CN104730175B - A kind of automatic vacuum Split liquid chromatographic apparatus and control method thereof - Google Patents
A kind of automatic vacuum Split liquid chromatographic apparatus and control method thereof Download PDFInfo
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- 238000000034 method Methods 0.000 title claims abstract description 32
- 239000007788 liquid Substances 0.000 title claims abstract description 21
- 238000010828 elution Methods 0.000 claims abstract description 14
- 238000006243 chemical reaction Methods 0.000 claims abstract description 4
- 239000012530 fluid Substances 0.000 claims description 39
- 230000006835 compression Effects 0.000 claims description 29
- 238000007906 compression Methods 0.000 claims description 29
- 238000011049 filling Methods 0.000 claims description 19
- 239000000945 filler Substances 0.000 claims description 18
- 239000003480 eluent Substances 0.000 claims description 14
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 12
- 239000000741 silica gel Substances 0.000 claims description 12
- 229910002027 silica gel Inorganic materials 0.000 claims description 12
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- 238000012545 processing Methods 0.000 claims description 6
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- 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 claims description 3
- 238000007689 inspection Methods 0.000 claims description 3
- 239000003456 ion exchange resin Substances 0.000 claims description 3
- 229920003303 ion-exchange polymer Polymers 0.000 claims description 3
- 238000004237 preparative chromatography Methods 0.000 claims description 2
- 238000000151 deposition Methods 0.000 claims 1
- 238000000926 separation method Methods 0.000 abstract description 21
- 239000012071 phase Substances 0.000 abstract description 15
- 238000003817 vacuum liquid chromatography Methods 0.000 abstract description 15
- 229940079593 drug Drugs 0.000 abstract description 5
- 239000003814 drug Substances 0.000 abstract description 5
- 229930014626 natural product Natural products 0.000 abstract description 2
- 239000007790 solid phase Substances 0.000 abstract description 2
- 239000003921 oil Substances 0.000 description 20
- 239000003208 petroleum Substances 0.000 description 11
- 238000011068 loading method Methods 0.000 description 10
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- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 8
- 238000001514 detection method Methods 0.000 description 8
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- 240000006079 Schisandra chinensis Species 0.000 description 7
- 238000004440 column chromatography Methods 0.000 description 7
- 235000013399 edible fruits Nutrition 0.000 description 7
- 238000004128 high performance liquid chromatography Methods 0.000 description 7
- 150000001875 compounds Chemical class 0.000 description 6
- SRCZQMGIVIYBBJ-UHFFFAOYSA-N ethoxyethane;ethyl acetate Chemical compound CCOCC.CCOC(C)=O SRCZQMGIVIYBBJ-UHFFFAOYSA-N 0.000 description 6
- 238000005516 engineering process Methods 0.000 description 5
- 238000002360 preparation method Methods 0.000 description 5
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- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 3
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
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- 239000000463 material Substances 0.000 description 3
- 239000012259 ether extract Substances 0.000 description 2
- 239000010720 hydraulic oil Substances 0.000 description 2
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- 230000003287 optical effect Effects 0.000 description 2
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- GBMDVOWEEQVZKZ-UHFFFAOYSA-N methanol;hydrate Chemical compound O.OC GBMDVOWEEQVZKZ-UHFFFAOYSA-N 0.000 description 1
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Abstract
A kind of automatic vacuum Split liquid chromatographic apparatus and control method thereof, belong to the technical field of chromatogram arrangement.Logic control system is organically combined by this device with liquid storage and fraction collection system, complete to go out pneumatic control valve and 1 and enter the 3 break-makes conversions going out the combination of the valve such as pneumatic control valve liquid storage and fraction collection system enter 1 more by controlling cylinder moving direction, constitute the automation control system of Separation of Natural Products, by elution program set in advance, the gradient elution and the cut that achieve flowing phase are collected automatically, it is achieved the intelligent manipulation that vacuum liquid-chromatography apparatus is isolated and purified to natural drug.This separator uses the chromatographic column combination of different size and solid-phase media, have separative efficiency height, labour intensity is low, the cost that saves manpower and time, the feature of Automated condtrol continuous prodution, can realize the separation quick, effective, intelligent to complex samples such as natural drugs.
Description
Technical field
The present invention relates to a kind of automatic vacuum Split liquid chromatographic apparatus and control method thereof, belong to the technology of chromatogram arrangement
Field.
Background technology
Although traditional chromatogram arrangement can reach isolated and purified purpose, but often there is disengaging time length, consumption
The defects such as quantity of solvent is big, relatively costly or separative efficiency is low, product purity is inadequate, and active ingredient is often tediously long, numerous
Many pillar layer separation, solvent concentration and sample transfer operations incur loss or structure changes, make the researcher must not
Do not strengthen inventory in the hope of obtaining preferable result.Therefore, separation preparation facilities efficient, high-throughout is always research field
Close point of contact.
