CN108017530B - Method for continuously separating coenzyme Q10 from mushroom dregs - Google Patents
Method for continuously separating coenzyme Q10 from mushroom dregs Download PDFInfo
- Publication number
- CN108017530B CN108017530B CN201711317608.7A CN201711317608A CN108017530B CN 108017530 B CN108017530 B CN 108017530B CN 201711317608 A CN201711317608 A CN 201711317608A CN 108017530 B CN108017530 B CN 108017530B
- Authority
- CN
- China
- Prior art keywords
- coenzyme
- raffinate
- moving bed
- simulated moving
- eluent
- 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.)
- Active
Links
- ACTIUHUUMQJHFO-UPTCCGCDSA-N coenzyme Q10 Chemical compound COC1=C(OC)C(=O)C(C\C=C(/C)CC\C=C(/C)CC\C=C(/C)CC\C=C(/C)CC\C=C(/C)CC\C=C(/C)CC\C=C(/C)CC\C=C(/C)CC\C=C(/C)CCC=C(C)C)=C(C)C1=O ACTIUHUUMQJHFO-UPTCCGCDSA-N 0.000 title claims abstract description 83
- ACTIUHUUMQJHFO-UHFFFAOYSA-N Coenzym Q10 Natural products COC1=C(OC)C(=O)C(CC=C(C)CCC=C(C)CCC=C(C)CCC=C(C)CCC=C(C)CCC=C(C)CCC=C(C)CCC=C(C)CCC=C(C)CCC=C(C)C)=C(C)C1=O ACTIUHUUMQJHFO-UHFFFAOYSA-N 0.000 title claims abstract description 81
- 235000017471 coenzyme Q10 Nutrition 0.000 title claims abstract description 81
- 229940110767 coenzyme Q10 Drugs 0.000 title claims abstract description 80
- 238000000034 method Methods 0.000 title claims abstract description 46
- 235000001674 Agaricus brunnescens Nutrition 0.000 title claims abstract description 17
- 239000003480 eluent Substances 0.000 claims abstract description 28
- 239000007788 liquid Substances 0.000 claims abstract description 21
- 239000003960 organic solvent Substances 0.000 claims abstract description 15
- 239000000287 crude extract Substances 0.000 claims abstract description 11
- 239000012527 feed solution Substances 0.000 claims abstract description 8
- 239000002904 solvent Substances 0.000 claims abstract description 8
- 238000001035 drying Methods 0.000 claims abstract description 7
- 238000001914 filtration Methods 0.000 claims abstract description 7
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 45
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 claims description 27
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 24
- 239000000047 product Substances 0.000 claims description 18
- 230000005526 G1 to G0 transition Effects 0.000 claims description 16
- 238000000926 separation method Methods 0.000 claims description 15
- IMNFDUFMRHMDMM-UHFFFAOYSA-N N-Heptane Chemical compound CCCCCCC IMNFDUFMRHMDMM-UHFFFAOYSA-N 0.000 claims description 14
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 claims description 12
- 239000000203 mixture Substances 0.000 claims description 12
- 239000007787 solid Substances 0.000 claims description 11
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 10
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 10
- 238000000605 extraction Methods 0.000 claims description 8
- 239000002245 particle Substances 0.000 claims description 8
- 238000001816 cooling Methods 0.000 claims description 7
- XDTMQSROBMDMFD-UHFFFAOYSA-N Cyclohexane Chemical compound C1CCCCC1 XDTMQSROBMDMFD-UHFFFAOYSA-N 0.000 claims description 6
- TVMXDCGIABBOFY-UHFFFAOYSA-N octane Chemical compound CCCCCCCC TVMXDCGIABBOFY-UHFFFAOYSA-N 0.000 claims description 6
- 239000011148 porous material Substances 0.000 claims description 6
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical group O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 5
- 239000003208 petroleum Substances 0.000 claims description 5
- 239000000741 silica gel Substances 0.000 claims description 5
- 229910002027 silica gel Inorganic materials 0.000 claims description 5
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 4
- WYURNTSHIVDZCO-UHFFFAOYSA-N Tetrahydrofuran Chemical compound C1CCOC1 WYURNTSHIVDZCO-UHFFFAOYSA-N 0.000 claims description 4
- PNEYBMLMFCGWSK-UHFFFAOYSA-N aluminium oxide Inorganic materials [O-2].[O-2].[O-2].[Al+3].[Al+3] PNEYBMLMFCGWSK-UHFFFAOYSA-N 0.000 claims description 4
- 125000004432 carbon atom Chemical group C* 0.000 claims description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 3
- ZMXDDKWLCZADIW-UHFFFAOYSA-N N,N-Dimethylformamide Chemical compound CN(C)C=O ZMXDDKWLCZADIW-UHFFFAOYSA-N 0.000 claims description 2
- 239000012065 filter cake Substances 0.000 claims description 2
- YLQBMQCUIZJEEH-UHFFFAOYSA-N tetrahydrofuran Natural products C=1C=COC=1 YLQBMQCUIZJEEH-UHFFFAOYSA-N 0.000 claims description 2
- 238000005406 washing Methods 0.000 claims description 2
- 238000011084 recovery Methods 0.000 abstract description 7
- 238000010924 continuous production Methods 0.000 abstract description 2
- 238000004458 analytical method Methods 0.000 description 9
- 230000008569 process Effects 0.000 description 9
- 235000019441 ethanol Nutrition 0.000 description 8
- 239000000284 extract Substances 0.000 description 8
- 238000004519 manufacturing process Methods 0.000 description 7
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 6
- 238000010898 silica gel chromatography Methods 0.000 description 6
- 238000000855 fermentation Methods 0.000 description 5
- 230000004151 fermentation Effects 0.000 description 5
- 239000012535 impurity Substances 0.000 description 5
