CN100444920C - Simulated moving bed chromatographic focusing - Google Patents
Simulated moving bed chromatographic focusing Download PDFInfo
- Publication number
- CN100444920C CN100444920C CNB2003801035972A CN200380103597A CN100444920C CN 100444920 C CN100444920 C CN 100444920C CN B2003801035972 A CNB2003801035972 A CN B2003801035972A CN 200380103597 A CN200380103597 A CN 200380103597A CN 100444920 C CN100444920 C CN 100444920C
- Authority
- CN
- China
- Prior art keywords
- district
- liquid
- displacement
- chromatographic column
- discharge pipe
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Images
Landscapes
- Treatment Of Liquids With Adsorbents In General (AREA)
Abstract
A continuous simulated moving bed chromatographic focusing process is disclosed for separating and concentrating multiple components from a complex mixture. Said process comprises a number of zones that are connected in series. An eluent flows unidirectionally along the zones and decreases its desorption strength by sequential dilutions zone after zone. Each zone is detachably installed with a chromatographic column and the columns are periodically relocated one zone upstream. Multiple components of a sample are introduced into a downstream zone, absorbed on the column in the zone, brought zone by zone upstream by column relocations, and selectively desorbed from different zones to get separations. A component desorbed from one zone is reabsorbed by the column in the adjacent downstream zone and is again brought back to said zone by column relocation said component is thus retained between said zone and said adjacent downstream zone and is accumulated by continuous sample feeding and repeatedly column relocation.
Description
Technical field
The present invention relates to a kind of method of using the simulation moving-bed structure of multizone to carry out continuous chromatography.More precisely be a kind of focus on separate and enriched mixture in different single composition in the invention in zone separately.
Background technology
Chromatography can be divided into dual mode: chromatography mode and continuous chromatography mode in batches.Most conventional chromatography takes in batches the chromatography mode to carry out.Usually, chromatography separates sample in batches in batches.The separation process of each batch constitutes by steps such as identical last sample, wash-out and fraction collection, chromatographic column regeneration.The advantage of chromatography is to be used for the separate complex compound in batches.But for to guarantee good separating effect, applied sample amount should be lower than 1% of chromatographic column cumulative volume.This is one of chromatography mode defective in process of production in batches.Owing to finishing back next batch separation fully, last batch of separation process just can begin again, so another shortcoming of chromatography mode is to need the separation of cost plenty of time obtain quantitative product in batches.Simultaneously, limited chromatographic column length has hindered the separation of analogue compounds, and this is the another one shortcoming of chromatography in batches.
Be different from chromatography in batches, continuous chromatography is a process that goes up sample, wash-out, fraction collection continuously.The injection of sample, wash-out and fraction collection carry out simultaneously.Thus, increased applied sample amount; Improved production efficiency; Eluant, eluent and filler have been saved.Simulated moving bed chromatography system (hereinafter to be referred as the SMB system) is typical continuous chromatography.Fig. 1 has shown the course of work of SMB system.As shown in the figure, the SMB system is made up of several districts, and these districts are connected into a cover can be by the circular loop of liquid.Chromatographic column be placed in each district and can be simultaneously in the same way from move to next district when the proparea.Two inlets and two liquid outlets are arranged respectively in this loop.Import F is used to supply with sample, and import D is as the input stripping liquid.Outlet R is as collecting weak adsorption component (raffinate), and outlet E is as collecting strong adsorption component (extract).Liquid flow direction is opposite with the chromatographic column moving direction in the loop.Sample is by the continuous injected system of import F.Weak adsorption component is come out by wash-out from outlet R, and strong adsorption component is retained in the chromatographic column, along with chromatographic column is distinguished to moving to the next one along the back washing off-square.Final strong adsorption component is come out by wash-out from outlet E.The separation that is applied to two constituents mixts of SMB system success, as: separation mirror isomers, sucrose and non-sugar ingredient, dimethyl benzene and isomer thereof.Outstanding high yield makes chromatography have more the major advantage that economy is the SMB technology.Yet it can only be used to separate two constituents mixts the characteristic limitations of SMB technology, and can not be used to a plurality of compositions in complex mixture of separation and purification simultaneously.Even be used to once in a while extract some compositions from complex mixture, it must be the strongest or the most weak also requiring the adsorption capacity of this composition on filler to compare with the adsorption capacity of other composition, otherwise the SMB system can't extract it from mixture.
The limitation of SMB system applies comes from its separation principle.A composition in the mixture may move along both direction in the SMB system.One is the stripping liquid wash-out, and composition (along the wash-out direction) downstream moves, two for chromatographic column reset drive composition upstream (back washing off-square to) mobile.When the chromatographic column travel frequency is fast, all become branch's upstream direction to move.If the adsorption capacity difference of two compositions on chromatographic column, when they by simultaneously from import F injected system the time, they have different translational speeds.If the travel frequency of the flow velocity of stripping liquid and chromatographic column is suitably regulated, then the one-tenth branch upstream direction of high adsorption capacity moves and comes out from outlet E wash-out, and the weak composition downstream of adsorption capacity moves and comes out from outlet R wash-out.
But when also being mixed with other composition in this mixture, they will move and its pollution the most at last along with the composition of high adsorption capacity or with the composition a little less than the adsorption capacity.By the same token, for the separate complex mixture more multi-region and the more also not success of trial of multiple exit are set in the SMB system.When the adsorption capacity of certain composition in the complex mixture when not being strong or the most weak, other component separation that the SMB system equally can not be similar to adsorption capacity with it comes.Therefore, be the separate complex mixture, separation principle must be different with the principle of SMB system.
SMB chromatography process is a dilution.The concentration of composition behind the purifying in cut be its in sample original content 1/tens.Be cut and the saving eluent that obtains high concentration, several patents have in the past been used gradient elution in the SMB system.These patents comprise: United States Patent (USP) (patent No.: US 4031155, US 4031156); U.S. Patent application (20020017492, WO 0033943).The way of gradient elution is: utilizes stripping liquid and sample solvent desorption ability different in chromatographic column, produces two desorb intensity along the wash-out direction, thereby with gradient mode sample separation composition.This can be saved stripping liquid by the improvement of patent protection, and makes its concentration in raw sample of concentration ratio of extract in the cut higher.Yet the principle of typical SMB system has been continued to use in these several inventions, so still can only separate two composition samples.Use the purpose of gradient elution to be that the composition that upstream direction is moved is come out by wash-out from system faster, saves stripping liquid simultaneously.Same gradient elution mode is also described in following article: Dorota Antos et al. (Chemical Engineering Science, VOL.56, (2001) 6667-6682), Thomas.B.Jensen et al. (Journal of Chromatography A, 873 (2000) 149-162), Stefanie Abel etal. (Journal of Chromatography A, 944 (2002) 23-29), Joukje Houwing et al. (Journalof Chromatography A, 944 (2002) 189-201), Dorota Antos et al. (Journal ofChromatography A, 944 (2002) 77-91), Joukje Houwing et al. (Journal of ChromatographyA, 952 (2002) 85-98).United States Patent (USP) (patent No.: US 6069289) discloses another one and has used two kinds of stripping liquids to carry out the process of gradient elution.This method can be used for separation and contain three kinds of mixture of ingredients.This patent focuses on three kinds of compositions in the separating mixture how, the low concentration problem of cut is not improved.Because the production process of this patent requires to contain 2.2-10 that the volume of the cut of extract is necessary for sample volume on the sample doubly.Also be that the concentration of separating composition in the tails is 0.1-0.45 times of concentration in raw sample.Simultaneously, this patent has also adopted typical SMB system principle.
In sum, for separating a complex mixture, the continuous chromatography process must be used the principle that is different from typical SMB system.This system should be able to the separate complex mixture, should be a low consumed continuous process of high production, effectively to save filler and eluant, eluent, higher productive rate is arranged, and can obtain the cut concentration higher than raw sample concentration.
