CN112639462A - Simulated moving bed mode chromatographic separation method and simulated moving bed mode chromatographic separation system - Google Patents

Simulated moving bed mode chromatographic separation method and simulated moving bed mode chromatographic separation system Download PDF

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CN112639462A
CN112639462A CN201980056741.2A CN201980056741A CN112639462A CN 112639462 A CN112639462 A CN 112639462A CN 201980056741 A CN201980056741 A CN 201980056741A CN 112639462 A CN112639462 A CN 112639462A
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eluent
section
supply port
port
flowing
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CN112639462B (en
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冈田一夫
荻野修大
佐藤康平
鹤田正树
宫岛俊树
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Organo Corp
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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • 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
    • B01D15/18—Selective adsorption, e.g. chromatography characterised by constructional or operational features relating to flow patterns
    • B01D15/1814—Selective adsorption, e.g. chromatography characterised by constructional or operational features relating to flow patterns recycling of the fraction to be distributed
    • B01D15/1821—Simulated moving beds
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • 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
    • B01D15/18—Selective adsorption, e.g. chromatography characterised by constructional or operational features relating to flow patterns
    • B01D15/1814—Selective adsorption, e.g. chromatography characterised by constructional or operational features relating to flow patterns recycling of the fraction to be distributed
    • B01D15/1821—Simulated moving beds
    • B01D15/185—Simulated moving beds characterized by the components to be separated
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/26—Conditioning of the fluid carrier; Flow patterns
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/26—Conditioning of the fluid carrier; Flow patterns
    • G01N30/38—Flow patterns
    • G01N30/46—Flow patterns using more than one column

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Abstract

A method of simulated moving bed chromatography which uses a circulation system in which a plurality of unit packed columns packed with an adsorbent are connected in series via pipes and are connected in a ring shape, wherein a weakly adsorptive component, a strongly adsorptive component and a middle adsorptive component having an intermediate adsorption between the two components are separated from a raw liquid by using 2 or more kinds of eluents, wherein a raw liquid supply port (F), 2 or more kinds of eluent supply ports (D) corresponding to the 2 or more kinds of eluents, a weak adsorptive component extraction port (A) containing the weakly adsorptive component, a middle adsorptive component extraction port (B) containing the middle adsorptive component and a strong adsorptive component extraction port (C) containing the strongly adsorptive component are provided in the pipes of the circulation system, the positions of the raw liquid supply port (F), the extraction port (A), the extraction port (B), and the extraction port (C) are set to a specific relationship.

Description

Simulated moving bed mode chromatographic separation method and simulated moving bed mode chromatographic separation system
Technical Field
The present invention relates to a simulated moving bed chromatography separation method and a simulated moving bed chromatography separation system.
Background
In the chromatographic separation by the simulated moving bed method, a plurality of unit packed columns (hereinafter, also simply referred to as "packed columns" or "columns") packed with an adsorbent having selective adsorption ability for a specific component among 2 or more components contained in a stock solution are connected in series via a pipe, and the packed column at the most downstream part and the packed column at the most upstream part are connected to form an annular circulation system. The raw liquid and the eluent are supplied to the circulation system, and the component having a high moving speed (weakly adsorptive component), the component having a low moving speed (strongly adsorptive component) and the component having an intermediate moving speed (moderately adsorptive component) in the circulation system are extracted from different positions, respectively, and then the raw liquid supply position, the eluent supply position, the extraction position of the weakly adsorptive component, the extraction position of the moderately adsorptive component and the extraction position of the strongly adsorptive component are moved in the fluid circulation direction of the circulation system while maintaining a predetermined positional relationship. By repeating this operation, a treatment operation of a moving bed capable of continuously supplying the stock solution was simulated.
Patent document 1 discloses a method of continuously separating 3 or more components having different affinities for an adsorbent by repeating a step of extracting a medium-adsorbable component while supplying an eluent and a stock solution, and a step of extracting a weak-adsorbable component and a strong-adsorbable component while supplying an eluent, in a series of modified simulated moving bed apparatuses.
As represented by the technique described in patent document 1, in conventional chromatographic separation by a general simulated moving bed method, basically 1 kind of eluent is used. Therefore, when a raw liquid containing a component having strong adsorption to the adsorbent and a raw liquid containing a component which is likely to cause tailing (a phenomenon of broadening the concentration distribution) are supplied to the circulation system, a large amount of eluent is required to be used to desorb (separate) these components. The use of a large amount of eluent causes an increase in the cost for concentrating the extract, and also reduces the production amount per adsorbent of the target purified product.
On the other hand, in the chromatographic separation by the simulated moving bed method, it has also been reported that 2 or more kinds of eluents are used. For example, patent document 2 describes that a high separation performance is achieved with a small adsorbent amount by using a first eluent having a weak desorption force and a second eluent having a strong desorption force and by combining the supply timing of these eluents, the raw liquid and the extraction timing of the weakly adsorptive component, the intermediate adsorptive component and the strongly adsorptive component in a specific manner.
Documents of the prior art
Patent document
Patent document 1: japanese patent No. 1998860
Patent document 2: japanese patent No. 4606092
Disclosure of Invention
Problems to be solved by the invention
Since the chromatographic separation by the simulated moving bed method can continuously obtain a target purified product with high purity, application to the medical field and the like has also been studied. For example, in the production of antibody pharmaceuticals, in addition to the target antibody, fragments of the antibody produced by cleavage or the like of the antibody, which do not function sufficiently, and aggregates of the antibody which are aggregated and enlarged, are produced in the extract or culture solution of antibody-producing cultured cells. Generally, the fragment has a small number of interaction sites with the adsorbent and has weak adsorption to the adsorbent. In contrast, the agglomerates are strongly adsorptive to the adsorbent. Therefore, when the simulated moving bed chromatography is applied to the purification of an antibody drug, it is necessary to separate the target antibody as a middle-adsorptive component having middle-adsorptive properties with respect to the adsorbent. On the other hand, for both weakly adsorptive components and strongly adsorptive components, sufficient removal at a high removal rate is required.
In addition, in practical use of such chromatographic separation, it is also important to reduce the amount of adsorbent required as much as possible to improve the separation efficiency and to reduce the cost.
However, as a result of studies on chromatographic separations by conventional simulated moving bed methods, such as the techniques described in the above patent documents, the present inventors have found that it is difficult to sufficiently achieve the above object.
Accordingly, the present invention is directed to a chromatographic separation method using a simulated moving bed system, and an object of the present invention is to provide a chromatographic separation method capable of separating a component to be purified from a raw liquid with high purity using a smaller amount of an adsorbent.
Another object of the present invention is to provide a chromatographic separation system suitable for carrying out the above-described chromatographic separation method.
Means for solving the problems
As a result of intensive studies in view of the above-mentioned problems, the present inventors have found that the above-mentioned problems can be solved by using 2 or more kinds of eluents and setting a raw liquid supply port, a strongly adsorbable component extraction port, a medium adsorbable component extraction port and a weakly adsorbable component extraction port in a circulation system to a specific positional relationship in a chromatographic separation method using a simulated moving bed system. Further, the present invention has been completed based on these findings.
The above object of the present invention is achieved by the following means.
〔1〕
A simulated moving bed mode chromatographic separation method comprises the following steps: separating weakly adsorptive components, strongly adsorptive components and intermediate adsorptive components contained in a raw liquid with respect to an adsorbent by using 2 or more kinds of eluents, the adsorbability of the intermediate adsorptive component being between that of the weakly adsorptive component and that of the strongly adsorptive component, with a circulation system in which a plurality of unit packed columns packed with the adsorbent are connected in series via pipes and connected in a ring shape,
the piping of the circulation system is provided with a raw liquid supply port F, 2 or more eluent supply ports D corresponding to the 2 or more eluents, a drawing port a for weakly adsorbing components containing the weakly adsorbing components, a drawing port B for moderately adsorbing components containing the moderately adsorbing components, and a drawing port C for strongly adsorbing components containing the strongly adsorbing components, and the positions of the raw liquid supply port F, the drawing port a, the drawing port B, and the drawing port C are set as follows (a) to (C):
(a) the extraction port B is provided downstream of the raw liquid supply port F with at least 1 unit packed column interposed therebetween;
(b) the extraction port C is provided in a pipe having the raw liquid supply port F, or the extraction port C is provided on the upstream side of the raw liquid supply port F with at least 1 unit packed column interposed therebetween;
(c) the extraction port A is provided in a pipe having the extraction port B, or the extraction port A is provided downstream of the extraction port B with at least 1 unit packed column interposed therebetween,
the chromatographic separation method comprises a step of repeating the following steps (A) and (B) in this order:
[ step (A) ]
Simultaneously or separately supplying a raw solution and 2 or more kinds of eluents from the raw solution supply port F and the 2 or more kinds of eluent supply ports D, respectively, and simultaneously or separately extracting a weakly adsorptive component, a moderately adsorptive component, and a strongly adsorptive component from the extraction port a, the extraction port B, and the extraction port C, respectively;
[ step (B) ]
And (D) after the end of the step (a), shifting the raw liquid supply port F, the eluent supply port D, the extraction port a, the extraction port B, and the extraction port C to the downstream side while maintaining a relative positional relationship.
〔2〕
The simulated moving bed chromatography separation method according to [ 1], wherein the step (A) comprises a plurality of substeps including a substep of supplying the stock solution and a substep of not supplying the stock solution.
〔3〕
The simulated moving bed chromatography separation method according to [ 1] or [ 2], wherein the extraction port C is provided downstream of an eluent supply port D1 through which an eluent D1 having the highest desorption force among 2 or more eluents is supplied, at least 1 unit packed column is arranged between the eluent supply port D1 and the extraction port C, and in the step (A), the same amount of strongly adsorbable component as the supply amount of the eluent D1 is extracted from the extraction port C during the supply of the eluent D1.
〔4〕
The simulated moving bed-type chromatographic separation method according to any one of [ 1] to [ 3], wherein the extraction port B is provided downstream of an eluent supply port D2 through which an eluent D2 having a second highest desorption force among 2 or more types of eluents is supplied, at least 1 unit packed column is disposed between the eluent supply port D2 and the extraction port B, and in the step (A), a period of time is provided during which the same amount of middle-adsorption components as the supply amount of the eluent D2 is extracted from the extraction port B during the supply of the eluent D2.
〔5〕
The simulated moving bed chromatography separation method according to any one of [ 1] to [ 4], wherein 4 to 6 kinds of eluents having different desorption forces are used.
〔6〕
The simulated moving bed-based chromatographic separation method according to any one of [ 1] to [ 5], wherein the circulation system has 4 or more unit packed columns, the circulation system is divided into 4 sections 1 to 4 continuous in a circular ring shape from the upstream side to the downstream side so that each section has at least 1 unit packed column, and the substeps (A1-1), (A2-1) and (A3-1) described below are performed in the step (A) using the 2 or more eluents,
< substep (A1-1) >)
An eluent D-I is supplied from the eluent supply port D-I with the upstream end of the zone 1 being the eluent supply port D-I, a strongly adsorptive component is extracted from the extraction port C with the downstream end of the zone 1 being the extraction port C, an eluent D-II is supplied from the eluent supply port D-II with the upstream end of the zone 2 being the eluent supply port D2, a stock solution is supplied from the stock solution supply port F with the upstream end of the zone 3 being the stock solution supply port F, a weakly adsorptive component is extracted from the extraction port A with the downstream end of the zone 4 being the extraction port A,
the desorption force of the eluent flowing in the section 1 is the strongest,
the desorption force of the eluent flowing through the section 2 is weaker than that of the eluent flowing through the section 1,
making the desorption force of the eluent flowing in the section 3 and the section 4 weaker than that of the eluent flowing in the section 2;
< substep (A2-1) >)
Supplying the eluent D-I from the eluent supply port D-I, extracting a strongly adsorptive component from the extraction port C, supplying the eluent D-II from the eluent supply port D-II, supplying the eluent D-III from the eluent supply port D-III with the upstream end of the section 3 as the eluent supply port D-III, extracting a weakly adsorptive component from the extraction port A,
the desorption force of the eluent flowing in the section 1 is the strongest,
the desorption force of the eluent flowing through the section 2 is weaker than that of the eluent flowing through the section 1,
making the desorption force of the eluent flowing in the section 3 and the section 4 weaker than that of the eluent flowing in the section 2;
< substep (A3-1) >)
Supplying the eluent D-I from the eluent supply port D-I, extracting a strongly adsorptive component from the extraction port C, supplying the eluent D-II from the eluent supply port D-II, extracting a moderately adsorptive component from the extraction port B with the downstream end of the zone 3 as the extraction port B, supplying the eluent D-IV from the eluent supply port D-IV with the upstream end of the zone 4 as the eluent supply port D-IV, and extracting a weakly adsorptive component from the extraction port A,
the desorption force of the eluent flowing in the section 1 is the strongest,
the desorption force of the eluent flowing through the sections 2 and 3 is made weaker than that of the eluent flowing through the section 1,
the desorption force of the eluent flowing through the zone 4 is made weaker than the desorption force of the eluents flowing through the zones 2 and 3.
