CN105664527A - Continuous chromatographic separation device and method using device - Google Patents
Continuous chromatographic separation device and method using device Download PDFInfo
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- CN105664527A CN105664527A CN201610057186.3A CN201610057186A CN105664527A CN 105664527 A CN105664527 A CN 105664527A CN 201610057186 A CN201610057186 A CN 201610057186A CN 105664527 A CN105664527 A CN 105664527A
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- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
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- B01D15/00—Separating processes involving the treatment of liquids with solid sorbents; Apparatus therefor
- B01D15/08—Selective adsorption, e.g. chromatography
- B01D15/10—Selective adsorption, e.g. chromatography characterised by constructional or operational features
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Abstract
The invention discloses a continuous chromatographic separation device and a method using the device. The device comprises a sample infusion pump, a system infusion pump, chromatographic columns, flow path selection valves and a detector, wherein the sample infusion pump is firstly connected with a sample flow path selection valve, then, the sample flow path selection valve is connected in parallel with flow path selection valves in the number equal to that of the chromatographic columns, each flow path selection valve is connected one corresponding chromatographic column, the tail ends of the chromatographic columns are connected with the flow path selection valves, the flow path selection valves connected to the front ends of the chromatographic columns are connected with the flow path selection valves which are not connected with the tail ends of the chromatographic columns, the flow path selection valves connected with the tail ends of the chromatographic columns are connected with two flow path selection valves simultaneously, and the two flow path selection valves are connected with the detector; the system infusion pump is firstly connected with a system flow path selection valve, and then, the system flow path selection valve is connected in parallel with the flow path selection valves connected to the front ends of the chromatographic columns.
Description
Technical field
The present invention relates to technical field of analytical chemistry, particularly relate to a kind of continuous chromatography segregation apparatus and method thereof.
Background technology
In bio-pharmaceuticals technological process, in process upstream, the cultivation of biological sample have employed perfusion training method, the continual output product of energy, and the separating technology in current downstream still adopts off-line type chromatographic isolation, tomographic system is adopted to carry out single sample introduction separation, each loading is subject to chromatographic column load quantitative limitation, and the filler utilization rate in chromatographic column is not high. In order to provide applied sample amount, big chromatographic column can only be adopted, filler make consumption big, produce equipment investment costly, and owing to the loading number of times of filler is restricted, be accomplished by discarding after fixed number of times, the utilization rate causing filler is not high, and production cost is high.
Summary of the invention
For solving chromatographic separation device of the prior art and method Problems existing, the invention provides a kind of continuous chromatography segregation apparatus and method thereof, being applied in the industry such as pharmacy, chemical industry, the protein example that can realize perfusion is cultivated carries out continuous sample introduction, and continues to export target product.
The present invention adopts the following technical scheme that
The continuous chromatography segregation apparatus of the present invention, including sample infusion pump, system infusion pump, chromatographic column, flow path selection valve and detector, the number of sample infusion pump is one, and the number of system infusion pump is one or two, and the number of chromatographic column is two or more, and the number of detector is two, first sample infusion pump connects a sample flow path selector valve, then the parallel connection of sample flow path selector valve and the equal numbers of flow path selection valve of chromatographic column, a chromatographic column it is connected more respectively with the equal numbers of flow path selection valve of chromatographic column, chromatographic column end connects a flow path selection valve respectively, the flow path selection valve being connected with chromatographic column front end connects with other flow path selection valve not being connected with chromatographic column end respectively, the flow path selection valve that chromatographic column end connects is simultaneously connected with two flow path selection valve respectively, then the two flow path selection valve connects a detector respectively, first system infusion pump connects a system flow path selection valve, then the flow path selection valve that system flow path selection valve parallel connection chromatographic column front end connects.
Described sample infusion pump is plunger displacement pump or membrane pump, and plunger displacement pump is used for laboratory process optimization or technique amplification stage, and membrane pump is used for the large-scale production stage.
Described flow path selection valve can be rotating selector valve or multiple-way valve group, and flow path selection valve is 5 mouthfuls, 4 position valve, 7 mouthfuls, 6 position valve, 9 mouthfuls, 8 position valve or 11 mouthfuls, 10 positions valve, it is achieved one of them common mouth can only communicate with other mouth; Rotating selector valve is rotated by the rotor in motor-driven valve, it is ensured that middle common mouth can only communicate with other a mouth, reaches the effect that stream selects. Multiple-way valve group is made up of multiple two logical diaphragm valves by electromagnetism or pneumatic actuation, it is achieved any one common connection end mouth communicates with other connectivity ports. Reach the effect that stream selects. Electric rotating magnet valve is usually used in laboratory process optimization or technique amplification stage, and multiple-way valve group is usually used in technique and amplifies or the large-scale production stage.
