CN106117033A - A kind of concurrently separating prepares high-purity coenzyme Q10 and the technique of reduced coenzyme Q 10 - Google Patents
A kind of concurrently separating prepares high-purity coenzyme Q10 and the technique of reduced coenzyme Q 10 Download PDFInfo
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- CN106117033A CN106117033A CN201610485022.0A CN201610485022A CN106117033A CN 106117033 A CN106117033 A CN 106117033A CN 201610485022 A CN201610485022 A CN 201610485022A CN 106117033 A CN106117033 A CN 106117033A
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- C07—ORGANIC CHEMISTRY
- C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
- C07C46/00—Preparation of quinones
- C07C46/10—Separation; Purification; Stabilisation; Use of additives
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- C07—ORGANIC CHEMISTRY
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- C07C41/00—Preparation of ethers; Preparation of compounds having groups, groups or groups
- C07C41/01—Preparation of ethers
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Abstract
The invention discloses a kind of concurrently separating and prepare high-purity coenzyme Q10 and the technique of reduced coenzyme Q 10, carry out seepage pressure effects with dreg for raw material and must contain the percolate of coenzyme Q10 and reduced coenzyme Q 10, major part coenzyme Q10 is gone out by Crystallization Separation, and crystalline mother solution is reduced to obtain reducing solution, reducing solution is through being obtained by extraction the extract containing reduced coenzyme Q 10, extract is carried out crystallization treatment and isolates reduced coenzyme Q 10, finally given purity reach more than 98% high-purity coenzyme Q10 and purity reach more than 98% high-purity reduced coenzyme Q 10, total recovery is more than 94%, whole simple and reliable process, easily operation, it is easily achieved, parameter is easy to control.
Description
Technical field
The invention belongs to technical field of chemical engineering, concurrently separate preparation high-purity coenzyme Q10 and reduction particularly to one
The technological process of type coenzyme Q10.Specifically, it is to utilize the combinations such as seepage pressure effects, crystallization purifying, reduction reaction, extract and separate
Separation circuit, separates and prepares high-purity coenzyme Q10 and reduced coenzyme Q 10 fine work.
Background technology
Coenzyme Q compounds is a kind of fat-soluble quinones being widely present nature, and exist in human body is auxiliary
Enzyme Q10, its standard named 2,3-dimethoxy-5-methyl-6-isopentene group in the last of the ten Heavenly stems-benzoquinone.Its structural formula is as follows:
Coenzyme Q10 has removing free radical and antioxidation, enhancing body immunity and antitumor, reinforcement Cardiac Power and increasing
The most mental and the regulation physiological function such as blood fat.Therefore coenzyme Q10 medically has preferably application, is primarily used to treat the heart
Angiopathy, chronic hepatitis, breast carcinoma, encephalitis B, nervous system disease and skin diseases etc..On the other hand, coenzyme
Q10 is also widely used in food, health product and cosmetics.Along with the further investigation to coenzyme Q10 function, it is doctor
Medicine, cosmetics, food and field of health care products all will have more wide application prospect.
Reduced coenzyme Q 10 is the reduzate of coenzyme Q10, and the 1,4-benzoquinone construction recovery of coenzyme Q10 is hydroquinone knot
Structure.Its structural formula is as follows:
The protected cardiovascular of coenzyme Q10 and slow down aging etc. act on, but its can not directly absorbed by the body and utilize, and
It is to need first to be converted into reduced coenzyme Q 10.Reduced coenzyme Q 10 has higher removing free radical and antioxygen than coenzyme Q10
Change, strengthen body immunity and antitumor, reinforcement Cardiac Power and strengthen mental and regulate the physiological functions such as blood fat, can apply
In pharmaceuticals, cosmetics, food and field of health care products.The more important thing is, with advancing age, coenzyme Q10 is turned by human body
The ability of chemical conversion reduced coenzyme Q 10 can reduce, and therefore human body directly supplements reduced coenzyme Q 10 and is more beneficial for it and effectively inhales
Receive, utilize.Therefore highly purified coenzyme Q10 is prepared in separation and reduced coenzyme Q 10 is the most crucial.
