CN104730175A - Automatic vacuum liquid-chromatography separator and control method thereof - Google Patents
Automatic vacuum liquid-chromatography separator and control method thereof Download PDFInfo
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Abstract
The invention belongs to the technical field of chromatography devices and relates to an automatic vacuum liquid-chromatography separator and a control method thereof. A logical control system and a liquid storage and fraction collection system are organically combined, on-off conversion of valve sets, such as multiple-in one-out pneumatic control valve and a one-in three-out pneumatic control valve in the liquid storage and fraction collection system is achieved by controlling the moving direction of a cylinder; the automatic control system for separation of natural products is formed; gradient elution of a moving phase and automatic collection of fractions is achieved through a pre-set elution program; intelligent control of the automatic vacuum liquid-chromatography separator for separation and purification of natural medicines is achieved. The automatic vacuum liquid-chromatography separator uses a combination of chromatographic columns different in specification and solid media, has the advantages that separation efficiency is high, labor intensity is low, labor and time cost is saved and automatic control and continuous production are allowed, and is suitable for quick, efficient and intelligent separation of the complex samples, such as natural medicines.
Description
Technical field
The present invention relates to a kind of automatic vacuum Split liquid chromatographic apparatus and control method thereof, belong to the technical field of chromatogram arrangement.
Background technology
Although traditional chromatogram arrangement can reach the object of separation and purification, but often there is the defects such as disengaging time is long, consumption quantity of solvent is large, cost is higher or separation efficiency is low, product purity is inadequate, and effective constituent often incurs loss in tediously long, various pillar layer separation, solvent concentration and sample jump operation or structure changes, researchist is made to have to strengthen inventory in the hope of obtaining desirable result.Therefore, efficient, high-throughout separation preparation facilities is the point of contact, pass of research field always.
Vacuum liquid chromatography (Vacuum Liquid Chromatography, VLC) to be suggested as a kind of novel, easy, effective isolation technics in mid-term in 20th century and to be applied to the purification and separation of natural organic-compound and volume organic reaction product
[Aust.J.chem.1977,30,1859], its essence is the integrated application of column chromatography, Preparative TLC chromatogram and vacuum filtration technology
[chromatogram .1990,8(5) 313].VLC is different from traditional Column chromatography techniques, because the latter carries out continuously when wash-out, can not be interrupted in operation, and VLC is when carrying out solvent elution, after eluant, eluent is all extracted out under reduced pressure after post, change solvent again, proceed the collection of cut next time, its principle is similar to the repeatedly expansion of thin-layer chromatography.Compared with traditional column chromatography, VLC has the features such as device is simple, amount of filler is few, the running time is short, organic solvent consumption is little, quantity of sample handling is large, and has been successfully applied to the preparative separation of diversified compound
[J.Natl.prod.1986,49,892; J.Natl.prod.1986,49,934; Mountain Western Medicine S University journal .1999,30,41].
But also there is many weak points as a kind of inexpensive efficient, simple and efficient isolation technics in VLC: chromatographic column filler filling does not closely cause post bed less stable, theoretical cam curve is low, chromatogram column efficiency is uncontrollable; Particularly feed flow, vacuum elution, the step such as to drain, and gradient cyclic process need manually be carried out, thus consumes a large amount of time and human cost.Therefore, how to reduce manual operation to greatest extent, realize automatic partition from, to raise the efficiency, to break through the bottleneck that classic method is formed, seem extremely urgent.
Summary of the invention
In order to solve the problems such as the unstable and manpower consumption of the chromatogram column efficiency that exists in traditional VLC technology is high, the invention provides a kind of automatic vacuum Split liquid chromatographic apparatus, with efficiently, serialization can prepare compound.