Summary of the invention
Be the separate complex mixture, the focusing unit (hereinafter to be referred as the F unit) that simulated moving bed chromatographic focusing system (hereinafter to be referred as SMBCF) forms be designed to key features of the present invention.The F unit constitutes by being marked as F (1) several districts to F (f), and these districts are connected on together, and a chromatographic column is installed in each district.The main eluent of system enters from F (f) district and flows out from F (1) district.When main eluent flow out from a district, when entering contiguous next one district, system modifies to reduce its desorption ability main eluent.At least comprise several districts in the F unit, the desorption ability of the eluent that wherein passes through differs from one another.F (1) district and F (f) district can adjacently put, and make these districts that are cascaded form an annulus.But may not having liquid between F (1) district and F (f) district exchanges.Chromatographic column in all districts all periodically flows direction from moving to adjacent region when the proparea against main eluent simultaneously.Sample mixture injects from a certain district in downstream, be adsorbed on the chromatographic column in this district, and along with chromatographic column back washing off-square to move met a district by a district band upstream.Each composition in the sample in same district not successively under the desorb, thereby separated.The composition that is focused is present in F (i+1) district that the F (i) that contains low desorb intensity eluent distinguished and contained higher desorb intensity eluent dynamically.The weak composition of absorption affinity is entered F (i) district by main eluent downstream wash-out in F (i+1) district, and moves into mixture in F (i) district with chromatographic column converge and be adsorbed on again in the chromatographic column in F (i) district from the catchment.Composition comes to this and is accumulated and along with moving of chromatographic column is brought into again in F (i+1) district.Because what chromatographic column did not stop moves and the continuous sample introduction of system, be focused just accumulating gradually between F (i) and F (i+1) distinguish of composition.If between F (i) and F (i+1) district, from main eluent, collect a small size cut, can obtain the high concentration composition that a purifying is crossed at every turn.
The present invention has realized with this target of continuous chromatography method separate complex mixture, and this target almost is not reached with existing chromatography method.Method of the present invention can effectively increase productivity, reduce the processing procedure after production consumption, increase product yield and purity, the simplification chromatography.Contrast chromatography mode in batches, mode of the present invention can effectively be saved chromatography media, solvent and disengaging time, and can effectively improve productive rate.With the comparison of SMB chromatography mode, the present invention afterwards obtains to have superiority aspect the high concentration composition with separating in the separation of complex mixture.
Description of drawings
Fig. 1 is the diagram of classical SMB chromatography process.
Fig. 2 a, 2b are first kind of assembling mode diagram of the present invention.
Fig. 3 is second kind of assembling mode diagram of the present invention.
Fig. 4 is the third assembling mode diagram of the present invention.
Fig. 5 is the 4th kind of assembling mode diagram of the present invention.
Fig. 6 is the 5th kind of assembling mode diagram of the present invention.
Fig. 7 is the 6th kind of assembling mode diagram of the present invention.
Fig. 8 is an improvement diagram of liquid stream collection method.
Fig. 9 is another improvement diagram of liquid stream collection method.
The specific embodiment
The present invention uses the focusing unit that comprises several districts to realize that continuous chromatography focuses on.Shown in accompanying drawing 2a, there are a chromatographic column liquid feeding end interface 1 and a chromatographic column outlet end interface 2 in each district.Interface 1 is connected with ascending pipe 3, and leaves opening 5 on ascending pipe 3.Equally, interface 2 is connected with discharge pipe 4, and leaves opening 6 on discharge pipe 4.Chromatographic column 7 is connected between interface 1 and the interface 2 by dismountable.Chromatographic column 7 can be filled mutually by any chromatography is fixing, comprising: positive, anti-phase, ion exchange phase, affinity phase, chelating phase, chirality phase, ligand exchange phase, molecular sieve mutually with organic polymer mutually.Chromatographic column 7 also can be filled mutually by monolithic.Interface 1 and interface 2 can be made of any connected mode, and these connected modes not only will allow liquid stream in ascending pipe 3, chromatographic column 7 and discharge pipe 4 intermediate flow, and also will guarantee does not have fluid seepage when mobile when chromatographic column 7 is rapid in the focal zone.Using the separating liquid rotary valve is the connected mode of typical interface 1 and interface 2.Opening 5 is used for carrying out liquid stream with proximity with opening 6 and is connected.Ascending pipe 3 is used for the injection of liquid stream.Discharge pipe 4 is used for the discharge of liquid stream.The injection of liquid stream and discharge all are subjected to a device control.But duration, volume and speed that this device regulator solution stream injects and discharges.
Accompanying drawing 2b has showed the operating process of the present invention.This process comprises following operation:
1.F the unit is used for chromatofocusing.The F unit comprises f district (f is the integer greater than 4).These are distinguished by continuous lining up, and with the sequence notation of F (f) to F (1).Wherein F represents the F unit, the position of a district of the digitized representation in the bracket in the F unit.F (f) district is defined as upstream extremity, and F (1) district is defined as downstream.Opening 6 in opening 5 in a certain district on the ascending pipe and the adjacent upstream on the discharge pipe links to each other, and constitutes passage 8.Because F (f) district and F (1) district are the two ends of F unit, institute thinks the discharge pipe discharge that makes main eluent enter and distinguish from F (1) from the ascending pipe in F (f) district, can be with closure of openings or the cancellation on above-mentioned two pipes.
2. in the F unit, select at least 4 ascending pipes and at least 2 discharge pipes to be used for injection and the discharge that liquid flows, seal remaining ascending pipe and discharge pipe.Set the composition and the flow velocity of the liquid stream that passes through in each selected ascending pipe and the discharge pipe, thereby in the F unit, form the desorb gradient of an expection.This gradient can be: ionic strength gradient, close ester gradient, PH gradient, chelating gradient, hydrogen bond gradient, affinity gradient, chirality gradient and other any chromatography gradient.Not chosen ascending pipe recited above and discharge pipe also can be open-minded again arbitrarily on demand, and close branch branch ostium on it simultaneously to finish liquid communication.
3. carrying out continuous chromatography focuses on
(1) by selected ascending pipe and discharge pipe, with flow velocity and the composition of setting, continuously and injection simultaneously and discharge following liquid:
Inject the main eluent that high desorb intensity is arranged from the ascending pipe in F (f) district;
Inject sample liquid from least one selected ascending pipe;
Inject the focusing liquid of low desorb intensity from remaining selected ascending pipe.Focus on liquid and mix, constitute the wash-out that a lower eluent of desorb intensity is used for this district from the ascending pipe injection in a certain district and with the eluent that elutes from adjacent upstream.The desorb gradient produces with the method exactly.Be the strongest desorb intensity of F (f) Qu Zhongyou, the most weak desorb intensity of F (1) Qu Zhongyou;
Collect corpusculum hydrops stream from selected discharge pipe, and collect main eluent from the discharge pipe in F (1) district;
(2) periodically reset chromatographic column in F (f) district to F (1) district, and make all the other chromatographic column upstream direction move a district at synchronization to carry out continuous chromatography focusing.Now focusing principle is set forth as follows: a mixture is injected the F unit by the ascending pipe from a certain district, some compositions in the mixture have very weak adsorption capacity in the chromatographic column in this district, so they are entered into the catchment by the eluent wash-out in this district.Other composition in the mixture then is adsorbed on the chromatographic column, and the band that is met a district by a district along with the replacement of chromatographic column upstream.Again because the difference of desorb intensity in the same district not, thus each composition got off by the desorb of selectivity, thereby be able to and other component separation.One in a certain district the composition under the desorb brought back to adjacent catchment by eluent, be adsorbed in (mixture that contains new injection in this chromatographic column) on the chromatographic column again at that.This composition is because the replacement next time of chromatographic column, brought back to again in the district that it was once got off by desorb.Along with the replacement repeatedly of continuous last sample and chromatographic column, this composition just is maintained between two above-mentioned districts, thereby with other component separation and concentrated.If collect corpusculum hydrops stream from the discharge pipe in above-mentioned district, just can obtain the high concentration composition that purifying is crossed.Same, those in above-mentioned two districts not the composition under the desorb will be brought to farther upstream, remain in the district that is fit to separately, with other component separation and focus on.Opposite, will be by the downstream wash-out by the composition of aforementioned two district's absorption, keep in the same way and focus on suitable separately position.
