfones, and service temperature is 40 ℃, and working pressure 0.1Mpa adds suitable water dialysis, obtains filtrate 95L.The filtrate clarification.
2. above-mentioned filtrate is inserted and is carried out uf processing in the ultrafiltration storage tank, filter membrane molecular weight specification 300
The form of film is a tubular fibre, and material is a polysulfones, and service temperature is 40 ℃, and working pressure 0.1Mpa with an amount of pure water rinsing trapped fluid, obtains trapped fluid 17.1L, and right acid anhydride concentration is 16.3%; Obtain filtrate 108L.Trapped fluid HPLC detects collection of illustrative plates and shows no fructose and oligosaccharides peak.Single, oligosaccharide concentration 3.2% such as fructose in the filtrate.
3. above-mentioned 2 filtrates are carried out two-stage ultrafiltering and are handled ultra-filtration membrane molecular weight specification 20
The form of film is a rolling, material is the improvement polysulfones, service temperature is 40 ℃, working pressure 0.7Mpa, with an amount of pure water rinsing trapped fluid, concentrate then, obtain trapped fluid 13.4L trapped fluid, list, oligosaccharides relative content such as fructose reaches below 0.5% in the trapped fluid, wherein single, oligosaccharide concentration 25.1% such as fructose; Filtrate 134L discards.
4. above-mentioned 3 trapped fluids heating adds hydrochloric acid and relaxes degraded, and the dextran molecule amount is adjusted in degraded, detects with HPLC and follows the tracks of detection molecules amount and distribution thereof, obtains molecular weight M
W=11000 adjust treatment solution, regulate PH to 6 with NaOH during end, add 1% (W/V) gac, filter, and obtain clarifying colourless filtrate.N content＜50ppm.
5. the molecular weight after above-mentioned 4 the decolouring is adjusted treatment solution and is carried out the ultrafiltration stage treatment, ultra-filtration membrane molecular weight specification 100
, service temperature is 40 ℃, working pressure 0.5Mpa, and the dextran dry-matter of ultrafiltration dialysis to 89.5% leaches, and obtains trapped fluid 5.6L, dextran concentration 5.2%; Obtain filtrate 60.8L, dextran concentration 4.1%; HPLC detects and the GPC software analysis shows: filtrate M
W=10500.
6. above-mentioned 5 filtrate is carried out the ultrafiltration stage treatment, ultra-filtration membrane molecular weight specification 20
, the form of film is a tubular fibre, and service temperature is 40 ℃, and working pressure 0.5Mpa, ultrafiltration dialysis have 15% dextran dry-matter to leach, and obtain trapped fluid 14.5L, dextran concentration 14.6%, HPLC detects and the GPC software analysis shows: trapped fluid M
W=10200.
7. above-mentioned 6 trapped fluid carries out the ion exchange column desalination, and activated carbon decolorizing is handled, and carries out spraying drying after concentrating, and obtains Dextran 10 powder 1810g.
Embodiment 3
(1) fermentation is a substrate with sucrose, through dextran bacterial classification L.N.1226-a fermentation, again through suitable pretreated fermented liquid, puts and carries out uf processing, filter membrane specification 10 in the ultrafiltration storage tank
4 
, the form of film is a tubular fibre, and material can be the improvement polysulfones, and service temperature is 60 ℃, and working pressure 0.3Mpa adds suitable water dialysis, obtains filtrate 95L, the filtrate clarification.