- 238000004811 liquid chromatography Methods 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- 238000004587 chromatography analysis Methods 0.000 description 4
- 230000000813 microbial effect Effects 0.000 description 4
- 239000012071 phase Substances 0.000 description 4
- 238000001291 vacuum drying Methods 0.000 description 4
- 238000002425 crystallisation Methods 0.000 description 3
- 230000008025 crystallization Effects 0.000 description 3
- 239000000945 filler Substances 0.000 description 3
- 238000012856 packing Methods 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- -1 quinone compound Chemical class 0.000 description 3
- 238000000638 solvent extraction Methods 0.000 description 3
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 229910052739 hydrogen Inorganic materials 0.000 description 2
- 239000001257 hydrogen Substances 0.000 description 2
- 230000014759 maintenance of location Effects 0.000 description 2
- 239000011259 mixed solution Substances 0.000 description 2
- 230000007935 neutral effect Effects 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 239000011347 resin Substances 0.000 description 2
- 229920005989 resin Polymers 0.000 description 2
- 238000007127 saponification reaction Methods 0.000 description 2
- 238000001179 sorption measurement Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 238000000194 supercritical-fluid extraction Methods 0.000 description 2
- NWUYHJFMYQTDRP-UHFFFAOYSA-N 1,2-bis(ethenyl)benzene;1-ethenyl-2-ethylbenzene;styrene Chemical compound C=CC1=CC=CC=C1.CCC1=CC=CC=C1C=C.C=CC1=CC=CC=C1C=C NWUYHJFMYQTDRP-UHFFFAOYSA-N 0.000 description 1
- YHQXBTXEYZIYOV-UHFFFAOYSA-N 3-methylbut-1-ene Chemical group CC(C)C=C YHQXBTXEYZIYOV-UHFFFAOYSA-N 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical group [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 150000001721 carbon Chemical group 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 239000002537 cosmetic Substances 0.000 description 1
- 239000012043 crude product Substances 0.000 description 1
- 125000003493 decenyl group Chemical group [H]C([*])=C([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000009792 diffusion process Methods 0.000 description 1
- 238000007599 discharging Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000010812 external standard method Methods 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 235000013373 food additive Nutrition 0.000 description 1
- 239000002778 food additive Substances 0.000 description 1
- 210000002064 heart cell Anatomy 0.000 description 1
- JYVHOGDBFNJNMR-UHFFFAOYSA-N hexane;hydrate Chemical compound O.CCCCCC JYVHOGDBFNJNMR-UHFFFAOYSA-N 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 239000003456 ion exchange resin Substances 0.000 description 1
- 229920003303 ion-exchange polymer Polymers 0.000 description 1
- 210000003292 kidney cell Anatomy 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 210000005229 liver cell Anatomy 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- AZQWKYJCGOJGHM-UHFFFAOYSA-N para-benzoquinone Natural products O=C1C=CC(=O)C=C1 AZQWKYJCGOJGHM-UHFFFAOYSA-N 0.000 description 1
- 238000005325 percolation Methods 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 238000004237 preparative chromatography Methods 0.000 description 1
- 238000005086 pumping Methods 0.000 description 1
- 238000001953 recrystallisation Methods 0.000 description 1
- NPCOQXAVBJJZBQ-UHFFFAOYSA-N reduced coenzyme Q9 Natural products COC1=C(O)C(C)=C(CC=C(C)CCC=C(C)CCC=C(C)CCC=C(C)CCC=C(C)CCC=C(C)CCC=C(C)CCC=C(C)CCC=C(C)C)C(O)=C1OC NPCOQXAVBJJZBQ-UHFFFAOYSA-N 0.000 description 1
- 238000007670 refining Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000029058 respiratory gaseous exchange Effects 0.000 description 1
- 230000002000 scavenging effect Effects 0.000 description 1
- 239000012798 spherical particle Substances 0.000 description 1
- 239000007858 starting material Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 229940035936 ubiquinone Drugs 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C46/00—Preparation of quinones
- C07C46/10—Separation; Purification; Stabilisation; Use of additives
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Organic Low-Molecular-Weight Compounds And Preparation Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses a method for continuously separating coenzyme Q10 from mushroom dregs, which comprises the following steps: (1) dissolving a crude extract of coenzyme Q10 in a nonpolar organic solvent to prepare a feed solution; (2) continuously introducing a feed liquid and an eluent into the simulated moving bed chromatographic system, and continuously collecting raffinate from a raffinate port of the simulated moving bed chromatographic system; (3) and (3) concentrating the raffinate obtained in the step (2) under reduced pressure, dissolving again, and crystallizing, filtering and drying to obtain a refined coenzyme Q10 product with the purity of more than 98%. The method has the characteristics of high yield, high recovery rate, low solvent consumption and continuous production, and is suitable for industrial large-scale popularization and application.