〔7〕
The simulated moving bed-based chromatographic separation method according to any one of [ 1] to [ 5], wherein the circulation system has 4 or more unit packed columns, the circulation system is divided into 4 sections 1 to 4 continuous in a circular ring shape from the upstream side to the downstream side so that each section has at least 1 unit packed column, and the following substeps (A1-2), (A2-2) and (A3-2) are performed in the step (A) using the 2 or more eluents:
< substep (A1-2) >)
The upstream end of the zone 1 is used as an eluent supply port D-II, an eluent D-II is supplied from the eluent supply port D-II, the upstream end of the zone 3 is used as the raw liquid supply port F, a raw liquid is supplied from the raw liquid supply port F, the downstream end of the zone 4 is used as the extraction port A, and the weakly adsorptive component is extracted from the extraction port A,
the desorption force of the eluent flowing through the zone 1 and the zone 2 is maximized,
making the desorption force of the eluent flowing in the section 3 and the section 4 weaker than that of the eluent flowing in the section 1 and the section 2;
< substep (A2-2) >)
An eluent D-I is supplied from the eluent supply port D-I with the upstream end of the zone 1 being the eluent supply port D-I, a strongly adsorptive component is extracted from the extraction port C with the downstream end of the zone 1 being the extraction port C, an eluent D-II is supplied from the eluent supply port D-II with the upstream end of the zone 2 being the eluent supply port D-II, an eluent D-III is supplied from the eluent supply port D-III with the upstream end of the zone 3 being the eluent supply port D-III, and a weakly adsorptive component is extracted from the extraction port A,
the desorption force of the eluent flowing in the section 1 is the strongest,
the desorption force of the eluent flowing through the section 2 is weaker than that of the eluent flowing through the section 1,
making the desorption force of the eluent flowing in the section 3 and the section 4 weaker than that of the eluent flowing in the section 2;
< substep (A3-2) >)
Supplying the eluent D-I from the eluent supply port D-I, extracting a strongly adsorbable component from the extraction port C, supplying the eluent D-II from the eluent supply port D-II in the substep (A2-2), extracting a moderately adsorbable component from the extraction port B with the downstream end of the zone 3 as the extraction port B, supplying the eluent D-IV from the eluent supply port D-IV with the upstream end of the zone 4 as the eluent supply port D-IV, and extracting a weakly adsorbable component from the extraction port A,
the desorption force of the eluent flowing in the section 1 is the strongest,
the desorption force of the eluent flowing through the sections 2 and 3 is made weaker than that of the eluent flowing through the section 1,
the desorption force of the eluent flowing through the zone 4 is made weaker than the desorption force of the eluents flowing through the zones 2 and 3.
〔8〕
The simulated moving bed-based chromatographic separation method according to any one of [ 1] to [ 5], wherein the circulation system has 5 or more unit packed columns, the circulation system is divided into 5 sections 1 to 5 continuous in a circular ring shape from the upstream side to the downstream side so that each section has at least 1 unit packed column, and the following substeps (A1-3), (A2-3) and (A3-3) are performed in the step (A) using the 2 or more eluents:
< substep (A1-3) >)
An eluent D-II is supplied from the eluent supply port D-II with the upstream end of the zone 1 being the eluent supply port D-II, a stock solution is supplied from the stock solution supply port F with the upstream end of the zone 3 being the stock solution supply port F, an eluent D-III is supplied from the eluent supply port D-III with the upstream end of the zone 4 being the eluent supply port D-III, and a weakly adsorptive component is extracted from the extraction port A with the downstream end of the zone 5 being the extraction port A,
the desorption force of the eluent flowing through the zone 1 and the zone 2 is maximized,
the desorption force of the eluent flowing through the zone 3 is made equal to or weaker than the desorption force of the eluent flowing through the zones 1 and 2,
making the desorption force of the eluent flowing in the section 4 and the section 5 weaker than that of the eluent flowing in the section 3;
< substep (A2-3) >)
An eluent D-I is supplied from the eluent supply port D-I with the upstream end of the zone 1 being the eluent supply port D-I, a strongly adsorptive component is extracted from the extraction port C with the downstream end of the zone 1 being the extraction port C, an eluent D-II is supplied from the eluent supply port D-II with the upstream end of the zone 2 being the eluent supply port D-II, the eluent D-III is supplied from the eluent supply port D-III, and a weakly adsorptive component is extracted from the extraction port A,
the desorption force of the eluent flowing in the section 1 is the strongest,
the desorption force of the eluent flowing through the sections 2 and 3 is made weaker than that of the eluent flowing through the section 1,
making the desorption force of the eluent flowing in the section 4 and the section 5 weaker than that of the eluent flowing in the section 2 and the section 3;
< substep (A3-3) >)
Supplying the eluent D-I from the eluent supply port D-I, extracting a strongly adsorbable component from the extraction port C, supplying the eluent D-II from the eluent supply port D-II in the substep (A2-3), extracting a moderately adsorbable component from the extraction port B with the downstream end of the zone 4 as the extraction port B, supplying the eluent D-IV from the eluent supply port D-IV with the upstream end of the zone 5 as the eluent supply port D-IV, and extracting a weakly adsorbable component from the extraction port A,
the desorption force of the eluent flowing in the section 1 is the strongest,
the desorption force of the eluent flowing through the zone 2, the zone 3 and the zone 4 is weaker than that of the eluent flowing through the zone 1,
the desorption force of the eluent flowing through the zone 5 is made weaker than the desorption force of the eluent flowing through the zones 2, 3 and 4.
〔9〕
The simulated moving bed-based chromatographic separation method according to any one of [ 1] to [ 5], wherein the circulation system has 7 or more unit packed columns, the circulation system is divided into 5 sections 1 to 5 continuous in a circular ring shape from the upstream side to the downstream side so that each section has at least 1 unit packed column, and the following substeps (A1-4), (A2-4) and (A3-4) are performed in the step (A) using the 2 or more eluents:
< substep (A1-4) >)
An eluent D-II is supplied from the eluent supply port D-II with the upstream end of the zone 1 being the eluent supply port D-II, a stock solution is supplied from the stock solution supply port F with the upstream end of the zone 3 being the stock solution supply port F, an eluent D-III is supplied from the eluent supply port D-III with the upstream end of the zone 4 being the eluent supply port D-III, and a weakly adsorptive component is extracted from the extraction port A with the downstream end of the zone 5 being the extraction port A,
the desorption force of the eluent flowing through the zone 1 and the zone 2 is maximized,
the desorption force of the eluent flowing through the zone 3 is made equal to or weaker than the desorption force of the eluent flowing through the zones 1 and 2,
making the desorption force of the eluent flowing in the section 4 and the section 5 weaker than that of the eluent flowing in the section 3;
< substep (A2-4) >)
An eluent D-I is supplied from the eluent supply port D-I with the upstream end of the zone 1 being the eluent supply port D-I, a strongly adsorptive component is extracted from the extraction port C with the downstream end of the zone 1 being the extraction port C, an eluent D-II is supplied from the eluent supply port D-II with the upstream end of the zone 2 being the eluent supply port D-II, an eluent D-IV is supplied from the eluent supply port D-IV with the upstream end of the zone 4 being the eluent supply port D-IV, and a weakly adsorptive component is extracted from the extraction port A,
the desorption force of the eluent flowing in the section 1 is the strongest,
the desorption force of the eluent flowing through the sections 2 and 3 is made weaker than that of the eluent flowing through the section 1,
making the desorption force of the eluent flowing in the section 4 and the section 5 weaker than that of the eluent flowing in the section 2 and the section 3;
< substep (A3-4) >)
Supplying the eluent D-I from the eluent supply port D-I, extracting a strongly adsorbable component from the extraction port C, supplying the eluent D2 from the eluent supply port D2 in the substep (A2-4), extracting a moderately adsorbable component from the extraction port B with the downstream end of the zone 4 as the extraction port B, supplying the eluent D-V from the eluent supply port D-V with the upstream end of the zone 5 as the eluent supply port D-V, and extracting a weakly adsorbable component from the extraction port A,
the desorption force of the eluent flowing in the section 1 is the strongest,
the desorption force of the eluent flowing through the zone 2, the zone 3 and the zone 4 is weaker than that of the eluent flowing through the zone 1,
the desorption force of the eluent flowing through the zone 5 is made weaker than the desorption force of the eluent flowing through the zones 2, 3 and 4.
〔10〕
The simulated moving bed-based chromatographic separation method according to any one of [ 1] to [ 5], wherein the circulation system has 5 or more unit packed columns, the circulation system is divided into 5 sections 1 to 5 continuous in a circular ring shape from the upstream side to the downstream side so that each section has at least 1 unit packed column, and the following substeps (A1-5), (A2-5) and (A3-5) are performed in the step (A) using the 2 or more eluents:
< substep (A1-5) >)
The raw liquid is supplied from the raw liquid supply port F with the upstream end of the zone 3 being the raw liquid supply port F, the eluent D-III is supplied from the eluent supply port D-III with the upstream end of the zone 4 being the eluent supply port D-III, and the weakly adsorptive component is extracted from the extraction port A with the downstream end of the zone 5 being the extraction port A,
the desorption force of the eluent flowing in the section 3 is the strongest,
making the desorption force of the eluent flowing in the section 4 and the section 5 weaker than that of the eluent flowing in the section 3;
< substep (A2-5) >)
An eluent D-I is supplied from the eluent supply port D-I with the upstream end of the zone 1 being the eluent supply port D-I, a strongly adsorptive component is extracted from the extraction port C with the downstream end of the zone 1 being the extraction port C, an eluent D-II is supplied from the eluent supply port D-II with the upstream end of the zone 2 being the eluent supply port D-II, the eluent D-III is supplied from the eluent supply port D-III, and a weakly adsorptive component is extracted from the extraction port A,
the desorption force of the eluent flowing in the section 1 is the strongest,
the desorption force of the eluent flowing through the sections 2 and 3 is made weaker than that of the eluent flowing through the section 1,
making the desorption force of the eluent flowing in the section 4 and the section 5 weaker than that of the eluent flowing in the section 2 and the section 3;
< substep (A3-5) >)
Supplying the eluent D-I from the eluent supply port D-I, extracting a strongly adsorptive component from the extraction port C, supplying the eluent D-II from the eluent supply port D-II, extracting a moderately adsorptive component from the extraction port B with the downstream end of the zone 4 as the extraction port B, supplying the eluent D-IV from the eluent supply port D-IV with the upstream end of the zone 5 as the eluent supply port D-IV, and extracting a weakly adsorptive component from the extraction port A,
the desorption force of the eluent flowing in the section 1 is the strongest,
the desorption force of the eluent flowing through the zone 2, the zone 3 and the zone 4 is weaker than that of the eluent flowing through the zone 1,
the desorption force of the eluent flowing through the zone 5 is made weaker than the desorption force of the eluent flowing through the zones 2, 3 and 4.