Coenzyme Q10 and reduced coenzyme Q 10 are widely present in animal and plant body, such as Semen sojae atricolor, Fish, Carnis Sus domestica, Carnis caprae seu ovis, beef
And milk etc..At present, the source of coenzyme Q10 and reduced coenzyme Q 10, in addition to natural origin, also includes chemosynthesis and micro-
Biological fermentation process etc..In natural origin, the content of coenzyme Q10 and reduced coenzyme Q 10 is relatively low, and microbe fermentation method leads to
Cross cultivation high-efficiency strain, the yield of coenzyme Q10 and reduced coenzyme Q 10 can be greatly improved, be more suitable for produce coenzyme Q10 and
The source of reduced coenzyme Q 10.In microbial fermentation product, there is coenzyme Q10 and reduced coenzyme Q 10, therefore the most simultaneously
Utilize microbial fermentation product separation to prepare highly purified coenzyme Q10 and reduced coenzyme Q 10 thus incite somebody to action both fully in addition
Utilization is of great practical significance and economic worth.
In prior art, predominantly microbial fermentation product extracts in separation coenzyme Q10, or microbial fermentation product and carry
Take separating reducing type coenzyme Q10, and the coenzyme Q10 in tunning and reduced coenzyme Q 10 are not extracted separation simultaneously, cause
Coenzyme Q10 therein or reduced coenzyme Q 10 are wasted with the form of impurity.The Chinese patent of Publication No. CN103819326A
A kind of method that document discloses ultrasonication extraction, chromatography purification prepares coenzyme Q10.This technique can prepare coenzyme Q10,
But the not mentioned process to reduced coenzyme Q 10 contained in tunning, causes the waste to it.Publication No.
The Chinese patent literature of CN103740777A discloses a kind of selection-breeding, cultivates a kind of reduced coenzyme Q 10 bacterial strain, and ferments from it
The method extracting separating reducing type coenzyme Q10 in product, but the not mentioned place to the coenzyme Q10 contained by possibility in tunning
Reason, causes the waste to it.
Therefore, existing extraction, purification coenzyme Q10 or the technique of reduced coenzyme Q 10 from microbial fermentation product, all
Only focusing on one of which, another product is then taken as impurity and discards, and causes the significant wastage of active substance.So that
On the basis of prior art, combine, simplify, optimize technique, search out that a kind of technique is simple, low cost, high yield from micro-life
Thing tunning separates preparation high-purity coenzyme Q10 and the method for reduced coenzyme Q 10.
Summary of the invention
The invention provides a kind of concurrently separating and prepare high-purity coenzyme Q10 and the technique of reduced coenzyme Q 10, final
Having arrived the high-purity coenzyme Q10 of purity more than 98% and the high-purity reduced coenzyme Q 10 of purity more than 98%, total recovery is
More than 94%, whole simple and reliable process, easily operate, it is easy to accomplish, parameter is easy to control.
A kind of separation prepares high-purity coenzyme Q10 and the technique of reduced coenzyme Q 10, comprises the steps:
(1) seepage pressure effects: dreg carries out seepage pressure effects, collects containing coenzyme Q10 and the percolate of reduced coenzyme Q 10.
Step (1) described seepage pressure effects process is as follows:
Dreg is loaded percolation post, in percolation post, adds normal hexane, soak 90～120min at 5～30 DEG C, be incubated shape
Under state, percolation post carries out percolation operation, collect exit containing coenzyme Q10 and the percolate of reduced coenzyme Q 10, and simultaneously
Flow velocity with 0.5～3BV/h adds extractant in percolation post continuously, until the volume of the percolate collected is 4～6BV.
Preferably, the volume of the Extraction solvent added during immersion is 0.6～1BV.
In step (1) with normal hexane extraction at 5～30 DEG C, coenzyme Q10 and reduced coenzyme Q 10 are at such a temperature
In normal hexane, dissolubility is relatively big, and in dreg, other compositions dissolubility wherein is relatively small, at a temperature of being therefore somebody's turn to do, and coenzyme
The extraction ratio of Q10 and reduced coenzyme Q 10 is high, and the content of impurity is low, thus is improving coenzyme Q10 and reduced coenzyme Q 10
Its content in extract is improved while extraction ratio.The extraction ratio of coenzyme Q10 can reach more than 95%, reduced coenzyme
The extraction ratio of Q10 can reach more than 85%, and in percolate, the mass percent of coenzyme Q10 is 65%～75% (with butt
On the basis of), the mass percent of reduced coenzyme Q 10 is 4%～10% (on the basis of butt), contains auxiliary in extracting solution simultaneously
The impurity such as enzyme Q10 isomer, remaining coenzyme Q kind material.At a temperature of most preferably operating at 10 DEG C, use normal hexane extraction.