As preferred technical scheme one: the number of system infusion pump is one, the number of chromatographic column is three, and the number of flow path selection valve is ten, and described flow path selection valve is 7 mouthfuls, 6 position valve or 7 mouthfuls of valve groups. sample infusion pump connects sample flow path selector valve, sample flow path selector valve is a flow path selection valve in parallel respectively, No. two flow path selection valve and No. three flow path selection valve, a number flow path selection valve, No. two flow path selection valve and No. three flow path selection valve connect a chromatographic column respectively, No. two chromatographic columns and No. three chromatographic columns, a number chromatographic column, No. two chromatographic columns and No. three chromatographic columns connect No. four flow path selection valve respectively, No. five flow path selection valve and No. six flow path selection valve, a number flow path selection valve is simultaneously connected with No. five flow path selection valve and No. six flow path selection valve, No. two flow path selection valve are simultaneously connected with No. four flow path selection valve and No. six flow path selection valve, No. three flow path selection valve are simultaneously connected with No. four flow path selection valve and No. five flow path selection valve, No. four flow path selection valve, No. five flow path selection valve and No. six flow path selection valve are simultaneously connected with No. seven flow path selection valve and No. eight flow path selection valve respectively, No. seven flow path selection valve connect sample detector, No. eight flow path selection valve connection system detectors, system infusion pump connection system flow path selection valve, system flow path selection valve is a flow path selection valve in parallel respectively, No. two flow path selection valve and No. three flow path selection valve.
As preferred technical scheme two: the number of system infusion pump is one, the number of chromatographic column is four, and the number of flow path selection valve is 12, and described flow path selection valve is 7 mouthfuls, 6 position valve or 7 mouthfuls of valve groups, and this programme is better than technical scheme one.
Assembly of the invention is utilized to carry out the specifically comprising the following steps that of method of continuous chromatography separation
A () sample infusion pump connects whole chromatographic column and sample detector by flow path selection valve, to chromatographic column sample detection;
B () system infusion pump connects one of them or more than one chromatographic column and system detector by flow path selection valve, the chromatographic column first connected is rinsed; Sample infusion pump connects remaining chromatographic column and detector by flow path selection valve simultaneously, carries out sample detection;
C () system infusion pump connects, by flow path selection valve, chromatographic column and another detector that step (b) is rinsed, carry out eluting, regeneration and reequilibrate; Sample infusion pump connects remaining chromatographic column and detector by flow path selection valve simultaneously, carries out sample detection;
D then remaining chromatographic column is rinsed by () successively, eluting, regeneration and reequilibrate; And utilize rinse, eluting, chromatographic column after regeneration and reequilibrate carry out sample detection, while circulation realizes the uninterrupted loading to sample continuously, continuous print collects the target substance after separating, the object that the staggered object due to the outflow of outer water reason collecting elution stage and overload loading stream are worn simultaneously.
The device described in preferred technical scheme one is utilized to carry out the specifically comprising the following steps that of method of continuous chromatography separation
A. the first step a: chromatographic column is carried out loading, sample infusion pump connects the common mouth of sample flow path selector valve, sample flow path selector valve is switched to No. 2 mouths, No. 2 mouths of sample flow path selector valve and No. 1 mouth of a flow path selection valve connect, a number flow path selection valve is switched to No. 1 mouth, the common mouth of a number flow path selection valve connects the entrance of a chromatographic column, the outlet of a number chromatographic column is connected with the common mouth of No. four flow path selection valve, No. four flow path selection valve are switched to No. 1 mouth, No. 1 mouth of No. four flow path selection valve and No. 2 mouths of No. two flow path selection valve are connected, No. two flow path selection valve are switched to No. 2 mouths, the common mouth of No. two flow path selection valve connects the entrance of No. two chromatographic columns, the outlet of No. two chromatographic columns is connected with the common mouth of No. five flow path selection valve, No. five flow path selection valve are switched to No. 2 mouths, No. 1 mouth of No. five flow path selection valve and No. 2 mouths of No. three flow path selection valve are connected, the common mouth of No. three flow path selection valve connects the entrance of No. three chromatographic columns, the outlet of No. three chromatographic columns is connected with the common mouth of No. six flow path selection valve, No. six flow path selection valve are switched to No. 2 mouths, No. 2 mouths of No. six flow path selection valve connect No. 5 mouths of No. seven flow path selection valve, No. seven flow path selection valve are switched to No. 5 mouths, the common mouth of No. seven flow path selection valve connects sample detector,
B. second step a: chromatographic column is rinsed, system infusion pump in turn switches on system flow path selection valve, a flow path selection valve, a chromatographic column, No. four flow path selection valve, No. three flow path selection valve, No. three chromatographic columns, No. six flow path selection valve, No. eight flow path selection valve and system detector, mobile phase from system infusion pump is rinsed, and No. three chromatographic columns are caught in a chromatographic column due to the object of residual in void volume; Sample infusion pump in turn switches on sample flow path selector valve, No. two flow path selection valve, No. two chromatographic columns, No. five flow path selection valve, No. seven flow path selection valve and sample detector simultaneously, to No. two chromatographic column loadings;
C. the 3rd step: system infusion pump in turn switches on system flow path selection valve, a number flow path selection valve, a number chromatographic column, No. four flow path selection valve, No. eight flow path selection valve and system detector, a number chromatographic column is carried out eluting by system infusion pump, regeneration and reequilibrate, sample infusion pump in turn switches on sample flow path selector valve simultaneously, No. two flow path selection valve, No. two chromatographic columns, No. five flow path selection valve, No. three flow path selection valve, No. three chromatographic columns, No. six flow path selection valve, No. seven flow path selection valve and sample detector, sample infusion pump is to No. two chromatographic column loadings, No. three chromatographic columns can be caught in No. two chromatographic columns owing to loading crosses the object that current-carrying is worn,
D. the 4th step: system infusion pump in turn switches on system flow path selection valve, No. two flow path selection valve, No. two chromatographic columns, No. five flow path selection valve, a flow path selection valve, a chromatographic column, No. four flow path selection valve, No. eight flow path selection valve and system detector, No. two chromatographic columns are rinsed so that a chromatographic column can be caught in No. two chromatographic columns due to the object of residual in void volume; Sample infusion pump in turn switches on sample flow path selector valve, No. three flow path selection valve, No. three chromatographic columns, No. six flow path selection valve, No. seven flow path selection valve and sample detector simultaneously, and sample infusion pump is to No. three chromatographic column loadings;
E. the 5th step: system infusion pump in turn switches on system flow path selection valve, No. two flow path selection valve, No. two chromatographic columns, No. five flow path selection valve, No. eight flow path selection valve and system detector, and No. two chromatographic columns are carried out eluting, regeneration and reequilibrate by system infusion pump;Sample infusion pump in turn switches on sample flow path selector valve, No. three flow path selection valve, No. three chromatographic columns, No. six flow path selection valve, a flow path selection valve, a chromatographic column, No. four flow path selection valve, No. seven flow path selection valve and sample detector simultaneously, sample infusion pump is to No. three chromatographic column loadings so that a chromatographic column can be caught in No. three chromatographic columns owing to loading crosses the object that current-carrying is worn;
F. the 6th step: system infusion pump in turn switches on system flow path selection valve, No. three flow path selection valve, No. three chromatographic columns, No. six flow path selection valve, No. two flow path selection valve, No. two chromatographic columns, No. five flow path selection valve, No. eight flow path selection valve and system detector, and No. three chromatographic columns are rinsed; Sample infusion pump in turn switches on sample flow path selector valve, a flow path selection valve, a chromatographic column, No. four flow path selection valve, No. seven flow path selection valve and sample detector simultaneously, and sample infusion pump is to a chromatographic column loading;
G. the 7th step: system infusion pump in turn switches on system flow path selection valve, No. three flow path selection valve, No. three chromatographic columns, No. six flow path selection valve, No. eight flow path selection valve and system detector, and No. three chromatographic columns are carried out eluting, regeneration and reequilibrate; Sample infusion pump in turn switches on sample flow path selector valve, a flow path selection valve, a chromatographic column, No. four flow path selection valve, No. two flow path selection valve, No. two chromatographic columns, No. five flow path selection valve, No. seven flow path selection valve and sample detector simultaneously, sample infusion pump is to a chromatographic column loading so that No. two chromatographic columns can be caught in a chromatographic column owing to loading crosses the object that current-carrying is worn;
The continuous of step A～G circulates, it is possible to achieve while the uninterrupted loading to sample, and continuous print collects the target substance after separating, the object that the staggered object due to the outflow of outer water reason collecting elution stage and overload loading stream are worn simultaneously.
The positive effect of the present invention is as follows:
The continuous chromatography segregation apparatus of the present invention and method thereof not only can realize the continuous sample introduction of biological sample and separate purification, upstream and downstream technique can be connected, realize continuously uninterrupted separation, production efficiency is provided, and can avoid owing to the void volume of chromatographic column is wasted when rinsing, thus providing the response rate of bio-separation.
The continuous chromatography segregation apparatus of the present invention can realize the chromatographic column that the requirement according to technique connects 2 to 10. And have the advantage that
1) combine with upstream perfusion culture process, it is possible to sample is also easily separated purification by continuous sample introduction, continues to export target product. Improve the production efficiency of 2～4 times.
2) stream when can catch overload loading is worn so that chromatographic column can be fully loaded with loading, and filler is fully used, and reduces the cost of production 30～50%, can catch chromatographic column due to the chromatographic column void volume sample of outflow when cleaning, improve the response rate.
Accompanying drawing explanation
Fig. 1 be the present invention continuous chromatography segregation apparatus in the schematic diagram of the 45 mouthfuls of flow path selection valve in position.
Fig. 2 be the present invention continuous chromatography segregation apparatus in the schematic diagram of the 67 mouthfuls of flow path selection valve in position.
Fig. 3 be the present invention continuous chromatography segregation apparatus in the schematic diagram of the 89 mouthfuls of flow path selection valve in position.
Fig. 4 is the schematic diagram of the continuous chromatography segregation apparatus of the embodiment of the present invention 1.
Fig. 5 is the schematic diagram of the continuous chromatography segregation apparatus of the embodiment of the present invention 2.
1 is sample infusion pump, 2 is system infusion pump, 3 is sample detector, 4 is system detector, 5 is sample flow path selector valve, 6 is system flow path selection valve, 7 is a flow path selection valve, 8 is No. two flow path selection valve, 9 is No. three flow path selection valve, 10 is No. four flow path selection valve, 11 is No. five flow path selection valve, 12 is No. six flow path selection valve, 13 is No. seven flow path selection valve, 14 is No. eight flow path selection valve, 15 is No. nine flow path selection valve, 16 is No. ten flow path selection valve, C1 is a chromatographic column, C2 is No. two chromatographic columns, C3 is No. three chromatographic columns, C4 is No. four chromatographic columns.
Detailed description of the invention
In conjunction with the following examples, the present invention is described in further detail.
Embodiment 1
As shown in Figure 4, in the continuous chromatography segregation apparatus of the present invention, the number of system infusion pump is one, and the number of chromatographic column is three, and the number of flow path selection valve is ten, and described flow path selection valve is 7 mouthfuls, 6 position valve.
Sample infusion pump 1 connects sample flow path selector valve 5, sample flow path selector valve 5 is a flow path selection valve 7 in parallel respectively, No. two flow path selection valve 8 and No. three flow path selection valve 9, a number flow path selection valve 7, No. two flow path selection valve 8 and No. three flow path selection valve 9 connect a chromatographic column C1 respectively, No. two chromatographic column C2 and No. three chromatographic column C3, a number chromatographic column C1, No. two chromatographic column C2 and No. three chromatographic column C3 connect No. four flow path selection valve 10 respectively, No. five flow path selection valve 11 and No. six flow path selection valve 12, a number flow path selection valve 7 is simultaneously connected with No. five flow path selection valve 11 and No. six flow path selection valve 12, No. two flow path selection valve 8 are simultaneously connected with No. four flow path selection valve 10 and No. six flow path selection valve 12, No. three flow path selection valve 9 are simultaneously connected with No. four flow path selection valve 10 and No. five flow path selection valve 11, No. four flow path selection valve 10, No. five flow path selection valve 11 and No. six flow path selection valve 12 are simultaneously connected with No. seven flow path selection valve 13 and No. eight flow path selection valve 14 respectively, No. seven flow path selection valve 13 connect sample detector 3, No. eight flow path selection valve 14 connection system detectors 4, system infusion pump 2 connection system flow path selection valve 6, system flow path selection valve 6 is a flow path selection valve 7 in parallel respectively, No. two flow path selection valve 8 and No. three flow path selection valve 9.
Assembly of the invention carries out specifically comprising the following steps that of the method for continuous chromatography separation
A. the first step: a chromatographic column C1 is carried out loading, sample infusion pump 1 connects the common mouth of sample flow path selector valve 5, sample flow path selector valve 5 is switched to No. 2 mouths, No. 2 mouths of sample flow path selector valve 5 and No. 1 mouth of a flow path selection valve 7 connect, a number flow path selection valve 7 is switched to No. 1 mouth, the common mouth of a number flow path selection valve 7 connects the entrance of a chromatographic column C1, the outlet of a number chromatographic column C1 is connected with the common mouth of No. four flow path selection valve 10, No. four flow path selection valve 10 are switched to No. 1 mouth, No. 1 mouth of No. four flow path selection valve 10 and No. 2 mouths of No. two flow path selection valve 8 are connected, No. two flow path selection valve 8 are switched to No. 2 mouths, the common mouth of No. two flow path selection valve 8 connects the entrance of No. two chromatographic column C2, the outlet of No. two chromatographic column C2 is connected with the common mouth of No. five flow path selection valve 11, No. five flow path selection valve 11 are switched to No. 2 mouths, No. 1 mouth of No. five flow path selection valve 11 and No. 2 mouths of No. three flow path selection valve 9 are connected, the common mouth of No. three flow path selection valve 9 connects the entrance of No. three chromatographic column C3, the outlet of No. three chromatographic column C3 is connected with the common mouth of No. six flow path selection valve 12, No. six flow path selection valve 12 are switched to No. 2 mouths, No. 2 mouths of No. six flow path selection valve 12 connect No. 5 mouths of No. seven flow path selection valve 13, No. seven flow path selection valve 13 are switched to No. 5 mouths, the common mouth of No. seven flow path selection valve 13 connects sample detector 3,
B. second step: a chromatographic column C1 is rinsed, system infusion pump 2 in turn switches on system flow path selection valve 7, chromatographic column C1 of 6, flow path selection valve, No. four flow path selection valve 9, No. three chromatographic column C3 of 10, No. three flow path selection valve, 12, No. eight flow path selection valve 14 of No. six flow path selection valve and system detector 4, water from system infusion pump 2 is rinsed, and No. three chromatographic column C3 catch in a chromatographic column C1 due to the object of residual in void volume;Sample infusion pump 1 in turn switches on sample flow path selector valve 8, No. two chromatographic column C2 of 5, No. two flow path selection valve, 11, No. seven flow path selection valve 13 of No. five flow path selection valve and sample detector 3 simultaneously, to No. two chromatographic column C2 loadings;
C. the 3rd step: system infusion pump 2 in turn switches on system flow path selection valve 6, a number flow path selection valve 7, a number chromatographic column C1, No. four flow path selection valve 10, No. eight flow path selection valve 14 and system detector 4, a number chromatographic column C1 is carried out eluting by system infusion pump 2, regeneration and reequilibrate, sample infusion pump 1 in turn switches on sample flow path selector valve 5 simultaneously, No. two flow path selection valve 8, No. two chromatographic column C2, No. five flow path selection valve 11, No. three flow path selection valve 9, No. three chromatographic column C3, No. six flow path selection valve 12, No. seven flow path selection valve 13 and sample detector 3, sample infusion pump 1 is to No. two chromatographic column C2 loadings, No. three chromatographic column C3 can be caught in No. two chromatographic column C2 owing to loading crosses the object that current-carrying is worn,