Technical scheme of the present invention is: a kind of automatic vacuum Split liquid chromatographic apparatus, comprise liquid-way system and logic control system, logic control system is provided with signal lamp, touch-screen and button on panel, in casing, be provided with PLC logic controller and solenoid valve, PLC logic controller comprises 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 adopts fluid reservoir to be connected to enter 1 more and go out pneumatic control valve through fluid reservoir arm, enter 3 through 1 again to go out pneumatic control valve and connect the first chromatographic column, the second chromatographic column, the 3rd chromatographic column respectively by arm in chromatographic column, then enter 1 through 3 successively by chromatographic column lower branch line and go out that pneumatic control valve, 1 enters to have more pneumatic control valve, gatherer arm is connected to gatherer; Described first chromatographic column, the second chromatographic column, the 3rd chromatographic column are provided with oil cylinder, and described gatherer is connected with vacuum pump; The solenoid valve that described logic control system controls by gas piping with enter 1 more and go out pneumatic control valve, 1 and enter 3 and go out pneumatic control valve, 3 and enter 1 and go out pneumatic control valve, 1 and enter to have more between pneumatic control valve and oil cylinder and be connected; Described central processing unit executive routine and storage data, automatic control process is controlled, importation is collection signal from switch, output then controls the direction of motion of oil cylinder by oil cylinder control module, and central processing unit enters 1 by break-make modular converter and solenoid control more and goes out pneumatic control valve, 1 and enter 3 and go out pneumatic control valve, 3 and enter 1 and go out the break-make conversion that pneumatic control valve, 1 enters to have more pneumatic control valve.
A kind of method of work of automatic vacuum Split liquid chromatographic apparatus comprises the following steps:
(1), before operation, routine inspection is carried out to logic control system, checks that whether equipment state is normal; First select automatically to run button at switch board guidance panel, its position is rotated to manual mode, and manual pilot lamp is lighted; Rotate afterwards to automated manner, automatic pilot lamp is lighted; Opening power, enters the homepage of logic control system, carries out experiment parameter setting;
(2) flushing line: inject initial mobile phase in fluid reservoir after, regulate successively and enter 1 more and go out pneumatic control valve, 1 and enter 3 and go out pneumatic control valve, 3 and enter 1 and go out pneumatic control valve and 1 and enter to have more pneumatic control valve, pipeline and the first chromatographic column, the second chromatographic column, the 3rd chromatographic column are rinsed;
(3) load chromatographic column: experimentally need to select preparative chromatography column dimension, carry out the filling of filler under vacuum, after filler filling, " chromatogram hydraulic pressure " button in button click makes dynamic axial compression column move down compacting chromatographic column; After compression, " stopping " button in button click, chromatographic column filling is complete;
(4) sample dress post: sample to be separated is added chromatographic column, the mobile phase configured is injected in fluid reservoir, button selectes chromatographic column, sample is joined in selected chromatographic column, enter 3 by 1 again to go out pneumatic control valve and 3 and enter the switching that 1 goes out pneumatic control valve, chromatographic column used and 1 is entered 3 and go out pneumatic control valve, 3 and enter 1 and go out pneumatic control valve and be communicated with;
(5) chromatographic column sample elution: going out by entering 1 switching that pneumatic control valve and 1 enters to have more pneumatic control valve more, connecting the first fluid reservoir, chromatographic column, the first gatherer, carrying out vacuum elution, until mobile phase is drained in chromatographic column, without eluent in pipeline; Reduce Trigger Logic control system without eluent or vacuum tightness in pipeline, logic control system switching is entered 1 more and is gone out pneumatic control valve and 1 and enter to have more pneumatic control valve, closes the path of the first fluid reservoir to the first gatherer; Going out by entering 1 the switching that pneumatic control valve and 1 enters to have more pneumatic control valve more, connecting the path of the second fluid reservoir to the second gatherer, repeat said process, until complete whole wash-out, logic control system sends settling signal;
(6), after being separated, each eluting fraction reclaims in the gatherer of correspondence.
The specification of described first chromatographic column, the second chromatographic column, the 3rd chromatographic column is 150mm × 280mm, 100mm × 280mm, 80mm × 280mm.