Raw material dreg is soaked 90～120min, makes dreg the most swelling, during percolation, need to ensure that dreg can be completely soaked
In Extraction solvent.While collecting percolate, in percolation post, add Extraction solvent under keeping warm mode continuously, thus protecting
Reduce solvent consumption while card coenzyme Q10 and reduced coenzyme Q 10 high extraction, and reduce the energy consumption of recycling design.Continuously
The Extraction solvent flow velocity added is preferably 0.5～3BV/h.In this flow rates, both can ensure that Extraction solvent and dreg had
Sufficiently time of contact, improve the utilization rate of solvent, production capacity can be taken into account again.
In order to improve the utilization ratio of solvent in this step, solvent can be recycled in the following manner: collect
To percolate be condensed into solid-state crude extract during, reclaim isolated solvent, its next time percolation during can conduct
Fresh solvent reuses.
(2) crystallization: obtain solid crude product, by described solid-state except solvent processes by concentrated for step (1) gained percolate
Crude product dissolves, and decrease temperature crystalline is filtrated to get coenzyme Q10 fine work and crystalline mother solution.
Step (2) described crystallization process is as follows:
At 20～70 DEG C, the solid crude product being concentrated to give is dissolved in organic solvent, obtains settled solution, progressively
Being cooled to-5～5 DEG C, crystallisation by cooling 6～filter after 24 hours, gained filtrate i.e. crystalline mother solution, gained filter cake is with appropriate cold ethanol
Washing, is vacuum dried the coenzyme Q10 fine work obtaining purity more than 98% for 6～12 hours at 20～40 DEG C.
Preferably, organic solvent described in this step be methanol, ethanol, acetone, ethyl acetate, normal hexane, normal heptane or
The mixture of any two kinds, the ratio of its addition and solid crude product is (40～80) mL 1g.If organic solvent addition mistake
Low, coenzyme Q10 crystallization purity is too low；If organic solvent addition is too high, coenzyme Q10 productivity is too low, it is therefore desirable to conservative control
The addition of organic solvent, to take into account purity and the productivity of coenzyme Q10.
Organic solvent more preferably acetone used, addition and solid crude product ratio are preferably 60mL:1g.
Preferably, this step crystallization temperature is-5～0 DEG C, and crystallization time is 6～12 hours.If crystallization temperature is too low or knot
Brilliant overlong time, the coenzyme Q10 purity obtained is too low, therefore wants conservative control crystallization temperature and time.This step crystallization temperature
More preferably 0 DEG C, crystallization time more preferably 6h.
Of a relatively high owing to extracting the content of coenzyme Q10 in the solid-state crude extract obtained, therefore this step is by crystallization point
Separate out in extract major part coenzyme Q10, suitably reduce the yield of coenzyme Q10 and ensure that the pure of the coenzyme Q10 that crystallization obtains
Degree.Remainder coenzyme Q10 is stayed in mother solution, is further processed it by subsequent step.
(3) reduction: carry out solvent displacement by mother liquid obtained for step (2), then mix with reducing agent, reduce to obtain reducing solution；
Step (3) described reduction process is as follows:
By the mother liquid obtained 5～20mg/mL solution being replaced as organic solvent A, reducing agent is configured to the 3～8mg/ of solvent B
ML solution, by the mixing of two kinds of solution equal-volumes, under 15～30 DEG C and atmosphere of inert gases, stirring reaction 1～5 hour, obtains reduction
Liquid.
Preferably, during described in this step, organic solvent A is ethanol, normal hexane, normal heptane and petroleum ether any one；Mother solution
The concentration more preferably 5～20mg/mL of organic solvent A solution, most preferably 10mg/mL.
Preferably, during solvent B described in this step is water, second alcohol and water/alcohol mixture any one.
Further preferably water, with organic solvent split-phase, thus can be directly separated part aqueous impurity after reduction reaction.
Preferably, described in this step, reducing agent is arbitrary in being sodium borohydride, ferrum, ascorbic acid and sodium dithionite
Kind.