Vacuum liquid chromatogram (Vacuum Liquid Chromatography, VLC) is new as one in mid-term in 20th century
Type, simplicity, the technology of efficiently separating be suggested and be applied to natural organic-compound and the purifying of volume organic reaction product and
Separate [Aust.J.chem.1977,30,1859], its essence is column chromatography, prepare combining of thin-layer chromatography and vacuum filtration technology
Conjunction application [chromatogram .1990,8(5) 313].VLC is different from traditional Column chromatography techniques, because the latter is to enter continuously when wash-out
Row, will not be interrupted in operation, and VLC is when carrying out solvent wash-out, by eluant, eluent after post under reduced pressure all
After extraction, then changing solvent, proceed the collection of cut next time, its principle is similar to the repeatedly expansion of thin-layer chromatography.With
Traditional column chromatography is compared, and VLC has that device is simple, amount of filler is few, the operating time is short, organic solvent consumption is little, sample
The features such as product treating capacity is big, and be successfully applied to variation compound preparative separation [J.Natl.prod.1986,49,892;
J.Natl.prod.1986,49,934;Mountain Western Medicine S University journal .1999,30,41].
But, VLC there is also many weak points as a kind of inexpensive efficient, simple and efficient isolation technics: chromatographic column
Filler filling the most closely causes post bed less stable, theoretical cam curve is low, chromatogram column efficiency is uncontrollable；Particularly feed flow, vacuum
The step such as elute, drain, and gradient cyclic process need to manually be carried out, thus consume substantial amounts of time and human cost.Cause
This, reduce the most to greatest extent manual operation, realize automatic partition from, to improve efficiency, to break through the bottle that formed of conventional method
Neck, it appears extremely urgent.
Summary of the invention
For problem, the present invention such as solve that chromatogram column efficiency present in traditional VLC technology is unstable and manpower consumption is high
Provide a kind of automatic vacuum Split liquid chromatographic apparatus, with efficiently, serialization can prepare compound.
The technical scheme is that a kind of automatic vacuum Split liquid chromatographic apparatus, including liquid-way system and logic control
System processed, logic control system is provided with signal lamp, touch-screen and button on panel, is provided with PLC logic controller in casing
And magnetic valve, PLC logic controller comprise central processing unit, button information module, positional information module, break-make modular converter,
Oil cylinder control module and signal control module；Described liquid-way system employing fluid reservoir is connected to enter 1 through fluid reservoir arm more and goes out
Pneumatic control valve, then through 1 enter 3 go out pneumatic control valve by arm in chromatographic column connect respectively the first chromatographic column, the second chromatographic column,
3rd chromatographic column, then sequentially passed through 3 to enter 1 to go out pneumatic control valve by chromatographic column lower branch line, 1 enter to have more pneumatic control valve, collector
Arm is connected to collector；Described first chromatographic column, the second chromatographic column, the 3rd chromatographic column are provided with oil cylinder, described collector with
Vavuum pump connects；The magnetic valve that described logic control system controls by gas piping with enter 1 to go out pneumatic control valve more, 1 enter 3 and go out
Pneumatic control valve, 3 enter 1 to go out pneumatic control valve, 1 enter to have more between pneumatic control valve and oil cylinder and be attached；Described central authorities process
Device performs program and storage data, is controlled automatic control process, and importation gathers signal from switch, exports part
Then controlled the direction of motion of oil cylinder by oil cylinder control module, central processing unit controls many by break-make modular converter and magnetic valve
Enter 1 go out pneumatic control valve, 1 enter 3 go out pneumatic control valve, 3 enter 1 to go out pneumatic control valve, 1 break-make entering to have more pneumatic control valve turns
Change.