Description
Technical Field
The invention belongs to the technical field of chemical separation, and particularly relates to a method for continuously separating coenzyme Q10 from mushroom dregs.
Background
Coenzyme Q10, also called ubiquinone, is a fat-soluble quinone compound, mainly exists in heart, liver and kidney cells of animals, has a chemical name of 2- (3,7,11,15,19,23,27,31,35, 39-decamethyl-2, 6,10,14,18,22,26,30,34, 38-forty decenyl) -5, 6-dimethoxy-3-methyl-p-benzoquinone, and a molecular formula of C-tetrafluoquinone59H90O4Molecular weight 863.34. Coenzyme Q10 has important physiological functions of scavenging free radicals, improving intracellular respiration, enhancing immunity and the like, has continuously expanded market demand, and is widely applied to the fields of medicines, cosmetics, food additives and the like.
The production method of the coenzyme Q10 mainly comprises a chemical synthesis method, an animal and plant cell culture method and a microbial fermentation method, wherein the microbial fermentation method has the advantages of high process stability, easiness in large-scale production, simplicity in operation, high biological activity of products, easiness in absorption and the like, and is a research hotspot for the production of the coenzyme Q10 at present. The fermentation liquor prepared by the microbial fermentation method is centrifuged, filtered, freeze-dried and crushed to obtain bacterial dregs, a coenzyme Q10 crude extract is obtained by extraction, and a high-purity coenzyme Q10 product is obtained by further purification treatment. The existing extraction method comprises a solvent extraction method, a saponification method and a supercritical fluid extraction method, and then the crude product of the coenzyme Q10 is further purified by combining the technologies of silica gel column chromatography, recrystallization and the like. However, the crude extract of coenzyme Q10 mainly contains coenzyme Q analogues with different numbers of isopentene units on side chains, and the separation difficulty is higher.
CN103819326A discloses a method for refining coenzyme Q10 by ultrasonic crushing, organic solvent extraction, silica gel column chromatography and crystallization in turn. CN101429108A discloses a method for purifying coenzyme Q10 by sequentially extracting with absolute ethyl alcohol, water and n-hexane, performing silica gel column chromatography and crystallizing. CN102391092A discloses a method for extracting mushroom dregs by supercritical carbon dioxide, and then obtaining coenzyme Q10 with the purity of more than 99.5 percent by silica gel column chromatography and crystallization. CN101987815A discloses a method for preparing coenzyme Q10 with purity of more than 98% by combining adsorption resin and silica gel column chromatography. These methods all require silica gel column chromatography, and although high-purity coenzyme Q10 can be obtained, the above methods are all batch operation processes, and have problems of large organic solvent usage, small preparation amount, low silica gel utilization rate and the like, so that the process is not economical.
The simulated moving bed chromatography is a preparative chromatography technology with the most industrialized prospect at present, and four inlets and outlets, such as an eluent inlet, a feeding liquid inlet, an extract liquid outlet, a raffinate liquid outlet and the like, divide all chromatographic columns into four regions with different flow rates and respectively undertake different functions. The device simulates the countercurrent movement of an eluent and a stationary phase by timing switching of four inlet and outlet materials, thereby realizing the continuity of feeding and discharging. The mixed solution containing the strongly adsorbed component and the eluent is continuously collected at the extract outlet, and the mixed solution containing the weakly adsorbed component and the eluent is continuously collected at the raffinate outlet. On the one hand, this operation allows continuous feeding and therefore high productivity; on the other hand, the eluent is recycled, so that the consumption of the solvent is less, and the cost of large-scale preparation can be reduced. The target component with high purity can be obtained by designing appropriate flow rates for each zone.