The filler of described first chromatographic column, the second chromatographic column and the 3rd chromatographic column is 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 the integral valve that pneumatic control valve is rotating switching type or the combination valve be made up of multiple stop valve, switching power is electric power or pressurized air more.Switching command is time command, pressure instruction or light instruction; Time command is the process time that provides according to preliminary experiment and artificially sets, and pressure instruction comes from pressure transducer, and light instruction comes from optical sensor.The chromatographic column of this device is dynamic axial compression chromatographic column.Dynamic axial compression chromatographic column can ensure stability and the Gao Zhuxiao of chromatographic column bed.
In this device, automated intelligent mode is mechanical type, logic circuit type or electronic type.Logic control system is primarily of small-sized programmable sequence PLC logic controller (MicroPLC) (i.e. siemens PLC) and eight cun of man-machine interface (HMI) compositions.In logic control system, Siemens S7-200PLC logic controller comprises a central processor CPU and 5 expansion modules (button information 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 executive routine and stores data, to control the automatic control task of native system or process; Input and output are reference mark of system, and importation is collection signal from switch, and output is then by drive magnetic by-pass valve control switch and cylinder movement direction; 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 control valve valve group break-make change; Button information modules A gathers button information, positional information module B gathers the positional information of valve combination, break-make modular converter C is responsible for the switching of valve, oil cylinder control module D controls dynamic axial compression chromatographic column, signal control module E control signal lamp, as hummer during system malfunctions will send sound and light alarm.
The vacuum liquid-phase chromatographic column of this tripping device adopts dynamic axial compression chromatography piece-rate system, it is by pumping unit, integrated package or valve combination, fuel tank, electrical box and chromatographic column combine, rotated by driven by motor oil pump, pump is bought oil after oil suction from oil pump, be the pressure energy of hydraulic oil by changes mechanical energy, hydraulic oil realizes direction by integrated package (or valve combination), pressure, in the oil cylinder being transferred to hydraulic machinery through external pipeline after flow regulation or oil motor, thus control the conversion in hydraulic motor direction, the size of strength and the speed of speed, promote the acting of various hydraulic machinery.It loads under the driving of gas drive hydraulic power, and by carrying out compression to a certain degree to chromatographic column bed fillers, ensure continuous, all even compactedness of filler, chromatographic column is by the function realizing filling post, maintenance post pressure and unloading post that moves up and down of piston simultaneously, piston periphery is equipped with the O-ring seal of particular design, while permission piston freely up and down slides, can keep higher sealing pressing again.It is compared with traditional chromatographic column, have employed the chromatographic column type of feed of dynamic axial compression, by carrying out compression to a certain degree to chromatographic column bed fillers under the driving of gas drive hydraulic power, ensure that the Gao Zhuxiao of dynamic axial compression chromatographic column and superior separating property; In addition, the system disposition chromatographic column of three kinds of different sizes, can form multicolumn separation system, can carry out lock out operation during use according to material properties and quality choice appropriate gauge; Separating filler of different nature (comprising silica gel, C8, C18, bonded silica gel, ion exchange resin etc.) can be loaded in chromatographic column, be separated the experiment demand of object with satisfied difference.
Beneficial effect of the present invention: logic control system and liquid storage and fraction collection system organically combine by this device, complete to go out pneumatic control valve and 1 and enter to entering 1 in liquid storage and fraction collection system more the break-makes conversion that 3 go out the combination of the valve such as pneumatic control valve by control cylinder direction of motion, form the automation control system of Separation of Natural Products, by the elution program preset, the gradient elution and the cut that achieve mobile phase are collected automatically, realize vacuum liquid-chromatography apparatus and manipulate the intellectuality of natural drug separation and purification.This tripping device adopts the chromatographic column combination of different size and solid-phase media, have that separation efficiency is high, labour intensity is low, the feature of the cost that saves manpower and time, Automated condtrol continuous prodution, can realize being separated the quick, effective, intelligent of the complex samples such as natural drug.
Accompanying drawing explanation
Fig. 1 is a kind of structural representation of automatic vacuum liquid-chromatography apparatus.