Further preferably sodium dithionite, reducing agent can preferably be dissolved in water, reducing agent oxidation product energy solvent
Water directly removes, does not produce new impurity.
Preferably, described in this step, reduction reaction is carried out at 15～30 DEG C, and noble gas is nitrogen.Atmosphere of inert gases
Lower reaction, can guarantee that the reduced coenzyme Q 10 that reduction obtains will not be the most oxidized, thus ensure that the efficiency of reduction reaction.
Preferably, in the B solution of reducing agent described in this step, the addition of reducing agent is the 1～6 of required theoretical hydrogen equivalent
Times.If reducing agent addition is too low, cause coenzyme Q10 can not be reduced into reduced coenzyme Q 10 completely, if reducing agent addition
Too high, cause the waste of reducing agent.Therefore the addition of conservative control reducing agent, takes into account the efficiency of reduction reaction and economic effect
Benefit.
Further preferably in the B solution of reducing agent, the addition of reducing agent is 2～5 times of required theoretical hydrogen equivalent.
Owing to there is a certain amount of coenzyme Q10 and reduced coenzyme Q 10 in crystalline mother solution simultaneously, but both content are the most not
Height, therefore overwhelming majority coenzyme Q10 is reduced into reduced coenzyme Q 10, reduced coenzyme Q 10 by adding reducing agent by this step
Become target product.By processing further of reducing solution is isolated reduced coenzyme Q 10.
(4) extraction: concentrated by step (3) gained reducing solution, extracts through organic solvent, obtains extract.
Preferably, in this step, reducing solution is concentrated into 50～700mg/mL；More preferably 200～400mg/mL.
Preferably, during described in this step, organic solvent is methanol, acetonitrile, DMF and dimethyl sulfoxide
Any one, consumption be concentrate after 0.5～1.5 times of reducing solution volume.
The further preferred N,N-dimethylformamide of organic solvent described in this step.
After standing 10～60min layerings after extraction terminates in this step, isolated is biphase.Can be by overwhelming majority reduced form
Coenzyme Q10 is extracted in extraction phase, stays in raffinate phase, preferably by coenzyme Q10 by a small amount of unreduced coenzyme Q10 simultaneously
Separate with reduced coenzyme Q 10.The raffinate containing a small amount of coenzyme Q10 and reduced coenzyme Q 10 obtained after extraction is incorporated to crystallization
Mother solution reduces, it is to avoid coenzyme Q10 and reduced coenzyme Q 10 loss.
(5) crystallization: step (4) gained extract is concentrated, crystallisation by cooling after organic solvent dissolving, filtration, dried
After reduced coenzyme Q 10 fine work.
Step (5) described crystallization process is as follows:
It is dissolved in organic solvent at 20～50 DEG C after extract is concentrated, obtains settled solution, be progressively cooled to-10
～0 DEG C, crystallisation by cooling 6～filtration after 24 hours, gained crystal is vacuum dried at 20～40 DEG C with after appropriate cold washing with alcohol
Obtain reduced coenzyme Q 10 fine work.
Crystalline mother solution may be incorporated in extract, carries out extracting, crystallization treatment, to improve the response rate of reduced coenzyme Q 10.
Preferably, organic solvent described in this step is methanol, acetone, ethanol, acetonitrile or the mixture of any two kinds, adds
The ratio of amount and solids after concentration is (20～50) L:1kg.Described organic solvent is most preferably ethanol, and addition is most preferably
30L:1kg.
Preferably, this step organic solvent adds a certain amount of acid, is beneficial to reduced coenzyme Q 10 and keeps in atmosphere
Stable.Described acid is preferably hydrochloric acid, and addition is 100mmol hydrogen chloride/kg ethanol.After obtaining crystalline product, volatile acid is more
It is prone to remove, it is to avoid acid residual.
Preferably, in this step, cooling extent is 0.5～3 DEG C/min, and crystallization temperature is-10～0 DEG C, further preferably
For-5～0 DEG C；Most preferably 0 DEG C.
Explicans specific explanations involved in the present invention is as follows:
BV: bed volume, is packed into percolation post by dreg wet method, and dreg is in the volume under free accumulation state.