Disclosure of Invention
Aiming at the defects of the method, the invention provides the method for continuously separating the coenzyme Q10 from the mushroom dregs, and the method has the advantages of simple process, large product preparation amount, high purity, less solvent consumption and low production cost.
The method separates coenzyme Q10 from the mushroom dregs by a simulated moving bed chromatography and crystallization combined method, the method takes a filler with a surface rich in polar groups as a stationary phase, adjusts the type and the proportion of an eluent, designs proper flow rate and switching time of each area, can realize continuous separation of coenzyme Q10 and impurities by adopting the simulated moving bed chromatography, and then crystallizes and purifies the coenzyme Q10 obtained by separation.
A method for continuously separating coenzyme Q10 from mushroom dregs comprises the following steps:
(1) dissolving a crude extract of coenzyme Q10 in a nonpolar organic solvent to prepare a feed solution;
(2) continuously introducing a feed liquid and an eluent into the simulated moving bed chromatographic system, and continuously collecting raffinate from a raffinate port of the simulated moving bed chromatographic system;
the simulated moving bed chromatographic system consists of 4-32 chromatographic columns filled with a fixed phase, and comprises four zones, wherein each zone is formed by connecting 1-8 chromatographic columns in series. The zones can be connected in series or disconnected, and an isocratic operation mode or a gradient operation mode can be adopted. Presetting operation parameters such as flow, switching time, switching times, column temperature and the like of each area, continuously pumping the feed liquid and the eluent, and continuously collecting raffinate rich in coenzyme Q10 from a raffinate port after the system reaches a steady state.
Before a simulated moving bed chromatographic system operates, a wet column packing method is adopted to pack stationary phase particles into a chromatographic column, a column packing solvent is n-hexane or petroleum ether, symmetry experiments are carried out on the pressure, the column efficiency, the solute retention time, the separation degree and the total porosity of each column, and the performance indexes of each chromatographic column are ensured to be consistent. The separable zones were preliminarily determined according to "trigonometric theory", and the flow rates and switching times of the zones were adjusted until complete separation of coenzyme Q10 from the impurities was achieved.
(3) Concentrating the raffinate obtained in the step (2) to remove the solvent, adding an organic solvent at 20-60 ℃ until the solid is just dissolved, cooling to-5 ℃, cooling and crystallizing for 12-36 hours, filtering, washing a filter cake with water, and drying in vacuum at 20-40 ℃ to obtain a refined coenzyme Q10 product with the purity of more than 98%.
The crude extract of coenzyme Q10 in the step (1) is extracted from the mushroom dregs obtained by microbial fermentation, specifically, the method described in the patent application with the publication number of CN101314782A, CN101619330A or CN105886562A can be referred to culture the strain, and the fermented liquid is filtered, dried and crushed to obtain the mushroom dregs; the extraction method for extracting coenzyme Q10 crude extract from mushroom residue is percolation extraction, organic solvent extraction, alcohol-base saponification or supercritical fluid extraction, and can be specifically disclosed in patent application CN106146278A, CN101381747A, CN102391092A or CN 104694613A.
The nonpolar organic solvent is one or a mixture of any two of n-hexane, cyclohexane, n-heptane, n-octane and petroleum ether. These hydrophobic organic solvents have a higher solubility for the crude coenzyme Q10 extract.
The total concentration of the feeding liquid is 5-500 g/L, and the total concentration is preferably 50-300 g/L. If the feed concentration is too low, the production capacity is reduced and the process economy is reduced; if the feeding concentration is too high, the complete separation area is obviously reduced, the difficulty of designing the operation conditions is increased, and the separation difficulty is increased.
The stationary phase of the simulated moving bed chromatographic system is polar macroporous adsorption resin, ion exchange resin, silica gel or alumina. The stationary phases are rich in polar groups such as hydroxyl groups and the like, can form hydrogen bonds with carbon groups in coenzyme Q class analogs, and identify the micro structural difference between the analogs according to the difference of the acting force of the hydrogen bonds.
The stationary phase is spherical particles with uniform particle size, uniform pore diameter and high mechanical strength.
The particle size of the stationary phase is controlled to be 5-200 μm, and more preferably 10-100 μm. If the particle size is too large, the column efficiency is reduced, which is not beneficial to the separation of coenzyme Q10 and impurities; if the particle size is too small, the column pressure is too high, which is not favorable for operation.