(2) one-level membrane sepn: the filtrate in the above-mentioned steps (1) inserted carry out uf processing, filter membrane specification 10 in the ultrafiltration storage tank
4 
, the form of film is a tubular fibre, and material can be the improvement polysulfones, and service temperature is 60 ℃, working pressure 0.3Mpa, with an amount of pure water rinsing trapped fluid, the filtrate clarification;
(3) secondary membrane sepn: with ultra-filtration membrane the ultrafiltration membrane treatment filtrate in the above-mentioned steps (2) is carried out dextran and separate with list, oligosaccharides and other non-polymers such as fructose, separate dialysis simultaneously, obtain " filtrate " and " trapped fluid ", dextran is present in trapped fluid, ultra-filtration membrane molecular weight specification 10
3 
The form of film is a tubular type, and material is a polysulfones, and service temperature is 60 ℃, working pressure 3.5Mpa;
(4) molecular weight degradation is handled: with suitable sulfuric acid heating hydrolysis method the solution that contains dextran in the step (3) is carried out molecular weight degradation and handle, follow the tracks of detection molecules amount and distribution thereof, obtaining molecular-weight average is 9.4 * 10
3Molecular weight adjust treatment solution;
(5) decolouring purifying: the gac that in the decolouring container, adds 0.5% (W/V), heating, after molecular weight adjustment treatment solution is regulated pH to 6 in the step (4), decolour, filter then, remove nitrogenous class impurity such as free protein and can be obtained clarifying colourless " decolouring filtrate " by other impurity of charcoal absorption;
(6) three, level Four membrane sepn classification: with 1000
The molecular weight of three grades of films of ultrafiltration after to decolouring adjust treatment solution and carry out molecular-weight gradation, make wherein relatively that the macromole part is trapped in three grades of membrane sepn, small molecules partly filters relatively, to remove macromole part, filtrate M
W9.0 * 10
3About relative small molecules partly use ultra-filtration membrane molecular weight specification 50
The level Four film carry out classification, to remove small molecules, hold back M
W9.8 * 10
3About trapped fluid; The form of film is flat, and material is a polyethersulfone, and working pressure is 3.5Mpa, and service temperature is 60 ℃;
(7) from handing over desalting and purifying: the filtrate in the above-mentioned steps (6) is concentrated with nanofiltration membrane, perhaps with evaporation concentration to 10% concentration, then by the yin, yang ion exchange column, remove remaining salt and charged all kinds of impurity, obtain " from handing over solution ", " from handing over solution " after the ion-exchange requires chlorine ion concentration below 0.15%; PH=5.5; Below the heavy metal 8ppm;
(8) the smart filter of decolouring: " from handing over solution " that obtain in the step (7) added 0.5% (W/W) gac heating decolouring once more in suitable container, be further purified refining purpose, by the microporous membrane fine filter sterilizing and remove particulate matter to reach;
(9) spraying drying: by spraying drying powder-forming, and control moisture content below 7% promptly.
Embodiment 4
(1) fermentation is a substrate with sucrose, through dextran bacterial classification L.N.1226-a fermentation, again through suitable pretreated fermented liquid, puts and carries out uf processing, filter membrane specification 300 in the ultrafiltration storage tank
, the form of film is a tubular type, and material is a polyethersulfone, and service temperature is 10 ℃, and working pressure 0.7Mpa adds suitable water dialysis, obtains filtrate, the filtrate clarification;
(2) one-level membrane sepn: the filtrate in the above-mentioned steps (1) inserted carry out uf processing, filter membrane specification 300 in the ultrafiltration storage tank
, the form of film is a tubular type, and material is a polyethersulfone, and service temperature is 10 ℃, working pressure 0.7Mpa, with an amount of pure water rinsing trapped fluid, the filtrate clarification;
(3) secondary membrane sepn: with nanofiltration membrane the nanofiltration membrane treatment filtrate in the above-mentioned steps (2) is carried out dextran and separate with list, oligosaccharides and other non-polymers such as fructose, separate dialysis simultaneously, obtain " filtrate " and " trapped fluid ", dextran is present in trapped fluid, nanofiltration membrane molecular weight specification 250.The form of film is a rolling, and material is the improvement polysulfones, and service temperature is 10 ℃, working pressure 0.7Mpa;
(4) molecular weight degradation is handled: with suitable nitric acid heating hydrolysis method the solution that contains dextran in the step (3) is carried out molecular weight degradation and handle, follow the tracks of detection molecules amount and distribution thereof, obtaining molecular-weight average is 9 * 10
3Molecular weight adjust treatment solution;
(5) decolouring purifying: the gac that in the decolouring container, adds 2% (W/V), heating, after molecular weight adjustment treatment solution is regulated pH to 6 in the step (4), decolour, filter then, remove nitrogenous class impurity such as free protein and can be obtained clarifying colourless " decolouring filtrate " by other impurity of charcoal absorption;
(6) three, level Four membrane sepn classification: with 10
3 The molecular weight of three grades of films of ultrafiltration after to decolouring adjust treatment solution and carry out molecular-weight gradation, make wherein relatively that the macromole part is trapped in three grades of membrane sepn, small molecules partly filters relatively, to remove macromole part, filtrate M
W9.8 * 10
3About relative small molecules partly use ultra-filtration membrane molecular weight specification 50
The level Four film carry out classification, to remove small molecules, hold back M
W10.8 * 10
3About trapped fluid; The form of film is a rolling, and material is the improvement polysulfones, and working pressure is 3.5Mpa, and service temperature is 60 ℃;
(7) from handing over desalting and purifying: the filtrate in the above-mentioned steps (6) is concentrated with nanofiltration membrane, perhaps with evaporation concentration to 30% concentration, then by the yin, yang ion exchange column, remove remaining salt and charged all kinds of impurity, obtain " from handing over solution ", " from handing over solution " after the ion-exchange requires chlorine ion concentration below 0.15%; PH=6; Below the heavy metal 8ppm;
(8) the smart filter of decolouring: " from handing over solution " that obtain in the step (7) added 2% (W/W) gac heating decolouring once more in suitable container, be further purified refining purpose, by the microporous membrane fine filter sterilizing and remove particulate matter to reach;
(9) spraying drying: by spraying drying powder-forming, and control moisture content below 7% promptly.