Fig. 2 is the HPLC figure that a kind of automatic vacuum liquid-chromatography apparatus is separated fruit of Chinese magnoliavine petroleum ether extract.
Fig. 3 is the TLC figure that a kind of automatic vacuum liquid-chromatography apparatus is separated fruit of Chinese magnoliavine S5 component.
Fig. 4 is the HPLC figure that a kind of automatic vacuum liquid-chromatography apparatus is separated 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, solenoid 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, 5, oil cylinder, 6, first chromatographic column, 6a, second chromatographic column, 6b, 3rd chromatographic column, 6c, arm in chromatographic column, 6d, chromatographic column lower branch line, 7, 3 enter 1 goes out pneumatic control valve, 8, 1 enters to have more pneumatic control valve, 9, gatherer, 9a, gatherer arm, 10, vacuum pump, 11, gas piping, 21, first fluid reservoir, 22, second fluid reservoir, 91, first gatherer, 92, second gatherer, 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.
Embodiment
Be described the preferred embodiments of the present invention below in conjunction with accompanying drawing, the preferred embodiment that this place is set forth, only for instruction and explanation of the present invention, is not intended to limit the present invention.
Fig. 1 shows the structure of this device, mainly comprises gradient mobile phase liquid-storage system, dynamic axial compression chromatographic column system, fraction collection system and logic control system four part.Its stream is: 10 fluid reservoir 2(can hold the different eluent of 10 kinds of gradients respectively) outlet respectively with enter 1 10 entrances going out pneumatic control valve 3 more 1 and be connected; Enter 1 outlet going out pneumatic control valve 3 to enter 3 and go out pneumatic control valve 4 entrance with 11 and be connected, 1 enters the chromatographic column (as 150mm × 280mm, 100mm × 280mm, 80mm × 280mm) that 3 outlets going out pneumatic control valve 4 do not wait with three diameters respectively connects more; The outlet of chromatographic column is entered 13 entrances going out pneumatic control valve 7 respectively and is connected with 13,3 enter 1 outlet going out pneumatic control valve 7 is connected with 1 entrance entering to have more pneumatic control valve 8, and 1 outlet entering to have more pneumatic control valve 8 is connected with the gatherer 9 of 10 kinds of eluents respectively; The gatherer 9 of eluent 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 more and be connected with vacuum pump 10; The stream of gatherer 9 connecting chromatographic column and eluent installs 10 pressure transducers or optical sensor (as ultraviolet or infrared detection pond).Go out the automatic switchover that pneumatic control valve 3 and 1 enters to have more pneumatic control valve 8 realize automated procedure by entering 1 more.
A kind of course of work 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, ensures gas input, and is connected with device by vacuum pump.Opening power, metering-in control system key frame, red round dot is that switch cuts out, and green round dot is switch opens; Red triangle is for stopping, and green triangle is work;
2. flushing line: can select to run manually or automatically (manually: when only supplying equipment debugging, equipment maintenance, have corresponding human users; Automatic: to be operated by laboratory technician or process engineer when equipment is normal).Main below the automatic method of operation to be illustrated.All enter the time of gatherer according to each mobile phase, each container vacuum-pumping time is set in " setting parameter ", click setting parameter and confirm, select corresponding chromatographic column, switch board guidance panel is selected automatically to run button, and rotated to automated manner, automatic pilot lamp is lighted.
3, post loading is filled: after quality selectes suitable chromatographic column model and separating filler per sample, load under vacuum.Be example with " dry column-packing, dry method loading ": as selected chromatographic column, click " compression ", then dynamic axial compression column moves down, and translational speed regulates by variable valve, variable valve then compresses the increase of post speed to the right, and then speed slows down left.Meanwhile, note the registration of oil pressure table, general maximum pressure controls at about 5MPa.After compression, click " stopping " button.