The method of work of a kind of automatic vacuum Split liquid chromatographic apparatus comprises the following steps:
(1), before operation, logic control system is carried out routine inspection, checks that equipment state is the most normal；First controlling
Cabinet guidance panel selects automatically to run button, and its position rotates to manual mode, and manual indicator lamp is lighted；Rotate afterwards to certainly
Flowing mode, automatic pilot lamp is lighted；Turn on the power, enter the homepage of logic control system, carry out experiment parameter setting；
(2) flushing line: after injecting initial flowing mutually in fluid reservoir, more regulation is entered 1 to go out pneumatic control valve, 1 is entered 3 successively
Go out pneumatic control valve, 3 enter 1 and go out pneumatic control valve and 1 and enter to have more pneumatic control valve, to pipeline and the first chromatographic column, the second chromatogram
Post, the 3rd chromatographic column are rinsed；
(3) filling chromatographic column: need to select preparative chromatography column dimension according to experiment, carry out filler under vacuum
Filling, after filler filling, " chromatogram hydraulic pressure " button clicked in button makes dynamic axial compression column move down compacting look
Spectrum post；After compression, clicking on " stopping " button in button, chromatographic column filling is complete；
(4) sample dress post: sample to be separated is added chromatographic column, injects the flowing phase configured, button in fluid reservoir
Selected chromatographic column, joins sample in selected chromatographic column, then enters 3 by 1 and go out pneumatic control valve and 3 and enter 1 and go out pneumatic control valve
Switching, enter chromatographic column used with 13 to go out pneumatic control valve, 3 enter 1 and go out pneumatic control valve and connect；
(5) chromatographic column sample elution: go out pneumatic control valve and 1 enter to have more the switching of pneumatic control valve by entering 1 more, connects
First fluid reservoir, chromatographic column, the first collector, carry out vacuum elution, until flowing is drained in chromatographic column, without washing in pipeline
De-liquid；Reducing without eluent or vacuum in pipeline and trigger logic control system, logic control system switching is entered 1 more and is gone out pneumatic control
Valve processed and 1 enters to have more pneumatic control valve, closes the path of the first fluid reservoir to the first collector；Go out pneumatic control by entering 1 more
Valve and 1 enters to have more the switching of pneumatic control valve, connects the path of the second fluid reservoir to the second collector, repeats said process, directly
To completing whole wash-out, logic control system has sent signal；
(6), after having separated, each eluting fraction reclaims in corresponding collector.
Described first chromatographic column, the second chromatographic column, the specification of the 3rd chromatographic column are 150mm × 280mm, 100mm ×
280mm, 80mm × 280mm.
The filler of described first chromatographic column, the second chromatographic column and the 3rd chromatographic column selected from silica gel, C8, C18, bonded silica gel or
Ion exchange resin.
Enter 1 to go out pneumatic control valve and 1 and enter to have more integral valve that pneumatic control valve is rotating switching type or by multiple cut-offs more
The combination valve of valve composition, switching power is electric power or compressed air.Switching command is time command, pressure instruction or light instruction；
Time command is the process time provided according to preliminary experiment and is manually set, and pressure instruction comes from pressure sensor, light instruction source
In optical sensor.The chromatographic column of this device is dynamic axial compression chromatographic column.Dynamic axial compression chromatographic column can ensure chromatographic column
The stability of bed and Gao Zhuxiao.
In this device, automated intelligent mode is mechanical type, logic circuit type or electronic type.Logic control system is mainly by little
Type programmable PLC logic controller (MicroPLC) (i.e. siemens PLC) and eight cun of man-machine interface (HMI) compositions.Logic
In control system, Siemens S7-200PLC logic controller includes that (button is believed for a central processor CPU and 5 expansion modules
Breath modules A, positional information module B, break-make modular converter C, oil cylinder control module D and signal control module E), CPU is responsible for execution
Program and storage data, in order to native system is automatically controlled task or process is controlled；Input and output are the control of system
Point, importation gathers signal from switch, and output part is then by DM control valve door switch and cylinder movement side
To；CPU be responsible for control enter more 1 go out pneumatic control valve, 1 enter 3 go out pneumatic control valve, 3 enter 1 go out pneumatic control valve, 1 enter to have more pneumatic
The break-make conversion of control valve valve group；Button information modules A gathers button information, and positional information module B gathers the position of valve combination
Information, break-make modular converter C is responsible for the switching of valve, and oil cylinder control module D controls dynamic axial compression chromatographic column, and signal controls mould
Block E control signal lamp, as during system malfunctions, buzzer will send sound and light alarm.
The vacuum liquid-phase chromatographic column of this separator uses dynamic axial compression chromatography piece-rate system, and it is by pump installation, collection
Become block or valve combination, fuel tank, electrical box and chromatographic column to combine, rotated by driven by motor oil pump, make pump inhale from oil pump
Buying oil after oil, by the pressure energy that changes mechanical energy is hydraulic oil, hydraulic oil realizes direction, pressure by integrated package (or valve combination)
After power, Flow-rate adjustment in external pipeline is transferred to oil cylinder or the oil motor of hydraulic machinery, thus control the change in hydraulic motor direction
Change, the size of strength and the speed of speed, promote various hydraulic machinery to do work.It fills under the driving of gas drive hydraulic power
Fill out, and by chromatographic column bed fillers is carried out a certain degree of compression, it is ensured that the compactness continuous, the most even of filler, look simultaneously
Spectrum post realizes filling post, maintaining post to press and unload the function of post by the up and down motion of piston, and piston periphery is equipped with particular design
Sealing ring, while allowing piston freely up and down to slide, higher sealing pressure can be kept again.It and traditional chromatographic column phase
Ratio, have employed the chromatographic column type of feed of dynamic axial compression, by filling out chromatographic column bed under the driving of gas drive hydraulic power
Material carries out a certain degree of compression, it is ensured that the Gao Zhuxiao of dynamic axial compression chromatographic column and superior separating property；It addition, be
System is equipped with the chromatographic column of three kinds of different sizes, can form multicolumn separation system, can be according to material properties and quality sorting during use
Select appropriate gauge and carry out lock out operation；Chromatographic column can be loaded separating filler of different nature and (include silica gel, C8, C18, bonding
Silica gel, ion exchange resin etc.), with the satisfied different experiment demands separating purpose.