Fig. 3 has shown another concrete form of the present invention.Fig. 3 has increased a R unit that is used for chromatographic column regeneration on the basis of Fig. 2 a, 2b.This improvement is the extension to Fig. 2 a, 2b form.The R unit comprises r district (r is an integer).The sequence notation of these districts from R (r) to R (1), and with the identical connected mode connection in Fig. 2 b F unit.The R unit is placed in the upstream of Fig. 2 b F unit, makes R (1) district adjacent with F (f) district.As setting up a flow channel, can seal or remove the opening 6 on the discharge pipe 4 during the opening 5 on the ascending pipe 3 and R (1) are distinguished in R (r) district from the ascending pipe in R (r) district to the discharge pipe in R (1) district.If in the R unit, there is not extra liquid stream to inject and collection process, the preferably ascending pipe of sealing except that R (r) district in and whole ascending pipes and the discharge pipe the discharge pipe in R (1) district, or use and do not have the alternative branch of the pipeline branch ostium formation fluid course that divides branch ostium.Improvement to process shown in Fig. 2 b also comprises:
(1) injection and the discharge of additional liquid stream: inject regenerated liquid continuously and receive this regenerated liquid from the discharge pipe in R (1) district from the ascending pipe in R (r) district.
(2) improvement that chromatographic column is reset: the chromatographic column in the R (r) that periodically resets the simultaneously district is to F (1) district, and the chromatographic column in replacement F (f) district is to R (1) district, with remaining chromatographic column from be reset to corresponding adjacent upstream district when the proparea.
The described process of Fig. 3 is to remove to be adsorbed on the impurity on the chromatographic column in F (f) district securely.Regenerated liquid (regeneration solution) has than the stronger desorption ability of eluent in F (f) district.When chromatographic column when F (f) district is reset to the R unit, regenerated liquid can be from chromatographic column the aforesaid impurity of flush away.In some cases, the R unit can comprise a district, and regenerated liquid enters and discharges from discharge pipe from the ascending pipe in this district.In the R unit, use the advantage in a plurality of districts to be to save regenerated liquid.
Because chromatographic column can be reset to F (1) district in the F unit from the R the R unit (r) district, so this process will be brought the regenerated liquid of high desorption ability into F (1) district, that is to say, have the liquid of high eluting power will temporarily be present in F (1) district along with finishing that each chromatographic column is reset.If lower and stable this condition of desorb intensity is to the vital words of wash-out in F (1) district, above-mentioned situation will influence the application of native system.Form shown in Figure 4 has promptly solved this problem.Fig. 5 has increased the balance that a B unit carries out chromatographic column on the basis of Fig. 3.The B unit comprises b district (b is an integer).The sequence notation of these districts from B (b) to B (1), and with the identical connected mode connection in Fig. 3 F unit.The B unit is placed in the upstream of Fig. 3 R unit, makes B (1) district adjacent with R (r) district.As setting up a flow channel, can seal or remove the opening 6 on the discharge pipe 4 during the opening 5 on the ascending pipe 3 and B (1) are distinguished in B (b) district from the ascending pipe in B (b) district to the discharge pipe in B (1) district.If in the B unit, there is not extra liquid stream to inject and collection process, the preferably ascending pipe of sealing except that B (b) district in and whole ascending pipes and the discharge pipe the discharge pipe in B (1) district, or use and do not have the alternative branch of the pipeline branch ostium formation fluid course that divides branch ostium.Improvement to process shown in Figure 3 also comprises:
(1) injection and the discharge of additional liquid stream: inject equilibrium liquid continuously and receive this equilibrium liquid from the discharge pipe in B (1) district from the ascending pipe in B (b) district.
(2) improvement that chromatographic column is reset: the chromatographic column in the B (b) that periodically resets the simultaneously district is distinguished to F (1); Chromatographic column in replacement F (f) district is distinguished to R (1); Chromatographic column in replacement R (r) district is distinguished to B (1).Remaining chromatographic column is reset to corresponding adjacent upstream district from working as the proparea.
In some cases, the B unit can comprise a district, and equilibrium liquid enters and discharges from discharge pipe from the ascending pipe in this district.In the B unit, use the advantage in a plurality of districts to be to save equilibrium liquid.
Different with this method of sample on the ascending pipe in a certain district from the F unit shown in Fig. 2 a, the 2b, 3 and 4, beyond having shown from the F unit, goes up Fig. 5 the method for sample.Fig. 5 is adding a S unit that is used for sample on Fig. 4 basis.The S unit comprises s district (s is an integer).The sequence notation of these districts from S (s) to S (1), and with the identical connected mode connection in Fig. 4 F unit.The S unit is placed in the upstream of B unit among Fig. 4, makes S (1) district adjacent with B (b) district.As setting up a flow channel, can seal or remove the opening 6 on the discharge pipe 4 during the opening 5 on the ascending pipe 3 and S (1) are distinguished in S (s) district from the ascending pipe in S (s) district to the discharge pipe in S (1) district.If in the S unit, there is not extra liquid stream to inject and collection process, the preferably ascending pipe of sealing except that S (s) district in and whole ascending pipes and the discharge pipe the discharge pipe in S (1) district, or use and do not have the alternative branch of the pipeline branch ostium formation fluid course that divides branch ostium.Improvement to process shown in Figure 4 also comprises:
(1) injection and the discharge process of additional liquid stream: inject sample liquid continuously and flow out liquid from the discharge pipe reception in S (1) district from the ascending pipe in S (s) district.In the F unit, there is not the injection of sample.
(2) improvement that chromatographic column is reset: the chromatographic column in the R (r) that periodically resets the simultaneously district is distinguished to B (1); Chromatographic column in replacement B (b) district is distinguished to S (1); Chromatographic column in replacement S (s) district is distinguished to F (1); Chromatographic column in replacement F (f) district is distinguished to R (1).Remaining chromatographic column is reset to corresponding adjacent upstream district from working as the proparea.
In some cases, the S unit can comprise a district, and sample liquid is injected from the ascending pipe in this district, discharges from discharge pipe.In the S unit, use the advantage in a plurality of districts to be to inject a large amount of samples and to prevent sample loss.
When not having impurity absorption in F (f) district and require to keep lower and stable eluotropic strength in F (1) district, form shown in Figure 6 just can satisfy this condition.Form shown in Figure 6 is the simplification to form shown in Figure 5.B in Fig. 6 (1) district is adjacent with F (f) district, has removed the R unit among Fig. 5.Improvement to process shown in Figure 5 also comprises:
(1) to the injection of liquid stream and the improvement of discharge process: cancelled the injection and the discharge of regenerated liquid.
(2) improvement that chromatographic column is reset: the chromatographic column in the F (f) that periodically resets the simultaneously district is distinguished to B (1); Chromatographic column in replacement B (b) district is distinguished to S (1); Chromatographic column in replacement S (s) district is distinguished to F (1).Remaining chromatographic column is reset to corresponding adjacent upstream district from working as the proparea.
Same, from Fig. 6, can remove the B unit.Different unit further constitutes other combining form, has more than to be limited to several that the present invention exemplifies.
If in some applications, do not have impurity in the eluate (flow out fluid) in F (1) district and do not have then to make up shown in Fig. 2 b and can be modified to combination shown in Figure 7 on the chromatographic column of impurity absorption in F (f) district.That is: the opening 6 during the opening 5 on the ascending pipe 3 is distinguished with F (1) in F (f) district among Fig. 2 b on the discharge pipe 4 connects and composes flow channel.Liquid flow path direction control device 10 is installed in the above-mentioned flow channel, makes liquid stream flow back to F (f) district from F (1) district.Above-mentioned liquid flow path direction control device 10 comprises: check valve, fluid pump and other are used to form the conventional equipment of one-way flow.The advantage of Fig. 7 is to save eluent.
With Fig. 2 a, 2b to continuous liquid collection mode shown in Fig. 7 different be that Fig. 8 has shown a non-continuous process by liquid stream displacement carrying out fraction collection in the F unit.The displacement of liquid stream allows the special time in operating process selectively to collect cut.Collection process can carry out also can carrying out once after chromatographic column replacement several times after each chromatographic column is reset.Liquid stream substitute mode among Fig. 8 is collected for liquid elasticity is provided, and obtains to have advantage aspect the purer product how.As shown in Figure 8, the displacement of liquid stream betides F (i) district (i is an integer less than f) of F unit.Triple valve 11 is installed in the branch location of ascending pipe and discharge pipe in F (i) district.In the routine operation of Fig. 7, triple valve is in full-gear at Fig. 2.In order to carry out the displacement of liquid stream, routine operation is discontinuous.And following operation bidirectional can take place:
(1) in the F unit, suspends injection and the discharge that liquid flows at least simultaneously.