4, wash-out is separated: be mainly used in the continuous wash-out that mobile phase ratio is determined, for " dry column-packing dry method loading ", after loading, after fluid reservoir 2 injects suitable mobile phase, select the automatic method of operation.Select " optimum configurations " in the one row button of left side, set the pumpdown time of 10 fluid reservoirs 2 respectively, unit is min, minimumly be not less than 1min, after having set, as selected " chromatographic column 1 ", click failure reset, then bring into operation from No. 0, and click " device start ", valve combination is automatically switched according to setting-up time, and mobile phase continuously flows in chromatographic column, efflux after separation, by entering fraction collection system after 3 groups of valves and 10 groups of Vavle switching, realizes automatically being separated.Click " key frame ", " valve work timing " hurdle display device working time, unit is second.The emptying time of the current vacuum-pumping container of valve working time set and display.When end of run, all controlled valves become redness, and in receiving flask, blue bands no longer flashes.
Embodiment 1:
Before operation, the gas piping that enters of equipment is connected with air pump device, after having gas to enter, vacuum pump is connected with device.Select automatically to run button at the switch board guidance panel of automatic vacuum liquid-chromatography apparatus, its position is rotated to manual mode, and manual pilot lamp is lighted; Rotate afterwards to automated manner, automatic pilot lamp is lighted.Inject initial mobile phase (sherwood oil) in fluid reservoir 2 after, opening power, enters logic control system 1 homepage, regulate the switch of 10 groups of valves and 3 groups of valves successively, select the automatic method of operation (knob button SA), click start button in HMI touch-screen 1b, carry out flushing of pipeline.After flushing, carry out the filling of filler and sample successively: wherein under vacuum condition, filler is 300-400 order column chromatography silica gel (dry column-packing), and sample is 80g fruit of Chinese magnoliavine petroleum ether extraction position (dry method loading); After filling, " chromatogram hydraulic pressure " is clicked at the control system page, be 150 mm × 280 mm in the first chromatographic column 6(chromatographic column specification) to click " compression " button under item and make dynamic axial compression column move down with compacting chromatographic column, oil pressure table maximum pressure controls at about 5MPa; After compression, click " stopping " button, chromatographic column filling is complete.In fluid reservoir, injecting the mobile phase configured, (the equal wash-out of each ratio 2 times, injects fluid reservoir 2 successively for petroleum ether-ethyl acetate=95:5,90:10,80:20,60:40,0:100 gradient elution, each wash-out 1L; Select " automatically running " mode at the control system page, in " optimum configurations ", the valve of each fluid reservoir of setting combines switching time is 30min; After completing setting, click " failure reset " under the first chromatographic column 6, then program brings into operation from the first fluid reservoir 21, click " device start " afterwards, separable programming starts automatic operation, mobile phase in fluid reservoir 2 flow in the first chromatographic column 6 one by one, realizes continuous print ON-LINE SEPARATION under vacuum conditions by Automated condtrol; Now return homepage, the working time (unit is second) of equipment can be checked at " valve work timing " place; When all controlled valves become redness, and when in receiving flask 9, blue bands no longer flashes, detachment process terminates, and whole detachment process is 5h consuming time only.After separation completes, HPLC is carried out to the eluting fraction in each fraction collector and analyzes detection, final acquisition 6 elution fractions (S1 to S6).