Beneficial effects of the present invention: logic control system is organically combined by this device with liquid storage and fraction collection system, logical
Cross control cylinder moving direction to complete to go out pneumatic control valve and 1 and enter 3 and go out pneumatic control liquid storage and fraction collection system enter 1 more
The break-make conversion of the valve combinations such as valve, constitutes the automation control system of Separation of Natural Products, by elution program set in advance,
The gradient elution and the cut that achieve flowing phase are collected automatically, it is achieved vacuum liquid-chromatography apparatus is isolated and purified to natural drug
Intelligent manipulation.This separator uses the chromatographic column combination of different size and solid-phase media, has separative efficiency height, work by force
Spend low, the cost that saves manpower and time, the feature of Automated condtrol continuous prodution, can realize complex samples such as natural drugs
Separation quick, effective, intelligent.
Accompanying drawing explanation
Fig. 1 is the structural representation of a kind of automatic vacuum liquid-chromatography apparatus.
Fig. 2 is the HPLC figure that a kind of automatic vacuum liquid-chromatography apparatus separates fruit of Chinese magnoliavine petroleum ether extract.
Fig. 3 is the TLC figure that a kind of automatic vacuum liquid-chromatography apparatus separates fruit of Chinese magnoliavine S5 component.
Fig. 4 is the HPLC figure that a kind of automatic vacuum liquid-chromatography apparatus separates fruit of Chinese magnoliavine S5 component.
In figure: 1, logic control system, 1a, signal lamp, 1b, touch-screen, 1c, button, 1d, PLC logic controller, 1e,
Magnetic valve, 2, fluid reservoir, 2a, fluid reservoir arm, 3, enter 1 more and go out pneumatic control valve, 4,1 enters 3 goes out pneumatic control valve, and 5, oil cylinder,
6, the first chromatographic column, 6a, the second chromatographic column, 6b, the 3rd chromatographic column, arm in 6c, chromatographic column, 6d, chromatographic column lower branch line, 7,3
Entering 1 and go out pneumatic control valve, 8,1 enters to have more pneumatic control valve, and 9, collector, 9a, collector arm, 10, vavuum pump, 11, gas
Pipeline, the 21, first fluid reservoir, the 22, second fluid reservoir, the 91, first collector, the 92, second collector, CPU, central processing unit,
A, button information module, B, positional information module, C, break-make modular converter, D, oil cylinder control module, E, signal control module.
Detailed description of the invention
Illustrating the preferred embodiments of the present invention below in conjunction with accompanying drawing, preferred embodiment set forth herein is only used for
The description and interpretation present invention, is not intended to limit the present invention.
Fig. 1 shows the structure of this device, mainly include gradient flow phase liquid-storage system, dynamic axial compression chromatographic column system
System, fraction collection system and logic control system four part.Its stream is: 10 fluid reservoir 2(can accommodate 10 kinds of gradients not respectively
With eluent) outlet respectively with enter 1 10 entrances going out pneumatic control valve 3 more 1 and be connected；Enter 1 more and go out pneumatic control valve
The outlet of 3 is entered 3 and is gone out pneumatic control valve 4 entrance and be connected with 11,1 enter 3 go out the outlet of pneumatic control valve 4 respectively with three diameters
The chromatographic column (such as 150mm × 280mm, 100mm × 280mm, 80mm × 280mm) not waited connects；The outlet of chromatographic column is respectively with 1
Individual 3 enter 13 entrances going out pneumatic control valve 7 connects, and 3 enter 1 outlet and 1 going out pneumatic control valve 7 enters to have more pneumatic control valve 8
Entrance connect, 1 outlet entering to have more pneumatic control valve 8 is connected with the collector 9 of 10 kinds of eluents respectively；The receipts of eluent
Storage 9 respectively with enter 1 entrance going out pneumatic control valve 8 more one and be connected, enter 1 outlet going out pneumatic control valve 8 and vavuum pump more
10 connect；10 pressure sensors or optical sensor are installed (such as purple connecting chromatographic column on the stream of the collector 9 of eluent
Outer or infrared detection pond).Go out pneumatic control valve 3 and 1 and enter to have more the automatic switchover of pneumatic control valve 8 by entering 1 more and realize automatically
Process.