The aperture of the stationary phase is controlled to be 5-100 nm, and the preferred aperture is 10-50 nm. If the aperture of the filler is too small, the coenzyme Q10 molecules are not easy to enter the inside of the pore channel, so that the separation effect is reduced; if the pore diameter of the filler is too large, the diffusion in the pores is slow, and the mass transfer resistance is large.
The eluent is one or a mixture of any two of N-hexane, cyclohexane, N-heptane, N-octane, petroleum ether, acetonitrile, ethyl acetate, tetrahydrofuran, dimethyl sulfoxide, N-dimethylformamide and monohydric alcohol with the carbon atom number of 1-4. Further preferred eluents are mixtures of n-hexane and ethyl acetate.
Preferably, the volume percentage of the ethyl acetate in the eluent is 1-20%. If the proportion of the ethyl acetate is too high, the polarity of the mobile phase is strong, and the separation degree of the coenzyme Q10 and impurities is poor; if the proportion of ethyl acetate is too low, the polarity of the mobile phase is weak, the retention time of the coenzyme Q10 in the system is too long, and the system is difficult to stabilize.
The size of the chromatographic column of the simulated moving bed chromatographic system is 5-500 mm in diameter and 50-1000 mm in length, and the preferred size is 10-100 mm in diameter and 100-500 mm in length. If the size of the chromatographic column is too small, the production capacity is low; if the size of the chromatographic column is too large, the packing is difficult to fill, the wall effect is obvious, and the separation capability of the chromatographic column is reduced.
The operation parameters of the simulated moving bed chromatographic system are controlled as follows: the flow rate of the eluent is 1-1000 mL/min, the flow rate of the feeding liquid is 1-100 mL/min, the flow rate of the extraction liquid is 1-100 mL/min, the flow rate of the raffinate is 1-100 mL/min, and the switching time is 1-50 min. The switching time is further preferably 3-10 min, and if the switching time is too short, the switching valve is easy to damage; if the switching time is too long, the system is difficult to reach steady state.
The separation temperature of the simulated moving bed chromatographic system is 0-60 ℃, and the preferable temperature is 20-50 ℃. If the temperature is too low, the solubility of the coenzyme Q10 in the solvent is obviously reduced, and the concentration of the feeding liquid is limited; if the temperature is too high, coenzyme Q10 is easily oxidized and deteriorated during the separation process.
The organic solvent is one or a mixture of any two of monohydric alcohol with 1-4 carbon atoms, acetonitrile, acetone, ethyl acetate, n-hexane and n-heptane.
The volume-mass ratio of the added amount of the organic solvent to the solid obtained after the concentration of the raffinate is 20-80L/kg.
Compared with the prior art, the invention has the following advantages:
1. the continuous chromatographic technique is adopted, so that the utilization rate of the stationary phase is improved, and the consumption of the stationary phase is reduced.
2. The continuous production of the coenzyme Q10 is realized, the production process is full-automatic, the labor intensity is low, and the production cost is low.
3. The purity of the coenzyme Q10 obtained by the purification method of the invention reaches more than 98%, the yield is high, the solvent consumption is low, and the method is suitable for large-scale popularization and application.
Drawings
FIG. 1 is a liquid chromatogram of a coenzyme Q10 product obtained in example 1 of the present invention.
Detailed Description
In order to further understand the present invention, the following will specifically describe a method for continuously separating coenzyme Q10 from mushroom dregs, which is provided by the present invention, with reference to the following examples, but the present invention is not limited to these examples, and the insubstantial modifications and adaptations made by those skilled in the art under the core teaching of the present invention still fall within the scope of the present invention.
The following examples of simulated moving bed devices using German CESP C9116 (Noll, Germany) equipped with a multiport rotary valve, which can be connected to 16 chromatographic columns at most, each zone of chromatographic columns of the same number, in 1~4 changes; the device is provided with 4S-100 liquid phase pumps, wherein the flow rate of a feed pump is 0-10 mL/min, and the flow rates of an eluent pump, an extraction liquid pump and an extraction raffinate pump are 0-50 mL/min. Eluent is injected between zone 4 and zone 1, feed solution is injected between zone 2 and zone 3, coenzyme Q10 is collected at the raffinate outlet between zone 3 and zone 4, and impurity 10 is collected at the extract outlet between zone 1 and zone 2. At each switching time (note: this switching time is adjustable), the column switches one position in the opposite direction of the eluent flow.
In the following embodiments of the invention, the determination of the coenzyme Q10 content is performed according to the method described in Chinese pharmacopoeia, and the analysis conditions of the liquid chromatography are as follows: waters Atlantis T3 analytical column (250mm × 4.6mm,5 μm), methanol-absolute ethanol (1:1) as mobile phase, flow rate 1mL/min, sample amount 20 μ L, ultraviolet detector as detector, and detection wavelength 275 nm.