(2) switch three-way valve is closed the fluid exchange between the discharge pipe in the ascending pipe in F (i) district and F (i+1) district; Close the fluid exchange between the ascending pipe in the discharge pipe in F (i) district and F (i-1) district.Form the flow channel of a discharge pipe of distinguishing from the ascending pipe in F (i) district to F (i) as shown in Figure 8 with this.
(3) ascending pipe from F (i) district injects a replacement fluid, and collects this displacement liquid by the discharge pipe in F (i) district.
(4) switch three-way valve is got back to full-gear, restarts Fig. 2 to operating process shown in Figure 7.
Occur in conventional liq with shown in Figure 8 different, shown in Figure 9 liquid displacement and flow when carrying out, rather than occur in conventional liq flow suspend after.As shown in Figure 9, triple valve 11 is placed in the branch location of F (i) district ascending pipe.Triple valve is in full-gear during routine operation, and can be switched to a position and close fluid exchange between the discharge pipe in the ascending pipe in F (i) district and F (i+1) district, allow to collect whole liquid of coming from F (i+1) district with this from the discharge pipe in F shown in Figure 9 (i+1) district.Simultaneously, the focusing liquid that should inject from the ascending pipe in F (i) district is replaced by a displacement liquid, and this displacement liquid has and the identical desorb intensity of eluent during F (i) distinguishes.After liquid displacement, this displacement liquid is gained routine and is focused on liquid, and triple valve is switched back full-gear, and routine operation is restarted again.Above process has the advantage identical with process shown in Figure 8.
The present invention can realize chromatographic column reset process simultaneously with the similar separating liquid rotary valve of the described technology of United States Patent (USP) (US, the patent No. is 3706812) with one.This process can be realized by the separating liquid rotary valve of two rotations synchronously equally: a shunting rotary valve is installed in ascending pipe one end and connects as injecting, another is installed in discharge pipe one end as discharging connection, makes the injection of liquid stream and discharge to take place on the flow channel between two adjacent regions simultaneously.The replacement of chromatographic column also can realize by a mechanical process.This process comprises: suspend in the time of liquid flow; Chromatographic column is from breaking away from when the proparea; Reinstall chromatographic column restarting to adjacent upstream and liquid flow.Any method that is used for the chromatographic column replacement all can be used for this process of the present invention.
The present invention is the different of separation principle with former simulated moving bed chromatography system (being designated hereinafter simply as SMB) technology major different.The separation principle of SMB chromatography is that two groups or two kinds of compounds are pushed to upstream district and downstream district respectively from last sample district, and receives diluted composition respectively in these two districts.Separation principle of the present invention is heterogeneity to be retained in the given zone and from these distinguish the composition of collecting after concentrating.Because the difference of separation principle makes the present invention have many and the visibly different characteristics of prior art.
First characteristics of the present invention are: form eluent desorb gradient at the focusing unit that is connected (F unit).In theory, the eluent in each district all has desorb intensity separately.Each district all can keep a specific composition.The desorb intensity level that presents in focusing unit is many more, just has more composition energy separated.Between these districts, the effect of the more little separation of difference of desorb intensity is good more.Even, still multiple desorb strength grade need be arranged in focusing unit from complex mixture, separating under a kind of situation of composition.For example: a target component is focused between F (i) district and F (i+1) district, if the eluotropic strength difference between these two districts is excessive, then target component can be polluted by other composition in this complex mixture; If the eluotropic strength difference between these two districts is too small, then target component can be diffused into beyond these two districts, and is overlapping with other composition, weakens focussing force.For target component and other component separation are opened and made it to be accumulated between F (i) district and F (i+1) district, in the catchment in F (i) district, should there be an eluotropic strength to be lower than the eluent in F (i) district at least; At least in the upstream in F (i+1) district, should there be another eluotropic strength to be higher than the eluent in F (i+1) district simultaneously.This has just determined that each contiguous area has at least the gradient of four different eluotropic strengths to exist in the focusing unit, to carry out chromatofocusing of the present invention.Requirement to eluotropic strength number in the focusing unit is one of feature of technology before the present invention obviously is different from.Although the technology before a lot of has been used gradient separations in the SMB system, in gradient, only there are two desorb intensity and just are used for binary and separate.
Shown in Fig. 2 b, a composition is focused between two adjacent focal zone F (i) and F (i+1) dynamically.This composition should be adsorbed in the chromatographic column in F (i) district, and when this chromatographic column was reset to F (i+1) district of upstream from F (i) district, this composition should be got off by desorb from chromatographic column.Owing to this reason, the eluotropic strength of eluent must be lower than the eluotropic strength of eluent in F (i+1) district in F (i) district.Also because of same reason, the focusing liquid of a low eluotropic strength should be introduced in F (i) district and F (i+1) flow channel between distinguishing.Because of this composition by after desorb is got off from F (i+1) district again wash-out get back to F (i) district, so the collection of this composition also should occur in the flow channel between above-mentioned two districts.Both having focused on the injection of liquid and the collection of composition is to occur on the same flow channel that connects between F (i) district and F (i+1) district.The liquid that technology before all is carried out for the formation gradient injects with collecting and all occurs on the different flow channels.This is the present invention and the former obvious difference of technology.
" focusing " speech meaning in the present invention is for keeping and accumulation.In order to obtain purifying
, the composition of high concentration, what the volume that composition is collected should be relative is little.Should be from a collected cut volume of given zone less than 50% of the effluent volume that passes through this district.This point obviously is different from United States Patent (USP) 6069289 disclosed technology (although this patent has been used two kinds of eluents in the SMB system).As patent 6069289 is described: " flow velocity of second stripping liquid should be raw material inject flow velocity 2.5-12 doubly.The flow velocity of collecting second cut approximately is 90% of the second stripping liquid flow velocity.Although " the 6069289 disclosed specification requirements protections of this patent be to separate a mixture to become three parts, be not that the binary of other patented technology is separated, it still all is different from the present invention fully in mechanism and operation.
Compared with prior art the present invention has many advantages.It is a high yield, low consumption, continuous chromatography process; It has saved a large amount of chromatographic stuffings and solvent; Handle the required time again after having saved separation and having separated.Technology of the present invention can accumulate the composition of low concentration in a certain district, when the concentration of this composition began to raise, pollutant wherein can be squeezed away from this district.In this way, the present invention can provide purer product.
The present invention has very high purifying productive rate equally.In the LC of routine, a certain composition is eluted in several cuts usually, but has only 1 to 2 cut that gratifying purity is arranged.Other cut is then by contaminating impurity.Be in economic consideration and abandon these cuts usually, also therefore greatly reduce productive rate.In the present invention, required composition from and only be collected from that the highest district of purity, contaminated that part of composition still is retained in the system in adjacent region, is further purified and focuses on that the highest district of purity up to them.Therefore the amount of composition is not suffered a loss.
The present invention can use in the laboratory, also can use in industrial processes.Instrument based on focusing principle of the present invention can be combined and be used for the laboratory and separate with industrial.Generally, any simulation moving-bed design that can be used for chromatofocusing process of the present invention all belongs to scope of the present invention.
Example 1: the chromatography of gradient in batches of pig serum
Be filled to the chromatographic column of internal diameter 4.6mm, length 180mm with DEAE-Sepharose ion-exchange resin (particle size 50 μ m are from Pharmacia company).Sample is that 10mM KCl pellicle filters pig serum.Filling 1ml sample is used KCl solution (concentration is from 0.01M to 0.4M) the elution chromatography post of 30ml linear gradient then in chromatographic column, collect 30 cuts.In the Laemmli Laemmli buffer system Laemmli with 10% non-these cuts of modified polyacrylamide gel electrophoresis analysis.The albumin of maximum appears among the cut 14-20.Weak bands of a spectrum (called after composition B) come out with albumin common wash-out in cut 17 and cut 18.
