CN105131091B - A method of preparing capreomycin sulfate using capreomycin zymotic fluid - Google Patents
A method of preparing capreomycin sulfate using capreomycin zymotic fluid Download PDFInfo
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Abstract
The present invention relates to a kind of method preparing capreomycin sulfate using capreomycin zymotic fluid, this method is that capreomycin zymotic fluid decolourizes by pretreatment, plate-frame filtering, dilution, adjusting pH, macroporous resin adsorption, crystallizes and be dried to obtain capreomycin sulfate.The present invention realizes that bulk pharmaceutical chemicals go capreomycin sulfate stabilization, efficient production.This method is utilized simultaneously, the extract yield of capreomycin sulfate can be improved, shorten the production cycle, reduces cost, improves yield.
Description
Technical field
The invention belongs to antibiotic extraction, synthesis technical fields, and capreomycin zymotic fluid system is utilized more particularly to a kind of
The method of standby capreomycin sulfate.
Background technology
Capreomycin (CPM) is the ring type polypeptide class antibiotic generated by curling streptomycete fermentation, similar by 4 structures
Reactive compound --- IA, IB, IIA and IIb is formed, and the content of wherein IA and IB account for 80%~90%, is the master for playing curative effect
Ingredient.CPM is made generally in sulfate, and clinical application is in treating tuberculosis field.
Currently, being reported according to pertinent literature, the country prepares there are mainly two types of the production technologies of bulk pharmaceutical chemicals capreomycin sulfate：
Technique one：Mainly by capreomycin zymotic fluid, acidified, ion exchange, activated carbon decolorizing, film concentrate, are micro-
Membrane filtration and spray drying obtain its esters product, existing main problem：
1) extraction cost is high.Expensive oxalic acid has been used in extraction process, and use membrane filtration technique and from
Sub- switching technology leads to extraction cost height.
2) the extraction process period is long, has been more than 70h.
3) extract yield is low, generally 70~75%.
Technique two：Capreomycin is extracted using membrane filtering method, wherein micro-filtration membrane material is ceramic membrane, is used for zymotic fluid
Processing；Nanofiltration membrane material is rolling poly (ether sulfone) film, is concentrated for capreomycin exquisiteness liquid；Ultrafiltration membrane material is PS membrane, is used for
Remove the bacterial endotoxin in concentrate；Capreomycin is obtained using drying process with atomizing.Its existing main problem is production
Period is long, and membrane filter system need to periodically carry out care and maintenance, lead to production cost height.
Invention content
The purpose of the present invention is that the defect for overcoming the above-mentioned prior art, provides one kind and effectively improving extract yield, drop
Low cost shortens the production cycle, realizes the side that capreomycin sulfate is prepared using capreomycin zymotic fluid for stablizing, efficiently producing
Method.
The technical solution taken to achieve the above object is：
A method of preparing capreomycin sulfate using capreomycin zymotic fluid, it is characterised in that its processing step is：
Capreomycin zymotic fluid is pre-processed first, then plate-frame filtering, the potency that gained filtrate is diluted to capreomycin exists
Filtrate pH to 5.0~6.0 is adjusted after 7000~8000u/ml, is decolourized with macroporous resin adsorption, collects potency ＞ 4000u/ml's
Adsorption bleaching filtrate crystallizes, dry；
Above-mentioned preprocessing process is：
1) at 30~50r/min of speed of agitator, capreomycin zymotic fluid pH to 5.0~5.5 first is adjusted with phosphoric acid solution,
Stir 40~60min, after continue with phosphorus acid for adjusting pH to 4.0~4.5, stir 30~60min, then pH is adjusted with sodium carbonate liquor
To 7.5~8.0,20~40min is stirred；
2) at 100~120r/min of speed of agitator, microbial flocculant (JT-613) is added, it will after 1~3min of stirring
Rotating speed is down to 30~50r/min, and 40~60min is stood after stirring 20~30min；
3) filter aid is added, stirs 20~40min.
After plate-frame filtering, filter residue in sheet frame is washed with water, collects water lotion, when potency unit is less than 500 μ/ml, stops water
It washes, filtrate and water lotion is mixed.
Filtrate after plate-frame filtering adjusts pH with a concentration of 10~30% sulfuric acid.