Embodiment 5
(1) fermentation is a substrate with sucrose, through dextran bacterial classification L.N.1226-a fermentation, again through suitable pretreated fermented liquid, puts and carries out uf processing, filter membrane specification 10 in the ultrafiltration storage tank
4 
, the form of film is a rolling, and material is a polysulfones, and service temperature is 80 ℃, and working pressure 1.5Mpa adds suitable water dialysis, obtains filtrate, the filtrate clarification;
(2) nanofiltration membrane separation: with nanofiltration membrane the filtrate in the above-mentioned steps (1) is carried out dextran and separate with list, oligosaccharides and other non-polymers such as fructose, separate dialysis simultaneously, obtain " filtrate " and " trapped fluid ", dextran is present in trapped fluid, nanofiltration membrane specification 1000
, service temperature is 80 ℃, working pressure 3.5Mpa;
(3) ultra-filtration membrane separates: the filtrate in the above-mentioned steps (2) is inserted adopted tubular membrane to carry out uf processing, filter membrane specification 10 in the ultrafiltration storage tank
4 , the form of film is a rolling, and material is a polysulfones, and service temperature is 80 ℃, working pressure 1.5Mpa, with an amount of pure water rinsing trapped fluid, the filtrate clarification;
(4) molecular weight degradation is handled: after the solution process that contains dextran in the step (3) is concentrated, carry out molecular weight degradation with suitable citric acid heating hydrolysis method again and handle, follow the tracks of detection molecules amount and distribution thereof, obtaining molecular-weight average is 1.05 * 10
4Molecular weight adjust treatment solution;
(5) decolouring purifying: the gac that in the decolouring container, adds 0.5% (W/V), heating, after molecular weight adjustment treatment solution is regulated pH to 6 in the step (4), decolour, filter then, remove nitrogenous class impurity such as free protein and can be obtained clarifying colourless " decolouring filtrate " by other impurity of charcoal absorption;
(6) three, level Four membrane sepn classification: with 100
The molecular weight of three grades of films of ultrafiltration after to decolouring adjust treatment solution and carry out molecular-weight gradation, make wherein relatively that the macromole part is trapped in three grades of membrane sepn, small molecules partly filters relatively, to remove macromole part, filtrate M
W9.2 * 10
3About relative small molecules partly use ultra-filtration membrane molecular weight specification 20
The level Four film carry out classification, to remove small molecules, hold back M
W10.5 * 10
3About trapped fluid; The form of film is a tubular type, and material is a polysulfones, and working pressure is 0.5Mpa, and service temperature is 80 ℃;
(7) from handing over desalting and purifying: the filtrate in the above-mentioned steps (6) is concentrated with nanofiltration membrane, perhaps with evaporation concentration to 10% concentration, then by the yin, yang ion exchange column, remove remaining salt and charged all kinds of impurity, obtain " from handing over solution ", " from handing over solution " after the ion-exchange requires chlorine ion concentration below 0.15%; PH=5.5; Below the heavy metal 8ppm;
(8) the smart filter of decolouring: " from handing over solution " that obtain in the step (7) added 0.5% (W/W) gac heating decolouring once more in suitable container, be further purified refining purpose, by the microporous membrane fine filter sterilizing and remove particulate matter to reach;
(9) spraying drying: by spraying drying powder-forming, and control moisture content promptly getting a kind of Dextran 10 below 7%, the weight-average molecular weight of this Dextran 10 is 9000～11000, its structural formula is as follows:
Described Dextran 10 is a kind of white powder, and odorless is tasteless; Easily molten, colourless in the normal-temperature water, clarification, characteristic reaction is positive reaction, and specific optical rotation is+180 °～+ 190 °, muriate≤0.15%, weight loss on drying≤7%, heavy metal≤8ppm, residue on ignition≤0.3%.