The method for calculating the purity of the coenzyme Q10 comprises the following steps: a small amount of the collected coenzyme Q10 product was dissolved in absolute ethanol and diluted to prepare a solution containing about 0.2mg of coenzyme Q10 per 1mL, and the purity was calculated as the peak area by an external standard method.
The calculation method of the purity and the recovery rate of the invention is as follows:
purity ═ the mass of coenzyme Q10 in the product ÷ the total mass of the product × 100%
The recovery rate is the mass of coenzyme Q10 in the product ÷ the mass of coenzyme Q10 in the starting material × 100%.
Example 1
Dissolving crude extract of coenzyme Q10 in n-hexane to obtain feed solution with solid concentration of 50g/L, wherein the content of coenzyme Q10 is about 62.3%.
The simulated moving bed is provided with 8 chromatographic columns, and the size is 1cm multiplied by 25 cm; the stationary phase is silica gel with the particle size of 45 mu m and the aperture of 10 nm; the eluent is a mixture of normal hexane and ethyl acetate, wherein the volume percentage of the ethyl acetate is 10%; the operation temperature is 30 ℃; the operating parameters are optimally determined as: eluent flow rate of 16mL/min, feed flow rate of 2mL/min, extract flow rate of 9.5mL/min, raffinate flow rate of 8.5mL/min, and switching time of 5 min. After 32 consecutive switches, the system reached equilibrium and a coenzyme Q10 rich solution was collected from the raffinate outlet. Analysis showed that the coenzyme Q10 content of the raffinate was 95.6%.
Concentrating the raffinate into solid, adding ethanol at 50 ℃ until the ethanol is completely dissolved, wherein the solid-liquid ratio is 1:50, gradually reducing the temperature to 5 ℃, cooling and crystallizing for 24h, filtering, and drying in a vacuum drying oven at 30 ℃ for 24h to obtain a refined coenzyme Q10 product. The purity of the coenzyme Q10 product is 99.2% by liquid chromatography analysis, and the recovery rate of the whole process is 95.5%.
Example 2
Dissolving crude extract of coenzyme Q10 in n-hexane to obtain feed solution with solid concentration of 100g/L, wherein the content of coenzyme Q10 is about 68.7%.
The simulated moving bed is provided with 8 chromatographic columns, and the size is 1cm multiplied by 25 cm; the stationary phase is silica gel with the particle size of 20 mu m and the pore diameter of 22 nm; the eluent is a mixture of normal hexane and ethanol, wherein the volume percentage of the ethanol is 5%; the operation temperature is 30 ℃; the operating parameters are optimally determined as: eluent flow rate of 15.4mL/min, feed flow rate of 1.5mL/min, extract flow rate of 8.9mL/min, raffinate flow rate of 8.0mL/min, and switching time of 4 min. After 32 consecutive switches, the system reached equilibrium and a coenzyme Q10 rich solution was collected from the raffinate outlet. Analysis showed that the coenzyme Q10 content of the raffinate was 96.5%.
Concentrating the raffinate into solid, adding ethyl acetate at 30 ℃ until the raffinate is completely dissolved, wherein the solid-liquid ratio is 1:20, gradually reducing the temperature to 0 ℃, cooling, crystallizing for 24 hours, filtering, and drying in a vacuum drying oven at 30 ℃ for 24 hours to obtain a refined coenzyme Q10 product. The purity of the coenzyme Q10 product is 99.2% by liquid chromatography analysis, and the recovery rate of the whole process is 95.9%.
Example 3
Dissolving crude extract of coenzyme Q10 in n-hexane to obtain feed solution with solid concentration of 80g/L, wherein the content of coenzyme Q10 is about 70.3%.
The simulated moving bed is provided with 16 chromatographic columns, and the size is 1cm multiplied by 15 cm; the stationary phase is neutral alumina with 200-300 meshes; the eluent is a mixture of normal hexane and ethyl acetate, wherein the volume percentage of the ethyl acetate is 10%; the operation temperature is 40 ℃; the operating parameters are optimally determined as: eluent flow rate of 3mL/min, feed flow rate of 1.5mL/min, extract flow rate of 2.2mL/min, raffinate flow rate of 2.3mL/min, and switching time of 10 min. After 48 consecutive switching, the system reached equilibrium and a coenzyme Q10 rich solution was collected from the raffinate outlet. Analysis showed that the coenzyme Q10 content of the raffinate was 95.2%.