The solution of solid concentration 100mg/mL, refers to that every milliliter of solution contains 100mg solid when being concentrated into mass conservation
Thing, owing to solution composition is complicated, has certain insoluble matter, refers to the gross mass of whole system, bag when calculating gross mass
Include and suspend or the quality of precipitation.
Butt refers to the quality of the solid that solution obtains after drying, the quality summation of the whole solutes in ie in solution.
Theoretical hydrogen equivalent refers in reduction reaction, the amount of required reducing agent active hydrogen during complete dihydrocoenzyme Q10.
The technique of the present invention uses low temperature percolation to replace solvent leaching method to extract coenzyme Q10 from dreg and reduced form is auxiliary
Enzyme Q10, greatly reduces solvent consumption and the waste liquid output of extraction process, uses crystallization first to isolate most of coenzyme Q10,
The waste of reduced coenzyme Q 10 is avoided, in simple process by reduction and extract and separate coenzyme Q10 and reduced coenzyme Q 10
Concurrently separate with realization under conditions of low cost and prepare highly purified coenzyme Q10 and reduced coenzyme Q 10, it is thus achieved that high total receipts
Rate.
The present invention use percolation extract coenzyme Q10 and reduced coenzyme Q 10 from dreg, with conventional solvent leaching
Method is compared, and remains Concentraton gradient maximum inside and outside raw material, can accelerate dissolution mass transfer, significantly improve and carry during percolation
Taking speed, reduce solvent consumption, make coenzyme Q10 extraction rate reached to more than 95%, the extraction ratio of reduced coenzyme Q 10 can reach
More than 85%, in percolate, the mass percent (being as the criterion with butt) of coenzyme Q10 is between 65%～75%, and reduced form is auxiliary
The mass percent (being as the criterion with butt) of enzyme Q10 is between 4%～10%.
The present invention uses crystallization, first by the most of coenzyme Q10 Crystallization Separation in solid-state crude extract, suitably reduces coenzyme
The yield of Q10 and ensure that the high-purity of the coenzyme Q10 that crystallization obtains.Remainder coenzyme Q10 is stayed in mother solution, by follow-up
It is further processed by step.
The present invention, by remaining coenzyme Q10 in reduction reaction reduction crystalline mother solution, is translated into reduced coenzyme
Q10, reduced coenzyme Q 10 becomes target product.Also can be by less reduction originally while rationally having converted residue coenzyme Q10
Separate to type coenzyme Q10 high-purity.
The present invention uses the reduced coenzyme Q 10 after extract and separate reduction and a small amount of coenzyme Q10, and will obtain after extraction
Raffinate containing coenzyme Q10 and reduced coenzyme Q 10 is incorporated to crystalline mother solution and reduces, it is to avoid coenzyme Q10 and reduced coenzyme
Q10 loses.
And use primary crystallization to obtain purity to reach the reduced coenzyme Q 10 fine work of more than 98%, and will mother solution be also after crystallization
Enter in extract, carry out extracting, crystallization treatment, improve the yield of reduced coenzyme Q 10.
Generally speaking, the present invention above five processes is combined extract from dreg, purify, separate coenzyme Q10 and
The technology of reduced coenzyme Q 10, hinge structure is had the advantage that as follows:
The process route of the present invention is short, and to producing, equipment requirements is low, simple to operate；Each solvent in this process route
Reclaiming, therefore in this technique, most raw material can be repeatedly utilized so that；Reduction step effectively make use of content originally relatively low
Reduced coenzyme Q 10, has prepared highly purified coenzyme Q10 and reduced coenzyme Q 10 simultaneously；Raffinate and crystalline mother solution
The total recovery of the technical process that recycled is greatly improved.In this process route, technological process is simple, production cost is low, produce effect
Rate is high, it is easy to industrialization large-scale production.
Accompanying drawing explanation
Fig. 1 is the HPLC chromatogram that the mensuration sterling coenzyme Q10 concentration method set up according to the present invention obtains.
Fig. 2 is the HPLC chromatogram that the mensuration sterling reduced coenzyme Q 10 concentration method set up according to the present invention obtains.
Detailed description of the invention
Preparation high-purity coenzyme Q10 and the preparation method of reduced coenzyme Q 10 and test result is separated below in conjunction with a kind of
The present invention is further described.