The course of work of a kind of automatic vacuum liquid-chromatography apparatus is as follows:
1. device starts: be connected with air pump device by the air inlet pipeline of device, it is ensured that gas inputs, and by vavuum pump and dress
Put connection.Turning on the power, metering-in control system key frame, red round dot is closed for switch, and green round dot is that switch is opened；
Red triangle is for stopping, and green triangle is work；
2. flushing line: optional manually or automatically operation is (manually: when being intended for equipment debugging, equipment maintenance, have phase
Answer human users；Automatic: to be operated by laboratory technician or process engineer when equipment is normal).Mainly the automatic method of operation is entered below
Row illustrates.Fully enter the time of collector according to each flowing mutually, each container is set in " parameter setting " and takes out very
The empty time, clicking on parameter setting check, select corresponding chromatographic column, switch board guidance panel selects automatically to run button, is revolved
Going to automated manner, automatic pilot lamp is lighted.
3, dress post loading: after selecting suitable chromatographic column model and separating filler according to sample quality, under vacuum
Load.As a example by " dry column-packing, dry method loading ": as select chromatographic column, click on " compression ", then dynamic axial compression column to
Lower movement, translational speed can be by regulation valve regulation, and regulation valve the most then compresses post speed to be increased, the most then speed slows down.With
Time, notice that the registration of oil pressure table, general maximum pressure control at about 5MPa.After compression, click on " stopping " button.
4, wash-out separates: be mainly used in the continuous wash-out that mobile phase ratio determines, as a example by " dry column-packing dry method loading ",
After loading, after fluid reservoir 2 injects suitably flowing mutually, select the automatic method of operation.On the left of selection in a row button " parameter sets
Put ", set the pumpdown time of 10 fluid reservoirs 2 respectively, unit is min, minimum not less than 1min, after having set, such as choosing
Selecting " chromatographic column 1 ", click on failure reset, then bring into operation from No. 0, and click on " equipment startup ", valve combination is according to the time of setting
Automatically switching, flowing continuously flows in chromatographic column mutually, and the efflux after separation evaporates by entering after 3 groups of valves and 10 groups of Vavle switching
Divide collection system, it is achieved be automatically separated.Clicking on " key frame ", " valve work timing " hurdle display device working time, unit is
Second.The valve working time sets the emptying time showing current vacuum-pumping container.When end of run, all controlled valves become red
In look, and receiving flask, blue bands no longer flashes.
Embodiment 1:
Before operation, the entrance gas piping of equipment is connected with air pump device, by vavuum pump and device after having gas to enter
Connect.Switch board guidance panel at automatic vacuum liquid-chromatography apparatus selects automatically to run button, rotates in one's hands by its position
Flowing mode, manual indicator lamp is lighted；Rotating afterwards to automated manner, automatic pilot lamp is lighted.Fore flow is injected in fluid reservoir 2
After dynamic phase (petroleum ether), turn on the power, enter logic control system 1 homepage, regulate 10 groups of valves and the switch of 3 groups of valves successively,
Select the automatic method of operation (knob button SA), click on start button in HMI touch-screen 1b, carry out flushing of pipeline.Rinse complete
After, carrying out the filling of filler and sample under vacuum condition successively: wherein, filler is that (dry method fills 300-400 mesh column chromatography silica gel
Post), sample is 80g fruit of Chinese magnoliavine petroleum ether extraction position (dry method loading)；After filling, click on " look at the control system page
Spectrum hydraulic pressure ", be 150 mm × 280 mm in the first chromatographic column 6(chromatographic column specification) click on " compression " button under item and make dynamic axial
Compression post moves down to be compacted chromatographic column, and oil pressure table maximum pressure controls at about 5MPa；After compression, click on and " stop
Only " button, chromatographic column filling is complete.In fluid reservoir inject configured flowing phase (petroleum ether-ethyl acetate=95:5,90:
10,80:20,60:40,0:100 gradient elution, elutes 1L every time, and each ratio all elutes 2 times, is implanted sequentially fluid reservoir 2；In control
The system page processed selects " automatically run " mode, set in " parameter settings " valve of each fluid reservoir combine switching time as
30min；After completing to set, click under the first chromatographic column 6 " failure reset ", then program brings into operation from the first fluid reservoir 21,
Clicking on " equipment startup " afterwards, separable programming starts automatically to run, and the flowing in fluid reservoir 2 is flowed into the first chromatographic column mutually one by one
In 6, realized continuous print ON-LINE SEPARATION under vacuum conditions by Automated condtrol；Now return homepage, at " valve work meter
Time " place can check the operation time (unit is the second) of equipment；When all controlled valves become red, and blue bar in receiving flask 9
When band no longer flashes, separation process terminates, the only time-consuming 5h of whole separation process.After separation completes, in each fraction collector
Eluting fraction carries out HPLC and analyzes detection, 6 elution fractions (S1 to S6) of final acquisition.
