CN105859642A - Tetramethylpyrazine extraction and purification method - Google Patents
Tetramethylpyrazine extraction and purification method Download PDFInfo
- Publication number
- 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
- Authority
- CN
- China
- Prior art keywords
- tetramethylpyazine
- extraction
- purification
- supernatant
- tetramethylpyrazine
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 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
- 230000004151 fermentation Effects 0.000 claims abstract description 26
- 239000013078 crystal Substances 0.000 claims abstract description 16
- 239000012528 membrane Substances 0.000 claims abstract description 16
- 238000001223 reverse osmosis Methods 0.000 claims abstract description 12
- 239000007788 liquid Substances 0.000 claims description 25
- 238000001816 cooling Methods 0.000 claims description 17
- 238000005119 centrifugation Methods 0.000 claims description 16
- 238000002425 crystallisation Methods 0.000 claims description 16
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 12
- 239000000385 dialysis solution Substances 0.000 claims description 10
- 230000008569 process Effects 0.000 claims description 10
- 239000012530 fluid Substances 0.000 claims description 9
- 239000012535 impurity Substances 0.000 claims description 9
- 238000001728 nano-filtration Methods 0.000 claims description 9
- 230000000149 penetrating effect Effects 0.000 claims description 9
- 238000000108 ultra-filtration Methods 0.000 claims description 9
- 239000000243 solution Substances 0.000 claims description 8
- 238000010792 warming Methods 0.000 claims description 8
- 239000012141 concentrate Substances 0.000 claims description 7
- 238000004042 decolorization Methods 0.000 claims description 7
- 239000002245 particle Substances 0.000 claims description 6
- 238000001953 recrystallisation Methods 0.000 claims description 6
- 238000002156 mixing Methods 0.000 claims description 5
- 102000004169 proteins and genes Human genes 0.000 claims description 5
- 108090000623 proteins and genes Proteins 0.000 claims description 5
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 claims description 4
- 230000008025 crystallization Effects 0.000 claims description 4
- 238000001914 filtration Methods 0.000 claims description 4
- 238000004061 bleaching Methods 0.000 claims description 2
- 230000008595 infiltration Effects 0.000 abstract description 7
- 238000001764 infiltration Methods 0.000 abstract description 7
- 238000000926 separation method Methods 0.000 abstract description 3
- 229910052799 carbon Inorganic materials 0.000 abstract 1
- ROWKJAVDOGWPAT-UHFFFAOYSA-N Acetoin Chemical compound CC(O)C(C)=O ROWKJAVDOGWPAT-UHFFFAOYSA-N 0.000 description 20
- GFAZHVHNLUBROE-UHFFFAOYSA-N hydroxymethyl propionaldehyde Natural products CCC(=O)CO GFAZHVHNLUBROE-UHFFFAOYSA-N 0.000 description 10
- 239000000463 material Substances 0.000 description 10
- 239000000047 product Substances 0.000 description 8
- 241000193830 Bacillus <bacterium> Species 0.000 description 7
- 239000002243 precursor Substances 0.000 description 7
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 6
- 241000194108 Bacillus licheniformis Species 0.000 description 6
- 238000004519 manufacturing process Methods 0.000 description 6
- 230000001580 bacterial effect Effects 0.000 description 5
- KYQCOXFCLRTKLS-UHFFFAOYSA-N Pyrazine Chemical compound C1=CN=CC=N1 KYQCOXFCLRTKLS-UHFFFAOYSA-N 0.000 description 4
- 229910000831 Steel Inorganic materials 0.000 description 4
- 238000009825 accumulation Methods 0.000 description 4
- 238000006243 chemical reaction Methods 0.000 description 4
- 239000002994 raw material Substances 0.000 description 4
- 239000010959 steel Substances 0.000 description 4
- 238000003786 synthesis reaction Methods 0.000 description 4
- 244000063299 Bacillus subtilis Species 0.000 description 3
- 235000014469 Bacillus subtilis Nutrition 0.000 description 3
- 241000244365 Ligusticum sinense Species 0.000 description 3