〔11〕
The simulated moving bed-based chromatographic separation method according to any one of [ 1] to [ 5], wherein the circulation system has 5 or more unit packed columns, the circulation system is divided into 5 sections 1 to 5 continuous in a circular ring shape from the upstream side to the downstream side so that each section has at least 1 unit packed column, and the following substeps (A1-6), (A2-6) and (A3-6) are performed in the step (A) using the 2 or more eluents:
< substep (A1-6) >)
An eluent D-II is supplied from the eluent supply port D-II with the upstream end of the zone 1 being the eluent supply port D-II, the mid-adsorptive component is extracted from the extraction port B with the downstream end of the zone 3 being the extraction port B, the eluent D-IV is supplied from the eluent supply port D-IV with the upstream end of the zone 4 being the eluent supply port D-IV, and the weakly-adsorptive component is extracted from the extraction port A with the downstream end of the zone 5 being the extraction port A,
the desorption force of the eluent flowing through the zone 1, the zone 2 and the zone 3 is the strongest,
making the desorption force of the eluent flowing in the section 3 and the section 5 weaker than that of the eluent flowing in the section 1, the section 2 and the section 3;
< substep (A2-6) >)
An eluent D-I is supplied from the eluent supply port D-I with the upstream end of the zone 1 being the eluent supply port D-I, a strongly adsorptive component is extracted from the extraction port C with the downstream end of the zone 1 being the extraction port C, a stock solution is supplied from the stock solution supply port F with the upstream end of the zone 3 being the stock solution supply port F, an eluent D-III is supplied from the eluent supply port D-III with the upstream end of the zone 4 being the eluent supply port D-III, and a weakly adsorptive component is extracted from the extraction port A,
the desorption force of the eluent flowing in the section 1 is the strongest,
the desorption force of the eluent flowing through the section 3 is weaker than that of the eluent flowing through the section 1,
making the desorption force of the eluent flowing in the section 4 and the section 5 weaker than that of the eluent flowing in the section 3;
< substep (A3-6) >)
Supplying the eluent D-I from the eluent supply port D-I, extracting the strongly adsorptive component from the extraction port C, supplying the eluent D-II from the eluent supply port D-II with the upstream end of the section 2 as the eluent supply port D-II, supplying the eluent D-III from the eluent supply port D-III, extracting the weakly adsorptive component from the extraction port A,
the desorption force of the eluent flowing in the section 1 is the strongest,
the desorption force of the eluent flowing through the sections 2 and 3 is made weaker than that of the eluent flowing through the section 1,
the desorption force of the eluent flowing through the zone 4 and the zone 5 is made weaker than the desorption force of the eluent flowing through the zone 2 and the zone 3.
〔12〕
The simulated moving bed-based chromatographic separation method according to any one of [ 1] to [ 5], wherein the circulation system has 5 or more unit packed columns, the circulation system is divided into 5 sections 1 to 5 continuous in a circular ring shape from the upstream side to the downstream side so that each section has at least 1 unit packed column, and the following substeps (A1-7), (A2-7) and (A3-7) are performed in the step (A) using the 2 or more eluents:
< substep (A1-7) >)
An eluent D-I is supplied from the eluent supply port D-I with the upstream end of the zone 1 being the eluent supply port D-I, a strongly adsorptive component is extracted from the extraction port C with the downstream end of the zone 1 being the extraction port C, a stock solution is supplied from the stock solution supply port F with the upstream end of the zone 3 being the stock solution supply port F, an eluent D-III is supplied from the eluent supply port D-III with the upstream end of the zone 4 being the eluent supply port D-III, and a weakly adsorptive component is extracted from the extraction port A with the downstream end of the zone 5 being the extraction port A,
the desorption force of the eluent flowing in the section 1 is the strongest,
the desorption force of the eluent flowing through the section 3 is weaker than that of the eluent flowing through the section 1,
making the desorption force of the eluent flowing in the section 4 and the section 5 weaker than that of the eluent flowing in the section 3;
< substep (A2-7) >)
Supplying the eluent D-I from the eluent supply port D-I, extracting the strongly adsorptive component from the extraction port C, supplying the eluent D-II from the eluent supply port D-II with the upstream end of the section 2 as the eluent supply port D-II, supplying the eluent D-III from the eluent supply port D-III, extracting the weakly adsorptive component from the extraction port A,
the desorption force of the eluent flowing in the section 1 is the strongest,
the desorption force of the eluent flowing through the sections 2 and 3 is made weaker than that of the eluent flowing through the section 1,
making the desorption force of the eluent flowing in the section 4 and the section 5 weaker than that of the eluent flowing in the section 2 and the section 3;
< substep (A3-7) >)
Supplying the eluent D-I from the eluent supply port D-I, extracting a strongly adsorptive component from the extraction port C, supplying the eluent D-II from the eluent supply port D-II, extracting a moderately adsorptive component from the extraction port B with the downstream end of the zone 4 as the extraction port B, supplying the eluent D-IV from the eluent supply port D-IV with the upstream end of the zone 5 as the eluent supply port D-IV, and extracting a weakly adsorptive component from the extraction port A,
the desorption force of the eluent flowing in the section 1 is the strongest,
the desorption force of the eluent flowing through the zone 2, the zone 3 and the zone 4 is weaker than that of the eluent flowing through the zone 1,
the desorption force of the eluent flowing through the zone 5 is made weaker than the desorption force of the eluent flowing through the zones 2, 3 and 4.
〔13〕
A simulated moving bed type chromatographic separation system wherein a plurality of unit packed columns packed with an adsorbent are connected in series via a pipe and connected in a ring-like manner in a circulating system, wherein weakly adsorptive components, strongly adsorptive components and intermediate adsorptive components are separated from the adsorbent, the components being contained in a raw liquid, using 2 or more kinds of eluents, the adsorbability of the intermediate adsorptive component being between the adsorbability of the weakly adsorptive component and the adsorbability of the strongly adsorptive component,
the piping of the circulation system is provided with a raw liquid supply port F, 2 or more eluent supply ports D corresponding to the 2 or more eluents, a drawing port a for weakly adsorbing components containing the weakly adsorbing components, a drawing port B for moderately adsorbing components containing the moderately adsorbing components, and a drawing port C for strongly adsorbing components containing the strongly adsorbing components, and the positions of the raw liquid supply port F, the drawing port a, the drawing port B, and the drawing port C are set as follows (a) to (C):
(a) the extraction port B is provided downstream of the raw liquid supply port F with at least 1 unit packed column interposed therebetween;
(b) the extraction port C is provided in a pipe having the raw liquid supply port F, or the extraction port C is provided on the upstream side of the raw liquid supply port F with at least 1 unit packed column interposed therebetween;
(c) the extraction port A is provided in a pipe having the extraction port B, or the extraction port A is provided downstream of the extraction port B with at least 1 unit packed column interposed therebetween,
the chromatographic separation system has a unit for repeating the following steps (A) and (B) in this order:
[ step (A) ]
Simultaneously or separately supplying a raw solution and 2 or more kinds of eluents from the raw solution supply port F and the 2 or more kinds of eluent supply ports D, respectively, and simultaneously or separately extracting a weakly adsorptive component, a moderately adsorptive component, and a strongly adsorptive component from the extraction port a, the extraction port B, and the extraction port C, respectively;
[ step (B) ]
And (D) after the end of the step (a), shifting the raw liquid supply port F, the eluent supply port D, the extraction port a, the extraction port B, and the extraction port C to the downstream side while maintaining a relative positional relationship.
In the present specification, the terms "upstream" and "downstream" are used with respect to the direction of flow of the fluid in the circulation system. That is, with respect to a certain portion of the circulation system, "upstream side" means a side through which a fluid flows toward the certain portion, and "downstream side" means a side through which the fluid flows out from the certain portion.
In the present specification, the term "strongly adsorbable component" refers to a component having a strong adsorption force on an adsorbent out of a plurality of components contained in a stock solution, the term "weakly adsorbable component" refers to a component having a weak adsorption force on an adsorbent out of a plurality of components contained in a stock solution, and the term "moderately adsorbable component" refers to a component having a weak adsorption force on an adsorbent as compared with the strongly adsorbable component but having a strong adsorption force on an adsorbent as compared with the weakly adsorbable component. That is, the terms "strong adsorbability", "middle adsorbability", and "weak adsorbability" indicate the relative strength of the adsorption force of each component contained in the stock solution with respect to the adsorbent when the adsorption force is compared.
The "strongly adsorbable component", "the medium adsorbable component" and "the weakly adsorbable component" may be composed of a single component or a plurality of components, respectively. The adsorption forces of the plurality of components may be the same or different.
The groups of "strongly adsorbable component", "intermediate adsorbable component" and "weakly adsorbable component" for each component in the stock solution may be set as appropriate according to the purpose. In the case of 4 components in the raw liquid, the first 2 components in the order of strong to weak adsorption force on the adsorbent may be collectively positioned as a strong-adsorbable component, the third strong-adsorbable component on the adsorbent may be positioned as a medium-adsorbable component, and the weakest-adsorbable component on the adsorbent may be positioned as a weak-adsorbable component. Further, the component having the strongest adsorption force to the adsorbent may be determined as a strong adsorption component, the component having the second strongest adsorption force to the adsorbent and the component having the third strongest adsorption force may be combined and determined as a medium adsorption component, and the component having the weakest adsorption force to the adsorbent may be determined as a weak adsorption component. Further, the component having the strongest adsorption force to the adsorbent may be localized as a strong adsorption component, the component having the second strongest adsorption force to the adsorbent may be localized as a medium adsorption component, and the component having the third strongest adsorption force to the adsorbent and the component having the weakest adsorption force may be collectively localized as a weak adsorption component. When the stock solution contains 5 or more components, separation and purification by various groups can be performed similarly.
In the present invention, the "desorption force" of the eluent refers to the strength of the action of desorbing a component adsorbed on the adsorbent from the adsorbent.
Effects of the invention
According to the simulated moving bed chromatography method of the present invention, the amount of the adsorbent used can be suppressed, and the component to be purified in the raw liquid can be separated with high purity. The simulated moving bed chromatography separation system of the present invention can be suitably applied to the implementation of the simulated moving bed chromatography separation method of the present invention.
Drawings
Fig. 1 is a system diagram showing an example of a simulated moving bed type chromatography system according to the present invention.
Fig. 2 is a flowchart of each substep constituting step (a) in one embodiment of the simulated moving bed chromatography separation method of the present invention.
Fig. 3 is a flowchart showing the respective substeps of step (a) after step (a) shown in fig. 2 is completed and step (B) is performed.
Fig. 4 is a flowchart of each substep constituting step (a) in another embodiment of the simulated moving bed chromatography separation method of the present invention.
Fig. 5 is a flowchart showing the respective substeps of step (a) after step (a) shown in fig. 4 is completed, and step (B) is executed.
Fig. 6 is a flowchart of each substep constituting step (a) in still another embodiment of the simulated moving bed chromatography separation method of the present invention.
Fig. 7 is a flowchart showing the respective substeps of step (a) after step (a) shown in fig. 6 is completed, and step (B) is executed.
Fig. 8 is a flowchart of each substep constituting step (a) in still another embodiment of the simulated moving bed chromatography separation method of the present invention.
Fig. 9 is a flowchart showing the respective substeps of step (a) after step (a) shown in fig. 8 is completed, and step (B) is executed.
FIG. 10 is a flowchart showing the operation steps of the single column and discontinuous gradient in comparative examples 1 and 2.
FIG. 11 is a flowchart showing the operation steps of simulated mobile phase chromatography in comparative example 3.
FIG. 12 is a flowchart showing the operation procedure of the simulated mobile phase chromatography in comparative example 4.
Fig. 13 is a flowchart of each substep constituting step (a) in still another embodiment of the simulated moving bed chromatography separation method of the present invention.
Fig. 14 is a flowchart of the respective substeps of step (a) after step (a) shown in fig. 13 is completed, and step (B) is executed.
Fig. 15 is a flowchart of each substep constituting step (a) in still another embodiment of the simulated moving bed chromatography separation method of the present invention.
Fig. 16 is a flowchart showing the respective substeps of step (a) after step (a) shown in fig. 15 is completed, and step (B) is executed.
Fig. 17 is a flowchart of each substep constituting step (a) in still another embodiment of the simulated moving bed chromatography separation method of the present invention.
Fig. 18 is a flowchart showing the respective substeps of step (a) after step (a) shown in fig. 17 is completed, and step (B) is executed.
Detailed Description
A preferred embodiment of the simulated moving bed chromatography method of the present invention (hereinafter, also simply referred to as "the method of the present invention") will be described.
The method of the present invention is carried out by a circulation system in which a plurality of unit packed towers packed with an adsorbent are connected in series via pipes and connected in a ring shape. Circulation systems for use in the simulated moving bed system are known per se, and for example, refer to japanese patent laid-open No. 2009-36536, japanese patent laid-open No. 4606092, and the like.
The circulation system will be described below with reference to the drawings, but the present invention is not limited to these embodiments except for the ones defined in the present invention.
The drawings mentioned below are explanatory diagrams for easy understanding of the present invention, and the dimensions of the respective structures and the relative size relationship may be changed for convenience of explanation, and do not directly indicate the actual relationship. The shapes, relative positional relationships, and the like shown in the drawings are not limited to those shown in the drawings except for the matters defined in the present invention.
Conditions other than the conditions specified in the present invention, for example, the capacity of the packed column, the pipe inner cross-sectional area of the pipe, and the flow rate of the liquid supplied to the circulation system by the length, can be appropriately set according to the purpose.
In fig. 1, a preferred embodiment of a recycling system for the process of the invention is shown. The circulation system 100 shown in fig. 1 includes 4 (unit packed columns 10a, 10b, 10c, and 10d) unit packed columns (columns) packed with the adsorbent Ab, and the outlets of the respective unit packed columns are connected to the inlets of the adjacent unit packed columns via the pipes 1, and as a whole, the respective unit packed columns are connected in series.