The macroporous resin adsorption decolorization is：The filtrate of the plate-frame filtering of pH 5.0~6.0 is warming up to 30 first
~35 DEG C, macroporous absorbent resin equipment is entered with 12~15ml/min flow velocitys；Collect the filtrate through macroporous resin adsorption, flow velocity control
System is in 14~16ml/min, the potency ＞ 4000u/ml through macroporous resin adsorption filtrate, starts to collect filtrate.
The blade diameter length ratio 1 of the macroporous absorbent resin equipment:5~10.
The macroporous absorbent resin material is polyacrylate type polymer, and dosage is the 20~30 of activity in filtrate total hundred million
Times.
The crystallization process is, under the conditions of 50~60r/min of speed of agitator, 1~10 DEG C of temperature, acetone and sterling is added
Capreomycin sulfate solid continues 80~100min of stirring, is then allowed to stand 120~160min, solid matter sulphur is obtained after filtering
Sour capreomycin.
The dosage of the acetone is 5~10 times of adsorption bleaching filtrate volume, and sterling capreomycin sulfate solid dosage is
The 0.01~0.05% of adsorption bleaching filtrate volume.
The drying process is, enables bipyramid and mixes powder machine, adjusts inlet air temperature to 120~130 DEG C, temperature of outgoing air to 50~
80 DEG C, humidification powder is in charge door, after 8~12 mesh stainless steel mesh, with air-flow to cyclone separator, reaches sieving machine through sieving
Obtain capreomycin sulfate.
The preprocessing process is that the concentration of phosphoric acid is controlled 20~40%；The concentration of sodium carbonate is controlled 30~40%；
Microbial flocculant (JT-613) dosage is controlled in 60~70mg/L；Filter aid is diatomite or cellulose or perlite, dosage
It is the 1~5% of fermentating liquid volume.
The present invention technique effect be：
1 compared with technique one, and the yield of extraction has reached 85%, and yield improves 13.3%.
2 compared with technique one, replaces oxalic acid using phosphoric acid, reduces pretreatment cost.
3 compared with technique one, are respectively adopted macroporous absorbent resin and method for crystallising, instead of activated carbon decolorizing, ion exchange
With mocromembrane filtering technique, extraction process step is simplified, life cycle of the product is shortened, reduces production cost, improves production
The market competitiveness of product.
4 compared with technique two, simplifies capreomycin sulfate preparation process, reduces equipment maintenance expense, reduces
Extraction cost.The product yield of this patent is higher than technique two simultaneously, improves product competitiveness in the market.
5 prepare capreomycin sulfate using this patent, and final product quality meets the regulation of Chinese Pharmacopoeia.
Specific implementation mode
The present invention is explained with example, it should be understood that example is for illustrating rather than to this below
The limitation of invention.The scope of the present invention is determined with core content according to claims.
Capreomycin zymotic fluid source in following embodiments：
Capreomycin is to obtain capreomycin zymotic fluid by three grade fermemtation pattern using curling streptomycete, fermentation training
The carbon source supported in base can be glucose, wheat bran or starch, the main soybean cake powder of nitrogen source, fish meal or dried silkworm chrysalis meal etc..It is sent out
Ferment unit is in 11000u/ml or more.
The Hebei microbial flocculant JT-613 Jin Ti thermal insulation materials Co., Ltd in following embodiments.
Embodiment 1
Capreomycin zymotic fluid 10m3, potency 11896u/ml, total hundred million be 118.96.
1) it pre-processes：
20% phosphoric acid solution is added in 30r/min in speed of agitator control, adjusts pH to 5.0 first, stirs 40min, continues
20% phosphoric acid solution is added, adjusts pH to 4.0, stirs 30min, 30% sodium carbonate liquor is added, adjusts pH to 7.5, stirring
20min。
Speed of agitator control is in 100r/min, and rotating speed is down to 30r/ after microbial flocculant JT-613600g, 1min is added
Min stands 40min after stirring 20min.
Filter aid diatomite 10kg is added in zymotic fluid, stirs 20min.
2) plate-frame filtering
With plate-frame filtering zymotic fluid, clear filtrate 7.7m is collected3。
3) it dilutes
After the completion of filtering, filter residue in sheet frame should be washed.Water lotion is collected when potency unit is less than 500 μ/ml to stop
Sealing is washed.Filtrate and water lotion are mixed, the volume of filtrate is 15.9m3, the potency control of capreomycin is in 7052u/ml.