- 229910021529 ammonia Inorganic materials 0.000 description 3
- 230000008901 benefit Effects 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 238000000819 phase cycle Methods 0.000 description 3
- 239000000843 powder Substances 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- 239000002028 Biomass Substances 0.000 description 2
- 208000024172 Cardiovascular disease Diseases 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 2
- 244000068988 Glycine max Species 0.000 description 2
- 235000010469 Glycine max Nutrition 0.000 description 2
- PCNDJXKNXGMECE-UHFFFAOYSA-N Phenazine Natural products C1=CC=CC2=NC3=CC=CC=C3N=C21 PCNDJXKNXGMECE-UHFFFAOYSA-N 0.000 description 2
- 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
- 229930006000 Sucrose Natural products 0.000 description 2
- 150000003863 ammonium salts Chemical class 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 208000026106 cerebrovascular disease Diseases 0.000 description 2
- 230000006837 decompression Effects 0.000 description 2
- 238000004821 distillation Methods 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 230000002526 effect on cardiovascular system Effects 0.000 description 2
- 239000000284 extract Substances 0.000 description 2
- 239000008103 glucose Substances 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 230000000144 pharmacologic effect Effects 0.000 description 2
- 229950000845 politef Drugs 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- 210000000582 semen Anatomy 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- 239000005720 sucrose Substances 0.000 description 2
- 241000193755 Bacillus cereus Species 0.000 description 1
- 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
- 244000299461 Theobroma cacao Species 0.000 description 1
- 235000009470 Theobroma cacao Nutrition 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- 235000013334 alcoholic beverage Nutrition 0.000 description 1
- 229930013930 alkaloid Natural products 0.000 description 1
- 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
- 230000008485 antagonism Effects 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 235000015173 baked goods and baking mixes Nutrition 0.000 description 1
- 238000005452 bending Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 229910001424 calcium ion Inorganic materials 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 235000019504 cigarettes Nutrition 0.000 description 1
- DQLATGHUWYMOKM-UHFFFAOYSA-L cisplatin Chemical compound N[Pt](N)(Cl)Cl DQLATGHUWYMOKM-UHFFFAOYSA-L 0.000 description 1
- 229960004316 cisplatin Drugs 0.000 description 1
- 235000020965 cold beverage Nutrition 0.000 description 1
- 230000000052 comparative effect Effects 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000005265 energy consumption Methods 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 239000004744 fabric Substances 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 239000003205 fragrance Substances 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 231100000852 glomerular disease Toxicity 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 125000000623 heterocyclic group Chemical group 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 238000009434 installation Methods 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 235000013379 molasses Nutrition 0.000 description 1
- 231100000417 nephrotoxicity Toxicity 0.000 description 1
- 230000007694 nephrotoxicity Effects 0.000 description 1
- 239000011664 nicotinic acid Substances 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 230000036542 oxidative stress Effects 0.000 description 1
- 239000002304 perfume Substances 0.000 description 1
- 230000003449 preventive effect Effects 0.000 description 1
- 150000003216 pyrazines Chemical class 0.000 description 1
- 235000021067 refined food Nutrition 0.000 description 1
- 238000010992 reflux Methods 0.000 description 1
- 230000029058 respiratory gaseous exchange Effects 0.000 description 1