The outlet of the rearmost unit packed column (for example, unit packed column 10d) is connected to the inlet of the foremost unit packed column (for example, unit packed column 10a) via the pipe 1, and all the unit packed columns are connected in an annular shape (annular shape). With this structure, the fluid can be circulated in the circulation system 100. The shapes and sizes of the inside of the unit packed columns 10a to 10d and the packed amount of the adsorbent may be the same or different from each other. The shape, size, and amount of adsorbent packed in each packed column 10a to 10d are preferably equivalent to each other (preferably, the same shape, size, and amount of adsorbent packed in each packed column).
In the circulation system 100, a circulation pump P1 for circulating a fluid in the direction of the arrow may be disposed. The circulation pump P1 is preferably a fixed displacement pump. In the circulation system 100, the piping 1 between the adjacent 2 unit packed towers is provided with shutoff valves R1, R2, R3, and R4 capable of shutting off the flow of fluid to the unit packed tower on the downstream side.
Weakly adsorptive component extraction lines 2a, 2b, 2c, 2d for extracting a component containing a large amount of a weakly adsorptive component to the adsorbent Ab (referred to as "weakly adsorptive component to the adsorbent Ab" or simply as "weakly adsorptive component" in the present specification) are branched and arranged between the respective shutoff valves R1 to R4 and the outlets of the respective unit packed columns 10a to 10d located on the upstream side thereof, respectively. Weak adsorptive component extraction valves a1, a2, A3, and a4 are provided in the weak adsorptive component extraction lines 2a, 2b, 2c, and 2d, respectively, so as to be capable of opening and closing the weak adsorptive component extraction lines. The weakly adsorptive component extraction lines 2a, 2b, 2c and 2d are merged and collected in a weakly adsorptive component flow merging line 2J.
Similarly, intermediate adsorption component extraction lines 3a, 3b, 3c, and 3d for extracting a component containing a large amount of an intermediate adsorption component to the adsorbent Ab (referred to as "intermediate adsorption component to the adsorbent Ab" or simply as "intermediate adsorption component" in the present specification) are branched and disposed between the shutoff valves R1 to R4 and the outlets of the unit packed columns 10a to 10d located on the upstream side thereof. The intermediate adsorptive component extraction lines 3a, 3B, 3c and 3d are provided with intermediate adsorptive component extraction valves B1, B2, B3 and B4 which can open and close the intermediate adsorptive component extraction lines, respectively. The respective middle adsorptive component extraction lines 3a, 3b, 3c, 3d are merged and collected in one middle adsorptive component flow merging line 3J.
Similarly, strongly adsorbable component extraction lines 4a, 4b, 4c, 4d for extracting a component containing a large amount of strongly adsorbable components to the adsorbent Ab (referred to as "strongly adsorbable component to the adsorbent Ab" or simply as "strongly adsorbable component" in the present specification) are branched and arranged between the shutoff valves R1 to R4 and the outlets of the unit packed columns 10a to 10d located on the upstream side thereof. Strong adsorbable component extraction valves C1, C2, C3, and C4 that can open and close the strong adsorbable component extraction lines are provided in the strong adsorbable component extraction lines 4a, 4b, 4C, and 4d, respectively. The strongly adsorbable component extraction lines 4a, 4b, 4c, 4d merge together and are collected in one strongly adsorbable component flow-combining line 4J.
In step (a) described later, any one of the weakly adsorbable component extraction valves a1, a2, A3, a4 is in an open state. The connection portion between the weakly adsorptive component extraction line of the weakly adsorptive component extraction valve provided with the valve and the pipe 1 becomes the extraction port a of the weakly adsorptive component in the step (a) described later.
In step (a) described later, any one of the middle adsorptive component drawing valves B1, B2, B3, and B4 is opened. The connection part between the intermediate adsorptive component extraction line provided with the open valve and the pipe 1 serves as an extraction port B for the intermediate adsorptive component in step (a) to be described later.
In step (a) described later, any one of the strongly adsorbable component extraction valves C1, C2, C3, and C4 is in an open state. The portion of the pipe 1 connected to the strongly adsorbable component extraction line provided with the strongly adsorbable component extraction valve having this valve open becomes the extraction port C for the strongly adsorbable component in step (a) described later.
In the circulation system 100, a relief valve (or a relief valve), not shown, may be provided at an appropriate position in order to prevent an excessive pressure rise in the circulation system 100. Further, check valves T1, T2, T3 and T4 for preventing a reverse flow are preferably provided between the adjacent 2 unit packed columns.
As shown in fig. 1, the circulation system 100 is configured to be able to supply the raw liquid 7 stored in the raw liquid tank 6. In addition, the circulation system 100 is configured to be able to supply 2 or more kinds of eluents. In fig. 1, as an example, a mode of supplying 4 kinds of eluents is shown.
The raw liquid 7 is supplied through the raw liquid supply line 11 by a raw liquid supply pump P2 capable of controlling the supply flow rate. The raw liquid supply pump P2 is preferably a fixed displacement pump. As shown in fig. 1, the raw liquid supply line 11 is configured to be branched into 4 raw liquid supply branch lines 11a, 11b, 11c, and 11d, and the raw liquid can be supplied to the inlets of the unit packed towers 10a, 10b, 10c, and 10d via the raw liquid supply branch lines 11a, 11b, 11c, and 11d, respectively. Stock solution supply branch lines 11a, 11b, 11c, and 11d are provided with stock solution supply valves F1, F2, F3, and F4 that can be opened and closed, and the stock solution is supplied to a unit filling tower connected downstream thereof through the stock solution supply branch line having the open stock solution supply valve.
In step (a) described later, any one of the stock solution supply valves F1, F2, F3, and F4 is opened. The connection portion between the stock solution supply branch line of the stock solution supply valve provided with the valve and the pipe 1 is the stock solution supply port F in step (a) to be described later.
FIG. 1 shows the manner of supplying 4 kinds of eluents having different desorption forces. The eluent 9a stored in the eluent tank 8a is supplied to the eluent supply line 12 by an eluent supply pump P3 capable of controlling the supply flow rate. The eluent 9b stored in the eluent tank 8b is supplied to the eluent supply line 13 by an eluent supply pump P4 capable of controlling the supply flow rate. The eluent 9c stored in the eluent tank 8c is supplied to the eluent supply line 14 by an eluent supply pump P5 capable of controlling the supply flow rate. The eluent 9d stored in the eluent tank 8d is supplied to the eluent supply line 15 by an eluent supply pump P6 capable of controlling the supply flow rate.
The eluent supply pumps P3 to P6 are preferably quantitative pumps. As shown in fig. 1, the eluent supply line 12 is configured to be branched into 4 eluent supply branch lines 12a, 12b, 12c, 12d, and to be able to supply eluent to inlets of the unit packed columns 10a, 10b, 10c, 10d via the eluent supply branch lines 12a, 12b, 12c, 12 d. Eluent supply valves E1a, E2a, E3a, and E4a are provided to the eluent supply branch lines 12a, 12b, 12c, and 12d so as to be openable and closable, and the eluent is supplied to the unit packing columns connected downstream thereof through the eluent supply branch lines having the open eluent supply valves.
Similarly, the eluent supply line 13 is configured to be branched into 4 eluent supply branch lines 13a, 13b, 13c, 13d, the eluent supply line 14 is branched into 4 eluent supply branch lines 14a, 14b, 14c, 14d, the eluent supply line 15 is branched into 4 eluent supply branch lines 15a, 15b, 15c, 15d, and each eluent can be supplied to the inlet of each unit packed column 10a, 10b, 10c, 10 d.
Eluent supply valves E1b, E2b, E3b and E4b which can be opened and closed are provided in eluent supply branch lines 13a, 13b, 13c and 13d, eluent supply valves E1c, E2c, E3c and E4c which can be opened and closed are provided in eluent supply branch lines 14a, 14b, 14c and 14d, and eluent supply valves E1d, E2d, E3d and E4d which can be opened and closed are provided in eluent supply branch lines 15a, 15b, 15c and 15 d.
In step (a) described later, a connection portion between the eluent supply branch line provided with the open eluent supply valve and the pipe 1 is an eluent supply port D. In the method of the present invention, since 2 or more kinds of eluents are used, there are a plurality of eluent supply valves which are opened in step (a) described later. Therefore, in step (a) described later, the eluent supply ports D are present in the number (2 or more) corresponding to the type of eluent used.
Next, the operation of the circulation system when the method of the present invention is carried out by the circulation system will be described, but the present invention is not limited to these embodiments except for the fact that the present invention is defined in the present invention.
In the method of the present invention, the circulation system sets the positions of the raw liquid supply port F, the extraction port a for weakly adsorptive component, the extraction port B for medium-adsorptive component, and the extraction port C for strongly adsorptive component to satisfy the following relationships (a) to (C). That is, in the repetition of the steps (a) and (B) described later, the relative positional relationship among the dope supply port F, the weakly adsorbable component extraction port a, the medium adsorbable component extraction port B, and the strongly adsorbable component extraction port C always satisfies the above (a) to (C).
(a) The middle-adsorptive-component extraction port B was provided on the downstream side of the raw-liquid supply port F with at least 1 unit packed column therebetween.
(b) The strongly adsorbable component withdrawal port C is provided in a pipe having the raw liquid supply port F, or is provided on the upstream side of the raw liquid supply port F with at least 1 unit packed column therebetween.
Here, the phrase "the strongly adsorbable component withdrawal port C is provided in the piping having the raw liquid supply port F" means that no unit packed column is disposed between the strongly adsorbable component withdrawal port C and the raw liquid supply port F.
In the case of "the strongly adsorbable component extraction port C is provided in the pipe having the dope supply port F", the strongly adsorbable component extraction port C is provided on the upstream side of the same pipe from the dope supply port F. This applies to all relationships between the extraction port and the supply port provided in the same pipe. That is, in the circulation system, when a certain extraction port and a certain supply port are provided in the same pipe (when the extraction port and the supply port are not provided with a unit packed column interposed therebetween), the extraction port is disposed on the upstream side of the same pipe from the supply port. This is to prevent the supplied liquid from being drawn out through the draw-out port before the liquid reaches the unit packed column downstream thereof.
The above-mentioned (b) is preferably a mode in which the strongly adsorbable component withdrawal port C is provided upstream of the raw liquid supply port F with at least 1 unit packed column interposed therebetween.
(c) The weakly adsorptive component withdrawal port a is provided in a pipe having the intermediate adsorptive component withdrawal port B, or the weakly adsorptive component withdrawal port a is provided downstream of the intermediate adsorptive component withdrawal port B across at least 1 unit packed column.
Here, the phrase "the weakly adsorptive component withdrawal port a is provided in the pipe having the intermediate adsorptive component withdrawal port B" means that no unit packed column is disposed between the weakly adsorptive component withdrawal port a and the intermediate adsorptive component withdrawal port B.
The above-mentioned (c) is preferably a mode in which the weakly adsorptive component withdrawal port a is provided downstream of the intermediate adsorptive component withdrawal port B with at least 1 unit packed column interposed therebetween.
In the method of the present invention, the following steps (a) and (B) are sequentially repeated using the above circulation system.
[ step (A) ]
And simultaneously or separately supplying the raw solution and 2 or more kinds of eluents from the raw solution supply port F and the 2 or more kinds of eluent supply ports D, respectively, and simultaneously or separately extracting the weakly adsorptive component, the intermediate adsorptive component and the strongly adsorptive component from the weakly adsorptive component extraction port A, the intermediate adsorptive component extraction port B and the strongly adsorptive component extraction port C, respectively.
Here, in step (a), the sum of the amount of the raw liquid supplied from the raw liquid supply port F and the amount of the elution liquid supplied from the eluent supply port D coincides with the sum of the amount of the weakly adsorptive component withdrawn from the weakly adsorptive component withdrawal port a, the amount of the intermediate adsorptive component withdrawn from the intermediate adsorptive component withdrawal port B, and the amount of the strongly adsorptive component C withdrawn from the strongly adsorptive component withdrawal port C. That is, in a state where the liquid is supplied in the circulation system, the liquid is extracted from the circulation system in the same amount as the liquid.