Embodiment 1
(1) (wherein, coenzyme Q10 mass percent is 2.6%, and reduced coenzyme Q 10 mass percent is to weigh dreg
0.2%) 70g, wet method is packed in percolation post (Φ 2.0 × 35cm), and after filling uniformly, its stacking volume is about 100mL, 10 DEG C
Constant temperature soaks 2h so that it is the most swelling, opens percolation column outlet valve, and is continuously added to normal hexane, coutroi velocity from capital simultaneously
2.5～3.0mL/min, until the percolate volume collected is 540mL.
The percolate collected is as the material liquid of subsequent operation, and through analyzing, solid concentration is 4.8mg/mL, coenzyme Q10
Content is 69.5%, and extraction ratio is 98.9%, and reduced coenzyme Q 10 content is 4.6%, and extraction ratio is 85.4%.
(2) percolate concentrating removing solvent, at 30 DEG C, interpolation acetone is to being just completely dissolved, solid-to-liquid ratio 1:60, then
Temperature is progressively down to 0 DEG C, filters after crystallisation by cooling 6h, and by appropriate cold washing with alcohol.After fully draining, put into vacuum drying
30 DEG C of dry 6h in case, obtain 0.93g yellow coenzyme Q10 fine work, and purity is 98.0%, and the yield of this technique coenzyme Q10 is
51.6%.Mother solution after crystallization is for further processing.
(3) by after mother liquid obtained concentration, being configured to 10mg/mL solution with normal heptane, reducing agent sodium dithionite is prepared
Become 6.7mg/mL aqueous solution, at 25 DEG C, under nitrogen atmosphere, mother solution and reductant solution equal-volume mix and blend reaction 2h, must go back
Stock solution.Through analyzing, reduction reaction coenzyme Q10 conversion ratio 99.9%, reduced coenzyme Q 10 productivity 92.0%.Due to coenzyme Q10 and
Reduced coenzyme Q 10 is the most insoluble in water, therefore this technique reduction yield 92.0%.
(4) separate phase, reducing solution is concentrated into 300mg/mL, add the extraction of equivalent DMF.Extraction
After balance, analytical extraction liquid, reduced coenzyme Q 10 content is 82.5%, this technique reduced coenzyme response rate 87.7%.Extraction
Rear raffinate can further recycled be to reduction step, and the extraction stages response rate can calculate with 100%, this technique it is believed that
Substantially lose without reduced coenzyme Q 10.
(5) extract obtained is concentrated to dryness, and at 30 DEG C, interpolation ethanol is to being just completely dissolved, and adds appropriate hydrochloric acid
(100mmol hydrogen chloride/kg ethanol), solid-to-liquid ratio 1:30, then temperature is progressively down to 0 DEG C, filter after crystallisation by cooling 12h, and use
Appropriate cold washing with alcohol.After fully draining, put into 30 DEG C of dry 6h in vacuum drying oven, obtain 0.78g reduced coenzyme Q 10 essence
Product, purity is 98.0%.In dreg on the basis of coenzyme Q10 and reduced coenzyme Q 10, whole process overall yields is 87.2%.
Mother solution after crystallization can further recycled be to extract and separate step, and the crystallization stage response rate can calculate with 100%, and extraction
Taking that rear raffinate can further recycled be to reduction step, the extraction stages response rate can calculate with 100%, so that technique
Total recovery improves to 94.4%.
Embodiment 2
(1), (2) operational approach is with embodiment 1.
(3) by after mother liquid obtained concentration, being configured to 10mg/mL solution with normal heptane, reducing agent sodium dithionite is prepared
Become 5.0mg/mL aqueous solution, at 25 DEG C, under nitrogen atmosphere, mother solution and reductant solution equal-volume mix and blend reaction 2h, must go back
Stock solution.Through analyzing, reduction reaction coenzyme Q10 conversion ratio 92.6%, reduced coenzyme Q 10 productivity 86.2%.Due to coenzyme Q10 and
Reduced coenzyme Q 10 is the most insoluble in water, therefore this technique reduction yield 86.2%.
(4) separate phase, reducing solution is concentrated into 300mg/mL, add the extraction of equivalent DMF.Extraction
After balance, analytical extraction liquid, reduced coenzyme Q 10 content is 78.0%, this technique reduced coenzyme response rate 84.3%.Extraction
Rear raffinate can further recycled be to reduction step, and the extraction stages response rate can calculate with 100%, this technique it is believed that
Substantially lose without reduced coenzyme Q 10.