Concentrating the raffinate into solid, adding ethanol at 50 ℃ until the ethanol is completely dissolved, wherein the solid-liquid ratio is 1:50, gradually reducing the temperature to 5 ℃, cooling, crystallizing for 12h, filtering, and drying in a vacuum drying oven at 35 ℃ for 12h to obtain a refined coenzyme Q10 product. The purity of the coenzyme Q10 product is 98.2% by liquid chromatography analysis, and the recovery rate of the whole process is 94.5%.
Example 4
The crude extract of coenzyme Q10 was dissolved in cyclohexane to prepare a feed solution with a solid concentration of 150g/L, wherein the coenzyme Q10 content was about 72.7%.
The simulated moving bed is provided with 16 chromatographic columns, and the size is 1cm multiplied by 15 cm; the stationary phase is neutral alumina with 200-300 meshes; the eluent is a mixture of cyclohexane and methanol, wherein the volume percentage of the methanol is 5%; the operation temperature is 20 ℃; the operating parameters are optimally determined as: eluent flow rate of 7mL/min, feed flow rate of 2mL/min, extract flow rate of 4.8mL/min, raffinate flow rate of 4.2mL/min, and switching time of 5 min. After 48 consecutive switching, the system reached equilibrium and a coenzyme Q10 rich solution was collected from the raffinate outlet. Analysis showed that the coenzyme Q10 content of the raffinate was 95.0%.
Concentrating the raffinate into solid, adding acetone at 30 ℃ until the acetone is completely dissolved, wherein the solid-liquid ratio is 1:20, gradually reducing the temperature to 5 ℃, cooling and crystallizing for 24h, filtering, and drying in a vacuum drying oven at 35 ℃ for 12h to obtain a refined coenzyme Q10 product. The purity of the coenzyme Q10 product is 98.0% by liquid chromatography analysis, and the recovery rate of the whole process is 93.8%.
Claims (5)
1. A method for continuously separating coenzyme Q10 from mushroom dregs comprises the following steps:
(1) dissolving a crude extract of coenzyme Q10 in a nonpolar organic solvent to prepare a feed solution; the total concentration of the feeding liquid is 5-500 g/L;
(2) continuously introducing a feed liquid and an eluent into the simulated moving bed chromatographic system, and continuously collecting raffinate from a raffinate port of the simulated moving bed chromatographic system; the operation parameters of the simulated moving bed chromatographic system are controlled as follows: the separation temperature is 0-60 ℃, the flow rate of an eluent is 1-1000 mL/min, the flow rate of a feeding liquid is 1-100 mL/min, the flow rate of an extraction liquid is 1-100 mL/min, the flow rate of a raffinate is 1-100 mL/min, and the switching time is 1-50 min;
the simulated moving bed chromatographic system consists of 4-32 chromatographic columns filled with a fixed phase, and comprises four zones, wherein each zone is formed by connecting 1-8 chromatographic columns in series;
the size of a chromatographic column of the simulated moving bed chromatographic system is 10-100 mm in diameter and 100-500 mm in length;
the stationary phase of the simulated moving bed chromatographic system is silica gel or alumina, the particle size of the stationary phase is controlled to be 10-100 mu m, and the pore diameter is controlled to be 10-50 nm;
(3) concentrating the raffinate obtained in the step (2) to remove the solvent, adding an organic solvent to dissolve at the temperature of 20-60 ℃, cooling and crystallizing at the temperature of-5 ℃ for 12-36 h, filtering, washing a filter cake with water, and drying in vacuum at the temperature of 20-40 ℃ to obtain a refined coenzyme Q10 product with the purity of more than 98%;
the volume-mass ratio of the added amount of the organic solvent to the solid obtained after the concentration of the raffinate is 20-80L/kg.
2. The method for continuously separating the coenzyme Q10 from the mushroom dregs according to claim 1, wherein the nonpolar organic solvent is one or a mixture of any two of n-hexane, cyclohexane, n-heptane, n-octane and petroleum ether.
3. The method for continuously separating the coenzyme Q10 from the mushroom dregs according to claim 1, wherein the eluent is one or a mixture of any two of N-hexane, cyclohexane, N-heptane, N-octane, petroleum ether, acetonitrile, ethyl acetate, tetrahydrofuran, dimethyl sulfoxide, N-dimethylformamide and monohydric alcohol with 1-4 carbon atoms.
4. The method for continuously separating the coenzyme Q10 from the mushroom dregs according to claim 3, wherein the eluent is a mixture of n-hexane and ethyl acetate, and the volume percentage of the ethyl acetate is 1-20%.