More specifically, when a liquid is supplied from a certain supply port (X) and a liquid is extracted from an extraction port (Y) on the downstream side thereof, the amount of the liquid extracted from Y is the same as the amount of the liquid supplied from X when no liquid is supplied on the downstream side of X and on the upstream side of Y. In addition, when the liquid is supplied from the downstream side of X and the upstream side of Y, the amount of the liquid extracted from Y is the same as the sum of the amount of the liquid supplied from X and the amount of the liquid supplied from the upstream side of Y on the downstream side of X. For example, in the substep (a1-1) of fig. 2, the supply amount of the eluent supplied from the eluent supply port D1 is the same as the extraction amount of the strongly adsorbable component extracted from the strongly adsorbable component extraction port C. In the sub-step (a1-1), the sum of the supply amount of the eluent supplied from the eluent supply port D2 and the supply amount of the raw liquid supplied from the raw liquid supply port F is the same as the amount of the weakly adsorptive component extracted from the weakly adsorptive component extraction port a.
The above-mentioned "simultaneous supply or separate supply" means that supply is performed without providing a difference in time (without shifting the timing of supply), or supply is performed with providing a difference in time (with shifting the timing of supply). However, in the case where 2 or more supply ports for supplying 2 or more kinds of liquids are arranged in the same pipe in one step (a) (in the case where 2 or more supply ports are arranged without interposing a unit packed column in the pipe and different liquids are supplied from the 2 or more supply ports), the supply of the 2 or more kinds of liquids is not performed simultaneously. That is, in one step (a), the supply of the 2 or more kinds of liquids is performed as different substeps. Similarly, in the case where 2 or more extraction ports for extracting 2 or more components are arranged in the same piping in one step (a) (in the case where 2 or more extraction ports are arranged without interposing a unit packed column in piping and different components are extracted from the 2 or more extraction ports), the extraction of the 2 or more components is not performed simultaneously. That is, in one step (a), the extraction of the 2 or more components is performed as different substeps.
[ step (B) ]
And (C) after the completion of the step (a), transferring the raw liquid supply port F, the eluent supply port D, the weakly adsorptive component extraction port a, the intermediate adsorptive component extraction port B, and the strongly adsorptive component extraction port C to the downstream side while maintaining their relative positional relationship.
The transfer to the downstream side means an amount of 1 unit packed column to transfer the raw liquid supply port F, the eluent supply port D, the weakly adsorptive component extraction port a, the intermediate adsorptive component extraction port B, and the strongly adsorptive component extraction port C to the downstream side while maintaining their relative positional relationship.
For example, in the case where the portion of the pipe 1 connected to the stock solution supply branch line provided with the stock solution supply valve F1 in step (a) is the stock solution supply port F, the stock solution supply port F is shifted to the portion of the pipe 1 connected to the stock solution supply branch line provided with the stock solution supply valve F2 in step (B). The same applies to the eluent supply port D, the weakly adsorptive component extraction port a, the intermediate adsorptive component extraction port B, and the strongly adsorptive component extraction port C. The amount of 1 unit packed column transferred to the downstream side from each of the supply port and the extraction port can be adjusted by opening and closing various pumps and various valves disposed in the circulation system.
By the execution of step (B), in the next step (a) (referred to as step (a2)), in step (a) (referred to as step (a1)) before step (B), liquid supply and extraction are performed for one amount of each unit packed column on the downstream side of step (a1) in the same manner as performed for each unit packed column.
The step (a) is preferably composed of a plurality of substeps. In this case, in which substep the feed of the dope from the dope feed port F, the feed of each of 2 or more eluents from the 2 or more eluent feed ports D, the extraction of the weakly adsorptive component from the weakly adsorptive component extraction port a, the extraction of the intermediate adsorptive component from the intermediate adsorptive component extraction port B, and the extraction of the strongly adsorptive component from the strongly adsorptive component extraction port C are carried out may be appropriately set according to the purpose, within the range not to impair the effects of the present invention.
Among them, in the method of the present invention, the step (a) preferably includes a substep of supplying the stock solution and a substep of not supplying the stock solution. That is, in step (a), it is preferable that there be a time when the stock solution is supplied and a time when the stock solution is not supplied.
In step (a), 2 or more kinds of eluents are supplied. The supply port of the eluent having the strongest desorption force among the 2 or more kinds of eluents is preferably provided in the upstream side piping with respect to the strongly adsorbable component extraction port C with the unit packed column interposed therebetween. The supply port for the remaining eluent or the raw liquid supply port is preferably provided in the same pipe as the supply port for the eluent having the strongest desorption force, or is provided upstream of the supply port for the eluent having the strongest desorption force via the unit packed column. "provided in the same pipe as the supply port of the eluent having the strongest desorption force, or provided on the upstream side of the supply port of the eluent having the strongest desorption force across the unit packed column" means that the supply port of the eluent other than the strongest eluent or the raw liquid supply port is provided in any pipe between the pipe provided with the supply port of the eluent as a starting point and proceeding to the upstream side and reaching the pipe provided with the strongly adsorptive component extraction port C (in other words, the supply port of the eluent other than the strongest eluent or the raw liquid supply port is provided in any pipe between the pipe provided with the strongly adsorptive component extraction port C as a starting point and proceeding to the downstream side and reaching the pipe provided with the supply port of the strongest eluent; in other words, the case where 2 or more unit packed columns are provided between the supply port of the strongest eluent and the strongly adsorptive component extraction port C, no eluent supply port or raw liquid supply port is provided in the piping connecting the 2 or more unit packed columns). In this case, when there are 2 or more types of eluents other than the strongest eluent, a part of the 2 or more supply ports corresponding thereto may be provided in the same pipe, or may be provided in different pipes with a unit packed column interposed therebetween. Further, the eluent supply port and the raw liquid supply port other than the strongest eluent may be provided in the same pipe.
Further, it is a preferable embodiment of the present invention that 1 of the supply ports of the eluents other than the eluent having the strongest desorption force is provided in the same pipe as the supply port of the eluent having the strongest desorption force.
In the step (a), during the supply of the eluent having the strongest desorption force among the 2 or more kinds of eluents, it is preferable to extract the strongly adsorbable component in the same amount as the supply amount of the eluent having the strongest desorption force from the downstream side thereof. In this case, it is preferable that at least 1 unit packed column is disposed between the supply port of the eluent having the highest desorption force and the strongly adsorbable component extraction port located downstream thereof, and that no other supply port is present therebetween.
In the step (a), it is preferable to provide a time period (substep) during which the middle adsorptive component is extracted in the same amount as the supply amount of the eluent having the second highest desorption force from the downstream side while supplying the eluent having the second highest desorption force from among the 2 or more kinds of eluents. In this case, at least 1 unit packed column (preferably, a plurality of unit packed columns) is disposed between the supply port of the eluent having the second highest desorption force and the middle adsorptive component extraction port downstream thereof. Further, even if another supply port is provided between the pipe provided with the supply port of the eluent having the second highest desorption force and the pipe provided with the middle adsorptive component extraction port on the downstream side thereof, the liquid is not supplied from the other supply port during the period of the middle adsorptive component extraction.
During the implementation of step (a), it is preferred to withdraw the weakly adsorbing component at all times. Therefore, in the case where the step (a) is constituted by a plurality of substeps, it is also preferable that the weakly adsorptive component be extracted at all times among the plurality of substeps.
In the method of the present invention, it is preferable to use 3 or more kinds of eluents having different desorption forces from each other, more preferably 4 or more kinds of eluents having different desorption forces from each other, still more preferably 4 to 6 kinds of eluents having different desorption forces from each other, and particularly preferably 4 or 5 kinds of eluents having different desorption forces from each other.
The type of the eluent is not particularly limited, and is appropriately set in accordance with the relationship with the type of the adsorbent and the type of the component in the raw liquid. For example, when an ion exchange resin is used as the adsorbent, a plurality of eluents having different desorption forces can be prepared by changing the salt concentration of the eluent. For example, in the case of using a cation exchange resin, a plurality of eluents with varied NaCl concentrations may be used as 2 or more eluents.
Preferred embodiments of combinations of the sub-steps in the step (a) will be described below. The implementation of these embodiments can be performed using the system of fig. 1 or using a system of an objective implementation that is based on the system of fig. 1. In addition, the following embodiment is an example of the present invention, for example, from the viewpoint of relative desorption, 2 or more kinds of eluents positioned as the eluent d-1 described below may be prepared, and the kind of the eluent d-1 used may be changed between the substeps of supplying the eluent d-1. The same applies to the eluents d-II to d-V.
That is, in the present invention, in the case where the "eluent d-1" is referred to in an embodiment, in this embodiment, in the case where the "eluent d-1" is used in a different sub-step, the "eluents d-1" used in the different sub-steps may be the same as each other, or may be different from each other. The same applies to the eluents d-II to d-V.
Embodiment 1-
In embodiment 1, a circulation system having 4 or more unit packed columns is used. The circulation system is divided into 4 sections 1 to 4 which are continuous in an annular shape from the upstream side to the downstream side so that each section has at least 1 unit packed column. In addition, as the eluent, using the desorption force different 4 kinds of eluent d-I ~ d-IV.
In embodiment mode 1, the following substeps (A1-1), (A2-1) and (A3-1) are carried out in step (A).
In the present invention, "performing sub-steps X, Y and Z in step (a)" means that step (a) includes sub-steps X, Y and Z, and the order of performing sub-steps X, Y and Z may be set as appropriate within a range that does not impair the effect of the present invention. In addition, step (a) may include other substeps besides substeps X, Y and Z.
As a typical example of the method of "performing sub-steps X, Y and Z in step (a)", a method of sequentially performing sub-steps X, Y and Z as step (a) is mentioned, but the present invention is not limited to this method. That is, "performing sub-step X, Y and Z in step (a)" is not limited to the manner in which sub-step Y is performed immediately after sub-step X, sub-step Z is performed immediately after sub-step Y, step (B) is performed immediately after sub-step Z, and sub-step X is performed immediately after step (B). For example, in the mode including the above-described other substeps, other substeps (substeps other than substeps X, Y and Z) may be added before substep X (between substep (B) and substep X), between substeps X and Y, between substeps Y and Z, or between substep Z and substep (B) within a range of at least 1 without impairing the effect of the present invention. By adjusting the supply flow rate, eluent intensity, etc., the desired effect can sometimes be achieved even if additional sub-steps are introduced in addition to sub-step X, Y and Z, as will be readily understood by those skilled in the art in light of this disclosure.
< substep (A1-1) >)
An eluent D-I is supplied from the eluent supply port D-I with the upstream end of the zone 1 being an eluent supply port D-I, a strongly adsorptive component is extracted from the extraction port C with the downstream end of the zone 1 being a strongly adsorptive component extraction port C, an eluent D-II is supplied from the eluent supply port D-II with the upstream end of the zone 2 being an eluent supply port D-II, a stock solution is supplied from the stock solution supply port F with the upstream end of the zone 3 being a stock solution supply port F, a weakly adsorptive component is extracted from the extraction port A with the downstream end of the zone 4 being a weakly adsorptive component extraction port A,
the desorption force of the eluent flowing in the section 1 is the strongest,
the desorption force of the eluent flowing through the section 2 is weaker than that of the eluent flowing through the section 1,
the desorption force of the eluent flowing through the sections 3 and 4 is made weaker than that of the eluent flowing through the section 2.
In the substep (a1-1), the supply of each liquid and the extraction of each component described above are continuously performed (that is, in the substep (a1-1), all of the supply of each liquid and the extraction of each component described above are performed constantly). This is also true in each sub-step described below.
In the present invention or the present specification, the description of the strength of the desorption force of the eluent in each sub-step is the strength of the desorption force of the eluent in the sub-step, and is not the description of the strength of the desorption force of the eluent in a different sub-step. For example, in the case where the desorption force of the eluent flowing through the zone 1 is made the highest in one substep constituting the step (a) and the desorption force of the eluent flowing through the zone 1 is also made the highest in the other substeps constituting the step (a), the desorption force of the eluent flowing through the zone 1 in the one substep and the desorption force of the eluent flowing through the zone 1 in the other substeps may be the same or different.
< substep (A2-1) >)
Supplying the eluent D-I from the eluent supply port D-I, extracting a strongly adsorptive component from the extraction port C, supplying the eluent D-II from the eluent supply port D-II, supplying the eluent D-III from the eluent supply port D-III with the upstream end of the section 3 as the eluent supply port D-III, extracting a weakly adsorptive component from the extraction port A,
the desorption force of the eluent flowing in the section 1 is the strongest,
the desorption force of the eluent flowing through the section 2 is weaker than that of the eluent flowing through the section 1,
the desorption force of the eluent flowing through the sections 3 and 4 is made weaker than that of the eluent flowing through the section 2.
The eluent supply port D-III in the substep (A2-1) was provided in the same piping as the raw liquid supply port F in the substep (A1-1).