Claims (10)
1. a Dextran 10 is characterized in that, the weight-average molecular weight of this Dextran 10 is 9000～11000, and its structural formula is as follows:
2. a kind of Dextran 10 according to claim 1 is characterized in that, described Dextran 10 is a kind of white powder, and odorless is tasteless; Easily molten, colourless in the normal-temperature water, clarification, characteristic reaction is positive reaction, and specific optical rotation is+180 °～+ 190 °, muriate≤0.15%, weight loss on drying≤7%, heavy metal≤8ppm, residue on ignition≤0.3%.
3. the production technique of a Dextran 10 is characterized in that, this technology may further comprise the steps:
(1) fermentation is a substrate with sucrose, through dextran bacterial classification L.N.1226-a fermentation, again through suitable pretreated fermented liquid, put and carry out uf processing in the ultrafiltration storage tank, service temperature is 10～80 ℃, working pressure 0.1～1.5Mpa, add suitable water dialysis, obtain filtrate, the filtrate clarification;
(2) one-level membrane sepn: the filtrate in the above-mentioned steps (1) inserted carry out uf processing in the ultrafiltration storage tank, service temperature is 10 ℃～80 ℃, working pressure 0.1～1.5Mpa, and with an amount of pure water rinsing trapped fluid, the filtrate clarification;
(3) secondary membrane sepn: with ultra-filtration membrane or nanofiltration membrane the ultrafiltration membrane treatment filtrate in the above-mentioned steps (2) is carried out dextran and separate with list, oligosaccharides and other non-polymers such as fructose, separate dialysis simultaneously, obtain " filtrate " and " trapped fluid ", dextran is present in trapped fluid, service temperature is 10 ℃～80 ℃, working pressure 0.7～3.5Mpa;
(4) molecular weight degradation is handled: with suitable sour heating hydrolysis method the solution that contains dextran in the step (3) is carried out molecular weight degradation and handle, follow the tracks of detection molecules amount and distribution thereof, obtaining molecular-weight average is 9 * 10
3～1.1 * 10
4Molecular weight adjust treatment solution;
(5) decolouring purifying: the gac that in the decolouring container, adds 0.5～2% (W/V), heating, after molecular weight adjustment treatment solution is regulated pH to 6～7 in the step (4), decolour, filter then, remove nitrogenous class impurity such as free protein and can be obtained clarifying colourless " decolouring filtrate " by other impurity of charcoal absorption;
(6) three, level Four membrane sepn classification: with 100
～10
3 
The molecular weight of three grades of films of ultrafiltration after to decolouring adjust treatment solution and carry out molecular-weight gradation, make wherein relatively that the macromole part is trapped in three grades of membrane sepn, small molecules partly filters relatively, to remove macromole part, filtrate M
W8.8 * 10
3～10.5 * 10
3Between relative small molecules partly use ultra-filtration membrane molecular weight specification 20
～50
The level Four film carry out classification, to remove small molecules, hold back M
W9.5 * 10
3～11 * 10
3Between trapped fluid; Working pressure is 0.5～3.5Mpa, and service temperature is 10 ℃～80 ℃;
(7) from handing over desalting and purifying: the filtrate in the above-mentioned steps (6) is concentrated with nanofiltration membrane, perhaps with evaporation concentration to 10%～30% concentration, then by the yin, yang ion exchange column, remove remaining salt and charged all kinds of impurity, obtain " from handing over solution ", " from handing over solution " after the ion-exchange requires chlorine ion concentration below 0.15%; PH=4～7; Below the heavy metal 8ppm;
(8) the smart filter of decolouring: " from handing over solution " that obtain in the step (7) added 0.5%～2% (W/W) gac heating decolouring once more in suitable container, be further purified refining purpose, by the microporous membrane fine filter sterilizing and remove particulate matter to reach;
(9) spraying drying: by spraying drying powder-forming, and control moisture content below 7% promptly.
4. the production technique of a kind of Dextran 10 according to claim 3, it is characterized in that, the membrane sepn of uf processing is the tangential flow filtration that adopts in described step (1), (2), be that the feed liquid flow direction becomes tangential direction with the filter membrane face, leach direction and become vertical direction with film, the specification of film is 300
～10
4 
The form of film can be tubular fibre, tubular type or rolling, and material comprises polysulfones, improvement polysulfones, polyethersulfone.