D. the 4th step: system infusion pump 2 in turn switches on system flow path selection valve 8, No. two chromatographic column C2 of 6, No. two flow path selection valve, No. five flow path selection valve 7, chromatographic column C1 of 11, flow path selection valve, 10, No. eight flow path selection valve 14 of No. four flow path selection valve and system detector 4, No. two chromatographic column C2 are rinsed so that a chromatographic column C1 can catch in No. two chromatographic column C2 due to the object of residual in void volume; Sample infusion pump 1 in turn switches on sample flow path selector valve 9, No. three chromatographic column C3 of 5, No. three flow path selection valve, 12, No. seven flow path selection valve 13 of No. six flow path selection valve and sample detector 3 simultaneously, and sample infusion pump 1 is to No. three chromatographic column C3 loadings;
E. the 5th step: system infusion pump 2 in turn switches on system flow path selection valve 8, No. two chromatographic column C2 of 6, No. two flow path selection valve, 11, No. eight flow path selection valve 14 of No. five flow path selection valve and system detector 4, and No. two chromatographic column C2 are carried out eluting, regeneration and reequilibrate by system infusion pump 2; Sample infusion pump 1 in turn switches on sample flow path selector valve 9, No. three chromatographic column C3 of 5, No. three flow path selection valve, No. six flow path selection valve 7, chromatographic column C1 of 12, flow path selection valve, 10, No. seven flow path selection valve 13 of No. four flow path selection valve and sample detector 3 simultaneously, sample infusion pump 1 is to No. three chromatographic column C3 loadings so that a chromatographic column C1 can catch in No. three chromatographic column C3 owing to loading crosses the object that current-carrying is worn;
F. the 6th step: system infusion pump 2 in turn switches on system flow path selection valve 9, No. three chromatographic column C3 of 6, No. three flow path selection valve, No. six flow path selection valve 8, No. two chromatographic column C2 of 12, No. two flow path selection valve, 11, No. eight flow path selection valve 14 of No. five flow path selection valve and system detector 4, and No. three chromatographic column C3 are rinsed; Sample infusion pump 1 in turn switches on sample flow path selector valve 7, chromatographic column C1 of 5, flow path selection valve, 10, No. seven flow path selection valve 13 of No. four flow path selection valve and sample detector 3 simultaneously, and sample infusion pump 1 is to a chromatographic column C1 loading;
G. the 7th step: system infusion pump 2 in turn switches on system flow path selection valve 9, No. three chromatographic column C3 of 6, No. three flow path selection valve, 12, No. eight flow path selection valve 14 of No. six flow path selection valve and system detector 4, and No. three chromatographic column C3 are carried out eluting, regeneration and reequilibrate; Sample infusion pump 1 in turn switches on sample flow path selector valve 7, chromatographic column C1 of 5, flow path selection valve, No. four flow path selection valve 8, No. two chromatographic column C2 of 10, No. two flow path selection valve, 11, No. seven flow path selection valve 13 of No. five flow path selection valve and sample detector 3 simultaneously, sample infusion pump 1 is to a chromatographic column C1 loading so that No. two chromatographic column C2 can catch in a chromatographic column C1 owing to loading crosses the object that current-carrying is worn;
The continuous of step A～G circulates, it is possible to achieve while the uninterrupted loading to sample, and continuous print collects the target substance after separating, the object that the staggered object due to the outflow of outer water reason collecting elution stage and overload loading stream are worn simultaneously.
Embodiment 2
The present invention is continuously separated chromatogram arrangement connected mode as it is shown in figure 5, the method for work of this method employing is as follows:
A. sample infusion pump 1 is to a chromatographic column C1 loading. Connecting No. two chromatographic column C2 after a number chromatographic column C1, No. two chromatographic column C2 catch the stream after a chromatographic column C1 loading is transshipped and wear;
B. a chromatographic column C1 is rinsed by system infusion pump 2, and is simultaneously connected with No. four chromatographic column C4 so that No. four chromatographic column C4 can catch in a chromatographic column C1 due to the object of residual in void volume. Sample infusion pump 1 is to No. two chromatographic column C2 loadings simultaneously. Connecting No. three chromatographic column C3 after No. two chromatographic column C2, No. three chromatographic column C3 catch the stream after No. two chromatographic column C2 loadings are transshipped and wear;
C. a chromatographic column C1 is carried out eluting, regeneration and reequilibrate by system infusion pump 2, sample infusion pump 1 is to No. two chromatographic column C2 loadings simultaneously, and it is simultaneously connected with No. three chromatographic column C3 so that No. three chromatographic column C3 can catch in No. two chromatographic column C2 owing to loading crosses the object that current-carrying is worn;
D. No. two chromatographic column C2 are rinsed by system infusion pump 2, and are simultaneously connected with a chromatographic column C1 so that a chromatographic column C1 can catch in No. two chromatographic column C2 due to the object of residual in void volume. Sample infusion pump 1 is to No. three chromatographic column C3 loadings simultaneously. Connecting No. four chromatographic column C4 after No. three chromatographic column C3, No. four chromatographic column C4 catch the stream after No. three chromatographic column C3 loadings are transshipped and wear;
E. No. two chromatographic column C2 are carried out eluting, regeneration and reequilibrate by system infusion pump 2, sample infusion pump 1 is to No. three chromatographic column C3 loadings simultaneously, and it is simultaneously connected with No. four chromatographic column C4 so that No. four chromatographic column C4 can catch in No. three chromatographic column C3 owing to loading crosses the object that current-carrying is worn;
F. No. three chromatographic column C3 are rinsed by system infusion pump 2, and are simultaneously connected with No. two chromatographic column C2 so that No. two chromatographic column C2 can catch in No. three chromatographic column C3 due to the object of residual in void volume. Sample infusion pump 1 is to No. four chromatographic column C4 loadings simultaneously. Connecting a chromatographic column C1 after No. four chromatographic column C4, a chromatographic column C1 catches the stream after No. four chromatographic column C4 loadings are transshipped and wears;
G. No. three chromatographic column C3 are carried out eluting, regeneration and reequilibrate by system infusion pump 2, sample infusion pump 1 is to No. four chromatographic column C4 loadings simultaneously, and it is simultaneously connected with a chromatographic column C1 so that a chromatographic column C1 can catch in No. four chromatographic column C4 owing to loading crosses the object that current-carrying is worn;
H. No. four chromatographic column C4 are rinsed by system infusion pump 2, and are simultaneously connected with No. three chromatographic column C3 so that No. three chromatographic column C3 can catch in No. four chromatographic column C4 due to the object of residual in void volume. Sample infusion pump 1 is to a chromatographic column C1 loading simultaneously. Connecting No. two chromatographic column C2 after a number chromatographic column C1, No. two chromatographic column C2 catch the stream after a chromatographic column C1 loading is transshipped and wear;
I. No. four chromatographic column C4 are carried out eluting, regeneration and reequilibrate by system infusion pump 2, sample infusion pump 1 is to a chromatographic column C1 loading simultaneously, and it is simultaneously connected with No. two chromatographic column C2 so that No. two chromatographic column C2 can catch in a chromatographic column C1 owing to loading crosses the object that current-carrying is worn.
The continuous circulation of step A～I, it is possible to achieve while the uninterrupted loading to sample, continuous print collects the target substance after separating.The object that the staggered object due to the outflow of outer water reason collecting elution stage and overload loading stream are worn simultaneously, not only can improve the response rate, and can improve the utilization rate of chromatographic stuffing. Reduce production cost. Adopt 4 chromatographic columns, there is 1 passage that root pillar is carried out, takes off, regenerates and is balanced for another one passage while loading by wherein 1 chromatographic column, the object that stream when one of other two chromatographic columns is for catching loading is worn, other one is used for catching the object that the outer water of wash phase causes. So can avoid when occurring owing to when loading time is shorter than scavenging period, loading and cleaning can not carry out parallel.
Although an embodiment of the present invention has been shown and described, for the ordinary skill in the art, being appreciated that and these embodiments can be carried out multiple change, amendment, replacement and modification without departing from the principles and spirit of the present invention, the scope of the present invention be defined by the appended.
Claims (8)
1. a continuous chromatography segregation apparatus, described continuous chromatography segregation apparatus includes sample infusion pump, system infusion pump, chromatographic column, flow path selection valve and detector, it is characterized in that: the number of sample infusion pump is one, the number of system infusion pump is one or two, and the number of chromatographic column is two or more, and the number of detector is two, first sample infusion pump connects a sample flow path selector valve, then the parallel connection of sample flow path selector valve and the equal numbers of flow path selection valve of chromatographic column, a chromatographic column it is connected more respectively with the equal numbers of flow path selection valve of chromatographic column, chromatographic column end connects a flow path selection valve respectively, the flow path selection valve being connected with chromatographic column front end connects with other flow path selection valve not being connected with chromatographic column end respectively, the flow path selection valve that chromatographic column end connects is simultaneously connected with two flow path selection valve respectively, then the two flow path selection valve connects a detector respectively, first system infusion pump connects a system flow path selection valve, then the flow path selection valve that system flow path selection valve parallel connection chromatographic column front end connects.
2. continuous chromatography segregation apparatus as claimed in claim 1, it is characterized in that: described sample infusion pump is plunger displacement pump, membrane pump or peristaltic pump, plunger displacement pump or peristaltic pump are used for the large-scale production stage for laboratory process optimization or technique amplification stage, membrane pump or peristaltic pump.
3. continuous chromatography segregation apparatus as claimed in claim 1, it is characterized in that: described flow path selection valve can be rotating selector valve or multiple-way valve group, flow path selection valve is 5 mouthfuls, 4 position valve, 7 mouthfuls, 6 position valve, 9 mouthfuls, 8 position valve or 11 mouthfuls, 10 positions valve, it is achieved one of them common mouth can only communicate with other mouth; Rotating selector valve is rotated by the rotor in motor-driven valve, it is ensured that middle common mouth can only communicate with other a mouth, reaches the effect that stream selects; Multiple-way valve group is made up of multiple two logical diaphragm electromagnetic valves by electromagnetism or pneumatic actuation, it is achieved any one common connection end mouth communicates with other connectivity ports, reaches the effect that stream selects; Electric rotating magnet valve is usually used in laboratory process optimization or technique amplification stage, and diaphragm electromagnetic valve group is usually used in technique and amplifies or the large-scale production stage.
4. continuous chromatography segregation apparatus as claimed in claim 1, it is characterised in that: the number of system infusion pump is one, and the number of chromatographic column is three, and the number of flow path selection valve is ten, and described flow path selection valve is 7 mouthfuls, 6 position valve or 7 mouthfuls of valve groups.
5. continuous chromatography segregation apparatus as claimed in claim 4, it is characterized in that: sample infusion pump (1) connects sample flow path selector valve (5), sample flow path selector valve (5) is a flow path selection valve (7) in parallel respectively, No. two flow path selection valve (8) and No. three flow path selection valve (9), a number flow path selection valve (7), No. two flow path selection valve (8) and No. three flow path selection valve (9) connect a chromatographic column (C1) respectively, No. two chromatographic columns (C2) and No. three chromatographic columns (C3), a number chromatographic column (C1), No. two chromatographic columns (C2) and No. three chromatographic columns (C3) connect No. four flow path selection valve (10) respectively, No. five flow path selection valve (11) and No. six flow path selection valve (12), a number flow path selection valve (7) is simultaneously connected with No. five flow path selection valve (11) and No. six flow path selection valve (12), No. two flow path selection valve (8) are simultaneously connected with No. four flow path selection valve (10) and No. six flow path selection valve (12), No. three flow path selection valve (9) are simultaneously connected with No. four flow path selection valve (10) and No. five flow path selection valve (11), No. four flow path selection valve (10), No. five flow path selection valve (11) and No. six flow path selection valve (12) are simultaneously connected with No. seven flow path selection valve (13) and No. eight flow path selection valve (14) respectively, No. seven flow path selection valve (13) connect sample detector (3), No. eight flow path selection valve (14) connection system detector (4), system infusion pump (2) connection system flow path selection valve (6), system flow path selection valve (6) is a flow path selection valve (7) in parallel respectively, No. two flow path selection valve (8) and No. three flow path selection valve (9).
6. continuous chromatography segregation apparatus as claimed in claim 1, it is characterised in that: the number of system infusion pump is one, and the number of chromatographic column is four, and the number of flow path selection valve is 12, and described flow path selection valve is 7 mouthfuls, 6 position valve or 7 mouthfuls of valve groups.
7. the method that the continuous chromatography segregation apparatus that a kind utilizes described in any one of claim 1-3 carries out continuous chromatography separation, it is characterised in that: comprising the steps: of described method
A () sample infusion pump connects whole chromatographic column and sample detector by flow path selection valve, to chromatographic column sample detection;
B () system infusion pump connects one of them or more than one chromatographic column and system detector by flow path selection valve, the chromatographic column first connected is rinsed; Sample infusion pump connects remaining chromatographic column and detector by flow path selection valve simultaneously, carries out sample detection;
C () system infusion pump connects, by flow path selection valve, chromatographic column and another detector that step (b) is rinsed, carry out eluting, regeneration and reequilibrate; Sample infusion pump connects remaining chromatographic column and detector by flow path selection valve simultaneously, carries out sample detection;
D then remaining chromatographic column is rinsed by () successively, eluting, regeneration and reequilibrate; And utilize rinse, eluting, chromatographic column after regeneration and reequilibrate carry out sample detection, while circulation realizes the uninterrupted loading to sample continuously, continuous print collects the target substance after separating, the object that the staggered object due to the outflow of outer water reason collecting elution stage and overload loading stream are worn simultaneously.
8. the method that the device utilized described in claim 5 carries out continuous chromatography separation, it is characterised in that: specifically comprising the following steps that of described method
A. the first step a: chromatographic column (C1) is carried out loading, sample infusion pump (1) connects the common mouth of sample flow path selector valve (5), sample flow path selector valve (5) is switched to No. 2 mouths, No. 2 mouths of sample flow path selector valve (5) and No. 1 mouth of a flow path selection valve (7) connect, a number flow path selection valve (7) is switched to No. 1 mouth, the common mouth of a number flow path selection valve (7) connects the entrance of a chromatographic column (C1), the outlet of a number chromatographic column (C1) is connected with the common mouth of No. four flow path selection valve (10), No. four flow path selection valve (10) are switched to No. 1 mouth, No. 1 mouth of No. four flow path selection valve (10) and No. 2 mouths of No. two flow path selection valve (8) are connected, No. two flow path selection valve (8) are switched to No. 2 mouths, the common mouth of No. two flow path selection valve (8) connects the entrance of No. two chromatographic columns (C2), the outlet of No. two chromatographic columns (C2) is connected with the common mouth of No. five flow path selection valve (11), No. five flow path selection valve (11) are switched to No. 2 mouths, No. 1 mouth of No. five flow path selection valve (11) and No. 2 mouths of No. three flow path selection valve (9) are connected, the common mouth of No. three flow path selection valve (9) connects the entrance of No. three chromatographic columns (C3), the outlet of No. three chromatographic columns (C3) is connected with the common mouth of No. six flow path selection valve (12), No. six flow path selection valve (12) are switched to No. 2 mouths, No. 2 mouths of No. six flow path selection valve (12) connect No. 5 mouths of No. seven flow path selection valve (13), No. seven flow path selection valve (13) are switched to No. 5 mouths, the common mouth of No. seven flow path selection valve (13) connects sample detector (3),
B. second step a: chromatographic column (C1) is rinsed, system infusion pump (2) in turn switches on system flow path selection valve (6), a number flow path selection valve (7), a number chromatographic column (C1), No. four flow path selection valve (10), No. three flow path selection valve (9), No. three chromatographic columns (C3), No. six flow path selection valve (12), No. eight flow path selection valve (14) and system detector (4), mobile phase from system infusion pump (2) is rinsed, No. three chromatographic columns (C3) are caught in a chromatographic column (C1) due to the object of residual in void volume, sample infusion pump (1) in turn switches on sample flow path selector valve (5), No. two flow path selection valve (8), No. two chromatographic columns (C2), No. five flow path selection valve (11), No. seven flow path selection valve (13) and sample detector (3) simultaneously, to No. two chromatographic column (C2) loadings,
C. the 3rd step: system infusion pump (2) in turn switches on system flow path selection valve (6), a number flow path selection valve (7), a number chromatographic column (C1), No. four flow path selection valve (10), No. eight flow path selection valve (14) and system detector (4), a number chromatographic column (C1) is carried out eluting by system infusion pump (2), regeneration and reequilibrate, sample infusion pump (1) in turn switches on sample flow path selector valve (5) simultaneously, No. two flow path selection valve (8), No. two chromatographic columns (C2), No. five flow path selection valve (11), No. three flow path selection valve (9), No. three chromatographic columns (C3), No. six flow path selection valve (12), No. seven flow path selection valve (13) and sample detector (3), sample infusion pump (1) is to No. two chromatographic column (C2) loadings, No. three chromatographic columns (C3) can be caught in No. two chromatographic columns (C2) owing to loading crosses the object that current-carrying is worn,
D. the 4th step: system infusion pump (2) in turn switches on system flow path selection valve (6), No. two flow path selection valve (8), No. two chromatographic columns (C2), No. five flow path selection valve (11), a flow path selection valve (7), a chromatographic column (C1), No. four flow path selection valve (10), No. eight flow path selection valve (14) and system detector (4), No. two chromatographic columns (C2) are rinsed so that a chromatographic column (C1) can be caught in No. two chromatographic columns (C2) due to the object of residual in void volume; Sample infusion pump (1) in turn switches on sample flow path selector valve (5), No. three flow path selection valve (9), No. three chromatographic columns (C3), No. six flow path selection valve (12), No. seven flow path selection valve (13) and sample detector (3) simultaneously, and sample infusion pump (1) is to No. three chromatographic column (C3) loadings;
E. the 5th step: system infusion pump (2) in turn switches on system flow path selection valve (6), No. two flow path selection valve (8), No. two chromatographic columns (C2), No. five flow path selection valve (11), No. eight flow path selection valve (14) and system detector (4), and No. two chromatographic columns (C2) are carried out eluting, regeneration and reequilibrate by system infusion pump (2); Sample infusion pump (1) in turn switches on sample flow path selector valve (5), No. three flow path selection valve (9), No. three chromatographic columns (C3), No. six flow path selection valve (12), a flow path selection valve (7), a chromatographic column (C1), No. four flow path selection valve (10), No. seven flow path selection valve (13) and sample detector (3) simultaneously, sample infusion pump (1) is to No. three chromatographic column (C3) loadings so that a chromatographic column (C1) can be caught in No. three chromatographic columns (C3) owing to loading crosses the object that current-carrying is worn;
F. the 6th step: system infusion pump (2) in turn switches on system flow path selection valve (6), No. three flow path selection valve (9), No. three chromatographic columns (C3), No. six flow path selection valve (12), No. two flow path selection valve (8), No. two chromatographic columns (C2), No. five flow path selection valve (11), No. eight flow path selection valve (14) and system detector (4), and No. three chromatographic columns (C3) are rinsed;Sample infusion pump (1) in turn switches on sample flow path selector valve (5), a flow path selection valve (7), a chromatographic column (C1), No. four flow path selection valve (10), No. seven flow path selection valve (13) and sample detector (3) simultaneously, and sample infusion pump (1) is to chromatographic column (C1) loading;
G. the 7th step: system infusion pump (2) in turn switches on system flow path selection valve (6), No. three flow path selection valve (9), No. three chromatographic columns (C3), No. six flow path selection valve (12), No. eight flow path selection valve (14) and system detector (4), and No. three chromatographic columns (C3) are carried out eluting, regeneration and reequilibrate; Sample infusion pump (1) in turn switches on sample flow path selector valve (5), a flow path selection valve (7), a chromatographic column (C1), No. four flow path selection valve (10), No. two flow path selection valve (8), No. two chromatographic columns (C2), No. five flow path selection valve (11), No. seven flow path selection valve (13) and sample detector (3) simultaneously, sample infusion pump (1) is to chromatographic column (C1) loading so that No. two chromatographic columns (C2) can be caught in a chromatographic column (C1) owing to loading crosses the object that current-carrying is worn;
The continuous of step A～G circulates, it is possible to achieve while the uninterrupted loading to sample, and continuous print collects the target substance after separating, the object that when the staggered object due to the outflow of outer water reason collecting elution stage and overload loading, stream is worn simultaneously.
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