< substep (A3-1) >)
Supplying the eluent D-I from the eluent supply port D-I, extracting a strongly adsorptive component from the extraction port C, supplying the eluent D-II from the eluent supply port D-II, extracting a moderately adsorptive component from the extraction port B with the downstream end of the zone 3 being the moderately adsorptive component extraction port B, supplying the eluent D-IV from the eluent supply port D-IV with the upstream end of the zone 4 being the eluent supply port D-IV, and extracting a weakly adsorptive component from the extraction port A,
the desorption force of the eluent flowing in the section 1 is the strongest,
the desorption force of the eluent flowing through the sections 2 and 3 is weaker than that of the eluent flowing through the section 1,
the desorption force of the eluent flowing through the zone 4 is made weaker than the desorption force of the eluents flowing through the zones 2 and 3.
In the substep (a2-1), the desorption force of the eluent flowing through the zones 3 and 4 is preferably also the same as the desorption force of the eluent flowing through the zone 4 in the substep (A3-1).
As an example of the above sub-step (A1-1), a sub-step of carrying out the following sub-step (A1-1ex) can be cited, but the above sub-step (A1-1) is not limited to the sub-step (A1-1 ex).
< substep (A1-1ex) >)
The upstream end of the zone 1 was set as an eluent supply port D1, and among the 4 eluents, the eluent D1 having the highest desorption power was supplied from the eluent supply port D1,
the downstream end of the section 1 is set as a strongly adsorbable component extraction port C, from which the strongly adsorbable component is extracted,
the upstream end of the zone 2 was set as an eluent supply port D2, and the second strongest eluent D2 among the 4 eluents was supplied from the eluent supply port D2,
the upstream end of the block 3 is used as a stock solution supply port F, the stock solution is supplied from the stock solution supply port F,
the downstream end of the zone 4 is set as a weakly adsorptive component extraction port a, and the weakly adsorptive component is extracted from the extraction port a.
As an example of the above sub-step (A2-1), a sub-step of carrying out the following sub-step (A2-1ex) can be cited, but the above sub-step (A2-1) is not limited to the sub-step (A2-1 ex).
< substep (A2-1ex) >)
The eluent D1 was supplied from the eluent supply port D1,
a strongly adsorbable component is drawn out from the strongly adsorbable component draw-out port C,
the eluent D2 was supplied from the eluent supply port D2,
the upstream end of the zone 3 was set as an eluent supply port D3, and the eluent D3 having the weakest desorption force among the 4 eluents was supplied from the eluent supply port D3,
and extracting the weakly adsorptive component from the weakly adsorptive component extraction port A.
The eluent supply port D3 in the substep (A2-1ex) was provided in the same piping as the raw liquid supply port F in the substep (A1-1).
As an example of the above sub-step (A3-1), a sub-step of carrying out the following sub-step (A3-1ex) can be cited, but the above sub-step (A3-1) is not limited to the sub-step (A3-1 ex).
< substep (A3-1ex) >)
An eluent D1 was supplied from the eluent supply port D1,
a strongly adsorbable component is drawn out from the strongly adsorbable component draw-out port C,
the eluent D2 was supplied from the eluent supply port D2,
the downstream end of the zone 3 was set as a middle-adsorbable component extraction port B, from which the middle-adsorbable component was extracted,
the upstream end of the zone 4 was set as an eluent supply port D4, and the third strongest eluent D4 among the 4 eluents was supplied from the eluent supply port D4,
and extracting the weakly adsorptive component from the weakly adsorptive component extraction port A.
Taking a case where each section has 1 unit packed column as an example, a flowchart in the case where the above-described step (a) is sequentially performed by the above-described substeps (a1-1ex), (a2-1ex) and (A3-1ex) is shown in fig. 2. In fig. 2, the enclosing line of a square indicates 1 unit packed tower, and the numbers in the enclosing indicate the numbers (numbers in order from the left) of the unit packed towers.
After completion of step (A) in which the above substeps (A1-1ex), (A2-1ex) and (A3-1ex) are sequentially performed, step (B) is performed to transfer the raw liquid supply port F, the eluent supply port D, the weakly adsorptive component extraction port A, the intermediate adsorptive component extraction port B and the strongly adsorptive component extraction port C to the downstream side while maintaining their relative positional relationship, and FIG. 3 shows a flowchart in the case where the substeps (A1-1ex), (A2-1ex) and (A3-1ex) are sequentially performed. The unit packed towers arranged in the respective sections shown in fig. 2 are moved downstream one by one in fig. 3. In this case, the procedure starts with step (a) shown in fig. 2, and then step (B) is performed as 1 group, and by performing 4 groups, the procedure returns to the mode shown in fig. 2 again.
Embodiment 2
Embodiment 2 also uses a circulation system having 4 or more unit packed columns, as in embodiment 1. The circulation system is divided into 4 sections 1 to 4 which are continuous in an annular shape from the upstream side to the downstream side so that each section has at least 1 unit packed column. In addition, as the eluent, using the desorption force different 4 kinds of eluent d-I ~ d-IV.
In embodiment 2, as the step (a), the following substeps (a1-2), (a2-2) and (A3-2) are carried out in this order.
< substep (A1-2) >)
The upstream end of the zone 1 is used as an eluent supply port D-II, an eluent D-II is supplied from the eluent supply port D-II, the upstream end of the zone 3 is used as a stock solution supply port F, stock solution is supplied from the stock solution supply port F, the downstream end of the zone 4 is used as a weakly adsorptive component extraction port A, and the weakly adsorptive component is extracted from the extraction port A,
the desorption force of the eluent flowing in the sections 1 and 2 is the strongest,
the desorption force of the eluent flowing through the sections 3 and 4 is made weaker than the desorption force of the eluent flowing through the sections 1 and 2.
< substep (A2-2) >)
An eluent D-I is supplied from the eluent supply port D-I with the upstream end of the zone 1 being an eluent supply port D-I, a strongly adsorptive component is extracted from the extraction port C with the downstream end of the zone 1 being a strongly adsorptive component extraction port C, an eluent D-II is supplied from the eluent supply port D-II with the upstream end of the zone 2 being an eluent supply port D-II, an eluent D-III is supplied from the eluent supply port D-III with the upstream end of the zone 3 being an eluent supply port D-III, and a weakly adsorptive component is extracted from the extraction port A,
the desorption force of the eluent flowing in the section 1 is the strongest,
the desorption force of the eluent flowing through the section 2 is weaker than that of the eluent flowing through the section 1,
the desorption force of the eluent flowing through the sections 3 and 4 is made weaker than that of the eluent flowing through the section 2.
The eluent supply port D-I in the substep (A2-2) was provided in the same piping as the eluent supply port D-II in the substep (A1-2). The eluent supply port D-III is provided in the same pipe as the raw liquid supply port F.
The desorption force of the eluent flowing through the zone 1 in the substep (A2-2) is preferably stronger than the desorption force of the eluent flowing through the zone 1 in the substep (A1-2).
< substep (A3-2) >)
Supplying the eluent D-I from the eluent supply port D-I, extracting a strongly adsorptive component from the extraction port C, supplying the eluent D-II from the eluent supply port D-II in the substep (A2-2), extracting a moderately adsorptive component from the extraction port B with the downstream end of the zone 3 being the moderately adsorptive component extraction port B, supplying the eluent D-IV from the eluent supply port D-IV with the upstream end of the zone 4 being the eluent supply port D-IV, extracting a weakly adsorptive component from the extraction port A,
the desorption force of the eluent flowing in the section 1 is the strongest,
the desorption force of the eluent flowing through the sections 2 and 3 is weaker than that of the eluent flowing through the section 1,
the desorption force of the eluent flowing through the zone 4 is made weaker than the desorption force of the eluents flowing through the zones 2 and 3.
As an example of the above sub-step (A1-2), a sub-step of carrying out the following sub-step (A1-2ex) can be cited, but the above sub-step (A1-2) is not limited to the sub-step (A1-2 ex).
< substep (A1-2ex) >)
The upstream end of the zone 1 was set as an eluent supply port D2, and the second strongest eluent D2 among the 4 eluents was supplied from the eluent supply port D2,
the upstream end of the block 3 is used as a stock solution supply port F, the stock solution is supplied from the stock solution supply port F,
the downstream end of the zone 4 is set as a weakly adsorptive component extraction port a, and the weakly adsorptive component is extracted from the extraction port a.
As an example of the above sub-step (A2-2), a sub-step of carrying out the following sub-step (A2-2ex) can be cited, but the above sub-step (A2-2) is not limited to the sub-step (A2-2 ex).
< substep (A2-2ex) >)
The upstream end of the zone 1 was set as an eluent supply port D1, and among the 4 eluents, the eluent D1 having the highest desorption power was supplied from the eluent supply port D1,
the downstream end of the section 1 is set as a strongly adsorbable component extraction port C, from which the strongly adsorbable component is extracted,
the upstream end of the zone 2 was set as an eluent supply port D2, and the second strongest eluent D2 among the 4 eluents was supplied from the eluent supply port D2,
the upstream end of the zone 3 was set as an eluent supply port D3, and the eluent D3 having the weakest desorption force among the 4 eluents was supplied from the eluent supply port D3,
and extracting the weakly adsorptive component from the weakly adsorptive component extraction port A.
The eluent supply port D1 in this substep (A2-2ex) was provided in the same piping as the eluent supply port D2 in the substep (A1-2 ex). The eluent supply port D3 was provided in the same pipe as the raw liquid supply port F in the substep (a1-2 ex).
As an example of the above sub-step (A3-2), a sub-step of carrying out the following sub-step (A3-2ex) can be cited, but the above sub-step (A3-2) is not limited to the sub-step (A3-2 ex).
< substep (A3-2ex) >)
The eluent D1 was supplied from the eluent supply port D1,
a strongly adsorbable component is drawn out from the strongly adsorbable component draw-out port C,
the eluent D2 was supplied from the eluent supply port D2 in the substep (A2-2ex),
the downstream end of the zone 3 was set as a middle-adsorbable component extraction port B, from which the middle-adsorbable component was extracted,
the upstream end of the zone 4 was set as an eluent supply port D4, and the third strongest eluent D4 among the 4 eluents was supplied from the eluent supply port D4,
and extracting the weakly adsorptive component from the weakly adsorptive component extraction port A.
Taking a case where each section has 1 unit packed column as an example, a flowchart in the case where the above-described step (a) is sequentially performed by the above-described substeps (a1-2ex), (a2-2ex) and (A3-2ex) is shown in fig. 4. In fig. 4, the enclosing line of a square indicates 1 unit packed tower, and the number in the enclosure indicates the number of the unit packed tower.
After completion of step (A) in which the above substeps (A1-2ex), (A2-2ex) and (A3-2ex) are sequentially performed, the raw liquid supply port F, the eluent supply port D, the weakly adsorptive component extraction port A, the intermediate adsorptive component extraction port B and the strongly adsorptive component extraction port C are transferred to the downstream side while maintaining their relative positional relationship in step (B), and then, FIG. 5 shows a flowchart in the case where the substeps (A1-2ex), (A2-2ex) and (A3-2ex) are sequentially performed. The unit packed towers arranged in the respective sections shown in fig. 4 are moved downstream one by one in fig. 5. In this case, the procedure starts with step (a) shown in fig. 4, and then step (B) is performed as 1 group, and by performing 4 groups, the procedure returns to the mode shown in fig. 4 again.
Embodiment 3-
Embodiment 3 uses a circulation system having 5 or more unit packed columns. The circulation system is divided into 5 sections 1 to 5 continuous in an annular shape from the upstream side to the downstream side so that each section has at least 1 unit packed column. In addition, as the eluent, using the desorption force different 4 kinds of eluent d-I ~ d-IV.
In embodiment 3, as the step (a), the following substeps (a1-3), (a2-3) and (A3-3) are carried out in this order.
< substep (A1-3) >)
An eluent D-II is supplied from the eluent supply port D-II with the upstream end of the zone 1 being the eluent supply port D-II, a stock solution is supplied from the stock solution supply port F with the upstream end of the zone 3 being the stock solution supply port F, an eluent D-III is supplied from the eluent supply port D-III with the upstream end of the zone 4 being the eluent supply port D-III, and a weakly adsorptive component is extracted from the extraction port A with the downstream end of the zone 5 being the weakly adsorptive component extraction port A,
the desorption force of the eluent flowing in the sections 1 and 2 is the strongest,
the desorption force of the eluent flowing through the zone 3 is made equal to or weaker than the desorption force of the eluents flowing through the zones 1 and 2,
the desorption force of the eluent flowing through the sections 4 and 5 is weaker than that of the eluent flowing through the section 3.