5. The method for continuously separating the coenzyme Q10 from the mushroom dregs according to claim 1, wherein in the step (3), the organic solvent is one or a mixture of any two of monohydric alcohol with 1-4 carbon atoms, acetonitrile, acetone, ethyl acetate, n-hexane and n-heptane.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711317608.7A CN108017530B (en) | 2017-12-12 | 2017-12-12 | Method for continuously separating coenzyme Q10 from mushroom dregs |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711317608.7A CN108017530B (en) | 2017-12-12 | 2017-12-12 | Method for continuously separating coenzyme Q10 from mushroom dregs |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN108017530A CN108017530A (en) | 2018-05-11 |
| CN108017530B true CN108017530B (en) | 2020-10-13 |
Family
ID=62073186
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711317608.7A Active CN108017530B (en) | 2017-12-12 | 2017-12-12 | Method for continuously separating coenzyme Q10 from mushroom dregs |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108017530B (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110002985A (en) * | 2019-05-15 | 2019-07-12 | 丽珠集团(宁夏)制药有限公司 | One kind is from ubiquinone10Ubiquinone is isolated and purified in mother liquor10Method and ubiquinone10Crude product |
| CN110465114B (en) * | 2019-08-23 | 2021-08-20 | 内蒙古金达威药业有限公司 | Simulated moving bed continuous chromatography chromatographic system, application thereof and method for purifying coenzyme Q10 |
| CN112920035A (en) * | 2019-12-06 | 2021-06-08 | 中国科学院大连化学物理研究所 | Method for removing Q11 impurity in coenzyme Q10 by using preparation chromatography |
| CN115677468A (en) * | 2022-11-02 | 2023-02-03 | 广东润和生物科技有限公司 | Method for purifying coenzyme Q10 |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101233095A (en) * | 2005-06-10 | 2008-07-30 | 协和发酵工业株式会社 | Method of purifying ubiquinone-10 |
| CN101987815A (en) * | 2010-09-28 | 2011-03-23 | 华东理工大学 | Purification process for preparing high-purity coenzyme Q10 |
-
2017
- 2017-12-12 CN CN201711317608.7A patent/CN108017530B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101233095A (en) * | 2005-06-10 | 2008-07-30 | 协和发酵工业株式会社 | Method of purifying ubiquinone-10 |
| CN101987815A (en) * | 2010-09-28 | 2011-03-23 | 华东理工大学 | Purification process for preparing high-purity coenzyme Q10 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN108017530A (en) | 2018-05-11 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108017530B (en) | Method for continuously separating coenzyme Q10 from mushroom dregs | |
| CN111470953A (en) | Method for extracting and separating high-purity cannabidiol from low-content industrial cannabis sativa leaves | |
| CN103387489B (en) | Preparation method of high pure crocin and geniposide | |
| CN110465114B (en) | Simulated moving bed continuous chromatography chromatographic system, application thereof and method for purifying coenzyme Q10 | |
| CN112321408B (en) | Device and method for extracting vitamin K2 from microbial fermentation broth | |
| CN108084007B (en) | Method for separating coenzyme Q10 and coenzyme Q11 by simulated moving bed chromatography | |
| CN110041184A (en) | A kind of method of purification of vitamin menaquinone-7 | |
| WO2020015316A1 (en) | Method for extracting and purifying coenzyme q10 and coenzyme q10 prepared thereby | |
| CN102010387B (en) | Method for purifying orlistat | |
| CN1686988A (en) | Method for preparing high pure solanesol | |
| CN104418925B (en) | A method of preparing high-purity fidaxomicin | |
| CN101386614B (en) | Method for preparing epigallocatechin-3-gallate by resin adsorption method | |
| CN102558103A (en) | Method for separating and purifying Orlistat | |
| CN104788509B (en) | A kind of technique extracting preparation high-purity Raffinose from defatted wheat germ | |
| CN103570647A (en) | Method for preparing high-purity paclitaxel from taxus chinensis cell culture fluid | |
| CN105017367B (en) | A kind of method separating lanosterol and lanostenol | |
| CN109651301B (en) | Purification method of orlistat | |
| CN103254226B (en) | A kind of method of ADSORPTION IN A FIXED BED method separation and purification Yelkin TTS | |
| CN101353294A (en) | Separation and purification method of high-content resveratrol | |
| CN102381974A (en) | Method for separating and preparing caffeic tannic acid from honeysuckle by utilizing high speed countercurrent chromatography | |
| CN111135810B (en) | Preparation method of special chromatographic separation medium for cannabidiol separation | |
| CN105819444A (en) | Composite type activated carbon and application thereof in purifying tacrolimus | |
| CN104119421B (en) | Steroidal is hydroxylated the minimizing technology of impurity | |
| CN108047014B (en) | Method for extracting and separating coenzyme Q10 by using ionic liquid | |
| CN112390817B (en) | Method for salting out and extracting tacrolimus fermentation liquor |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |