CN104422735B - A kind of preparative cyclical chromatography device - Google Patents
A kind of preparative cyclical chromatography device Download PDFInfo
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- CN104422735B CN104422735B CN201310365011.5A CN201310365011A CN104422735B CN 104422735 B CN104422735 B CN 104422735B CN 201310365011 A CN201310365011 A CN 201310365011A CN 104422735 B CN104422735 B CN 104422735B
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Abstract
A kind of circulation preparative liquid chromatography device.This device can be applicable to the chromatographic isolation of target component in complex sample to be prepared, and its major advantage is, (1) on-line sample pretreatment and concentration, it is achieved the large volume loading under high post effect premise；(2) by the evolution of detached dowel with trapping column, it is achieved the multi-cycle separation purification of target component, have the advantages that circulation dead volume is little and post effect loss is little；(3) eluting final products are concentrated by trapping column, it is achieved separation, concentration integrated process.Function above is that the method for the absorption property by on-line control trapping column and Vavle switching realizes.Natural product, medicine are prepared by this device, the separation of chemical products, and especially to trace target component in mixture, prepared by the separation of labile element, and more traditional preparative hplc preparation method has a clear superiority in.
Description
Technical field
The present invention is complex sample separatory circulation preparative liquid chromatography device.
Background technology
In natural product, medical separation or chemical products separation preparation process, preparative liquid chromatography is the most normal
See and efficiently separate means, but in this technology application process, still with the presence of many problems: (1)
Target substance composition content in biased sample is the lowest, rather than the composition of target component is too much；(2) target
Composition is in the pretreatment or dried process of sample, owing to transfer process is loaded down with trivial details or because of to light, heat, oxygen
Lose etc. environmental factors instability；(3) sample solubility is poor, causes applied sample amount little, wastes solvent, or
Volume overload during big upper volume sample, affects separating effect；(4) sample is dissolved in and has the molten of strong eluting power
Loading in agent, causes peak shape and separating effect to be deteriorated；(5) containing a large amount of salt, non-targeted composition in sample
The impurity such as sugar, the sample pretreatment caused is loaded down with trivial details；(6) concentration of part is flowed during separating for several times again
Course of dissolution, or final products concentrate, dry run takes time and effort.Therefore, the breakthrough of problem above is sought
Become the key technology improving separation efficiency.
Prior art is often applicable only to the application of analytical type chromatograph, and practicality for preparative equipment
Not strong, and renovation technique generally requires great amount of investment.
Summary of the invention
Present invention aim at developing a set of, traditional chromatograph structural framing is built, it is possible to resolve above-mentioned
The preparative liquid chromatography system of problem.
A kind of circulation preparative liquid chromatography device, it is characterised in that by the tune to trapping column absorbability
Whole, and Vavle switching is to its change of position in system stream, it is achieved in sample pretreatment, large volume trapping
The concentration of sample, multi-cycle separation and target product, small size reclaim.This device at least should include that separates a color
Spectrum post (abbreviation detached dowel), trapping column, ten-way valve, large volume quantitative loop, a clematis stem
Valve, a liquid chromatography pump for preparative separation (being hereinafter called for short " preparing pump ") and one are for catching
The flowing of clustered column carries out the liquid chromatography pump (being hereinafter called for short " regulation pump ") of eluotropic strength adjustment mutually.
The method of attachment of ten-way valve following (with reference to accompanying drawing 1): two ten-way valve interfaces the most successively through prepare pump,
Three-way cut-off valve is connected with preparing pump mobile phase A, the 3rd interface emptying outlet of three-way cut-off valve；Two
3. 2. ten-way valve interface be connected with interface；4. two ten-way valve interfaces are connected with the stigma interface of detached dowel；
5. two ten-way valve interfaces are sequentially passed through and are connected with six-way valve by three-way cut-off valve, the 3rd of three-way cut-off valve
Individual interface is emptying outlet；6. two ten-way valve interfaces are connected with guard column stigma interface；Two ten-way valves connect
7. with 10. by quantitative loop mouth is connected；8. two ten-way valve interfaces are sample injection port, and 9. interface is vented.Six
The method of attachment of logical valve is as follows: 1. two six-way valve interfaces are connected with the post tail interface of guard column；Two clematis stem
2. valve interface is connected with ten-way valve by three-way cut-off valve, and the 3rd interface of three-way cut-off valve is emptying outlet；
3. the interface of two six-way valves is connected with the post tail interface of trapping column；4. the interface of two six-way valves is examined with chromatograph
The entrance surveying device is connected；5. the interface of two six-way valves is connected with the post tail interface of detached dowel；Two six-way valves
Interface be 6. connected with the stigma interface of trapping column by threeway, the other end of threeway successively with three-way cut-off valve,
Regulation pump, proportioning valve are connected with trapping column Mobile phase B, C, and the 3rd interface of three-way cut-off valve is for being vented out
Mouthful.Operational meaning and concrete grammar following (with reference to accompanying drawing 2): during application trapping column loading, trapping column is positioned at
Before detached dowel stigma, now chromatogram flow phase sequentially passes through separation pump, stop valve (drain closedowns), ten logical
Valve, quantitative loop, guard column, trapping column, six-way valve and stop valve (drain unlatching), the most weak eluting is strong
Degree Mobile phase B is adjusted by the flowing of trapping column mutually by regulation pump, plays sample concentration and pre-column
Effect (with reference to accompanying drawing 2-c)；During desorbing loading, before trapping column is positioned at detached dowel stigma, now separate pump stream
Dynamic phase A sequentially passes through separation pump, stop valve (drain closedown), quantitative loop, guard column, trapping column, cuts
Only valve (drain closedown) detached dowel and detector (with reference to accompanying drawing 2-a)；In sample separation process,
Separate pump mobile phase A sequentially pass through separation pump, stop valve (drain closedown), ten-way valve, detached dowel, six
Logical valve and detector, meanwhile guard column and trapping column by and be positioned at trapping pump and threeway cut-off in the way of connecting
After valve (drain closedown), they can be carried out clearly by trapping pump by pumping into the solvent C having by force eluting power
Washing, it is possible to be balanced them by pumping into weak eluting power solvent B, liquid (is put by three-way cut-off valve
Empty valve is opened) discharge (with reference to accompanying drawing 2-d)；When target substance flows out from detached dowel, by Vavle switching,
After trapping column is positioned at detached dowel post tail, separated flow sequentially passes through separation pump mutually, stop valve (drain closedown),
Ten-way valve, detached dowel, six-way valve, trapping column and detector, meanwhile, regulation pump by threeway and connects
In trapping column porch, inject weak eluting power Mobile phase B to system, strengthen the trapping ability (ginseng of trapping column
Examine accompanying drawing 2-e)；After target substance traps, by Vavle switching, detached dowel is made to be positioned at separation pump and detection
Between device, separate pump mobile phase A and sequentially pass through separation pump, ten-way valve, detached dowel, six-way valve and detector,
Complete other compositions and separate (with reference to accompanying drawing 2-b)；Above procedure can be repeated, and reaches the mesh of multi-cycle separation
's；After target component completes to separate, by strong elution flow phase C, regulation pump stream is carried out, now
Strong elution flow phase C is adjusted pump and is discharged system (with reference to accompanying drawing 2-f) by three-way cut-off valve (drain unlatching)；
During final trapping product eluting, trapping column is regulating between pump and detector, strong eluting flowing phase C
It is adjusted pump and flows through three-way cut-off valve (drain closedown), trapping column and detector successively, reclaim target component
(with reference to accompanying drawing 2-g).
Circulation preparative liquid chromatography device involved in the present invention, by separate pump, regulation pump, guard column,
Detached dowel, trapping column, quantitative loop, ten-way valve, six-way valve, stop valve, chromatographic detector and multidigit select
Valve forms.Wherein regulation pump is positioned at trapping column entrance, is in parallel with coming from the flowing separating pump, for right
The flowing flowing through trapping column carries out eluotropic strength regulation mutually.Switched by the combination of ten-way valve with six-way valve, cut
Only the on off state of valve is different, and the on off state of pump is different, can realize following several duty:
(a) trapping column loading: chromatogram flow phase sequentially passes through separation pump, quantitative loop, guard column, trapping
Post and stop valve, and be adjusted by the chromatogram flow phase of trapping column by regulation pump；
(b) trapping column eluting detached dowel loading: chromatogram flow phase sequentially pass through chromatogram pump, quantitative loop,
Guard column, trapping column, detached dowel and detector；
(c) guard column, cleaning and the balance of trapping column: chromatogram flow phase sequentially pass through separate pump, detached dowel,
Detector；Stop valve is opened, and regulation pump input is cleaned and balanced solvent, enters guard column and trapping column respectively
Row is reversely and forward eluting.
(d) quantitative loop loading: chromatogram flow phase sequentially passes through separation pump, detached dowel and detector, sample
Inject quantitative loop；
E () target component traps: chromatogram flow phase sequentially passes through separation pump, detached dowel, trapping column and inspection
Surveying device, chromatogram flow phase is adjusted by regulation pump in trapping column porch；
(a)～(e) process of repetition completes target component and again separates or multi-cycle separation.
(f) target component eluting: regulation pump directly rinses trapping column, and by detector monitors target component.
The present invention is through the coupling of six-way valve Yu ten-way valve, it is achieved flowing is passed through with same direction mutually all the time
Detached dowel and trapping column, intercolumniation dead volume is little, is prevented effectively from back-mixing in cyclic process and causes separating effect to damage
Lose, and extend the service life of trapping column.
The present invention regulates absorbability and the flowing of trapping column by the regulation different eluotropic strength solvent of pump input
The eluotropic strength of phase.
This trapping column of the present invention can be located at the diverse location everywhere in system stream, i.e. (1) point by Vavle switching
Before post, after (2) detached dowel, (3) are individually positioned in chromatographic system, be i.e. positioned at chromatogram pump and detector it
Between, (4) are positioned at outside chromatographic system.
Preparing pump is semi-preparative or preparative liquid chromatography pump；
Regulation pump is semi-preparative or preparative liquid chromatography pump, uses in the following two cases: (1) is catching
During collection, use when adding Mobile phase B in system；(2) when sample reclaims, in system, add second catch
Use during clustered column flowing phase C.
Detached dowel is semi-preparative or preparative high performance liquid chromatography post；
Guard column is the guard column consistent with detached dowel diameter dimension, built-in same or like with detached dowel character
Chromatograph packing material；
Trapping column is consistent with detached dowel diameter dimension, but is shorter in length than the chromatographic column of detached dowel, built-in with separate
The chromatograph packing material that post character is same or like.
Proportioning valve common phase chromatography-use proportioning valve, maybe can realize the valve gear of liquid chromatogram mobile phase switching.
Described two six-way valves are alternatively two eight and lead to valve or two ten-way valves；Described switching valve is one two ten
Logical valve, leads to valve with two six-way valves or two eight or two ten-way valves is used in conjunction, and completes the stream switching of necessity.
Can use large volume (asLiquid-phase chromatographic column can use capacity to be 5～10ml) quantitative loop,
On the premise of using trapping column sample to carry out online pretreatment and concentrates, application high power capacity quantitative loop is permissible
Realize bulk sample loading, without causing volume overload.
The chromatographic system that the present invention relates to passes through Vavle switching and the collaborative work of pump, can imitate under premise maintaining post,
Carry out online sample pretreatment and concentration, it is to avoid solvent effect, it is achieved large volume loading and multi-cycle separation,
It addition, target product can concentrate on post, use the eluent of small size high volatility, be greatly simplified
Sample post-processing step.Chromatogram arrangement involved in the present invention, is advantageously implemented the automatic partition of complex sample
From, not only take full advantage of the separation efficiency of detached dowel, simplify manpower that sample treatment brings, the time,
Energy resource consumption and sample loss, extend again the use of detached dowel under the dual function of guard column and trapping column
In the life-span, it is greatly saved separation costs.
Accompanying drawing explanation
Fig. 1 preparative cyclical chromatography structure drawing of device；
Fig. 2 is the structure of this chromatograph and respectively walks operation chart；
Fig. 3 is the application present invention, to the separating effect figure of meta-xylene in large volume xylene mixture sample；
Fig. 4 is the application present invention elution chromatography figure to the target component of final enrichment.
Detailed description of the invention
Below in conjunction with example, this chromatogram arrangement is further described, but does not constitute any limitation of the invention.
Its major advantage is, (1) on-line sample pretreatment and concentration, it is achieved under high post effect premise substantially
Long-pending loading；(2) by the evolution of detached dowel with trapping column, it is achieved the multi-cycle separation purification of target component,
Have the advantages that circulation dead volume is little and post effect loss is little；(3) concentrate eluting by trapping column finally to produce
Product, it is achieved separation, concentration integrated process.Function above is the absorption property by on-line control trapping column
And the method for Vavle switching realizes.Natural product, medicine are prepared by this device, the separation of chemical products,
Especially to trace target component in mixture, prepared by the separation of labile element, more traditional preparative hplc system
Preparation Method has a clear superiority in.
The operational approach of described device,
(a) trapping column loading: chromatogram flow phase sequentially passes through separation pump, quantitative loop, guard column, trapping column
And stop valve, and it is adjusted by the chromatogram flow phase of trapping column by regulation pump；
(b) trapping column eluting detached dowel loading: chromatogram flow phase sequentially pass through chromatogram pump, quantitative loop,
Guard column, trapping column, detached dowel and detector；
(c) guard column, cleaning and the balance of trapping column: chromatogram flow phase sequentially pass through separate pump, detached dowel,
Detector；Stop valve is opened, and regulation pump input is cleaned and balanced solvent, enters guard column and trapping column respectively
Row is reversely and forward eluting.
D () quantitative loop loading: chromatogram flow phase sequentially passes through separation pump, detached dowel and detector, sample is noted
Enter quantitative loop；
E () target component traps: chromatogram flow phase sequentially passes through separation pump, detached dowel, trapping column and detection
Device, chromatogram flow phase is adjusted by regulation pump in trapping column porch；
(a)～(e) process of repetition completes target component and again separates or multi-cycle separation.
(f) target component eluting: regulation pump directly rinses trapping column, and by detector monitors target component.
Embodiment: the isolation and purification of meta-xylene in xylene mixture
Experiment reagent: analytical pure dimethylbenzene (federal chemical reagent factory), chromatograph methanol (Tianjin Ke Miou),
Pure water (laboratory is made by oneself, millipore pure water system purification).
Laboratory sample: the methanol solution (V of dimethylbenzeneDimethylbenzene∶VMethanol=1: 1000)
Instrument condition:
Preparative liquid chromatography pump (two, containing three-way cut-off valve), three-way cut-off valve, threeway, liquid chromatograph
UV-detector and chromatographic signal sampling system acquisition are in big Liaanjiang county Shengong department.C10UW type type hand two ten
Logical flow channel switching valve, purchases in Valco company of the U.S..Two clematis stem streams of 7750-TPMV type automatic type are cut
Change valve to purchase in RHEODYNE company of the U.S..The filling of detached dowel, guard column and trapping column is by Dalian
Jiang Shengong department completes.
System connects:
As shown in Figure 1, two ten-way valve 3 the most separated pumps of interface, three-way cut-off valve and separate pump stream
Dynamic phase (A) is connected, the 3rd interface emptying outlet of three-way cut-off valve；2. and interface two ten-way valve 3 interfaces
3. it is connected；4. two ten-way valve 3 interfaces are connected with the stigma interface of detached dowel 5；Two ten-way valve 3 interfaces
5. sequentially passing through and be connected with six-way valve 13 by three-way cut-off valve 4, the 3rd interface of three-way cut-off valve 4 is for putting
Air Export；6. two ten-way valve 3 interfaces are connected with guard column 7 stigma interface；Two ten-way valve 3 interfaces 7. with
10. it is connected by quantitative loop 6；8. two ten-way valve 3 interfaces are sample injection port, and 9. interface is vented.
1. two six-way valve (13) interfaces are connected with the post tail interface of guard column 7；Two six-way valve 13 interfaces
2. being connected with ten-way valve 3 by three-way cut-off valve 4, the 3rd interface of three-way cut-off valve is emptying outlet；
3. the interface of two six-way valves 13 is connected with the post tail interface of trapping column；The interface of two six-way valves 13 4. with
The entrance of chromatographic detector 14 is connected；The interface of two six-way valves 13 5. with the post tail interface phase of detached dowel 5
Even；6. the interface of two six-way valves 13 is connected with the stigma interface of trapping column 8 by threeway 12, threeway 12
The other end successively with three-way cut-off valve 11, regulation pump 9, proportioning valve 10 and trapping column flowing phase (B), (C)
Being connected, the 3rd interface of three-way cut-off valve 11 is emptying outlet.
Chromatographic condition:
Guard column and trapping column are self-control chromatographic column (10i.d. × 10mm, chromatocrex, 10 μm, FUJI
Company), (10i.d. × 150mm, chromatocrex, 10 μm, Japan FUJI is public for self-control chromatographic column for detached dowel
Department).Separating pump flowing is 80% methanol mutually, and during unlatching, flow velocity is 4ml/min.Regulation pump flowing is water mutually,
During unlatching, flow velocity is 10ml/min.
Experiment one: contrast trapping column large volume loading method and the system separating effect of direct loading method
Experimental technique: on the premise of applied sample amount is 3ml, contrast application trapping column trapping loading and directly on
The peak shape situation of sample.
Directly loading experiment:
(1) change system as shown in Fig. 2-h, and carry out cleaning systems and balance, inject in quantitative loop
Preparation of samples loading；
(2) switching of valve position is put shown in 2-i figure, carries out sample introduction separation.
Trapping column trapping loading experiment:
(1) separating pump to open, regulation pump cuts out, and stop valve is closed.Valve position is as shown in accompanying drawing 2-a；Right
Cleaning systems and balance.
(2) separating pump to open, regulation pump cuts out, and stop valve is closed.Valve position is as shown in accompanying drawing 2-b；This
Time, 3ml sample can be injected quantitative loop, wait loading.
(3) separating pump to open, regulation pump is opened, and stop valve is closed.Valve position is as shown in accompanying drawing 2-c: this
Time chromatogram flow phase sequentially pass through separation pump 1, stop valve 2 (drain closedown), ten-way valve 3, quantitative loop 6,
Guard column 7, trapping column 8, six-way valve 13 and stop valve 4 (drain unlatching), the most weak eluotropic strength stream
Dynamic phase (B) is adjusted by the flowing of trapping column 8 mutually by regulation pump 9.Sample is carried mutually by flowing,
After guard column, it is brought into trapping column；Now (select owing to regulation pump carries out on-line control mutually to flowing
The Mobile phase B that eluotropic strength is weak), target substance is adsorbed in trapping column.
(4) separating pump to open, regulation pump cuts out, and stop valve is closed.Valve position is as shown in accompanying drawing 2-a: catch
Before clustered column 8 is positioned at detached dowel stigma, now separates pump flowing phase (A) and sequentially pass through separation pump 1, stop valve
2 (drain closedowns), quantitative loop 6, guard column 7, trapping column 8, stop valve 4 (drain closedown) point
From post 5 and detector 14.Sample is flowed and is quickly gone out from trapping column mutually, brings detached dowel into and carries out point
From；
Separating effect contrasts:
Separating effect is to such as accompanying drawing 2 " large volume dimethylbenzene sample trapping loading " and " large volume dimethylbenzene
The direct loading of sample " shown in, therefrom this it appears that trapping loading method is in large volume loading separation process
More traditional chromatograph has clear superiority.
Experiment two: multi-cycle separation effect examination under large volume Loaded samples
Experimental technique:
(1) separating pump to open, regulation pump cuts out, and stop valve is closed.Valve position is as shown in accompanying drawing 2-a；Right
Cleaning systems and balance.
(2) separating pump to open, donkey pump cuts out, and stop valve is closed.Valve position is as shown in accompanying drawing 2-b；This
Time, 3ml sample can be injected quantitative loop, wait loading.
(3) separating pump to open, regulation pump is opened, and stop valve is closed.Valve position is as shown in accompanying drawing 2-c: catch
Before clustered column is positioned at detached dowel stigma, now chromatogram flow phase sequentially passes through separation pump 1, stop valve 2 (drain
Close), ten-way valve 3, quantitative loop 6, guard column 7, trapping column 8, six-way valve 13 and stop valve 4 (emptying
Mouth is opened), the most weak eluotropic strength flowing phase (B) is by regulating pump 9 to by the flowing phase of trapping column 8
It is adjusted sample to be carried mutually by flowing, after guard column, is brought into trapping column；Now due to regulation pump
Flowing carries out on-line control (selecting the Mobile phase B that eluotropic strength is weak) mutually, and target substance is adsorbed on trapping
In post；
(4) separating pump to open, regulation pump cuts out, and stop valve is closed.Valve position is as shown in accompanying drawing 2-a: point
Separation pump 1, stop valve 2 (drain closedown), quantitative loop 6, protection is sequentially passed through from pump flowing phase (A)
Post 7, trapping column 8, stop valve 4 (drain closedown) detached dowel 5 and detector 14.Sample is flowed
Quickly go out from trapping column mutually (for ensure sample completely from trapping eluting, in this experiment, elution time is
3min), bring detached dowel into separate；；
(5) separating pump to open, regulation pump is opened, and stop valve is opened.Valve position is as shown in accompanying drawing 2-d: point
Sequentially pass through separation pump 1, stop valve 2 (drain closedown), ten-way valve 3 from pump flowing phase (A), separate
Post 5, six-way valve 13 and detector 14, chromatographic separation process continues；Meanwhile guard column 7 and trapping column
8 by and after being positioned at trapping pump 9 and three-way cut-off valve 11 (drain closedowns) in the way of connecting, trapping pump 9 can
By pumping into the solvent (C) having by force eluting power, guard column is carried out reverse cleaning, trapping column is carried out forward
Clean, it is possible to being balanced them by pumping into weak eluting power solvent (B), liquid is ended by threeway
Valve 4 (atmospheric valve unlatching) is discharged.
(6) separating pump to open, regulation pump cuts out, and stop valve is closed.Valve position is as shown in accompanying drawing 2-b；Color
Spectrum separation process continues.
(7) separating pump to open, regulation pump is opened, stopping.Valve position is as shown in accompanying drawing 2-e: work as mesh
Mark material flows out from detached dowel 5, by Vavle switching, after trapping column 8 is positioned at detached dowel 5 post tail, separates
Flowing sequentially passes through separation pump 1, stop valve 2 (drain closedown), ten-way valve 3, detached dowel 5, clematis stem mutually
Valve 13, trapping column 8 and detector 14, meanwhile, regulation pump 9 by threeway 12 and is connected in trapping column
Porch, injects weak eluting power flowing phase (B) to system, strengthens the trapping ability of trapping column 8；Pass through
Vavle switching, target component is rinsed to trapping column, and flowing is entered mutually by regulation pump by pumping into low-intensity solvent
Row regulation, target substance is adsorbed in trapping column.
(8) after target substance traps, by Vavle switching, detached dowel 5 is made to be positioned at separation pump 1 He
Between detector 14, separate pump flowing phase (A) sequentially pass through separation pump 1, ten-way valve 3, detached dowel 5,
Six-way valve 13 and detector 14, complete other compositions and separate；
(9) regulation pump cuts out, and stop valve is closed, and separates pump and opens (valve position is as shown in accompanying drawing 2-a),
Now it is adsorbed in the material in trapping column again to be brought into mutually by flowing detached dowel is circulated separation.
Operating procedure (4)～(9) can be by actual demand circulation operations, until peak purity reaches requirement, and
Final absorption is stored in trapping column.In this experiment, for the explanation separating power to target substance, make altogether
The operation of five multi-cycle separation, in effect such as accompanying drawing 2 " bulk sample one trap loading one multi-cycle separation one～
Five times " shown in.These five contrast experiment's figures can illustrate increasing along with multi-cycle separation number of times, meta-xylene
Purity can be effectively improved, when avoiding application traditional colour spectral method in the middle of required sample simultaneously
(such as concentration, the filtration etc.) process of process.
Experiment three: the target substance recovery experiment being enriched in trapping column
Experimental technique: repeat to test two, when for the last time by example enrichment in trapping column after, carry out following
Operation:
(1) separating pump 1 to close, regulation pump 9 is opened (selecting the solvent C that eluotropic strength is big), now
Strong elution flow phase (C) is adjusted pump 9 and is discharged system by three-way cut-off valve 11 (drain unlatching).This step
Purpose is for except the Weak solvent B in pump chamber with strong solvent A, as shown in Fig. 2-f.
(2) separating pump 1 to close, adjusting donkey pump 9 flow velocity is 2ml/min, and three-way cut-off valve is closed；
Shown in valve position such as " accompanying drawing 2-g ": trapping column 8 is regulating between pump 9 and detector 14, strong eluting
Flowing phase (C) is adjusted pump 9 and flows through three-way cut-off valve 11 (drain closedown), trapping column 8 and inspection successively
Survey device 14.Regulation pump pumps into the volatile organic solvent of a small amount of high intensity, is washed by the target substance in trapping column
Go out.
Final sample is recyclable in 1.2ml (2ml/min × 0.6min) methanol, contrasts general collection method
(as shown in accompanying drawing two, under conditions of 4ml/min 80% methanol is for flowing mutually, the peak of meta-xylene
Wide ＞ 1min, i.e. 4ml/min × 1min=4ml), it can be seen that concentration and recovery method involved in the present invention has
Efficiently, time, the advantage of the energy are saved.
Claims (5)
1. a preparative cyclical chromatography device, including one two ten-way valves (3), quantitative loop (6),
One two six-way valves (13), detached dowel (5), guard column (7), trapping column (8),
Prepare pump (1), a regulation pump (9) for one, a proportioning valve (10), a threeway (12), three
Three-way cut-off valve, a liquid chromatogram detector (14) and three kinds of liquid chromatogram mobile phases；
Two ten-way valve (3) interface the first three-way cut-off valves (2) the most successively, prepare pump (1) and prepare pump
Mobile phase A is connected, the 3rd interface emptying outlet of three-way cut-off valve；Two ten-way valve (3) interfaces 2. and
3. interface is connected；Two ten-way valve (3) interfaces 4. stigma interface with detached dowel (5) is connected；Two
By the second three-way cut-off valve (4) with six-way valve (13) interface 2. 5. ten-way valve (3) interface sequentially pass through
Being connected, the 3rd interface of the second three-way cut-off valve (4) is emptying outlet；Two ten-way valve (3) interfaces
6. it is connected with guard column (7) stigma interface；Two ten-way valve (3) interfaces 7. with 10. by quantitative loop (6)
It is connected；8. two ten-way valve (3) interfaces are sample injection port, and 9. interface is vented；
1. two six-way valve (13) interfaces are connected with the post tail interface of guard column (7)；Two six-way valves (13)
2. interface is connected with ten-way valve (3) by three-way cut-off valve (4), the 3rd interface of three-way cut-off valve
For emptying outlet；The interface of two six-way valves (13) is 3. connected with the post tail interface of trapping column；Two six
The interface of logical valve (13) 4. entrance with chromatographic detector (14) is connected；Two six-way valves (13)
5. interface is connected with the post tail interface of detached dowel (5)；The interface of two six-way valves (13) 6. passes through threeway
(12) the stigma interface with trapping column (8) is connected, the other end of threeway (12) successively with the 3rd threeway
Stop valve (11), regulation pump (9), proportioning valve (10) and first, second trapping column Mobile phase B, C
Being connected, the 3rd interface of three-way cut-off valve (11) is emptying outlet.
2. according to the device described in claim 1, it is characterised in that:
Preparing pump is semi-preparative or preparative liquid chromatography pump；
Regulation pump is semi-preparative or preparative liquid chromatography pump, uses in the following two cases: (1) exists
During trapping, use when adding Mobile phase B in system；(2) when sample reclaims, in system, the is added
Use during two trapping column flowing phases C.
3. according to the device described in claim 1, it is characterised in that:
Detached dowel (5) is semi-preparative or preparative high performance liquid chromatography post；
Guard column (7) is the guard column consistent with detached dowel diameter dimension, built-in identical with detached dowel character or
Close chromatograph packing material；
Trapping column (8) is consistent with detached dowel diameter dimension, but is shorter in length than the chromatographic column of detached dowel, built-in
The chromatograph packing material same or like with detached dowel character.
4. according to the device described in claim 1, it is characterised in that:
The common phase chromatography-use proportioning valve of proportioning valve (10), maybe can realize the valve dress of liquid chromatogram mobile phase switching
Put.
5. according to the device described in claim 1, it is characterised in that: using trapping column, sample is carried out
On the premise of line pretreatment and concentration, application high power capacity quantitative loop can realize bulk sample loading, and
Do not result in volume overload.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310365011.5A CN104422735B (en) | 2013-08-19 | 2013-08-19 | A kind of preparative cyclical chromatography device |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310365011.5A CN104422735B (en) | 2013-08-19 | 2013-08-19 | A kind of preparative cyclical chromatography device |
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| CN105938130B (en) * | 2016-06-30 | 2018-04-20 | 朱靖博 | It is a kind of to integrate separation method exploitation, the natural drug two dimension preparative chromatograph of on-line preconcentration and its method of work |
| CN108562678B (en) * | 2018-02-12 | 2023-09-22 | 大连博迈科技发展有限公司 | Three-dimensional liquid chromatographic separation system based on full online detection of same detector |
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