Example 2: simulated moving bed chromatographic focusing (hereinafter to be referred as SMBCF) separates pig serum
Form is formed the SMBCF system as shown in Figure 5.The F unit comprises 5 districts, and S unit, B unit, R unit respectively comprise a district.A chromatographic column (30mm * 4.6mm, length * internal diameter) is arranged in every district, fill with DEAE-Sepharose ion-exchange resin (50 μ m are from Pharmacia company).System operates as described in Figure 5 like that.Carrying out a chromatographic column in per 1.5 minutes resets.With handling pig serum as sample with example 1 identical mode, the composition of eluent and injection flow velocity are referring to table 1 in whole 8 districts.System carries out 40 times chromatographic column reset process.Each chromatographic column is reset, and collects whole eluates in S (1) district and F (1) district; Respectively collect 0 from F (2), F (3), F (4), F (5) district.The eluate of 025ml.The cut of collecting when under the Laemmli Laemmli buffer system Laemmli, using the 10th, 20,30,40 chromatographic columns of 10% non-modified polyacrylamide gel electrophoresis analysis to reset.Composition B is mainly focused in the cut in F (3) district and is presented very strong bands of a spectrum.The intensity of these bands of a spectrum is along with the increase of chromatographic column replacement number of times strengthens gradually.The bands of a spectrum of composition B are separated with albumin well.Albumin then focuses on outside the focal zone and is reproduced liquid and goes out at R (1) district wash-out.
Table 1
Example 3: by the pig serum SMBCF separation process of liquid stream displacement carrying out liquid stream collection
Form the SMBCF system as shown in Figure 5.The mode of use-case 2 is separated.But wherein the liquid in F (3) district is collected with liquid stream substitute mode shown in Figure 8 and is carried out.Per 5 chromatographic columns are reset and are collected a liquid.Each collection uses the NaCl solution 0.05ml of 0.17M as displacement liquid.Other liquid receives identical with method in the example 2.The cut of collecting when under the Laemmli Laemmli buffer system Laemmli, using the 10th, 20,30,40 chromatographic columns of 10% non-modified polyacrylamide gel electrophoresis analysis to reset.With example 2 relatively, the bands of a spectrum of composition B are stronger and purity is higher in the collected cut in F (3) district.
Example 4: the pig serum SMBCF separation process of carrying out with the mode of Fig. 2 b.
Shown in Fig. 2 b, form the SMBCF system.F unit in this system comprises 6 districts.Each district all comprises a chromatographic column (30mm * 4.6mm, length * internal diameter) of being filled by DEAE-Sepharose ion-exchange resin (50 μ m are from Pharmacia company).System operates according to the mode shown in Fig. 2 b, carries out a chromatographic column and resets in per 1.5 minutes.Mode in the sample pig serum use-case 1 is handled.Inject the liquid composition in 6 districts, F unit and inject flow velocity referring to table 2.System carries out 40 chromatographic columns altogether and resets.Each chromatographic column is reset, and collects whole eluates in F (1) district, and collects each 0.03ml of eluate in F (2), F (3), F (4), F (5), F (6) district.Under the Laemmli Laemmli buffer system Laemmli, use the 10th, 20,30,40 collected cuts of chromatographic columns replacement of 10% non-modified polyacrylamide gel electrophoresis analysis.The protein albumin of content maximum, most composition all focuses in the single district in serum.Albumin then is distributed between F (3) district and F (4) district.At this moment, the concentration of each composition approximately is 10 times of concentration in raw sample in the cut.
Table 2
Example 5: the monoclonal antibody SMBCF separation process of carrying out with the mode of Fig. 2 b
Shown in Fig. 2 b, form the SMBCF system.F unit in this system comprises 7 districts.Each district all comprises a chromatographic column (30mm * 4.6mm, length * internal diameter) of being filled by cation exchange resin (50 μ m).System operates according to the mode shown in Fig. 2 b, carries out a chromatographic column and resets in per 1.5 minutes.With the Chinese vole cell culture medium that contains monoclonal antibody as sample.Inject the liquid composition in 7 districts, F unit and inject flow velocity referring to table 3.System carries out 40 chromatographic columns altogether and resets.Each chromatographic column is reset, and collects whole eluates in F (1) district, and collect F (3), F (5), F (7) district eluate each 0.03ml。Under the Laemmli Laemmli buffer system Laemmli, use the 10th, 20,30,40 collected cuts of chromatographic columns replacement of 10% non-modified polyacrylamide gel electrophoresis analysis.The result shows that most impurity is present in the cut in F (1) district, F (3), F (7) district.In the collected cut in F (5) district, contain monoclonal antibody.In whole process, the concentration of the monoclonal antibody that the cut of collecting from F (5) district contains raises gradually, and purity is about 98%.Purity that it should be noted that antibody is carrying out and raise with whole process also.
Table 3
Example 6: the TSPG SMBCF separation process of carrying out with the mode of Fig. 2 b
Shown in Fig. 2 b, form the SMBCF system.F unit in this system comprises 6 districts.Each district all comprises a reversed phase chromatography post, and (from Phenomenex company, 30mm * 4.6mm), organic facies is the mixture (50: 50) of methyl alcohol and acetonitrile for silica gel C18,20 μ m.System operates according to the mode shown in Fig. 2 b, carries out a chromatographic column and resets in per 1.5 minutes.TSPG is dissolved in 20% methyl alcohol by 5% concentration, with this solution as sample.Inject the liquid composition in 6 districts, F unit and inject flow velocity referring to table 4.System carries out 20 chromatographic columns altogether and resets.Each chromatographic column is reset, and collects whole eluates in F (1) district, and collects each 0.03ml of eluate in F (2), F (3), F (4), F (5), F (6) district.Mixture (20: 80) with methyl alcohol and chloroform is a solvent, with silica gel tlc plate analysis the 10th and the 20th the collected cut of chromatographic column replacement, uses the iodine colour developing to judge the separation case of composition.Can see that protopanoxadiol focuses on F (6) district; The Protopanaxatriol focuses on F (5) district; Panaxoside Rh
1Focus on F (4) district; Panaxoside Rg
2Focus on F (2) and F (3) district.The concentration of ingredient all is higher than its concentration in raw sample in each cut.
Table 4
Claims (35)
1. one kind is used for the continuous method of separating and focusing on each composition of mixture, and the method comprises:
1) f district by arranged in sequence sets up a F unit that is used for chromatofocusing; Described f district is by the sequence notation from F (f) to F (1), f is the integer greater than 4, and F (f) district is defined as upstream extremity, and F (1) district is defined as downstream, all there are a chromatographic column liquid feeding end interface and chromatographic column outlet end interface in each district, removably connects a chromatographic column between two interfaces; This chromatographic column adopts ion exchange phase, the affinity phase, and the chelating phase, the chirality phase, the ligand exchange phase, the molecular sieve phase, monolith column monolith column, the condensate phase, positive, anti-phase or other any chromatography is fixing fills mutually; Chromatographic column liquid feeding end interface links to each other with ascending pipe, and chromatographic column outlet end interface links to each other with discharge pipe; Each ascending pipe and discharge pipe all have a branch branch ostium, and the branch branch ostium on a certain district ascending pipe links to each other with branch branch ostium on the adjacent region discharge pipe of upstream and forms flow channel between two adjacent regions; Branch branch ostium on branch branch ostium on F (f) the district ascending pipe and F (1) the district discharge pipe can be blocked or cancel, and enters the F unit and flows out the F unit from the discharge pipe that F (1) distinguishes from the ascending pipe in F (f) district to guarantee eluent;
2) selected at least 4 ascending pipes and at least 2 discharge pipes are used for the injection and the collection of liquid in the F unit, and have at least an ascending pipe and a discharge pipe on the flow channel to be selected simultaneously; Ascending pipe and discharge pipe that sealing is not selected, composition and flow velocity for each selected ascending pipe and discharge pipe formulation liquid, to form a continuous desorb gradient, this gradient is ionic strength gradient, close ester gradient, PH gradient, chelating gradient, hydrogen bond gradient, affinity gradient, chirality gradient or other any chromatography gradient; Described not chosen ascending pipe and discharge pipe do not contain the branch branch ostium, and are connected to form flow channel between adjacent region by these ascending pipes and discharge pipe;
3) in the F unit, carry out chromatofocusing
A) with pump by selected ascending pipe and discharge pipe, with the flow velocity set with form continuously and injection simultaneously and discharge the liquid of following composition:
Inject the eluent of high desorb intensity from the ascending pipe in F (f) district;
Inject sample liquid from least one selected ascending pipe;
Inject the focusing liquid that hangs down desorb intensity from remaining selected ascending pipe; Focusing on liquid injects and the eluent that elutes at branch's opening part and adjacent upstream mixes from the ascending pipe in a certain district, constitute the wash-out that a lower eluent of desorb intensity is used for this district, in the F unit, produce a desorb intensity desorb gradient from high to low of distinguishing F (1) district from F (f), in each district of F unit, at least 4 desorb strength levels are arranged;
Collect eluate from the discharge pipe in F (1) district, collect cut with flow velocity less than eluent flow rate 50% from the selected discharge pipe of remainder;
B) chromatographic column of periodically resetting in F (f) district is distinguished to F (1), and all the other chromatographic columns to the tight adjacent upstream of resetting simultaneously carries out continuous chromatography focusing, each composition in the sample is injected in certain district in downstream, be adsorbed in the chromatographic column in this district, along with upstream moving of a district distinguished in one of the replacement of chromatographic column, because the difference of desorb intensity in the same district not, each composition optionally never in the same district desorb get off, thereby it is separated from one another, got back to adjacent catchment from the composition that the desorb of a certain district is got off by wash-out, again be adsorbed in there on the chromatographic column that contains new injection sample, this composition is passed through aforesaid Na Yi district once more because chromatographic column is next time reset, and is retained and is accumulated between the catchment adjacent with it, this district owing to going up resetting repeatedly of sample and chromatographic column continuously.
2. by the described method of claim 1, wherein reset process is realized by liquid in district of separating liquid rotary valve conversion chromatographic column the time.
3. by the described method of claim 1, wherein reset process is by reinstalling described chromatographic column with the binding of chromatographic column and in the adjacent upstream district and realize from cutting off simultaneously when the proparea chromatographic column time.
4. by the described method of claim 1, wherein the process of at least one district's collection cut is finished by the displacement of following liquid stream from the F unit:
1) injection and the discharge of stop solution stream simultaneously in the F unit;
2) close flow channel between the discharge pipe in ascending pipe on the liquid stream displacement zone and adjacent upstream district, close the flow channel between the ascending pipe in discharge pipe on this liquid stream displacement zone and adjacent downstream district; The pass closed procedure of above-mentioned path is finished by any conventional blocking way that comprises switch and triple valve;
3) inject displacement liquid and collect displacement liquid from the discharge pipe in this district from the ascending pipe of displacement zone, the composition of the eluent in the composition of above-mentioned displacement liquid and claim 1 displacement zone is identical;
4) be communicated with flow channel again, restart the described operation of claim 1; Above-mentioned closing with opening process finished by the switching of triple valve.
5. by the described method of claim 1, wherein the process of at least one district's collection cut is undertaken by the displacement of following liquid stream from the F unit:
1) close flow channel between the discharge pipe in ascending pipe on the liquid stream displacement zone and adjacent upstream district, from then on upstream is collected cut simultaneously, and injects displacement liquid from the ascending pipe of displacement zone; Above-mentioned process of closing flow channel is finished by any conventional blocking way that comprises switch and triple valve; The composition of the eluent in the composition of above-mentioned displacement liquid and claim 1 displacement zone is identical;
2) path of open and close again stops the injection of displacement liquid simultaneously, restarts the described operation of claim 1; Described closing with opening process finished by the switching of a triple valve.
6. by the described method of claim 1, wherein the branch branch ostium on F (f) the district ascending pipe links to each other to form a liquid stream loop with the branch branch ostium that F (1) distinguishes on the discharge pipe, a liquid part that flows out from F (1) district receives through discharge pipe, another part then flows into F (f) district by the liquid loop, and this liquid stream loop is controlled with pump or check valve.
7. by the described method of claim 6, wherein reset process upstream moves a district and realizes by using the separating liquid rotary valve to switch liquid simultaneously chromatographic column the time.
8. by the described method of claim 6, wherein chromatographic column the time reset process finish be by use mechanical system simultaneously with chromatographic column from realizing when the proparea disconnects and is reinstalled in its adjacent upstream.
9. by the described method of claim 6, wherein the process of at least one district's collection cut is finished by the displacement of following liquid stream from the F unit:
1) injection and the discharge of stop solution stream simultaneously in the F unit;
2) close flow channel between the discharge pipe in ascending pipe on the liquid stream displacement zone and adjacent upstream district, close the flow channel between the ascending pipe in discharge pipe on this liquid stream displacement zone and adjacent downstream district; The pass closed procedure of above-mentioned path is finished by any conventional blocking way that comprises switch and triple valve;
3) inject displacement liquid and collect displacement liquid from the discharge pipe in this district from the ascending pipe of displacement zone, the composition of the eluent in the composition of above-mentioned displacement liquid and claim 6 displacement zone is identical;
4) be communicated with flow channel again, restart the described operation of claim 6; Above-mentioned closing with opening process finished by the switching of triple valve.
10. by the described method of claim 6, wherein the process of at least one district's collection cut is undertaken by the displacement of following liquid stream from the F unit:
1) close flow channel between the discharge pipe in ascending pipe on the liquid stream displacement zone and adjacent upstream district, from then on the discharge pipe in adjacent upstream district is collected cut simultaneously, and injects displacement liquid from the ascending pipe of displacement zone; Above-mentioned process of closing flow channel is finished by any conventional blocking way; The composition of the eluent in the composition of above-mentioned displacement liquid and claim 6 displacement zone is identical;
2) path of open and close again stops the injection of displacement liquid simultaneously, restarts the described operation of claim 6; Above-mentioned closing with opening process finished by the switching of triple valve.
11. by the described method of claim 1: r the additional district that connects with the connected mode of F unit in the claim 1 forms one and is used for the R unit that chromatographic column is regenerated, the sequence notation in R (1) district is distinguished in the R unit from R (r), r is an integer, the R unit is placed in the upstream of the described F of claim 1 unit, make R (1) district adjacent with F (f) district, the branch branch ostium on the ascending pipe in sealing or cancellation R (r) district and the discharge pipe in R (1) district is to form the flow channel of the discharge pipe of distinguishing from the ascending pipe in R (r) district to R (1); If in the R unit, there is not extra liquid stream to inject and discharge, the then ascending pipe of sealing in R (r) district and whole ascending pipes and the discharge pipe the discharge pipe in R (1) district, connect and divide branch ostium to form flow channel, perhaps cancellation divides branch ostium, replaces the branch branch ostium to carry out being connected of flow channel with ascending pipe and discharge pipe; Improvement to claim 1 also comprises:
(1) injection and the collection of additional liquid stream: inject regenerated liquid continuously and collect this regenerated liquid from the discharge pipe in R (1) district from the ascending pipe in R (r) district;
(2) improvement that chromatographic column is reset: the chromatographic column in the R (r) that periodically resets the simultaneously district is distinguished to F (1); Chromatographic column in replacement F (f) district is reset to corresponding adjacent upstream district with remaining chromatographic column from working as the proparea to R (1) district.
12. by the described method of claim 11, wherein reset process upstream moves a district and realizes by using the separating liquid rotary valve to switch liquid simultaneously chromatographic column the time.
13. by the described method of claim 11, wherein chromatographic column the time reset process finish be by use mechanical system simultaneously with chromatographic column from realizing when the proparea disconnects and is reinstalled in its adjacent upstream.
14. by the described method of claim 11, wherein the process of at least one district's collection cut is finished by the displacement of following liquid stream from the F unit:
1) injection and the discharge of stop solution stream simultaneously in the F unit at least:
2) close flow channel between the discharge pipe in ascending pipe on the liquid stream displacement zone and adjacent upstream district, close the flow channel between the ascending pipe in discharge pipe on this liquid stream displacement zone and adjacent downstream district: the pass closed procedure of above-mentioned path is finished by any conventional blocking way that comprises switch and triple valve;
3) inject displacement liquid and collect displacement liquid from the discharge pipe in this district from the ascending pipe of displacement zone, the composition of the eluent in the composition of above-mentioned displacement liquid and claim 11 displacement zone is identical:
4) be communicated with flow channel again, restart the described operation of claim 11.Above-mentioned closing with opening process finished by the switching of triple valve.
15. by the described method of claim 11, wherein the process of at least one district's collection cut is undertaken by the displacement of following liquid stream from the F unit:
1) close flow channel between the discharge pipe in ascending pipe on the liquid stream displacement zone and adjacent upstream district, from then on the discharge pipe in adjacent upstream district is collected cut simultaneously, and injects displacement liquid from the ascending pipe of displacement zone; Above-mentioned process of closing flow channel is finished by any conventional blocking way that comprises switch and triple valve; The composition of the eluent in the composition of above-mentioned displacement liquid and claim 11 displacement zone is identical;
2) path of open and close again stops the injection of displacement liquid simultaneously, restarts the described operation of claim 11; Above-mentioned closing with opening process finished by the switching of triple valve.
16. by the described method of claim 11: b the additional district that connects with the connected mode of F unit in the claim 11 forms one and is used for the B unit that chromatographic column is regenerated, the sequence notation in B (1) district is distinguished in the B unit from B (b), b is an integer, the B unit is placed in the upstream of the described R of claim 11 unit, make B (1) district adjacent with R (r) district, branch branch ostium on the ascending pipe in sealing or cancellation B (b) district and the discharge pipe in B (1) district is to form the flow channel of a discharge pipe of distinguishing from the ascending pipe in B (b) district to B (1), if in the R unit, there is not extra liquid stream to inject and discharge, the then ascending pipe of sealing in R (r) district and whole ascending pipes and the discharge pipe the discharge pipe in R (1) district, connect and divide branch ostium to form flow channel, perhaps cancellation divides branch ostium, replaces the branch branch ostium to carry out being connected of flow channel with ascending pipe and discharge pipe; Improvement to claim 11 also comprises:
1) injection and the collection of additional liquid stream: inject equilibrium liquid continuously and collect this equilibrium liquid from the discharge pipe in B (1) district from the ascending pipe in B (B) district.
2) improvement that chromatographic column is reset: the chromatographic column in the B (b) that periodically resets the simultaneously district is distinguished to F (1); Chromatographic column in replacement F (f) district is distinguished to R (1); Chromatographic column in replacement R (r) district is reset to corresponding adjacent upstream district with remaining chromatographic column from working as the proparea to B (1) district.
17. by the described method of claim 16, wherein reset process upstream moves a district and realizes by using the separating liquid rotary valve to switch liquid simultaneously chromatographic column the time.
18. by the described method of claim 16, wherein chromatographic column the time reset process finish be by use mechanical system simultaneously with chromatographic column from realizing when the proparea disconnects and is reinstalled in its adjacent upstream.
19. by the described method of claim 16, wherein the process of at least one district's collection cut is finished by the displacement of following liquid stream from the F unit:
1) injection and the discharge of stop solution stream simultaneously in the F unit at least;
2) close flow channel between the discharge pipe in ascending pipe on the liquid stream displacement zone and adjacent upstream district, close the flow channel between the ascending pipe in discharge pipe on this liquid stream displacement zone and adjacent downstream district; The pass closed procedure of above-mentioned path is finished by any conventional blocking way that comprises switch and triple valve;
3) inject displacement liquid and collect displacement liquid from the discharge pipe in this district from the ascending pipe of displacement zone, the composition of the eluent in the composition of above-mentioned displacement liquid and claim 1 displacement zone is identical;
4) be communicated with flow channel again, restart the described operation of claim 16; Above-mentioned closing with opening process finished by the switching of triple valve.
20. by the described method of claim 16, wherein the process of at least one district's collection cut is undertaken by the displacement of following liquid stream from the F unit:
1) close flow channel between the discharge pipe in ascending pipe on the liquid stream displacement zone and adjacent upstream district, from then on the discharge pipe in adjacent upstream district is collected cut simultaneously, and injects displacement liquid from the ascending pipe of displacement zone; Above-mentioned process of closing flow channel is finished by any usual manner; The composition of the eluent in the composition of above-mentioned displacement liquid and claim 16 displacement zone is identical;
2) path of open and close again stops the injection of displacement liquid simultaneously, restarts the described operation of claim 16; Above-mentioned closing with opening process finished by the switching of triple valve.
21. by the described method of claim 16: distinguish and form a S unit that is used for sample for additional s that links together with the connected mode of F unit in the claim 16, the sequence notation in S (1) district is distinguished in the S unit from S (s), s is an integer, the S unit is placed in the upstream of B unit, make S (1) district adjacent with B (b) district, branch branch ostium on the ascending pipe in sealing or cancellation S (s) district and the discharge pipe in S (1) district is to form the flow channel of a discharge pipe of distinguishing from the ascending pipe in S (s) district to S (1), if in the R unit, there is not extra liquid stream to inject and discharge, the then ascending pipe of sealing in R (r) district and whole ascending pipes and the discharge pipe the discharge pipe in R (1) district, connect and divide branch ostium to form flow channel, perhaps cancellation divides branch ostium, replaces the branch branch ostium to carry out being connected of flow channel with ascending pipe and discharge pipe; Improvement to claim 16 also comprises:
1) injection and the collection of additional liquid stream: inject sample liquid continuously and collect this sample liquid from the discharge pipe in S (1) district from the ascending pipe in S (s) district.
2) improvement that chromatographic column is reset: the chromatographic column in the R (r) that periodically resets the simultaneously district is distinguished to B (1); Chromatographic column in replacement B (b) district is distinguished to S (1); Chromatographic column in replacement S (s) district is distinguished to F (1); Chromatographic column in replacement F (f) district is reset to corresponding adjacent upstream district with remaining chromatographic column from working as the proparea to R (1) district.
22. by the described method of claim 21, wherein reset process upstream moves a district and realizes by using the separating liquid rotary valve to switch liquid simultaneously chromatographic column the time.
23. by the described method of claim 21, finishing of reset process is by using mechanical system simultaneously chromatographic column to be realized from working as the proparea disconnection and being reinstalled in its adjacent upstream in the time of its center pillar.
24. by the described method of claim 21, wherein the process of at least one district's collection cut is finished by the displacement of following liquid stream from the F unit:
1) injection and the discharge of stop solution stream simultaneously in the F unit at least;
2) close flow channel between the discharge pipe in ascending pipe on the liquid stream displacement zone and adjacent upstream district, close the flow channel between the ascending pipe in discharge pipe on this liquid stream displacement zone and adjacent downstream district: the pass closed procedure of above-mentioned path is finished by any conventional blocking way that comprises switch and triple valve;
3) inject displacement liquid and collect displacement liquid from the discharge pipe in this district from the ascending pipe of displacement zone, the composition of the eluent in the composition of above-mentioned displacement liquid and claim 1 displacement zone is identical:
4) be communicated with flow channel again, restart the described operation of claim 21: above-mentioned closing with opening process finished by the switching of triple valve.
25. power is by profit requirement 21 described methods, wherein the process of at least one district's collection cut is undertaken by the displacement of following liquid stream from the F unit:
1) closes flow channel between the discharge pipe in ascending pipe on the liquid stream displacement zone and adjacent upstream district, from then on the discharge pipe in adjacent upstream district is collected cut simultaneously, and injects displacement liquid from the ascending pipe of displacement zone: above-mentioned process of closing flow channel is finished by any conventional blocking type that comprises switch and triple valve: the composition of above-mentioned displacement liquid is identical with the composition of the eluent of claim 21 displacement zone;
2) path of open and close again stops the injection of displacement liquid simultaneously, and restart the described operation of claim 21: described closing with opening process finished by the switching of a triple valve.
26. by the described method of claim 21, wherein remove the R unit and make B (1) district and F (f) distinguish direct neighbor to separate, the improvement of claim 21 method is also comprised:
1) the regenerated liquid collection process of the regenerated liquid injection process of cancellation R (r) district feed tube and R (1) district discharge pipe;
2) chromatographic column of periodically resetting simultaneously in F (f) district is distinguished to B (1); Chromatographic column in replacement B (b) district is distinguished to S (1); Chromatographic column in replacement S (s) district is distinguished to F (1); Remaining chromatographic column is reset to corresponding adjacent upstream district from working as the proparea.
27. by the described method of claim 26, wherein reset process upstream moves a district and realizes by using the separating liquid rotary valve to switch liquid simultaneously chromatographic column the time.
28. by the described method of claim 26, wherein chromatographic column the time reset process finish be by use mechanical system simultaneously with chromatographic column from realizing when the proparea disconnects and is reinstalled in its adjacent upstream.
29. by the described method of claim 26, wherein the process of at least one district's collection cut is finished by the displacement of following liquid stream from the F unit:
1) injection and the discharge of stop solution stream simultaneously in the F unit at least;
2) close flow channel between the discharge pipe in ascending pipe on the liquid stream displacement zone and adjacent upstream district, close the flow channel between the ascending pipe in discharge pipe on this liquid stream displacement zone and adjacent downstream district; The pass closed procedure of above-mentioned path is finished by any conventional blocking way that comprises switch and triple valve;
3) inject displacement liquid and collect displacement liquid from the discharge pipe in this district from the ascending pipe of displacement zone, the composition of the eluent in the composition of above-mentioned displacement liquid and claim 1 displacement zone is identical;
4) be communicated with flow channel again, restart the described operation of claim 26; Above-mentioned closing with opening process finished by the switching of triple valve.
