CN105859642A - Tetramethylpyrazine extraction and purification method - Google Patents
Tetramethylpyrazine extraction and purification method Download PDFInfo
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- CN105859642A CN105859642A CN201610242785.2A CN201610242785A CN105859642A CN 105859642 A CN105859642 A CN 105859642A CN 201610242785 A CN201610242785 A CN 201610242785A CN 105859642 A CN105859642 A CN 105859642A
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- tetramethylpyazine
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- tetramethylpyrazine
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- 238000000746 purification Methods 0.000 title claims abstract description 28
- 238000000034 method Methods 0.000 title claims abstract description 27
- 238000000605 extraction Methods 0.000 title claims abstract description 24
- FINHMKGKINIASC-UHFFFAOYSA-N Tetramethylpyrazine Chemical compound CC1=NC(C)=C(C)N=C1C FINHMKGKINIASC-UHFFFAOYSA-N 0.000 title abstract description 20
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims abstract description 35
- 239000006228 supernatant Substances 0.000 claims abstract description 28
- 238000000855 fermentation Methods 0.000 claims abstract description 26
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- 239000013078 crystal Substances 0.000 claims abstract description 16
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- 238000001223 reverse osmosis Methods 0.000 claims abstract description 12
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- 238000000926 separation method Methods 0.000 abstract description 3
- 229910052799 carbon Inorganic materials 0.000 abstract 1
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- 244000063299 Bacillus subtilis Species 0.000 description 3
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- 244000068988 Glycine max Species 0.000 description 2
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- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 2
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- BHPQYMZQTOCNFJ-UHFFFAOYSA-N Calcium cation Chemical compound [Ca+2] BHPQYMZQTOCNFJ-UHFFFAOYSA-N 0.000 description 1
- 208000022461 Glomerular disease Diseases 0.000 description 1
- 239000005909 Kieselgur Substances 0.000 description 1
- 206010029155 Nephropathy toxic Diseases 0.000 description 1
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 1
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- 150000003797 alkaloid derivatives Chemical class 0.000 description 1
- WYTGDNHDOZPMIW-RCBQFDQVSA-N alstonine Natural products C1=CC2=C3C=CC=CC3=NC2=C2N1C[C@H]1[C@H](C)OC=C(C(=O)OC)[C@H]1C2 WYTGDNHDOZPMIW-RCBQFDQVSA-N 0.000 description 1
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- DQLATGHUWYMOKM-UHFFFAOYSA-L cisplatin Chemical compound N[Pt](N)(Cl)Cl DQLATGHUWYMOKM-UHFFFAOYSA-L 0.000 description 1
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- 125000000623 heterocyclic group Chemical group 0.000 description 1
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07D—HETEROCYCLIC COMPOUNDS
- C07D241/00—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings
- C07D241/02—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings not condensed with other rings
- C07D241/10—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings not condensed with other rings having three double bonds between ring members or between ring members and non-ring members
- C07D241/12—Heterocyclic compounds containing 1,4-diazine or hydrogenated 1,4-diazine rings not condensed with other rings having three double bonds between ring members or between ring members and non-ring members with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached to ring carbon atoms
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Separation Using Semi-Permeable Membranes (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention relates to a tetramethylpyrazine extraction and purification method. The method includes the following steps that firstly, a fermentation solution is heated, thalluses are centrifugally separated to obtain supernatant, and the supernatant is subjected to active carbon decoloration; secondly, the decolorized supernatant is filtered to obtain tetramethylpyrazine dialysate; thirdly, the tetramethylpyrazine dialysate is filtered and then concentrated through a reverse osmosis filter membrane to obtain an infiltration solution; fourthly, the infiltration solution is cooled and crystallized, the temperature is reduced, tetramethylpyrazine crystals are centrifugally separated, then the tetramethylpyrazine crystals are cooled and re-crystallized after redissolving, centrifugal separation is conducted, and high-purity tetramethylpyrazine is obtained. By means of the separation and extraction method, the purity of tetramethylpyrazine is high and is up to 99% or above, and purification efficiency is high and is up to 88% or above.
Description
Technical field
The invention belongs to bioengineering field, relate to the method for extraction and purification of a kind of tetramethylpyazine, particularly relate to a kind of from fermentation
The production technology of isolated and purified tetramethylpyazine in liquid.
Background technology
Tetramethylpyazine, molecular formula is C8H12N2, molecular weight 136.20, fusing point 84-86 DEG C, boiling point 190 DEG C, is a kind of normal
The heterocyclic nitrogen seen, is widely present in raw-food material, processed food and alcoholic beverage it is considered to be a class is important
Perfume compound, has the fragrance of baking, Semen arachidis hypogaeae, Semen coryli heterophyllae and cocoa, be usually used in baked goods, cold drink, meat, milk product,
The allotment of the essence such as Medicated cigarette.Except the purposes as flavour of food products additive, tetramethylpyazine is as the master of Chinese crude drug Rhizome of Ligusticum Sinense Oliv. Cv. Chuanxiong
Want active alkaloid composition, have been demonstrated to have the pharmacological action for the treatment of cardiovascular and cerebrovascular disease, and to the oxidative stress of cisplatin induction,
Apoptosis and nephrotoxicity have preventive effect.Along with going deep into of pharmacological research, the most as a kind of novel calcium ion antagonism
Agent is widely used in clinic, is large medication of circulation, nervus centralis, breathing and other system related disorders, is widely used in
The treatment of cardiovascular and cerebrovascular disease, respiratory system disease, renal glomerular disease etc..
The production method of tetramethylpyazine can be divided into biological synthesis process, directly extraction method and chemical synthesis three kinds from plant.Cut
The most at present, existing considerable amount of document and patent document report utilize Maillard reaction and Strecker degraded chemical method synthesis pyrazine
Compounds, but, generally there is severeer environmental issue in chemical synthesis, reaction condition is the most violent, equipment
Ask higher；Although the price of the pyrazine that chemosynthesis obtains is far below the former, but consumer is more likely to use " natural " or " raw
Thing synthesizes " pyrazine.The method the most therefrom extracting tetramethylpyazine in Ligusticum chuanxiong rhizome often faces Rhizoma Chuanxiong material of vegetable origin supply
Limited problem, and, tetramethylpyazine content in Rhizome of Ligusticum Sinense Oliv. Cv. Chuanxiong relatively low (about 0.075%) causes extraction process cost relatively
High and be difficulty with large-scale production.Along with people's raising day by day to green product demand, bionic means are utilized to produce
Tetramethylpyazine has the advantages such as Product Green is natural, low cost, reaction condition gentle, environmental pollution is little and is extensively closed by consumer
Note.
Chinese patent document CN101445786A (application number: 200810235366.1) discloses a plant height and produces tetramethylpyazine
Bacillus subtilis and the method for fermentation producing tetramethylpyrazine thereof, this bacterial strain can be with glucose, sucrose, molasses and soybean cake powder
For substrate, improve the yield of tetramethylpyazine with source precursor in a large amount of accumulation.Solve fermentable and produce tetramethylpyazine
Middle production concentration is low, need external source to add precursor and the low problem of precursor utilization rate.With sucrose soybean cake powder under the conditions of during substrate 37 DEG C
Shake-flask culture 120h can obtain the tetramethylpyazine of 4.08g/L.
Chinese patent document CN 101955980A (application number: 201010238685.5) discloses a kind of production tetramethylpyazine
Method and produce bacterial strain, specifically a kind of microorganism utilizes reducing sugar fermentation accumulation precursor acetoin and acetoin
Two-step method production technology with ammonia is synthesized tetramethylpyazine through non-enzymatic, belongs to technical field of bioengineering.Described microorganism
For bacillus subtilis (CCTCCNO:M208157), Bacillus licheniformis (CGMCCNO:3961) Bacillus licheniformis (CGMCCNO:
3962) and an arbitrary strain in Bacillus licheniformis (CGMCCNO:3963), above-mentioned bacterial strains utilizes reducing sugar fermentation to obtain Biomass
And source precursor acetoin in accumulating, then, acetoin and ammonia in fermentation system synthesize tetramethylpyazine through non-enzymic catalytic reaction.
Advantage is as follows: 1. obtain higher biomass, and acetoin accumulation is effectively improved (38-44g/L)；2. high concentration is interior
Source precursor acetoin and ammonia fast reaction can generate tetramethylpyazine (16-20g/L) under appropriate conditions；The most endogenous acetoin
Accumulation and in-situ fermentation environment, significantly improve the utilization rate (40.3%) of precursor.
Chinese patent document CN 102618587A (application number: 201210089617.6) discloses with distiller grains as raw material, utilizes
Fermentable produces bioflavours acetoin and the method for tetramethylpyazine, belongs to technical field of bioengineering.Adjusting through acidity
The fresh vinasse of joint supplements appropriate glucose 0～100g/kg, adds bacillus subtilis (CCTCCNO:M208157) or ground
After the inoculum that in clothing bacillus cereus (CGMCC3961,3962,3963), arbitrary strain is cultivated, 37 DEG C ferment 1～3 day, energy
Enough a large amount of generation bioflavours acetoin.On this basis, production is converted also by interpolation ammonium salt in above-mentioned tunning
Another kind of bioflavours tetramethylpyazine.Prominent advantage be utilize distiller grains for raw material, by whether it is selectable to add ammonium salt
Control to produce bioflavours acetoin or the method for tetramethylpyazine, both can selectively obtain the bioflavours of different high added value,
Prevent again distiller grains to pollute environment, improve the value of distiller grains.
Chinese patent document CN 105002115A (application number: 201510450768.3) disclose a kind of Bacillus licheniformis and
The Daqu (massive raw stater for alcholic liquor) prepared with it and the preparation method of this Daqu (massive raw stater for alcholic liquor), the Classification And Nomenclature of this bacterial strain is Bacillus licheniformis
(Bacilluslincheniformis), being preserved in China typical culture collection center, preserving number is CCTCCNO.M2015224,
And after being prepared zymocyte liquid, as functional bacterium solution, more functional bacterium solution is seeded on starter-making materials carries out fermentation process,
Obtaining Daqu (massive raw stater for alcholic liquor) product, this technological process is short, easily operates, and the Daqu (massive raw stater for alcholic liquor) obtained is compared with tradition Daqu (massive raw stater for alcholic liquor), its tetramethylpyazine
Content improve to more than 9 times.
Although method disclosed in above-mentioned patent document is helpful for improving the concentration of tetramethylpyazine in fermentation liquid, but from
In fermentation liquid, purification & isolation obtains high-purity tetramethylpyazine does not has play-by-play.And at Chinese patent document CN 102618587A
In, the tetramethylpyazine of description extracts from solid state fermentation culture thing, proposes to use water or soak with ethanol, ultrasonic 1h, then adopts
After being centrifuged or filtering, carrying out supernatant decompression distillation, collection fraction carries out crystallisation by cooling and obtains wet crystal, then carries out freezing dry
The dry powder crystal that obtains, ultrasonic, lyophilization and decompression distillation energy consumption that whole extraction process uses are big, are not suitable for scale
Application, and seriality is restricted, during tetramethylpyazine volatilize easily damaged mistake, thus cause relatively low tetramethylpyazine
The response rate.
Summary of the invention
For purification difficult point and the defect of existing fermentation producing tetramethylpyrazine, the present invention provides the extraction of a kind of tetramethylpyazine pure
Change method.
Term illustrates:
The fermentation liquid that the present invention is extracted is hay spore bacillus or the lichen bacillus ferments gained contains sending out of tetramethylpyazine
Ferment liquid；Wherein, hay spore bacillus or Bacillus licheniformis are existing bacterial strain, and sweat is all by prior art.
Technical scheme is as follows:
The method for extraction and purification of a kind of tetramethylpyazine, comprises the following steps that
(1) fermentation liquid is warming up to 70-80 DEG C, centrifugation thalline, obtain supernatant；Supernatant is maintained temperature 70-80 DEG C
Carry out decolorizing with activated carbon；
(2) by the supernatant liquid filtering after decolouring, tetramethylpyazine dialysis solution is obtained；
(3) by concentrating through reverse osmosis filter membrane after tetramethylpyazine dialysate filter, penetrating fluid is obtained again；
(4) penetrating fluid is carried out crystallisation by cooling, when being cooled to 10-15 DEG C, centrifugation tetramethylpyazine crystal, the most again will
Tetramethylpyazine crystal carries out cooling down recrystallization, centrifugation after dissolving in 70-90 DEG C, obtains high-purity tetramethylpyazine.
According to the invention it is preferred to, the device therefor that heated up by fermentation liquid in step (1) is pipe heat exchanger, more preferably
Inside and outside jacket heat-exchanger, the serpentine bend for the additional sleeve pipe of long bend pipe combines, and material is 316L rustless steel, and heat exchange area is so that sending out
Ferment liquid temp is warming up to 80 DEG C of calculating from 30 DEG C.Heating materials in chuck is the hot water of 90-95 DEG C；
Preferably, centrifugation thalline device therefor is centrifuge, and material is 316L rustless steel, further preferred disc-stack centrifuge
Machine, centrifugal rotational speed is 7000-10000 rev/min, and further preferred rotating speed is 8000 revs/min；
Preferably, decolorization is to use activated carbon to seal decolouring, it is further preferred that described activated carbon is pharmaceutical grade activated carbon
Powder, activated carbon content is the 1-1.5% of supernatant volume；The pressure of decolorization is 0.1-0.2MPa, and bleaching time is
20-40min, adjusting pH after decolouring is 7.0-8.0.
According to the invention it is preferred to, in step (2), filter process is: first filters with plate filter, then carries out ultrafiltration and go
Removal of impurity albumen, finally carries out nanofiltration and goes the removal of impurity further；
During it is further preferred that plate filter filters, laying kieselguhr in plate filter, described kieselguhr is particle diameter
Press the mixing of 3:1 mass ratio at the thin kieselguhr of 0.02-2 μm at thick kieselguhr and the particle diameter of 28-68 μm；Lay thickness collocation silicon
Diatomaceous earth, it is possible to extend filtration time, increases and filters clarity, prevent from blocking filter cloth space because the viscosity of medium is too high；
The material of plate filter screen plate is politef；
It is further preferred that the ultrafilter membrane molecular cut off of ultra-filtration process is 1000Dalton；
It is further preferred that the NF membrane molecular cut off of nanofiltration process is 200Dalton.
According to the invention it is preferred to, the temperature concentrated in step (3) is 60-70 DEG C, and before concentrating, solution is solution body after concentrating
Long-pending 2.5-3 times, takes concentration and carries out decrease temperature crystalline mutually after reverse osmosis concentration, reverse osmosis membrane maximum operating pressure is less than 0.8MPa.
Reverse osmosis filter membrane primarily serve the purpose of concentration penetrating fluid.
According to the invention it is preferred to, in step (4), cooling rate is lowered the temperature with 5-10 DEG C/h, until crystallization.Crystallisation by cooling
Carrying out in crystallizer, crystallizer is vertical mixing crystallizer, crystallizer sandwich, and centrifugal centrifuge is carried out, described centrifugal
Machine is cloth-bag type tripod pendulum type batch centrifugal.
The present invention contains separating-purifying the fermentation liquid of tetramethylpyazine from hay spore bacillus or the lichen bacillus ferments gained
Tetramethylpyazine, every contain the fermentation liquid of tetramethylpyazine through hay spore bacillus or the lichen bacillus ferments all can conduct
The raw material of the inventive method.
Beneficial effects of the present invention is as follows:
1, the purity of the process for separation and purification tetramethylpyazine of the present invention is high, and up to more than 99%, purification efficiency is high, and up to 88%
Above.
2, the inventive method be adapted to fermentable prepare tetramethylpyazine industrialization extract preparation, the inventive method automatization
Degree is high, and utilization rate of equipment and installations is high, and process is simple, and loss is few, and removal of impurity is high, and the product purity of preparation and the response rate are the most relatively
Traditional method improves a lot.
Detailed description of the invention
Below by specific embodiment, the present invention will be further described, but is not limited to this.
The raw materials used convenient source that is in embodiment, device therefor is conventional equipment.
Wherein: fermentation liquid is the fermentation liquid obtained with reference to method disclosed in Chinese patent document CN 102618587A.
Activated carbon is pharmaceutical grade active carbon powder.
Pipe heat exchanger is interior additional to buy or get by illegal means hot device, and for the bending combination of long pipe-in-pipe, material is 316L rustless steel, heat exchange area with
Broth temperature is made to be warming up to 80 DEG C of calculating from 30 DEG C；
Centrifugation thalline device therefor is disc centrifuge, and material is 316L rustless steel, and centrifugal rotational speed is 7000-10000
Rev/min；
Bleacher is for sealing pressure fermentation tank, and proof pressure is 0.2MPa, and fermentation tank is with pressure relief opening, and pressure relief opening is with recirculated water
Cooling and reflux device；
Plate filter is for being equipped with diatomaceous plate filter, and kieselguhr is particle diameter at the thick kieselguhr of 30-40 μm and particle diameter
Thin kieselguhr in 1-2 μm presses the mixing of 3:1 mass ratio；The material of plate filter screen plate is politef；
Ultrafilter membrane molecular cut off used by ultrafiltration system is 1000Dalton, and NF membrane molecular cut off used by nanofiltration is
200Dalton；
Crystallizer is vertical mixing crystallizer, crystallizer sandwich, and centrifuge used after crystallization is cloth-bag type tripod pendulum type batch centrifugal.
Embodiment 1
The method for extraction and purification of a kind of tetramethylpyazine, comprises the following steps that
(1) by fermentation liquid after pipe heat exchanger heat exchange, it is warming up to 70 DEG C, pumps into centrifuge through sanitary centrifugal pump, from
After heart separating thallus, obtain supernatant；Supernatant is entered the activated carbon decolorizing tank with return duct, seals decolouring 30min, dimension
Hold temperature 70-80 DEG C；Activated carbon content is the 1% of supernatant volume；The pressure of decolorization is 0.1-0.12MPa, decolouring decolouring
Rear adjustment pH is 7.0-8.0；
(2) supernatant after step (1) being decoloured, after plate filter filters, enters surge tank, subsequently into ultrafiltration system
System, after being filtered to remove foreign protein, after going the removal of impurity by nanofiltration further, it is thus achieved that tetramethylpyazine dialysis solution；
(3) after being concentrated in 60-65 DEG C by tetramethylpyazine dialysis solution in step (2) again after reverse osmosis filter membrane concentrates, infiltration is obtained
Liquid, takes concentrated phase after infiltration, and cycles of concentration is 2.5 times
(4) by step (3) carries out crystallisation by cooling in penetrating fluid entrance crystallisation by cooling tank, it is cooled to 15 with the speed of 10 DEG C/h
DEG C time, centrifugation tetramethylpyazine crystal, the most again by tetramethylpyazine crystal in 80 DEG C dissolve after carry out cool down recrystallization,
Centrifugation, obtains high-purity tetramethylpyazine.
The tetramethylpyazine purity that the present embodiment obtains is 99.1%, and purification efficiency is 89.4%, reaches pharmaceutical grade purity requirement.
Embodiment 2
The method for extraction and purification of a kind of tetramethylpyazine, comprises the following steps that
(1) by fermentation liquid after pipe heat exchanger heat exchange, it is warming up to 80 DEG C, pumps into centrifuge through sanitary centrifugal pump, from
After heart separating thallus, obtain supernatant；Supernatant is entered the activated carbon decolorizing tank with return duct, seals decolouring 40min, dimension
Hold temperature 80 DEG C；Activated carbon content is the 1.5% of supernatant volume；The pressure of decolorization is 0.12-0.15MPa, decolouring decolouring
Rear adjustment pH is 7.0-8.0；
(2) supernatant after step (1) being decoloured, after plate filter filters, enters surge tank, subsequently into ultrafiltration system
System, after being filtered to remove foreign protein, after going the removal of impurity by nanofiltration further, it is thus achieved that tetramethylpyazine dialysis solution；
(3) after being concentrated in 70-75 DEG C by tetramethylpyazine dialysis solution in step (2) again after reverse osmosis filter membrane concentrates, infiltration is obtained
Liquid；Concentrated solution concentrated phase cycles of concentration reaches 3 times
(4) by step (3) carries out crystallisation by cooling in penetrating fluid entrance crystallisation by cooling tank, it is cooled to 10 with the speed of 5 DEG C/h
DEG C time, centrifugation tetramethylpyazine crystal, the most again by tetramethylpyazine crystal in 70 DEG C dissolve after carry out cool down recrystallization,
Centrifugation, obtains high-purity tetramethylpyazine.
The tetramethylpyazine purity that the present embodiment obtains is 99.2%, and purification efficiency is 88.2%, and purity reaches pharma grade product and wants
Ask.
Embodiment 3
The method for extraction and purification of a kind of tetramethylpyazine, comprises the following steps that
(1) by fermentation liquid after pipe heat exchanger heat exchange, it is warming up to 75 DEG C, pumps into centrifuge through sanitary centrifugal pump, from
After heart separating thallus, obtain supernatant；Supernatant is entered the activated carbon decolorizing tank with return duct, seals decolouring 20min, dimension
Hold temperature 75 DEG C；Activated carbon content is the 1.2% of supernatant volume；The pressure of decolorization is 0.15-0.19MPa, and decolouring is de-
Adjusting pH after color is 7.0-8.0；
(2) supernatant after step (1) being decoloured, after plate filter filters, enters surge tank, subsequently into ultrafiltration system
System, after being filtered to remove foreign protein, after going the removal of impurity by nanofiltration further, it is thus achieved that tetramethylpyazine dialysis solution；
(3) after being concentrated in 55-60 DEG C by tetramethylpyazine dialysis solution in step (2) again after reverse osmosis filter membrane concentrates, infiltration is obtained
Liquid；Concentrated solution concentrated phase cycles of concentration reaches 2 times；
(4) by step (3) carries out crystallisation by cooling in penetrating fluid entrance crystallisation by cooling tank, it is cooled to 12 with the speed of 8 DEG C/h
DEG C time, centrifugation tetramethylpyazine crystal, the most again by tetramethylpyazine crystal in 90 DEG C dissolve after carry out cool down recrystallization,
Centrifugation, obtains high-purity tetramethylpyazine.
The tetramethylpyazine purity that the present embodiment obtains is 99.05%, and purification efficiency is 88.67%, reaches pharma grade product requirement.
Comparative example
The method for extraction and purification of a kind of tetramethylpyazine, comprises the following steps that
(1) by fermentation liquid after pipe heat exchanger heat exchange, it is warming up to 75 DEG C, pumps into centrifuge through sanitary centrifugal pump, from
After heart separating thallus, obtain supernatant；Supernatant is entered the activated carbon decolorizing tank with return duct, seals decolouring 20min, dimension
Hold temperature 75 DEG C；Activated carbon content is the 1.2% of supernatant volume；The pressure of decolorization is 0.1MPa, adjusts after decolouring decolouring
Whole pH is 5.0-6.0；
(2) supernatant after step (1) being decoloured, after plate filter filters, enters surge tank, subsequently into ultrafiltration system
System, after being filtered to remove foreign protein, after going the removal of impurity by nanofiltration further, it is thus achieved that tetramethylpyazine dialysis solution；
(3) after being concentrated in 50-55 DEG C by tetramethylpyazine dialysis solution in step (2) again after reverse osmosis filter membrane concentrates, infiltration is obtained
Liquid；Concentrated solution concentrated phase cycles of concentration reaches 2.5 times；
(4) by step (3) carries out crystallisation by cooling in penetrating fluid entrance crystallisation by cooling tank, it is cooled to 12 with the speed of 8 DEG C/h
DEG C time, centrifugation tetramethylpyazine crystal, the most again by tetramethylpyazine crystal in 90 DEG C dissolve after carry out cool down recrystallization,
Centrifugation, obtains high-purity tetramethylpyazine.