- 208000023504 respiratory system disease Diseases 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 238000012807 shake-flask culturing Methods 0.000 description 1
- 229910052710 silicon Inorganic materials 0.000 description 1
- 239000010703 silicon Substances 0.000 description 1
- 238000010563 solid-state fermentation Methods 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 210000004243 sweat Anatomy 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
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.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610242785.2A CN105859642B (en) | 2016-04-19 | 2016-04-19 | A kind of method for extraction and purification of Tetramethylpyrazine |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610242785.2A CN105859642B (en) | 2016-04-19 | 2016-04-19 | A kind of method for extraction and purification of Tetramethylpyrazine |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105859642A true CN105859642A (en) | 2016-08-17 |
| CN105859642B CN105859642B (en) | 2018-07-31 |
Family
ID=56632346
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610242785.2A Active CN105859642B (en) | 2016-04-19 | 2016-04-19 | A kind of method for extraction and purification of Tetramethylpyrazine |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105859642B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106800593A (en) * | 2017-01-09 | 2017-06-06 | 博瑞生物医药泰兴市有限公司 | A kind of method for purifying anidulafungin precursor compound |
| CN115386525A (en) * | 2022-10-26 | 2022-11-25 | 中粮营养健康研究院有限公司 | Bacillus subtilis, microbial inoculum, application and method for preparing tetramethylpyrazine |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1603313A (en) * | 2004-08-25 | 2005-04-06 | 长沙卷烟厂 | Method for separation purification of polyhydroxy-alkyl-pyrazine from suger amine reaction liquid |
| CN1803779A (en) * | 2006-01-24 | 2006-07-19 | 长沙卷烟厂 | Method for purifying polyhydroxy alkyl pyrazine compound from browning reaction liquor |
| CN101602735A (en) * | 2009-07-01 | 2009-12-16 | 南京泽朗医药科技有限公司 | A kind of preparation method of Ligustrazine |
| CN102344417A (en) * | 2010-08-06 | 2012-02-08 | 苏州瑞蓝博中药技术开发有限公司 | Extraction method for ligustrazine |
| CN102618587A (en) * | 2012-03-30 | 2012-08-01 | 江南大学 | Method for producing acetoin by vinasse fermentation and producing tetramethylpyrazine by acetoin transformation |
| CN104323996A (en) * | 2010-07-29 | 2015-02-04 | 湖南康都制药有限公司 | Ligustrazine phosphate lipidosome combined drug, and large-scale production process and application thereof |
-
2016
- 2016-04-19 CN CN201610242785.2A patent/CN105859642B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1603313A (en) * | 2004-08-25 | 2005-04-06 | 长沙卷烟厂 | Method for separation purification of polyhydroxy-alkyl-pyrazine from suger amine reaction liquid |
| CN1803779A (en) * | 2006-01-24 | 2006-07-19 | 长沙卷烟厂 | Method for purifying polyhydroxy alkyl pyrazine compound from browning reaction liquor |
| CN101602735A (en) * | 2009-07-01 | 2009-12-16 | 南京泽朗医药科技有限公司 | A kind of preparation method of Ligustrazine |
| CN104323996A (en) * | 2010-07-29 | 2015-02-04 | 湖南康都制药有限公司 | Ligustrazine phosphate lipidosome combined drug, and large-scale production process and application thereof |
| CN102344417A (en) * | 2010-08-06 | 2012-02-08 | 苏州瑞蓝博中药技术开发有限公司 | Extraction method for ligustrazine |
| CN102618587A (en) * | 2012-03-30 | 2012-08-01 | 江南大学 | Method for producing acetoin by vinasse fermentation and producing tetramethylpyrazine by acetoin transformation |
Non-Patent Citations (2)
| Title |
|---|
| 徐伟泉: "川芎嗪提取分离纯化及稳定性研究", 《北京化工大学硕士研究生学位论文》 * |
| 马双成,等: "川芎提取、纯化工艺条件的实验研究", 《中国中药杂志》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106800593A (en) * | 2017-01-09 | 2017-06-06 | 博瑞生物医药泰兴市有限公司 | A kind of method for purifying anidulafungin precursor compound |
| CN106800593B (en) * | 2017-01-09 | 2021-04-27 | 博瑞生物医药泰兴市有限公司 | Method for purifying anidulafungin precursor compound |
| CN115386525A (en) * | 2022-10-26 | 2022-11-25 | 中粮营养健康研究院有限公司 | Bacillus subtilis, microbial inoculum, application and method for preparing tetramethylpyrazine |
| CN115386525B (en) * | 2022-10-26 | 2023-01-31 | 中粮营养健康研究院有限公司 | Bacillus subtilis, microbial inoculum, application and method for preparing tetramethylpyrazine |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105859642B (en) | 2018-07-31 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103184246B (en) | Biosynthesis preparation method for L-Ergothioneine | |
| CN108752231B (en) | Method for extracting theanine from sweet tea and simultaneously extracting rubusoside and tea polyphenol | |
| CN102352396B (en) | Method of producing functional starch sugar through wheat starch | |
| CN101766289A (en) | Method for preparing high fructose corn syrup | |
| CN104086365B (en) | A kind of method of being prepared by antierythrite production mother liquor recycling to mixing sugar alcohol product | |
| CN101914054A (en) | Comprehensive method for extracting L-tryptophan from fermentation liquor | |
| CN103214533A (en) | Method for continuously preparing cordycepin and cordyceps polysaccharide by using membrane separation technology | |
| CN104789624B (en) | A kind of method of nisin fermentation co-production of lactic acid calcium | |
| CN107474088A (en) | A kind of extraction process for pleocidin industrialized production | |
| CN109320400B (en) | Method for extracting natural mannitol from waste liquid of mogroside production | |
| CN102050737B (en) | Method for extracting and purifying pleuromutilin | |
| CN104961839A (en) | Preparation method of specific pachyman formula granule | |
| CN108785341A (en) | A kind of preparation method of purslane extract | |
| CN105859642A (en) | Tetramethylpyrazine extraction and purification method | |
| CN102557970A (en) | Preparation method of anhydrous betaine | |
| CN108315380B (en) | Purple sweet potato anthocyanin synthesis and extraction method | |
| CN101979616B (en) | Method for producing erythritol by using broken rice | |
| CN110468169A (en) | A kind of method of corncob production L-arabinose | |
| CN106432387A (en) | Combined extraction method for rutin and polysaccharide in buckwheat seeds | |
| CN102070677B (en) | Method for refining raffinose | |
| CN103145602A (en) | Novel extraction and purification technology for 5-hydroxytryptophan | |
| CN105861570B (en) | A kind of production method of odorless antierythrite | |
| CN107382936A (en) | A kind of method that young fustic and Rhoifolin are extracted from Rhus succedanea | |
| CN107344930A (en) | A kind of method that young fustic is extracted from Rhus succedanea | |
| CN102875330A (en) | Method for preparing arabitol by flammulina velutipes |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| CB03 | Change of inventor or designer information | ||
| CB03 | Change of inventor or designer information |
Inventor after: Wang Tengfei Inventor after: Meng Wu Inventor after: Wang Ruiming Inventor after: Wang Jianbin Inventor after: Sui Songsen Inventor before: Wang Tengfei Inventor before: Meng Wu Inventor before: Wang Ruiming Inventor before: Wang Jianbin |
|
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CB03 | Change of inventor or designer information | ||
| CB03 | Change of inventor or designer information |
Inventor after: Meng Wu Inventor after: Wang Tengfei Inventor after: Wang Ruiming Inventor after: Wang Jianbin Inventor after: Sui Songsen Inventor before: Wang Tengfei Inventor before: Meng Wu Inventor before: Wang Ruiming Inventor before: Wang Jianbin Inventor before: Sui Songsen |
|
| CP03 | Change of name, title or address | ||
| CP03 | Change of name, title or address |
Address after: 250353 University Road, Changqing District, Ji'nan, Shandong Province, No. 3501 Patentee after: Qilu University of Technology Patentee after: Dongxiao Biotechnology Co.,Ltd. Address before: 250353 University Road, Changqing District, Ji'nan, Shandong Province, No. 3501 Patentee before: Qilu University of Technology Patentee before: ZHUCHENG DONGXIAO BIOTECHNOLOGY Co.,Ltd. |