< substep (A2-3) >)
An eluent D-I is supplied from the eluent supply port D-I with the upstream end of the zone 1 being an eluent supply port D-I, a strongly adsorptive component is extracted from the extraction port C with the downstream end of the zone 1 being a strongly adsorptive component extraction port C, an eluent D-II is supplied from the eluent supply port D-II with the upstream end of the zone 2 being an eluent supply port D-II, the eluent D-III is supplied from the eluent supply port D-III, and a weakly adsorptive component is extracted from the extraction port A,
the desorption force of the eluent flowing in the section 1 is the strongest,
the desorption force of the eluent flowing through the sections 2 and 3 is weaker than that of the eluent flowing through the section 1,
the desorption force of the eluent flowing through the sections 4 and 5 is weaker than that of the eluent flowing through the sections 2 and 3.
The desorption force of the eluent flowing through the zone 1 in the substep (a2-3) is preferably stronger than the desorption force of the eluent flowing through the zone 1 in the substep (a 1-3).
The eluent supply port D-I in the substep (A2-3) was provided in the same piping as the eluent supply port D-II in the substep (A1-3).
< substep (A3-3) >)
Supplying the eluent D-I from the eluent supply port D-I, extracting a strongly adsorptive component from the extraction port C, supplying the eluent D-II from the eluent supply port D-II in the substep (A2-3), extracting a moderately adsorptive component from the extraction port B with the downstream end of the zone 4 being the moderately adsorptive component extraction port B, supplying the eluent D-IV from the eluent supply port D-IV with the upstream end of the zone 5 being the eluent supply port D-IV, extracting a weakly adsorptive component from the extraction port A,
the desorption force of the eluent flowing in the section 1 is the strongest,
the desorption force of the eluent flowing through the zone 2, the zone 3 and the zone 4 is weaker than that of the eluent flowing through the zone 1,
the desorption force of the eluent flowing through the zone 5 is made weaker than the desorption force of the eluent flowing through the zones 2, 3 and 4.
As an example of the above sub-step (A1-3), a sub-step of carrying out the following sub-step (A1-3ex) can be cited, but the above sub-step (A1-3) is not limited to the sub-step (A1-3 ex).
< substep (A1-3ex) >)
The upstream end of the zone 1 was set as an eluent supply port D2, and the second strongest eluent D2 among the 4 eluents was supplied from the eluent supply port D2,
the upstream end of the block 3 is used as a stock solution supply port F, the stock solution is supplied from the stock solution supply port F,
the upstream end of the zone 4 was set as an eluent supply port D3, and the eluent D3 having the weakest desorption force among the 4 kinds of eluents was supplied from the eluent supply port D3,
the downstream end of the zone 5 is set as a weakly adsorptive component extraction port a, and the weakly adsorptive component is extracted from the extraction port a.
As an example of the above sub-step (A2-3), a sub-step of carrying out the following sub-step (A2-3ex) can be cited, but the above sub-step (A2-3) is not limited to the sub-step (A2-3 ex).
< substep (A2-3ex) >)
The upstream end of the zone 1 was set as an eluent supply port D1, and among the 4 eluents, the eluent D1 having the highest desorption power was supplied from the eluent supply port D1,
the downstream end of the section 1 is set as a strongly adsorbable component extraction port C, from which the strongly adsorbable component is extracted,
the upstream end of the zone 2 was set as an eluent supply port D2, and the second strongest eluent D2 among the 4 eluents was supplied from the eluent supply port D2,
the eluent D3 was supplied from the eluent supply port D3,
and extracting the weakly adsorptive component from the weakly adsorptive component extraction port A.
The eluent supply port D1 in this substep (A2-3ex) was provided in the same piping as the eluent supply port D2 in the substep (A1-3 ex).
As an example of the above sub-step (A3-3), a sub-step of carrying out the following sub-step (A3-3ex) can be cited, but the above sub-step (A3-3) is not limited to the sub-step (A3-3 ex).
< substep (A3-3ex) >)
The eluent D1 was supplied from the eluent supply port D1,
a strongly adsorbable component is drawn out from the strongly adsorbable component draw-out port C,
the eluent D2 was supplied from the eluent supply port D2 in the substep (A2-3ex),
the downstream end of the zone 4 was set as a middle-adsorbable component extraction port B, from which the middle-adsorbable component was extracted,
the upstream end of the zone 5 was set as an eluent supply port D4, and the third strongest eluent D4 among the 4 eluents was supplied from the eluent supply port D4,
and extracting the weakly adsorptive component from the weakly adsorptive component extraction port A.
Taking a case where each section has 1 unit packed column as an example, a flowchart in the case where the above-described step (a) is sequentially performed by the above-described substeps (a1-3ex), (a2-3ex), and (A3-3ex) is shown in fig. 6. In fig. 6, the enclosing line of a square indicates 1 unit packed tower, and the number in the enclosure indicates the number of the unit packed tower.
After completion of step (A) in which the above substeps (A1-3ex), (A2-3ex) and (A3-3ex) are sequentially performed, the raw liquid supply port F, the eluent supply port D, the weakly adsorptive component extraction port A, the intermediate adsorptive component extraction port B and the strongly adsorptive component extraction port C are transferred to the downstream side while maintaining their relative positional relationship in step (B), and then, FIG. 7 shows a flowchart in the case where the substeps (A1-3ex), (A2-3ex) and (A3-3ex) are sequentially performed. The unit packed towers arranged in the respective sections shown in fig. 6 are moved downstream one by one in fig. 7. In this case, the procedure starts with step (a) shown in fig. 6, and then step (B) is performed as 1 group, and the procedure is performed as 5 groups, thereby returning to the scheme shown in fig. 6 again.
Embodiment 4
Embodiment 4 uses a circulation system having 7 or more unit packed columns. The circulation system is divided into 5 sections 1 to 5 continuous in an annular shape from the upstream side to the downstream side so that each section has at least 1 unit packed column. Further, as the eluent, 5 kinds of eluents d-I to d-V different in desorption force were used.
In embodiment 4, as the step (a), the following substeps (a1-4), (a2-4) and (A3-4) are carried out in this order.
< substep (A1-4) >)
An eluent D-II is supplied from the eluent supply port D-II with the upstream end of the zone 1 being the eluent supply port D-II, a stock solution is supplied from the stock solution supply port F with the upstream end of the zone 3 being the stock solution supply port F, an eluent D-III is supplied from the eluent supply port D-III with the upstream end of the zone 4 being the eluent supply port D-III, and a weakly adsorptive component is extracted from the extraction port A with the downstream end of the zone 5 being the extraction port A,
the desorption force of the eluent flowing in the sections 1 and 2 is the strongest,
the desorption force of the eluent flowing through the zone 3 is made equal to or weaker than the desorption force of the eluents flowing through the zones 1 and 2,
the desorption force of the eluent flowing through the sections 4 and 5 is weaker than that of the eluent flowing through the section 3.
< substep (A2-4) >)
An eluent D-I is supplied from the eluent supply port D-I with the upstream end of the zone 1 being an eluent supply port D-I, a strongly adsorptive component is extracted from the extraction port C with the downstream end of the zone 1 being a strongly adsorptive component extraction port C, an eluent D-II is supplied from the eluent supply port D-II with the upstream end of the zone 2 being an eluent supply port D-II, an eluent D-IV is supplied from the eluent supply port D-IV with the upstream end of the zone 4 being an eluent supply port D-IV, and a weakly adsorptive component is extracted from the extraction port A,
the desorption force of the eluent flowing in the section 1 is the strongest,
the desorption force of the eluent flowing through the sections 2 and 3 is weaker than that of the eluent flowing through the section 1,
the desorption force of the eluent flowing through the sections 4 and 5 is weaker than that of the eluent flowing through the sections 2 and 3.
The desorption force of the eluent flowing through the zone 1 in the substep (a2-4) is preferably stronger than the desorption force of the eluent flowing through the zone 1 in the substep (a 1-4).
The eluent supply port D-I in the substep (A2-4) was provided in the same piping as the eluent supply port D-II in the substep (A1-4).
< substep (A3-4) >)
Supplying the eluent D-I from the eluent supply port D-I, extracting a strongly adsorptive component from the extraction port C, supplying the eluent D2 from the eluent supply port D2 in the substep (A2-4), extracting a moderately adsorptive component from the extraction port B with the downstream end of the zone 4 being the moderately adsorptive component extraction port B, supplying the eluent D-V from the eluent supply port D-V and extracting a weakly adsorptive component from the extraction port A with the upstream end of the zone 5 being the eluent supply port D-V,
the desorption force of the eluent flowing in the section 1 is the strongest,
the desorption force of the eluent flowing through the zone 2, the zone 3 and the zone 4 is weaker than that of the eluent flowing through the zone 1,
the desorption force of the eluent flowing through the zone 5 is made weaker than the desorption force of the eluent flowing through the zones 2, 3 and 4.
As an example of the above sub-step (A1-4), a sub-step of carrying out the following sub-step (A1-4ex) can be cited, but the above sub-step (A1-4) is not limited to the sub-step (A1-4 ex).
< substep (A1-4ex) >)
The upstream end of the zone 1 was set as an eluent supply port D2, and the second strongest eluent D2 among the 5 eluents was supplied from the eluent supply port D2,
the upstream end of the block 3 is used as a stock solution supply port F, the stock solution is supplied from the stock solution supply port F,
the upstream end of the zone 4 was set as an eluent supply port D3, and the eluent D3 having the weakest desorption force among the 5 kinds of eluents was supplied from the eluent supply port D3,
the downstream end of the zone 5 is set as a weakly adsorptive component extraction port a, and the weakly adsorptive component is extracted from the extraction port a.
In the substep (A3-3) described above, the desorption force of the eluent flowing through the zones 4 and 5 is also preferably the same as the desorption force of the eluent flowing through the zone 5 in the substep (A3-4) described above.
As an example of the above sub-step (A2-4), a sub-step of carrying out the following sub-step (A2-4ex) can be cited, but the above sub-step (A2-4) is not limited to the sub-step (A2-4 ex).
< substep (A2-4ex) >)
The upstream end of the zone 1 was set as an eluent supply port D1, and among the 5 eluents, eluent D1 having the strongest desorption force was supplied from the eluent supply port D1,
the downstream end of the section 1 is set as a strongly adsorbable component extraction port C, from which the strongly adsorbable component is extracted,
the upstream end of the zone 2 was set as an eluent supply port D2, and the second strongest eluent D2 among the 5 eluents was supplied from the eluent supply port D2,
the upstream end of the zone 4 was set as an eluent supply port D4, and the eluent D4 with the fourth strongest desorption force among the 5 kinds of eluents was supplied from the eluent supply port D4,
and extracting the weakly adsorptive component from the weakly adsorptive component extraction port A.
The eluent supply port D1 in this substep (A2-4ex) was provided in the same piping as the eluent supply port D2 in the substep (A1-4 ex).
As an example of the above sub-step (A3-4), a sub-step of carrying out the following sub-step (A3-4ex) can be cited, but the above sub-step (A3-4) is not limited to the sub-step (A3-4 ex).
< substep (A3-4ex) >)
The eluent D1 was supplied from the eluent supply port D1,
a strongly adsorbable component is drawn out from the strongly adsorbable component draw-out port C,
the eluent D2 was supplied from the eluent supply port D2 in the substep (A2-4ex),
the downstream end of the zone 4 was set as a middle-adsorbable component extraction port B, from which the middle-adsorbable component was extracted,
the upstream end of the zone 5 was set as an eluent supply port D5, and the third strongest eluent D5 among the 5 eluents was supplied from the eluent supply port D5,
and extracting the weakly adsorptive component from the weakly adsorptive component extraction port A.
FIG. 8 shows an example of the flow of the case where the above-described step (A) sequentially performs the above-described substeps (A1-4ex), (A2-4ex), and (A3-4 ex). In fig. 8, a square enclosure line indicates 1 unit packed tower, and the number in the enclosure indicates the number of the unit packed tower. The system shown in fig. 8 includes 7 unit packed towers, 1 unit packed tower in the section 1, 2 unit packed towers in the section 2, 2 unit packed towers in the section 3, 1 unit packed tower in the section 4, and 1 unit packed tower in the section 5.
After completion of step (A) in which the above substeps (A1-4ex), (A2-4ex) and (A3-4ex) are sequentially performed, the raw liquid supply port F, the eluent supply port D, the weakly adsorptive component extraction port A, the intermediate adsorptive component extraction port B and the strongly adsorptive component extraction port C are transferred to the downstream side while maintaining their relative positional relationship in step (B), and then, FIG. 9 shows a flowchart in the case where the substeps (A1-4ex), (A2-4ex) and (A3-4ex) are sequentially performed. The unit packed columns arranged in the respective sections shown in fig. 8 are shifted downstream by one unit packed column in fig. 9. In this case, the procedure starts with step (a) shown in fig. 8, and then step (B) is performed as 1 group, and the procedure is performed as 7 groups, thereby returning to the method shown in fig. 8 again.