Embodiment 2:
Before operation, the gas piping that enters of equipment is connected with air pump device, after having gas to enter, vacuum pump is connected with device.Select automatically to run button at the logic control system 1 of device, its position is rotated to manual mode, manual pilot lamp is lighted; Rotate afterwards to automated manner, automatic pilot lamp is lighted.Inject initial mobile phase (sherwood oil) in fluid reservoir 2 after, opening power, metering-in control system homepage, regulates the switch of 10 groups of valves and 3 groups of valves successively, selects the automatic method of operation, clicks start button in HMI touch-screen 1b, carries out flushing of pipeline.After flushing, be 100 mm × 280 mm to the second chromatographic column 6a(chromatographic column specification successively under vacuum condition) and the 3rd chromatographic column 6b(chromatographic column specification be 80 mm × 280 mm) carry out the filling of filler, wherein the second chromatographic column 6a filler is 300-400 order column chromatography silica gel, 3rd chromatographic column 6b filler is 20/45 μm of spherical silica gel, after filling, " chromatogram hydraulic pressure " is clicked, in " chromatographic column at the control system page
" click " compression " button under item and make dynamic axial compression column move down with compacting chromatographic column, oil pressure table maximum pressure controls at about 5MPa, after compression, and click " stopping " button, the 3rd chromatographic column 6b compression method is identical with the second chromatographic column 6a; After chromatographic column filling, wet method loading method is adopted to inject in the second chromatographic column 6a by fruit of Chinese magnoliavine S4 component, in fluid reservoir 2, inject mobile phase (petroleum ether-ethyl acetate=100:0,95:5,90:10,85:15,80:20 gradient elution configured simultaneously, each wash-out 0.5L, the equal wash-out of each ratio 2 times, injects fluid reservoir 2 successively); Select " automatically running " mode at the control system page, in " optimum configurations ", the valve of each fluid reservoir of setting combines switching time is 30min; After completing setting, click " failure reset " under the second chromatographic column 6a item, then program brings into operation from the first fluid reservoir 21, click " device start " afterwards, separable programming starts automatic operation, mobile phase in fluid reservoir flow in the second chromatographic column 6a one by one, realizes continuous print ON-LINE SEPARATION under vacuum conditions by Automated condtrol; Now return homepage, the working time (unit is second) of equipment can be checked at " valve work timing " place; When all controlled valves become redness, and when in receiving flask, blue bands no longer flashes, the second chromatographic column 6a detachment process terminates, and after carrying out analysis detection, obtains 5 elution fractions (S4-1 to S4-5) by TLC to each fraction sample; Afterwards using S4-3 component as further separate object, wet method loading, in the 3rd chromatographic column 6b, injects the mobile phase (petroleum ether-ethyl acetate=95:5 isocratic elution, each wash-out 0.5L that configure simultaneously in fluid reservoir 2, co-elute 10 times, injects fluid reservoir 2 successively); Select " automatically running " mode at the control system page, in " optimum configurations ", the valve of each fluid reservoir of setting combines switching time is 30min; After completing setting, click " failure reset " under the 3rd chromatographic column 6b item, then program brings into operation from No. 0 fluid reservoir, click " device start " afterwards, separable programming starts automatic operation, mobile phase in fluid reservoir flow in the 3rd chromatographic column 6b one by one, realizes continuous print ON-LINE SEPARATION under vacuum conditions by Automated condtrol; Now return homepage, the working time (unit is second) of equipment can be checked at " valve work timing " place; When all controlled valves become redness, and when in receiving flask, blue bands no longer flashes, the 3rd chromatographic column 6b detachment process terminates; So far the mask work of S4 component all completes, altogether 10h consuming time.Utilize TLC to carry out analysis to each cut to detect, result shows, and S4 component obtains 5 TLC single-point compounds altogether through secondary automatic vacuum liquid phase column chromatography for separation, detect the purity of wherein 4 compounds more than 85%, and the purity of Q, component R is greater than 90% through HPLC.
Embodiment 3:
Chromatographic column is loaded: after being filled to 150 mm × 280 mm chromatographic columns under taking 2kg silica gel (300-400 order) vacuum state, " chromatogram hydraulic pressure " is clicked at the control system page, under the first chromatographic column 6, click " compression " button makes dynamic axial compression column move down with compacting chromatographic column, and oil pressure table maximum pressure controls at about 5MPa; After compression, click " stopping " button, chromatographic column filling is complete.
Sample preparation and loading: take 1kg schisandra chinensis medicinal material pulverize after in 2L methyl alcohol refluxing extraction 3 times, filter and reduced pressure concentration acquisition medicinal extract 280g, by medicinal extract with after the dispersion of 1L 10% methanol-water, add equal-volume petroleum ether extraction, coextraction three times, merges organic phase solvent reduced pressure concentration and obtains fruit of Chinese magnoliavine petroleum ether extract 84g; By extract dispersing and dissolving in 500ml ethyl acetate, add 160g silica gel (300-400 order) dry method and mix sample, after solvent evaporated, open chromatographic column, sample is placed in uniformly chromatographic column bed top, click " chromatogram hydraulic pressure " at the control system page, under the first chromatographic column 6, click " compression " button compacting chromatographic column, complete loading process.