The tetramethylpyazine purity that the present embodiment obtains is 97%, and purification efficiency is 65.8%, reaches chemical pure product requirement.
Claims (10)
1. a method for extraction and purification for tetramethylpyazine, comprises the following steps that
(1) fermentation liquid is warming up to 70-80 DEG C, centrifugation thalline, obtain supernatant；Supernatant is maintained temperature 70-80 DEG C
Carry out decolorizing with activated carbon；
(2) by the supernatant liquid filtering after decolouring, tetramethylpyazine dialysis solution is obtained；
(3) by concentrating through reverse osmosis filter membrane after tetramethylpyazine dialysate filter, penetrating fluid is obtained again；
(4) penetrating fluid is carried out crystallisation by cooling, when being cooled to 10-15 DEG C, centrifugation tetramethylpyazine crystal, the most again will
Tetramethylpyazine crystal carries out cooling down recrystallization, centrifugation after dissolving in 70-90 DEG C, obtains high-purity tetramethylpyazine.
The method for extraction and purification of tetramethylpyazine the most according to claim 1, it is characterised in that will fermentation in step (1)
Liquid intensification device therefor is pipe heat exchanger.
The method for extraction and purification of tetramethylpyazine the most according to claim 1, it is characterised in that centrifugal in step (1)
Separating thallus device therefor is centrifuge, and centrifugal rotational speed is 7000-10000 rev/min.
The method for extraction and purification of tetramethylpyazine the most according to claim 1, it is characterised in that decolouring in step (1)
Process is to use activated carbon to seal decolouring；
Preferably, activated carbon content is the 1-1.5% of supernatant volume；
Preferably, the pressure of decolorization is 0.1-0.2MPa, and bleaching time is 20-40min.
The method for extraction and purification of tetramethylpyazine the most according to claim 1, it is characterised in that decolouring in step (1)
Rear adjustment pH is 7.0-8.0.
The method for extraction and purification of tetramethylpyazine the most according to claim 1, it is characterised in that filter in step (2)
Process is: first filter with plate filter, then carries out ultrafiltration and removes impurity protein, finally carries out nanofiltration and go the removal of impurity further.
The method for extraction and purification of tetramethylpyazine the most according to claim 6, it is characterised in that sheet frame in step (2)
When filter filters, plate filter is laid kieselguhr, described kieselguhr be particle diameter 28-68 μm thick kieselguhr with
Particle diameter presses the mixing of 3:1 mass ratio at the thin kieselguhr of 0.02-2 μm.
The method for extraction and purification of tetramethylpyazine the most according to claim 6, it is characterised in that ultrafiltration in step (2)
The ultrafilter membrane molecular cut off of process is 1000Dalton；
Preferably, the NF membrane molecular cut off of nanofiltration process is 200Dalton.
The method for extraction and purification of tetramethylpyazine the most according to claim 1, it is characterised in that concentrate in step (3)
Temperature be 60-70 DEG C, before concentrating solution be concentrate after 2.5-3 times of liquor capacity, take after reverse osmosis concentration to concentrate and drop mutually
Temperature crystallization, reverse osmosis membrane maximum operating pressure is less than 0.8MPa.
The method for extraction and purification of tetramethylpyazine the most according to claim 1, it is characterised in that cooling in step (4)
Speed is lowered the temperature with 5-10 DEG C/h, until crystallization.
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