Embodiment 2:
Before operation, the entrance gas piping of equipment is connected with air pump device, by vavuum pump and device after having gas to enter
Connect.Logic control system 1 at device selects automatically to run button, and its position rotates to manual mode, manual indicator lamp
Light；Rotating afterwards to automated manner, automatic pilot lamp is lighted.After injecting initial flowing phase (petroleum ether) in fluid reservoir 2, beat
Switch power supply, metering-in control system homepage, regulate 10 groups of valves and the switch of 3 groups of valves successively, select the automatic method of operation, click on
Start button in HMI touch-screen 1b, carries out flushing of pipeline.After flushing, successively to the second chromatographic column 6a(look under vacuum condition
Spectrum post specification be 100 mm × 280 mm) and the 3rd chromatographic column 6b(chromatographic column specification be 80 mm × 280 mm) carry out filler
Filling, wherein the second chromatographic column 6a filler is 300-400 mesh column chromatography silica gel, and the 3rd chromatographic column 6b filler is that 20/45 μm is spherical
Silica gel, after filling, clicks on " chromatogram hydraulic pressure " at the control system page, in " chromatographic column" click on " compression " button under item and make
Dynamic axial compression column moves down to be compacted chromatographic column, and oil pressure table maximum pressure controls after about 5MPa, compression,
Clicking on " stopping " button, the 3rd chromatographic column 6b compression method and the second chromatographic column 6a are identical；After chromatographic column filling, use wet
Fruit of Chinese magnoliavine S4 component is injected in the second chromatographic column 6a by method loading method, injects the flowing phase (stone configured in fluid reservoir 2 simultaneously
Oil ether-ethyl acetate=100:0,95:5,90:10,85:15,80:20 gradient elution, elutes 0.5L every time, and each ratio all elutes 2
Secondary, it is implanted sequentially fluid reservoir 2)；Select " automatically running " mode at the control system page, " parameter setting " sets each liquid storage
Tank valve combination switching time be 30min；Complete set after, click under the second chromatographic column 6a item " failure reset ", then program from
First fluid reservoir 21 brings into operation, and clicks on " equipment startup " afterwards, and separable programming starts automatically to run, the flowing phase in fluid reservoir
It is flowed into one by one in the second chromatographic column 6a, is realized continuous print ON-LINE SEPARATION by Automated condtrol under vacuum conditions；Now return
Return homepage, the operation time (unit is the second) of equipment can be checked at " valve work timing " place；When all controlled valves become red
When in look, and receiving flask, blue bands no longer flashes, the second chromatographic column 6a separation process terminates, by TLC to each fraction sample
After being analyzed detection, it is thus achieved that 5 elution fractions (S4-1 to S4-5)；Afterwards that S4-3 component is right as further separating
As, in wet method loading to the 3rd chromatographic column 6b, in fluid reservoir 2, inject the flowing phase (petroleum ether-ethyl acetate configured simultaneously
=95:5 isocratic elution, elutes 0.5L, co-elute 10 times every time, is implanted sequentially fluid reservoir 2)；Select " certainly at the control system page
Dynamic operation " mode, the valve setting each fluid reservoir in " parameter setting " combines switching time as 30min；After completing to set, click on
Under 3rd chromatographic column 6b item " failure reset ", then program brings into operation from No. 0 fluid reservoir, clicks on " equipment startup " afterwards, separates
Program starts automatically to run, and the flowing in fluid reservoir is flowed in the 3rd chromatographic column 6b, mutually one by one by Automated condtrol very
Continuous print ON-LINE SEPARATION is realized under dummy status；Now return homepage, when the operation of equipment can be checked by " valve work timing " place
Between (unit is the second)；When all controlled valves become red, and when in receiving flask, blue bands no longer flashes, the 3rd chromatographic column 6b
Separation process terminates；So far the mask work of S4 component is fully completed, altogether time-consuming 10h.Utilize TLC that each cut is analyzed inspection
Surveying, result shows, S4 component obtains 5 TLC single-point compounds altogether through secondary automatic vacuum liquid phase column chromatography for separation, examines through HPLC
The purity of survey wherein 4 compounds is more than 85%, and the purity of Q, component R is more than 90%.