Pre-process yield：94.1%.
4) pH is adjusted
Capreomycin filtrate pH to 5.0 is adjusted with 10% sulfuric acid.
5) macroporous resin adsorption is decolourized
Decolorizing resin 2379.2kg is added in resin filter plant, capreomycin sulfate solution is warming up to 30 DEG C, with 12ml/
Min flow velocitys enter macroporous absorbent resin equipment；The filtrate through macroporous resin adsorption is collected, flow control is in 14ml/min, through big
The potency ＞ 4000u/ml of hole resin adsorption filtrate start to collect filtrate, and the volume of filtrate is 15.8m3, the potency of capreomycin
Control is in 7011u/ml.
The yield of resin adsorption decoloration：98.8%.
6) it crystallizes
In 50r/min, temperature is controlled at 1 DEG C, and acetone 79m is added in filtrate for speed of agitator control3With sterling capreomycin sulfate
Mycin solid 1.58kg continues to stir 80min, is then allowed to stand 120min, solid matter capreomycin sulfate is obtained after filtering.
7) dry
It enables bipyramid and mixes powder machine, adjust inlet air temperature to 120~130 DEG C, temperature of outgoing air is to 50~80 DEG C, and humidification powder is in adding
Material mouth, after 8~12 mesh stainless steel mesh, with air-flow to cyclone separator, reaching sieving machine, through sieving to obtain capreomycin sulfate mould
Plain 109.7kg.
Total recovery：(109.7-1.58) × 0.94/ total hundred million=85.3%.
Embodiment 2
Capreomycin zymotic fluid 10m3, potency 11920u/ml, total hundred million be 119.2.
1) it pre-processes：
25% phosphoric acid solution is added in 35r/min in speed of agitator control, adjusts pH to 5.1 first, stirs 45min, continues
25% phosphoric acid solution is added, adjusts pH to 4.1, stirs 40min, 33% sodium carbonate liquor is added, adjusts pH to 7.6, stirring
25min。
Speed of agitator control is in 105r/min, and rotating speed is down to after microbial flocculant JT-613625g, 1.5min is added
35r/min stands 45min after stirring 23min.
Filter aid cellulose 20kg is added in zymotic fluid, stirs 25min.
2) plate-frame filtering
With plate-frame filtering zymotic fluid, clear filtrate 7.5m is collected3。
3) it dilutes
After the completion of filtering, filter residue in sheet frame should be washed.Water lotion is collected when potency unit is less than 500 μ/ml to stop
Sealing is washed.Filtrate and water lotion are mixed, the volume of filtrate is 15.4m3, the potency control of capreomycin is in 7361u/ml.
Pre-process yield：95.1%.
4) pH is adjusted
Capreomycin filtrate pH to 5.3 is adjusted with 15% sulfuric acid.
5) macroporous resin adsorption is decolourized
Decolorizing resin 2741.6kg is added in resin filter plant, capreomycin sulfate solution is warming up to 32 DEG C, with 13ml/
Min flow velocitys enter macroporous absorbent resin equipment；The filtrate through macroporous resin adsorption is collected, flow control is in 14.5ml/min, warp
The potency ＞ 4000u/ml of macroporous resin adsorption filtrate start to collect filtrate, and the volume of filtrate is 15.3m3, the effect of capreomycin
Valence is controlled in 7298u/ml.
The yield of resin adsorption decoloration：98.5%.
6) it crystallizes
In 52r/min, temperature is controlled at 3 DEG C, and acetone 91.8m is added in filtrate for speed of agitator control3It is rolled up with sterling sulfuric acid
Aspergillin solid 3.06kg continues to stir 85min, is then allowed to stand 130min, solid matter capreomycin sulfate is obtained after filtering.
7) dry
It enables bipyramid and mixes powder machine, adjust inlet air temperature to 120~130 DEG C, temperature of outgoing air is to 50~80 DEG C, and humidification powder is in adding
Material mouth, after 8~12 mesh stainless steel mesh, with air-flow to cyclone separator, reaching sieving machine, through sieving to obtain capreomycin sulfate mould
Plain 111.7kg
Total recovery：(111.76-3.06) × 0.94/ total hundred million=85.7%.