(5) extract obtained is concentrated to dryness, and at 30 DEG C, interpolation ethanol is to being just completely dissolved, and adds appropriate hydrochloric acid
(100mmol hydrogen chloride/kg ethanol), solid-to-liquid ratio 1:30, then temperature is progressively down to 0 DEG C, filter after crystallisation by cooling 12h, and use
Appropriate cold washing with alcohol.After fully draining, put into 30 DEG C of dry 6h in vacuum drying oven, obtain 0.74g reduced coenzyme Q 10 essence
Product, purity is 98.0%.In dreg on the basis of coenzyme Q10 and reduced coenzyme Q 10, whole process overall yields is 85.2%.
Mother solution after crystallization can further recycled be to extract and separate step, and the crystallization stage response rate can calculate with 100%, and extraction
Taking that rear raffinate can further recycled be to reduction step, the extraction stages response rate can calculate with 100%, so that technique
Total recovery improves to 91.8%.
Embodiment 3
(1), (2), (3), (4) operational approach are with embodiment 1.
(5) extract obtained concentrates, and at 30 DEG C, interpolation acetone is to being just completely dissolved, and adds appropriate hydrochloric acid
(100mmol hydrogen chloride/kg acetone), solid-to-liquid ratio 1:20, then temperature is progressively down to 5 DEG C, filter after crystallisation by cooling 12h, and use
Appropriate cold washing with alcohol.After fully draining, put into 30 DEG C of dry 6h in vacuum drying oven, obtain 0.69g reduced coenzyme Q 10 essence
Product, purity is 98.4%.In dreg on the basis of coenzyme Q10 and reduced coenzyme Q 10, whole process overall yields is 82.7%.
Mother solution after crystallization can further recycled be to extract and separate step, and the crystallization stage response rate can calculate with 100%, and extraction
Taking that rear raffinate can further recycled be to reduction step, the extraction stages response rate can calculate with 100%, so that technique
Total recovery improves to 94.4%.
Coenzyme Q10 and reduced coenzyme Q 10 method for measurement of concentration
Above example all uses following methods test coenzyme Q10 and the concentration of reduced coenzyme Q 10.
Using U.S.'s WATERS high performance liquid chromatograph to set up HPLC and analyze method, detector is UV-detector；Chromatograph
Post is: WATERS Atlantis T3column (250mm × 4.6mm, 5 μm)；Liquid inlet volume: 10 μ L；Flowing is methanol/ethanol mutually
(1:1,v/v)；Flow velocity: 1mL/min；Column temperature: 30 DEG C.
Each range of linearity:
Coenzyme Q10: 0～3.2mg/mL；Reduced coenzyme Q 10: 0～3.1mg/mL.
Each standard curve:
Coenzyme Q10: y=8.14322 × 106x；Reduced coenzyme Q 10: y=1.09537 × 106x.X peak area,
Y concentration.
After testing, the retention time of coenzyme Q10 is 22.779min (accompanying drawing 1), and the retention time of reduced coenzyme Q 10 is
(13.924min accompanying drawing 2).
The above is only the preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For Yuan, on the premise of without departing from the technology of the present invention principle, it is also possible to make some improvements and modifications, these improvements and modifications
Also should be regarded as protection scope of the present invention.
Claims (10)
1. one kind concurrently separates preparation high-purity coenzyme Q10 and the technique of reduced coenzyme Q 10, it is characterised in that include as follows
Step:
(1) dreg is carried out seepage pressure effects, collect containing coenzyme Q10 and the percolate of reduced coenzyme Q 10；
(2) solid crude product is processed to obtain by concentrated for step (1) gained percolate, cooling knot after being dissolved by described solid crude product
Crystalline substance, is filtrated to get coenzyme Q10 fine work and crystalline mother solution；
(3) carry out solvent displacement by mother liquid obtained for step (2), then mix with reducing agent, reduce to obtain reducing solution；
(4) extract through organic solvent after step (3) gained reducing solution being concentrated, obtain extract；
(5) adding organic solvent after being concentrated by step (4) gained extract to dissolve, after crystallisation by cooling, filtration, dried must be reduced
Type coenzyme Q10 fine work.