Embodiment 3:
Chromatographic column is loaded: weighs and fills to 150 mm × 280 mm chromatographic columns under 2kg silica gel (300-400 mesh) vacuum state
After, click on " chromatogram hydraulic pressure " at the control system page, under the first chromatographic column 6, click on " compression " button make dynamic axial compression
Post moves down to be compacted chromatographic column, and oil pressure table maximum pressure controls at about 5MPa；After compression, click on " stopping " and press
Button, chromatographic column filling is complete.
Sample preparation and loading: weigh refluxing extraction 3 times in 2L methyl alcohol after 1kg schisandra chinensis medicinal material is pulverized, filter and subtract
Pressure concentrates and obtains medicinal extract 280g, after being disperseed with 1L 10% methanol-water by medicinal extract, adds equal-volume petroleum ether extraction, coextraction three
Secondary, merge organic phase solvent reduced pressure concentration and obtain fruit of Chinese magnoliavine petroleum ether extract 84g；By extract in 500ml ethyl acetate
Dispersing and dissolving, adds 160g silica gel (300-400 mesh) dry method and mixes sample, after solvent evaporated, open chromatographic column, put uniformly by sample
In chromatographic column bed top, click on " chromatogram hydraulic pressure " at the control system page, under the first chromatographic column 6, click on " compression " button pressure
Real chromatographic column, completes loading process.
Gradient elution and eluent are collected: preparation petroleum ether-ethyl acetate flowing phase solution, and solvent ratios is followed successively by 90:
10,80:20 ... 20:80,10:90,0:100,1L all prepared by each gradient solvent, is added sequentially in fluid reservoir 2；Connect electricity
Source, button selectes the first chromatographic column 6, starts chromatogram arrangement, and separation process runs the most automatically；After elution process completes, device
It is automatically stopped.
Eluent detection and concentration: the eluting fraction in each eluent collector is carried out HPLC and analyzes detection, experiment knot
Fruit as shown in Figure 2, finally obtains 6 components.
Embodiment 4:
Chromatographic column is loaded: weighs and fills to 80 mm × 280 mm chromatographic columns under 700g silica gel (300-400 mesh) vacuum state
After, click on " chromatogram hydraulic pressure " at the control system page, under " the 3rd chromatographic column " item, click on " compression " button make dynamic axial pressure
Contracting post moves down to be compacted chromatographic column, and oil pressure table maximum pressure controls at about 5MPa；After compression, click on " stopping "
Button, chromatographic column filling is complete.
Sample preparation and loading: in Example 1, S5 component 5g is molten in 100ml petroleum ether-ethyl acetate=85:15 mixing
In liquid after ultrasonic dissolution, it is directly injected in sample cell.
Gradient elution and eluent are collected: preparation petroleum ether-ethyl acetate flowing phase solution, and solvent ratios is followed successively by 85:
15,81:19,73:27,65:35 and 55:45, each gradient solvent is all prepared 1L, is added 0.5L, be sequentially added in each container
In fluid reservoir 2；Switching on power, button selectes the 3rd chromatographic column 6b, starts chromatogram arrangement, and separation process runs the most automatically；When
After elution process completes, device is automatically stopped.
Eluent detection and concentration: the eluting fraction in each eluent collector 9 is carried out TLC and HPLC and analyzes detection,
Result shows can obtain 5 TLC single-point compound (as shown in Figure 3), warp through an automatic vacuum liquid phase column chromatography for separation
The purity of the HPLC each compound of detection is all more than 85%, and the purity of Q, component R is more than 90%(as shown in Figure 4).
Claims (5)
1. an automatic vacuum Split liquid chromatographic apparatus, including liquid-way system and logic control system (1), logic control system
(1) on panel, it is provided with signal lamp (1a), touch-screen (1b) and button (1c), in casing, is provided with PLC logic controller (1d)
With magnetic valve (1e), PLC logic controller (1d) comprises central processing unit (CPU), button information module (A), positional information mould
Block (B), break-make modular converter (C), oil cylinder control module (D) and signal control module (E)；It is characterized in that: described fluid path system
System uses fluid reservoir (2) to be connected to enter 1 through fluid reservoir arm (2a) to go out pneumatic control valve (3) more, then through 1 enter 3 go out pneumatic
Control valve (4) is connected the first chromatographic column (6), the second chromatographic column (6a), the 3rd chromatographic column respectively by arm in chromatographic column (6c)
(6b), then by chromatographic column lower branch line (6d) sequentially pass through 3 enter 1 go out pneumatic control valve (7), 1 enter to have more pneumatic control valve (8), receive
Storage arm (9a) is connected to collector (9)；In described first chromatographic column (6), the second chromatographic column (6a), the 3rd chromatographic column (6b)
Being provided with oil cylinder (5), described collector (9) is connected with vavuum pump (10)；The magnetic valve (1e) that described logic control system (1) controls
By gas piping (11) with enter 1 to go out pneumatic control valve (3) more, 1 enter 3 to go out pneumatic control valve (4), 3 enter 1 and go out pneumatic control valve
(7), 1 enters to have more and is attached between pneumatic control valve (8) and oil cylinder (5)；Described central processing unit (CPU) execution program and depositing
Storage data, are controlled automatic control process, and importation gathers signal from switch, and output part is then controlled by oil cylinder
Module (D) controls the direction of motion of oil cylinder (5), and central processing unit (CPU) is controlled by break-make modular converter (C) and magnetic valve (1e)
More system is entered 1 to go out pneumatic control valve (3), 1 is entered 3 to go out pneumatic control valve (4), 3 enters 1 to go out pneumatic control valve (7), 1 enters to have more pneumatic control
The break-make conversion of valve processed (8).