Embodiment 3
Capreomycin zymotic fluid 10m3, potency 12045u/ml, total hundred million be 120.45.
1) it pre-processes：
30% phosphoric acid solution is added in 40r/min in speed of agitator control, adjusts pH to 5.3 first, stirs 50min, continues
30% phosphoric acid solution is added, adjusts pH to 4.3, stirs 45min, 35% sodium carbonate liquor is added, adjusts pH to 7.7, stirring
30min。
Speed of agitator control is in 110r/min, and rotating speed is down to 40r/ after microbial flocculant JT-613650g, 2min is added
Min stands 50min after stirring 25min.
Filter aid perlite 30kg is added in zymotic fluid, stirs 30min.
2) plate-frame filtering
With plate-frame filtering zymotic fluid, clear filtrate 7.6m is collected3。
3) it dilutes
After the completion of filtering, filter residue in sheet frame should be washed.Water lotion is collected when potency unit is less than 500 μ/ml to stop
Sealing is washed.Filtrate and water lotion are mixed, the volume of filtrate is 15.3m3, the potency control of capreomycin is in 7516u/ml.
Pre-process yield：95.6%.
4) pH is adjusted
Capreomycin filtrate pH to 5.5 is adjusted with 20% sulfuric acid.
5) macroporous resin adsorption is decolourized
Decolorizing resin 3011.25kg is added in resin filter plant, and capreomycin sulfate solution is warming up to 33 DEG C, with
13.5ml/min flow velocitys enter macroporous absorbent resin equipment；The filtrate through macroporous resin adsorption is collected, flow control is in 15ml/
Min, the potency ＞ 4000u/ml through macroporous resin adsorption filtrate start to collect filtrate, and the volume of filtrate is 15.1m3, crimp mould
The potency of element is controlled in 7486u/ml.
The yield of resin adsorption decoloration：98.3%.
6) it crystallizes
In 55r/min, temperature is controlled at 5 DEG C, and acetone 113.25m is added in filtrate for speed of agitator control3With sterling sulfuric acid
Capreomycin solid 4.53kg continues to stir 90min, is then allowed to stand 140min, it is mould that solid matter capreomycin sulfate is obtained after filtering
Element.
7) dry
It enables bipyramid and mixes powder machine, adjust inlet air temperature to 120~130 DEG C, temperature of outgoing air is to 50~80 DEG C, and humidification powder is in adding
Material mouth, after 8~12 mesh stainless steel mesh, with air-flow to cyclone separator, reaching sieving machine, through sieving to obtain capreomycin sulfate mould
Plain 111.7kg.
Total recovery：(114.6-4.53) × 0.94/ total hundred million=85.9%.
Embodiment 4
Capreomycin zymotic fluid 10m3, potency 11900u/ml, total hundred million be 119.
1) it pre-processes：
35% phosphoric acid solution is added in 45r/min in speed of agitator control, adjusts pH to 5.4 first, stirs 55min, continues
35% phosphoric acid solution is added, adjusts pH to 4.4, stirs 50min, 37% sodium carbonate liquor is added, adjusts pH to 7.8, stirring
35min。
Speed of agitator control is in 115r/min, and rotating speed is down to after microbial flocculant JT-613675g, 2.5min is added
45r/min stands 55min after stirring 27min.
Filter aid diatomite 40kg is added in zymotic fluid, stirs 35min.
2) plate-frame filtering
With plate-frame filtering zymotic fluid, clear filtrate 7.8m is collected3。
3) it dilutes
After the completion of filtering, filter residue in sheet frame should be washed.Water lotion is collected when potency unit is less than 500 μ/ml to stop
Sealing is washed.Filtrate and water lotion are mixed, the volume of filtrate is 14.4m3, the potency control of capreomycin is in 7874u/ml.
Pre-process yield：95.3%.
4) pH is adjusted
Capreomycin filtrate pH to 5.7 is adjusted with 25% sulfuric acid.
5) macroporous resin adsorption is decolourized
Decolorizing resin 3213kg is added in resin filter plant, capreomycin sulfate solution is warming up to 34 DEG C, with 14ml/
Min flow velocitys enter macroporous absorbent resin equipment；The filtrate through macroporous resin adsorption is collected, flow control is in 15.5ml/min, warp
The potency ＞ 4000u/ml of macroporous resin adsorption filtrate start to collect filtrate, and the volume of filtrate is 14.2m3, the effect of capreomycin
Valence is controlled in 7825u/ml.