30. by the described method of claim 26, wherein the process of at least one district's collection cut is undertaken by the displacement of following liquid stream from the F unit:
1) close flow channel between the discharge pipe in ascending pipe on the liquid stream displacement zone and adjacent upstream district, from then on the discharge pipe in adjacent upstream district is collected cut simultaneously, and injects displacement liquid from the ascending pipe of displacement zone; Above-mentioned process of closing flow channel is finished by any conventional blocking way that comprises switch and triple valve; The composition of the eluent in the composition of above-mentioned displacement liquid and claim 1 displacement zone is identical;
2) path of open and close again stops the injection of displacement liquid simultaneously, restarts the described operation of claim 26; Above-mentioned closing with opening process finished by the switching of triple valve.
31. by the described method of claim 26, wherein remove the B unit and make S (1) district and F (f) district direct neighbor carry out separating process, the improvement of claim 26 method is also comprised:
(1) the equilibrium liquid collection process of the equilibrium liquid injection process of cancellation B (b) district feed tube and B (1) district discharge pipe;
(2) chromatographic column of periodically resetting simultaneously in F (f) district is distinguished to S (1); Chromatographic column in replacement S (s) district is reset to corresponding adjacent upstream district with remaining chromatographic column from working as the proparea to F (1) district.
32. by the described method of claim 31, wherein reset process upstream moves a district and realizes by using the separating liquid rotary valve to switch liquid simultaneously chromatographic column the time.
33. by the described method of claim 31, wherein chromatographic column the time reset process finish be by use mechanical system simultaneously with chromatographic column from realizing when the proparea disconnects and is reinstalled in its adjacent upstream.
34. by the described method of claim 31, wherein the process of at least one district's collection cut is finished by the displacement of following liquid stream from the F unit:
1) injection and the discharge of stop solution stream simultaneously in the F unit at least;
2) close flow channel between the discharge pipe in ascending pipe on the liquid stream displacement zone and adjacent upstream district, close the flow channel between the ascending pipe in discharge pipe on this liquid stream displacement zone and adjacent downstream district; The pass closed procedure of above-mentioned path is finished by any conventional blocking way that comprises switch and triple valve;
3) inject displacement liquid and collect displacement liquid from the discharge pipe in this district from the ascending pipe of displacement zone, the composition of the eluent in the composition of above-mentioned displacement liquid and claim 1 displacement zone is identical;
4) be communicated with flow channel again, restart the described operation of claim 31; Above-mentioned closing with opening process finished by the switching of triple valve.
35. by the described method of claim 31, wherein the process of at least one district's collection cut is undertaken by the displacement of following liquid stream from the F unit:
1) close flow channel between the discharge pipe in ascending pipe on the liquid stream displacement zone and adjacent upstream district, from then on the discharge pipe of upstream is collected cut simultaneously, and injects displacement liquid from the ascending pipe of displacement zone; Above-mentioned process of closing flow channel is finished by any conventional blocking way that comprises switch and triple valve; The composition of the eluent in the composition of above-mentioned displacement liquid and claim 1 displacement zone is identical;
2) path of open and close again stops the injection of displacement liquid simultaneously, restarts the described operation of claim 31; Above-mentioned closing with opening process finished by the switching of triple valve.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US43610402P | 2002-12-21 | 2002-12-21 | |
| US60/436,104 | 2002-12-21 | ||
| US10/375,987 | 2003-03-01 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1713944A CN1713944A (en) | 2005-12-28 |
| CN100444920C true CN100444920C (en) | 2008-12-24 |
Family
ID=35719200
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB2003801035972A Expired - Fee Related CN100444920C (en) | 2002-12-21 | 2003-12-09 | Simulated moving bed chromatographic focusing |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN100444920C (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101230080B (en) * | 2007-01-22 | 2011-08-17 | 李平亚 | simulated moving bed chromatography separation of 20(S) and 20(R)-ginsenoside Rg3 enantiomer |
| CN109091911B (en) * | 2018-09-04 | 2021-04-20 | 浙江大学宁波理工学院 | Double-column switching cycle chromatographic separation system and method for separating and concentrating target components from raw materials |
| CN111728867A (en) * | 2020-08-01 | 2020-10-02 | 河南方舟医疗器械有限公司 | Serum sterile preparation subpackaging output system and method |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3706812A (en) * | 1970-12-07 | 1972-12-19 | Universal Oil Prod Co | Fluid-solid contacting apparatus |
| US4031156A (en) * | 1975-03-07 | 1977-06-21 | Exxon Research And Engineering Company | Separation and recovery of ethylbenzene by dual desorbent composition and temperature separation processes |
| US4031155A (en) * | 1975-09-29 | 1977-06-21 | Exxon Research And Engineering Company | Dual desorbent composition and temperature separation processes |
| CN1126628A (en) * | 1994-06-17 | 1996-07-17 | 代科化学工业株式会社 | Simulated moving bed chromatographic separation process |
| US5565104A (en) * | 1994-04-06 | 1996-10-15 | Uop | Small scale simulated moving bed separation process |
-
2003
- 2003-12-09 CN CNB2003801035972A patent/CN100444920C/en not_active Expired - Fee Related
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3706812A (en) * | 1970-12-07 | 1972-12-19 | Universal Oil Prod Co | Fluid-solid contacting apparatus |
| US4031156A (en) * | 1975-03-07 | 1977-06-21 | Exxon Research And Engineering Company | Separation and recovery of ethylbenzene by dual desorbent composition and temperature separation processes |
| US4031155A (en) * | 1975-09-29 | 1977-06-21 | Exxon Research And Engineering Company | Dual desorbent composition and temperature separation processes |
| US5565104A (en) * | 1994-04-06 | 1996-10-15 | Uop | Small scale simulated moving bed separation process |
| CN1126628A (en) * | 1994-06-17 | 1996-07-17 | 代科化学工业株式会社 | Simulated moving bed chromatographic separation process |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1713944A (en) | 2005-12-28 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102050865B (en) | Method of purifying polypeptides by simulated moving bed chromatography | |
| KR960010366B1 (en) | Method of choromatographic separation | |
| CN101272838B (en) | Method and device for separating fractions of a mixture | |
| CN101143274B (en) | Process and device for simulated moving bed seperation with a reduced number of valves | |
| KR101908464B1 (en) | A process and device for simulated moving bed adsorption and separation with a reduced number of control valves | |
| CN101340960A (en) | Device for chromatographic separations | |
| CN103619432A (en) | Reduction in flushing volume in an adsorptive separation system | |
| CN101961565A (en) | Method for separating three-component mixture by simulated moving bed chromatography | |
| CN102215930A (en) | Separation process | |
| CN102151418A (en) | Separation method and device for simulated moving bed | |
| CN100444920C (en) | Simulated moving bed chromatographic focusing | |
| US6805799B2 (en) | Simulated moving bed chromatographic focusing | |
| US3992175A (en) | Method of and device for chromatographic separation of fluid mixtures into fractions | |
| CN201030247Y (en) | Sequential type simulation moving bed chromatogram device | |
| Zhong et al. | Simulated moving bed chromatography. Effects of axial dispersion and mass transfer under linear conditions | |
| CN103987433A (en) | System and process for recovering products using simulated-moving-bed adsorption | |
| EP3930866A1 (en) | Improvements in and relating to optimizing the operation of a chromatography system | |
| CN111068360A (en) | Combined chromatographic device and continuous flow chromatographic method | |
| CN103987434A (en) | System and process for recovering products using simulated-moving-bed adsorption | |
| JPH0639205A (en) | Liquid chromatographic separator for three components | |
| CN101961564B (en) | Five-zone simulated movable bed chromatographic separation system | |
| CN102631795B (en) | Chromatographic separation system of five-region simulated moving bed (SMB) and separation method thereof | |
| CN104023805B (en) | Moving-bed adsorption method is used to reclaim the system and method for product | |
| CN107185273A (en) | The method and its equipment of a kind of continuous chromatography | |
| JPH03134562A (en) | Dummy mobile layer type chromatographic separation apparatus |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20081224 Termination date: 20101209 |