5. according to the production technique of claim 3 or 5 described a kind of Dextran 10s, it is characterized in that the tubular fibre working pressure in described step (1), (2) working pressure is selected 0.1～0.3Mpa for use; The working pressure of rolling and tubular membrane is selected 0.7～1.5Mpa for use; Service temperature is 40～60 ℃.
6. the production technique of a kind of Dextran 10 according to claim 3, it is characterized in that, the membrane sepn of uf processing is the tangential flow filtration that adopts in the described step (3), be that the feed liquid flow direction becomes tangential direction with the filter membrane face, leach direction and become vertical direction with film, the specification of film is selected according to the molecular-weight average of dextran in the fermented liquid, and wherein the film specification of ultra-filtration membrane is a molecular weight 20
～10
3 
The molecular weight 100～1000 of nanofiltration membrane.
7. the production technique of a kind of Dextran 10 according to claim 3 is characterized in that, the form of the film of membrane sepn comprises tubular type or rolling in the described step (3), and material comprises polysulfones, improvement polysulfones, polyethersulfone etc., and service temperature is 40～60 ℃.
8. the production technique of a kind of Dextran 10 according to claim 3 is characterized in that, described step (2) (3) two step operation also can be undertaken by reversed sequence, but the ultrafiltration filtrate must just can be for further processing after concentrating.
9. the production technique of a kind of Dextran 10 according to claim 3 is characterized in that, the acid in the described step (4) comprises hydrochloric acid, sulfuric acid, nitric acid, citric acid.
10. the production technique of a kind of Dextran 10 according to claim 3, it is characterized in that, described step (6) membrane sepn is to adopt tangential flow filtration, the form of film comprises tubular fibre, flat board, tubular type or rolling, material comprises polysulfones, improvement polysulfones, polyethersulfone etc., and service temperature is 40～60 ℃.
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|---|---|---|---|
| CNA2006101476148A CN101205257A (en) | 2006-12-20 | 2006-12-20 | Dextran 10 and production technique thereof |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2006101476148A CN101205257A (en) | 2006-12-20 | 2006-12-20 | Dextran 10 and production technique thereof |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102978257A (en) * | 2012-12-20 | 2013-03-20 | 江苏久吾高科技股份有限公司 | Production method of dextranum |
| CN105153244A (en) * | 2015-08-04 | 2015-12-16 | 山东金朗生物科技有限公司 | Method for extracting sugar solution from waste liquid resulting from production of dextran macromolecule 1000 |
| CN105385721A (en) * | 2015-11-27 | 2016-03-09 | 山东金洋药业有限公司 | Method for producing dextran by directional fermentation |
| CN108640354A (en) * | 2018-06-01 | 2018-10-12 | 华北水利水电大学 | A kind of separation of heavy metal ions film device |
| CN113567370A (en) * | 2021-07-23 | 2021-10-29 | 广西南宁市桃源兽药厂 | Method for detecting content of sugar anhydride in iron dextran injection |
-
2006
- 2006-12-20 CN CNA2006101476148A patent/CN101205257A/en active Pending
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102978257A (en) * | 2012-12-20 | 2013-03-20 | 江苏久吾高科技股份有限公司 | Production method of dextranum |
| CN102978257B (en) * | 2012-12-20 | 2014-12-10 | 江苏久吾高科技股份有限公司 | Production method of dextranum |
| CN105153244A (en) * | 2015-08-04 | 2015-12-16 | 山东金朗生物科技有限公司 | Method for extracting sugar solution from waste liquid resulting from production of dextran macromolecule 1000 |
| CN105153244B (en) * | 2015-08-04 | 2018-10-09 | 山东金朗生物科技有限公司 | A method of it is produced from dextran macromolecular 1000 and extracts sugar juice in waste liquid |
| CN105385721A (en) * | 2015-11-27 | 2016-03-09 | 山东金洋药业有限公司 | Method for producing dextran by directional fermentation |
| CN108640354A (en) * | 2018-06-01 | 2018-10-12 | 华北水利水电大学 | A kind of separation of heavy metal ions film device |
| CN113567370A (en) * | 2021-07-23 | 2021-10-29 | 广西南宁市桃源兽药厂 | Method for detecting content of sugar anhydride in iron dextran injection |
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