The method of work of a kind of automatic vacuum Split liquid chromatographic apparatus the most according to claim 1, it is characterised in that bag
Include following steps:
(1), before operation, logic control system (1) is carried out routine inspection, checks that equipment state is the most normal；Enter back into logic control
The homepage of system processed (1), carries out experiment parameter setting；
(2) flushing line: after injecting initial flowing mutually in the fluid reservoir (2), successively regulation enter more 1 go out pneumatic control valve (3), 1
Enter 3 to go out pneumatic control valve (4), 3 enter 1 and go out pneumatic control valve (7) and 1 and enter to have more pneumatic control valve (8), to pipeline and the first chromatogram
Post (6), the second chromatographic column (6a), the 3rd chromatographic column (6b) are rinsed；
(3) filling chromatographic column: need to select preparative chromatography column dimension according to experiment, carry out the filling of filler under vacuum,
After filler filling, " chromatogram hydraulic pressure " button clicked in button (1c) makes dynamic axial compression column move down compacting chromatogram
Post；After compression, clicking on " stopping " button in button (1c), chromatographic column filling is complete；
(4) sample dress post: sample to be separated is added chromatographic column, in the inner flowing phase configured of injecting of fluid reservoir (2), button choosing
Determine chromatographic column, sample is joined in selected chromatographic column, then enter 3 by 1 and go out pneumatic control valve (4) and enter 1 go out pneumatic control valve with 3
(7) switching, enters chromatographic column used with 13 to go out pneumatic control valve (4), 3 enters 1 and go out pneumatic control valve (7) and connect；
(5) chromatographic column sample elution: go out pneumatic control valve (3) and 1 switching of entering have more pneumatic control valve (8) by entering 1 more, connect
Logical first fluid reservoir (21), chromatographic column, the first collector (91), carry out vacuum elution, until flowing is drained in chromatographic column,
Without eluent in pipeline；Reduce without eluent or vacuum in pipeline and trigger logic control system (1), logic control system (1)
Switching is entered 1 more and is gone out pneumatic control valve (3) and 1 and enter to have more pneumatic control valve (8), closes the first fluid reservoir (21) to the first collector
(91) path；Go out pneumatic control valve (3) and 1 switching of entering have more pneumatic control valve (8) by entering 1 more, connect the second liquid storage
Tank (22), to the path of the second collector (92), repeats said process, until completing whole wash-out, logic control system (1) is sent out
Go out signal；
(6), after having separated, each eluting fraction reclaims in corresponding collector (9).
A kind of automatic vacuum Split liquid chromatographic apparatus the most according to claim 1, it is characterised in that: described first chromatogram
The specification of post (6) is 150mm × 280mm, and the specification of the second chromatographic column (6a) is 100mm × 280mm, the 3rd chromatographic column (6b)
Specification is 80mm × 280mm.
A kind of automatic vacuum Split liquid chromatographic apparatus the most according to claim 1, it is characterised in that: described first chromatogram
Post (6), the second chromatographic column (6a) are silica gel or ion exchange resin with the filler of the 3rd chromatographic column (6b).
A kind of automatic vacuum Split liquid chromatographic apparatus the most according to claim 1, it is characterised in that: described first chromatogram
Bonded silica gel elected as by the filler of post (6), the second chromatographic column (6a) and the 3rd chromatographic column (6b).
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| CN110196302A (en) * | 2018-02-27 | 2019-09-03 | 中国石油化工股份有限公司 | Automatic flushing device used in a kind of column chromatography adsorbing separation |
| CN113915177B (en) * | 2021-09-16 | 2024-05-14 | 利穗科技(苏州)有限公司 | Electrohydraulic servo driving device and chromatographic equipment |
| CN115055168B (en) * | 2022-06-09 | 2023-05-26 | 珠海高新区维得力生物工程有限公司 | Preparation process applied to fructo-oligosaccharide chromatographic separation filler and artificial intelligent separation device |
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