Gradient elution and eluent are collected: preparation petroleum ether-ethyl acetate mobile phase solution, and solvent ratios is followed successively by 90:10,80:20 ... 20:80,10:90,0:100, each gradient solvent all prepares 1L, joins in fluid reservoir 2 successively; Switch on power, button selectes the first chromatographic column 6, and start chromatogram arrangement, detachment process just runs automatically; After elution process completes, device stops automatically.
Eluent detects and concentrates: carry out HPLC to the eluting fraction in each eluent gatherer and analyze detection, and experimental result as shown in Figure 2, finally obtains 6 components.
Embodiment 4:
Chromatographic column is loaded: after being filled to 80 mm × 280 mm chromatographic columns under taking 700g silica gel (300-400 order) vacuum state, " chromatogram hydraulic pressure " is clicked at the control system page, under " the 3rd chromatographic column " item, click " compression " button makes dynamic axial compression column move down with compacting chromatographic column, and oil pressure table maximum pressure controls at about 5MPa; After compression, click " stopping " button, chromatographic column filling is complete.
Sample preparation and loading: in Example 1, S5 component 5g after ultrasonic dissolution, directly injects sample cell in 100ml petroleum ether-ethyl acetate=85:15 mixed solution.
Gradient elution and eluent are collected: preparation petroleum ether-ethyl acetate mobile phase solution, solvent ratios is followed successively by 85:15,81:19,73:27,65:35 and 55:45, each gradient solvent all prepares 1L, adds 0.5L in each container, joins in fluid reservoir 2 successively; Switch on power, button selectes the 3rd chromatographic column 6b, and start chromatogram arrangement, detachment process just runs automatically; After elution process completes, device stops automatically.
Eluent detects and concentrates: carry out TLC and HPLC to the eluting fraction in each eluent gatherer 9 and analyze detection, result display can obtain 5 TLC single-point compounds (as shown in Figure 3) through an automatic vacuum liquid phase column chromatography for separation, detect the purity of each compound all more than 85% through HPLC, and the purity of Q, component R is greater than 90%(as shown in Figure 4).
Claims (4)
1. an automatic vacuum Split liquid chromatographic apparatus, comprise liquid-way system and logic control system (1), logic control system (1) is provided with signal lamp (1a), touch-screen (1b) and button (1c) on panel, in casing, be provided with PLC logic controller (1d) and solenoid valve (1e), PLC logic controller (1d) comprises central processing unit (CPU), button information module (A), positional information module (B), break-make modular converter (C), oil cylinder control module (D) and signal control module (E); It is characterized in that: described liquid-way system adopts fluid reservoir (2) to be connected to enter 1 more and go out pneumatic control valve (3) through fluid reservoir arm (2a), enter 3 through 1 again to go out pneumatic control valve (4) and connect the first chromatographic column (6), the second chromatographic column (6a), the 3rd chromatographic column (6b) respectively by arm in chromatographic column (6c), then enter 1 through 3 successively by chromatographic column lower branch line (6d) and go out that pneumatic control valve (7), 1 enters to have more pneumatic control valve (8), gatherer arm (9a) is connected to gatherer (9); Described first chromatographic column (6), the second chromatographic column (6a), the 3rd chromatographic column (6b) are provided with oil cylinder (5), and described gatherer (9) is connected with vacuum pump (10); The solenoid valve (1e) that described logic control system (1) controls by gas piping (11) with enter 1 more and go out pneumatic control valve (3), 1 and enter 3 and go out pneumatic control valve (4), 3 and enter 1 and go out pneumatic control valve (7), 1 and enter to have more between pneumatic control valve (8) and oil cylinder (5) and be connected; Described central processing unit (CPU) executive routine and storage data, automatic control process is controlled, importation is collection signal from switch, output then controls the direction of motion of oil cylinder (5) by oil cylinder control module (D), and central processing unit (CPU) controls enter 1 by break-make modular converter (C) and solenoid valve (1e) more and goes out pneumatic control valve (3), 1 and enter 3 and go out pneumatic control valve (4), 3 and enter 1 and go out the break-make that pneumatic control valve (7), 1 enters to have more pneumatic control valve (8) and change.