The yield of resin adsorption decoloration：98.0%.
6) it crystallizes
In 57r/min, temperature is controlled at 7 DEG C, and acetone 127.8m is added in filtrate for speed of agitator control3It is rolled up with sterling sulfuric acid
Aspergillin solid 5.68kg continues to stir 95min, is then allowed to stand 150min, solid matter capreomycin sulfate is obtained after filtering.
7) dry
It enables bipyramid and mixes powder machine, adjust inlet air temperature to 120~130 DEG C, temperature of outgoing air is to 50~80 DEG C, and humidification powder is in adding
Material mouth, after 8~12 mesh stainless steel mesh, with air-flow to cyclone separator, reaching sieving machine, through sieving to obtain capreomycin sulfate mould
Plain 111.7kg.
Total recovery：(114-5.68) × 0.94/ total hundred million=85.6%.
Embodiment 5
Capreomycin zymotic fluid 10m3, potency 11843u/ml, total hundred million be 118.43.
1) it pre-processes：
40% phosphoric acid solution is added in 50r/min in speed of agitator control, adjusts pH to 5.5 first, stirs 60min, continues
40% phosphoric acid solution is added, adjusts pH to 4.5, stirs 50min, 40% sodium carbonate liquor is added, adjusts pH to 8.0, stirring
40min。
Speed of agitator control is in 120r/min, and rotating speed is down to 50r/ after microbial flocculant JT-613700g, 3min is added
Min stands 60min after stirring 30min.
Filter aid cellulose 50kg is added in zymotic fluid, stirs 40min.
2) plate-frame filtering
With plate-frame filtering zymotic fluid, clear filtrate 7.6m is collected3。
3) it dilutes
After the completion of filtering, filter residue in sheet frame should be washed.Water lotion is collected when potency unit is less than 500 μ/ml to stop
Sealing is washed.Filtrate and water lotion are mixed, the volume of filtrate is 14.1m3, the potency control of capreomycin is in 7986u/ml.
Pre-process yield：95.1%.
4) pH is adjusted
Capreomycin filtrate pH to 6.0 is adjusted with 30% sulfuric acid.
5) macroporous resin adsorption is decolourized
Decolorizing resin 3553kg is added in resin filter plant, capreomycin sulfate solution is warming up to 35 DEG C, with 15ml/
Min flow velocitys enter macroporous absorbent resin equipment；The filtrate through macroporous resin adsorption is collected, flow control is in 16ml/min, through big
The potency ＞ 4000u/ml of hole resin adsorption filtrate start to collect filtrate, and the volume of filtrate is 13.9m3, the potency of capreomycin
Control is in 7939u/ml.
The yield of resin adsorption decoloration：98.0%.
6) it crystallizes
In 60r/min, temperature is controlled at 10 DEG C, and acetone 139m is added in filtrate for speed of agitator control3It is rolled up with sterling sulfuric acid
Aspergillin solid 6.95kg continues to stir 100min, is then allowed to stand 160min, it is mould that solid matter capreomycin sulfate is obtained after filtering
Element.
7) dry
It enables bipyramid and mixes powder machine, adjust inlet air temperature to 120~130 DEG C, temperature of outgoing air is to 50~80 DEG C, and humidification powder is in adding
Material mouth, after 8~12 mesh stainless steel mesh, with air-flow to cyclone separator, reaching sieving machine, through sieving to obtain capreomycin sulfate mould
Plain 111.7kg.
Total recovery：(114.3-6.95) × 0.94/ total hundred million=85.2%.