Concurrently separate preparation high-purity coenzyme Q10 and the technique of reduced coenzyme Q 10, its feature the most according to claim 1
Being, described in step (2), decrease temperature crystalline is: at 20～70 DEG C, and the solid crude product being concentrated to give is dissolved in organic solvent
In, progressively it is cooled to-5～5 DEG C, crystallisation by cooling 6～filtration after 24 hours, obtain the most described crystalline mother solution of filtrate, after Cake Wash
Described coenzyme Q10 fine work it is vacuum dried to obtain at 20～40 DEG C.
Concurrently separate preparation high-purity coenzyme Q10 and the technique of reduced coenzyme Q 10, its feature the most according to claim 2
Being, used in the process of cooling, organic solvent is methanol, ethanol, acetone, ethyl acetate, normal hexane, normal heptane or any
The mixture of two kinds, the ratio of organic solvent addition and solid crude product is (40～80) mL 1g.
Concurrently separate preparation high-purity coenzyme Q10 and the technique of reduced coenzyme Q 10, its feature the most according to claim 1
Being, by the mother liquid obtained 5～20mg/mL solution being replaced as organic solvent A in step (3), reducing agent is configured to the 3 of solvent B
～8mg/mL solution, by the mixing of two kinds of solution, under 15～30 DEG C and atmosphere of inert gases, stirring reaction 1～5 hour, obtains reduction
Liquid.
Concurrently separate preparation high-purity coenzyme Q10 and the technique of reduced coenzyme Q 10, its feature the most according to claim 4
Be, for parent solution solvents displacement organic solvent be A be any one in ethanol, normal hexane, normal heptane and petroleum ether；For joining
The solvent B putting reducing agent is any one in water, second alcohol and water/alcohol mixture；Reducing agent is sodium borohydride, ferrum, ascorbic acid
With in sodium dithionite any one；Noble gas is any one in nitrogen, argon and carbon dioxide.
Concurrently separate preparation high-purity coenzyme Q10 and the technique of reduced coenzyme Q 10, its feature the most according to claim 4
Being, the addition of described reducing agent is 1～6 times of theoretical hydrogen equivalent.
Concurrently separate preparation high-purity coenzyme Q10 and the technique of reduced coenzyme Q 10, its feature the most according to claim 1
Being, step (4) described extraction process is: gained reducing solution is concentrated into 50～700mg/mL, extracts with organic solvent, obtains extraction
Take liquid.
Concurrently separate preparation high-purity coenzyme Q10 and the technique of reduced coenzyme Q 10, its feature the most according to claim 7
Being, in extraction process, organic solvent used is any one in methanol, acetonitrile, DMF and dimethyl sulfoxide,
Consumption of organic solvent be concentrate after 0.5～1.5 times of reducing solution volume.
Concurrently separate preparation high-purity coenzyme Q10 and the technique of reduced coenzyme Q 10, its feature the most according to claim 1
Being, step (5) described crystallisation by cooling process is: be dissolved in organic solvent at 20～50 DEG C after being concentrated by extract, by
Step is cooled to-10～0 DEG C, crystallisation by cooling 6～filtration after 24 hours, is vacuum dried and obtains reduction after Cake Wash at 20～40 DEG C
Type coenzyme Q10 fine work.
Concurrently separate preparation high-purity coenzyme Q10 and the technique of reduced coenzyme Q 10, its feature the most according to claim 9
Being, in step (5), organic solvent used by crystallisation by cooling is methanol, acetone, ethanol, acetonitrile or the mixture of any two kinds；Have
The ratio of machine solvent adding amount and solids after concentration is (20～50) L:1kg, and adds appropriate amount of acid simultaneously.
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| CN111943827A (en) * | 2020-09-28 | 2020-11-17 | 陕西嘉禾药业有限公司 | Method for purifying coenzyme Q10 |
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| EP2289866A2 (en) * | 2001-07-13 | 2011-03-02 | Kaneka Corporation | Method of producing reduced coenzyme Q10 as oily product |
| CN102391092A (en) * | 2011-11-22 | 2012-03-28 | 杭州华东医药集团康润制药有限公司 | Method for preparing high-purity coenzyme Q10 in large scale |
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| EP2289866A2 (en) * | 2001-07-13 | 2011-03-02 | Kaneka Corporation | Method of producing reduced coenzyme Q10 as oily product |
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