2. the method for work of a kind of automatic vacuum Split liquid chromatographic apparatus according to claim 1, is characterized in that, comprise the following steps:
(1), before operation, routine inspection is carried out to logic control system (1), checks that whether equipment state is normal; Enter the homepage of logic control system (1) again, carry out experiment parameter setting;
(2) flushing line: inject initial mobile phase in fluid reservoir (2) after, regulate successively and enter 1 more and go out pneumatic control valve (3), 1 and enter 3 and go out pneumatic control valve (4), 3 and enter 1 and go out pneumatic control valve (7) and 1 and enter to have more pneumatic control valve (8), pipeline and the first chromatographic column (6), the second chromatographic column (6a), the 3rd chromatographic column (6b) are rinsed;
(3) chromatographic column is loaded: experimentally need to select preparative chromatography column dimension, carry out the filling of filler under vacuum, after filler filling, " chromatogram hydraulic pressure " button in button click (1c) makes dynamic axial compression column move down compacting chromatographic column; After compression, " stopping " button in button click (1c), chromatographic column filling is complete;
(4) sample dress post: sample to be separated is added chromatographic column, inject the mobile phase configured fluid reservoir (2) is inner, button selectes chromatographic column, sample is joined in selected chromatographic column, enter 3 by 1 again to go out pneumatic control valve (4) and 3 and enter the switching that 1 goes out pneumatic control valve (7), chromatographic column used and 1 is entered 3 and go out pneumatic control valve (4), 3 and enter 1 and go out pneumatic control valve (7) and be communicated with;
(5) chromatographic column sample elution: go out the switching that pneumatic control valve (3) and 1 enters to have more pneumatic control valve (8) by entering 1 more, connect the first fluid reservoir (21), chromatographic column, the first gatherer (91), carry out vacuum elution, until mobile phase is drained in chromatographic column, without eluent in pipeline; Trigger Logic control system (1) is reduced without eluent or vacuum tightness in pipeline, logic control system (1) switches to enter 1 more and go out pneumatic control valve (3) and 1 and enter to have more pneumatic control valve (8), closes the path of the first fluid reservoir (21) to the first gatherer (91); Go out by entering 1 the switching that pneumatic control valve (3) and 1 enters to have more pneumatic control valve (8) more, connect the path of the second fluid reservoir (22) to the second gatherer (92), repeat said process, until complete whole wash-out, logic control system (1) sends settling signal;
(6), after being separated, each eluting fraction reclaims in the gatherer (9) of correspondence.
3. a kind of automatic vacuum Split liquid chromatographic apparatus according to claim 1, it is characterized in that: the specification of described first chromatographic column (6) is 150mm × 280mm, the specification of the second chromatographic column (6a) is 100mm × 280mm, and the specification of the 3rd chromatographic column (6b) is 80mm × 280mm.
4. a kind of automatic vacuum Split liquid chromatographic apparatus according to claim 1, is characterized in that: described first chromatographic column (6), the second chromatographic column (6a) are selected from silica gel, C8, C18, bonded silica gel or ion exchange resin with the filler of the 3rd chromatographic column (6b).
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| CN107102538A (en) * | 2017-06-21 | 2017-08-29 | 利穗科技(苏州)有限公司 | A kind of flow correction method matched somebody with somebody in liquid layer analysis system |
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| 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 |
| CN115055168A (en) * | 2022-06-09 | 2022-09-16 | 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 intelligence separation device |
| 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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