Claims (9)
1. a kind of method preparing capreomycin sulfate using capreomycin zymotic fluid, it is characterised in that its processing step is：It is first
First capreomycin zymotic fluid is pre-processed, then plate-frame filtering, gained filtrate is diluted to the potency of capreomycin 7000
Filtrate pH to 5.0~6.0 is adjusted after~8000u/ml, is decolourized with macroporous resin adsorption, and the absorption of potency ＞ 4000u/ml is collected
Decoloration filtrate, crystallizes, dry；
Above-mentioned preprocessing process is：
1）At 30~50r/min of speed of agitator, first capreomycin zymotic fluid pH to 5.0~5.5, stirring are adjusted with phosphoric acid solution
40~60min, after continue with phosphorus acid for adjusting pH to 4.0~4.5, stir 30~60min, then with sodium carbonate liquor adjust pH to
7.5~8.0, stir 20~40min；
2）At 100~120r/min of speed of agitator, microbial flocculant is added, stir after 1~3min and rotating speed is down to 30~
50r/min stands 40~60min after stirring 20~30min；
3）Filter aid is added, stirs 20~40min；
Above-mentioned crystallization process is, under the conditions of 50~60r/min of speed of agitator, 1~10 DEG C of temperature, acetone and sterling sulfuric acid is added
Capreomycin solid continues 80~100min of stirring, is then allowed to stand 120~160min, and solid matter sulfuric acid volume is obtained after filtering
Aspergillin.
2. the method described in accordance with the claim 1 for preparing capreomycin sulfate using capreomycin zymotic fluid, it is characterized in that：Plate
After frame filtering, filter residue in sheet frame is washed with water, collects water lotion, when potency unit is less than 500 μ/ml, stops washing, by filtrate
It is mixed with water lotion.
3. the method described in accordance with the claim 1 for preparing capreomycin sulfate using capreomycin zymotic fluid, it is characterized in that：Plate
The filtered filtrate of frame adjusts pH with a concentration of 10~30% sulfuric acid.
4. the method described in accordance with the claim 1 for preparing capreomycin sulfate using capreomycin zymotic fluid, it is characterised in that
The macroporous resin adsorption decolorization is：The filtrate of the plate-frame filtering of pH 5.0~6.0 is warming up to 30~35 DEG C first, with
12~15ml/min flow velocitys enter macroporous absorbent resin equipment；Collect the filtrate through macroporous resin adsorption, flow control 14~
16ml/min, the potency ＞ 4000u/ml through macroporous resin adsorption filtrate start to collect filtrate.
5. the method for preparing capreomycin sulfate using capreomycin zymotic fluid according to claim 4, it is characterised in that
The blade diameter length ratio 1 of the macroporous absorbent resin equipment:5~10.
6. according to the method for preparing capreomycin sulfate using capreomycin zymotic fluid described in claim 1 or 4, feature exists
It is polyacrylate type polymer in the macroporous absorbent resin material, dosage is 20~30 times of activity in filtrate total hundred million.
7. the method described in accordance with the claim 1 for preparing capreomycin sulfate using capreomycin zymotic fluid, it is characterised in that
The dosage of the acetone is 5~10 times of adsorption bleaching filtrate volume, and sterling capreomycin sulfate solid dosage is adsorption bleaching
The 0.01~0.05% of filtrate volume.
8. the method described in accordance with the claim 1 for preparing capreomycin sulfate using capreomycin zymotic fluid, it is characterised in that
The drying process is to enable bipyramid to mix powder machine, adjusts inlet air temperature to 120~130 DEG C, temperature of outgoing air adds to 50~80 DEG C
Wet-milling is in charge door, after 8~12 mesh stainless steel mesh, with air-flow to cyclone separator, reaches sieving machine and obtains sulphur through sieving
Sour capreomycin.
9. the method described in accordance with the claim 1 for preparing capreomycin sulfate using capreomycin zymotic fluid, it is characterised in that
The preprocessing process is that the concentration of phosphoric acid is controlled 20~40%；The concentration of sodium carbonate is controlled 30~40%；Microbial flocculation
Agent dosage is controlled in 60~70mg/L；Filter aid is diatomite or cellulose or perlite, dosage be fermentating liquid volume 1~
5%。
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| CN105506042A (en) * | 2016-02-23 | 2016-04-20 | 华北制药集团新药研究开发有限责任公司 | Method for producing capreomycin by fermentation |
| CN105504023A (en) * | 2016-02-23 | 2016-04-20 | 华北制药集团新药研究开发有限责任公司 | Capreomycin preparation method |
| CN105622593B (en) * | 2016-02-25 | 2018-06-26 | 中国农业科学院蜜蜂研究所 | A kind of extracting method of fumidil |
| CN110483617A (en) * | 2019-09-17 | 2019-11-22 | 陕西绿盾环境工程研究院有限公司 | A kind of preparation method of high-content polyoxin original powder |
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