Embodiment 5
Embodiment 5 uses a circulation system having 5 or more unit packed columns. The circulation system is divided into 5 sections 1 to 5 continuous in an annular shape from the upstream side to the downstream side so that each section has at least 1 unit packed column. In addition, as the eluent, using the desorption force different 4 kinds of eluent d-I ~ d-IV.
In embodiment 5, as the step (a), the following substeps (a1-5), (a2-5) and (A3-5) are carried out in this order.
< substep (A1-5) >)
The raw liquid is supplied from the raw liquid supply port F with the upstream end of the zone 3 being the raw liquid supply port F, the eluent D-III is supplied from the eluent supply port D-III with the upstream end of the zone 4 being the eluent supply port D-III, the weakly adsorptive component is extracted from the extraction port A with the downstream end of the zone 5 being the weakly adsorptive component extraction port A,
the desorption force of the eluent flowing in the section 3 is the strongest,
the desorption force of the eluent flowing through the sections 4 and 5 is weaker than that of the eluent flowing through the section 3.
< substep (A2-5) >)
An eluent D-I is supplied from the eluent supply port D-I with the upstream end of the zone 1 being an eluent supply port D-I, a strongly adsorptive component is extracted from the extraction port C with the downstream end of the zone 1 being a strongly adsorptive component extraction port C, an eluent D-II is supplied from the eluent supply port D-II with the upstream end of the zone 2 being an eluent supply port D-II, the eluent D-III is supplied from the eluent supply port D-III, and a weakly adsorptive component is extracted from the extraction port A,
the desorption force of the eluent flowing in the section 1 is the strongest,
the desorption force of the eluent flowing through the sections 2 and 3 is weaker than that of the eluent flowing through the section 1,
the desorption force of the eluent flowing through the sections 4 and 5 is weaker than that of the eluent flowing through the sections 2 and 3.
< substep (A3-5) >)
Supplying the eluent D-I from the eluent supply port D-I, extracting a strongly adsorptive component from the extraction port C, supplying the eluent D-II from the eluent supply port D-II, extracting a moderately adsorptive component from the extraction port B with the downstream end of the zone 4 being the moderately adsorptive component extraction port B, supplying the eluent D-IV from the eluent supply port D-IV with the upstream end of the zone 5 being the eluent supply port D-IV, extracting a weakly adsorptive component from the extraction port A,
the desorption force of the eluent flowing in the section 1 is the strongest,
the desorption force of the eluent flowing through the zone 2, the zone 3 and the zone 4 is weaker than that of the eluent flowing through the zone 1,
the desorption force of the eluent flowing through the zone 5 is made weaker than the desorption force of the eluent flowing through the zones 2, 3 and 4.
The desorption force of the eluent flowing through the zone 1 in the substep (A3-5) is preferably the same as the desorption force of the eluent flowing through the zone 1 in the substep (A2-5) described above.
As an example of the above sub-step (A1-5), a sub-step of carrying out the following sub-step (A1-5ex) can be cited, but the above sub-step (A1-5) is not limited to the sub-step (A1-5 ex).
< substep (A1-5ex) >)
The upstream end of the block 3 is used as a stock solution supply port F, the stock solution is supplied from the stock solution supply port F,
the upstream end of the zone 4 was set as an eluent supply port D3, and the eluent D3 having the weakest desorption force among the 4 kinds of eluents was supplied from the eluent supply port D3,
the downstream end of the zone 5 is set as a weakly adsorptive component extraction port a, and the weakly adsorptive component is extracted from the extraction port a.
As an example of the above sub-step (A2-5), a sub-step of carrying out the following sub-step (A2-5ex) can be cited, but the above sub-step (A2-5) is not limited to the sub-step (A2-5 ex).
< substep (A2-5ex) >)
The upstream end of the zone 1 was set as an eluent supply port D1, and among the 4 eluents, the eluent D1 having the highest desorption power was supplied from the eluent supply port D1,
the downstream end of the section 1 is set as a strongly adsorbable component extraction port C, from which the strongly adsorbable component is extracted,
the upstream end of the zone 2 was set as an eluent supply port D2, and the second strongest eluent D2 among the 4 eluents was supplied from the eluent supply port D2,
the eluent D3 was supplied from the eluent supply port D3,
and extracting the weakly adsorptive component from the weakly adsorptive component extraction port A.
As an example of the above sub-step (A3-5), a sub-step of carrying out the following sub-step (A3-5ex) can be cited, but the above sub-step (A3-5) is not limited to the sub-step (A3-5 ex).
< substep (A3-5ex) >)
The eluent D1 was supplied from the eluent supply port D1,
a strongly adsorbable component is drawn out from the strongly adsorbable component draw-out port C,
the eluent D2 was supplied from the eluent supply port D2,
the downstream end of the zone 4 was set as a middle-adsorbable component extraction port B, from which the middle-adsorbable component was extracted,
the upstream end of the zone 5 was set as an eluent supply port D4, and the third strongest eluent D4 among the 4 eluents was supplied from the eluent supply port D4,
and extracting the weakly adsorptive component from the weakly adsorptive component extraction port A.
Taking a case where each section has 1 unit packed column as an example, a flowchart in the case where the above-described step (a) is sequentially performed by the above-described substeps (a1-5ex), (a2-5ex), and (A3-5ex) is shown in fig. 13. In fig. 13, the enclosing line of a square indicates 1 unit packed tower, and the number in the enclosure indicates the number of the unit packed tower.
After completion of step (A) in which the above substeps (A1-5ex), (A2-5ex) and (A3-5ex) are sequentially performed, the raw liquid supply port F, the eluent supply port D, the weakly adsorptive component extraction port A, the intermediate adsorptive component extraction port B and the strongly adsorptive component extraction port C are transferred to the downstream side while maintaining their relative positional relationship in step (B), and then, FIG. 14 shows a flowchart in the case where the substeps (A1-5ex), (A2-5ex) and (A3-5ex) are sequentially performed. The unit packed towers arranged in the respective sections shown in fig. 13 are moved downstream one by one in fig. 14. In this case, the procedure starts with step (a) shown in fig. 13, and then step (B) is performed as 1 group, and the procedure is performed as 5 groups, thereby returning to the mode shown in fig. 13 again.
Embodiment 6-
Embodiment 6 uses a circulation system having 5 or more unit packed columns. The circulation system is divided into 5 sections 1 to 5 continuous in an annular shape from the upstream side to the downstream side so that each section has at least 1 unit packed column. In addition, as the eluent, using the desorption force different 4 kinds of eluent d-I ~ d-IV.
In embodiment 6, as the step (a), the following substeps (a1-6), (a2-6) and (A3-6) are carried out in this order.
< substep (A1-6) >)
An eluent D-II is supplied from the eluent supply port D-II with the upstream end of the zone 1 being an eluent supply port D-II, the middle adsorptive component is extracted from the extraction port B with the downstream end of the zone 3 being a middle adsorptive component extraction port B, the eluent D-IV is supplied from the eluent supply port D-IV with the upstream end of the zone 4 being an eluent supply port D-IV, the weak adsorptive component is extracted from the extraction port A with the downstream end of the zone 5 being a weak adsorptive component extraction port A,
the desorption force of the eluent flowing through the zone 1, the zone 2 and the zone 3 is the strongest,
the desorption force of the eluent flowing through the zones 4 and 5 is made weaker than the desorption force of the eluent flowing through the zones 1, 2 and 3.
< substep (A2-6) >)
An eluent D-I is supplied from the eluent supply port D-I with the upstream end of the zone 1 being an eluent supply port D-I, a strongly adsorbable component is extracted from the extraction port C with the downstream end of the zone 1 being a strongly adsorbable component extraction port C, a stock solution is supplied from the stock solution supply port F with the upstream end of the zone 3 being the stock solution supply port F, an eluent D-III is supplied from the eluent supply port D-III with the upstream end of the zone 4 being an eluent supply port D-III, and a weakly adsorbable component is extracted from the extraction port A,
the desorption force of the eluent flowing in the section 1 is the strongest,
the desorption force of the eluent flowing through the section 3 is weaker than that of the eluent flowing through the section 1,
the desorption force of the eluent flowing through the sections 4 and 5 is weaker than that of the eluent flowing through the section 3.
The desorption force of the eluent flowing through the zone 1 in the substep (A2-6) is preferably stronger than the desorption force of the eluent flowing through the zone 1 in the substep (A1-6).
The eluent supply port D-I in the substep (A2-6) was provided in the same piping as the eluent supply port D-II in the substep (A1-6).
< substep (A3-6) >)
Supplying the eluent D-I from the eluent supply port D-I, extracting the strongly adsorptive component from the extraction port C, supplying the eluent D-II from the eluent supply port D-II with the upstream end of the section 2 as the eluent supply port D-II, supplying the eluent D-III from the eluent supply port D-III, extracting the weakly adsorptive component from the extraction port A,
the desorption force of the eluent flowing in the section 1 is the strongest,
the desorption force of the eluent flowing through the sections 2 and 3 is weaker than that of the eluent flowing through the section 1,
the desorption force of the eluent flowing through the sections 4 and 5 is weaker than that of the eluent flowing through the sections 2 and 3.
The desorption force of the eluent flowing through the zone 1 in the substep (A3-6) is preferably the same as the desorption force of the eluent flowing through the zone 1 in the substep (A2-6) described above.
As an example of the above sub-step (A1-6), a sub-step of carrying out the following sub-step (A1-6ex) can be cited, but the above sub-step (A1-6) is not limited to the sub-step (A1-6 ex).
< substep (A1-6ex) >)
The upstream end of the zone 1 was set as an eluent supply port D2, and the second strongest eluent D2 among the 4 eluents was supplied from the eluent supply port D2,
the downstream end of the zone 3 was set as a middle-adsorbable component extraction port B, from which the middle-adsorbable component was extracted,
the upstream end of the zone 4 was set as an eluent supply port D4, and the third strongest eluent D4 among the 4 eluents was supplied from the eluent supply port D4,
the downstream end of the zone 5 is set as a weakly adsorptive component extraction port a, and the weakly adsorptive component is extracted from the extraction port a.
As an example of the above sub-step (A2-6), a sub-step of carrying out the following sub-step (A2-6ex) can be cited, but the above sub-step (A2-6) is not limited to the sub-step (A2-6 ex).
< substep (A2-6ex) >)
The upstream end of the zone 1 was set as an eluent supply port D1, and among the 4 eluents, the eluent D1 having the highest desorption power was supplied from the eluent supply port D1,
the downstream end of the section 1 is set as a strongly adsorbable component extraction port C, from which the strongly adsorbable component is extracted,
the upstream end of the block 3 is used as a stock solution supply port F, the stock solution is supplied from the stock solution supply port F,
the upstream end of the zone 4 was set as an eluent supply port D3, and the eluent D3 having the weakest desorption force among the 4 kinds of eluents was supplied from the eluent supply port D3,
and extracting the weakly adsorptive component from the weakly adsorptive component extraction port A.
As an example of the above sub-step (A3-6), a sub-step of carrying out the following sub-step (A3-6ex) can be cited, but the above sub-step (A3-6) is not limited to the sub-step (A3-6 ex).
< substep (A3-6ex) >)
The eluent D1 was supplied from the eluent supply port D1,
a strongly adsorbable component is drawn out from the strongly adsorbable component draw-out port C,
the upstream end of the zone 2 was set as an eluent supply port D2, and the eluent D2 was supplied from the eluent supply port D2,
the eluent D3 was supplied from the eluent supply port D3,
and extracting the weakly adsorptive component from the weakly adsorptive component extraction port A.
Taking a case where each section has 1 unit packed column as an example, a flowchart in the case where the above-described step (a) is performed in the order of the above-described substeps (a1-6ex), (a2-6ex), and (A3-6ex) is shown in fig. 15. In fig. 15, the enclosing line of a square indicates 1 unit packed tower, and the number in the enclosure indicates the number of the unit packed tower.
After completion of step (A) in which the above substeps (A1-6ex), (A2-6ex) and (A3-6ex) are sequentially performed, the raw liquid supply port F, the eluent supply port D, the weakly adsorptive component extraction port A, the intermediate adsorptive component extraction port B and the strongly adsorptive component extraction port C are transferred to the downstream side while maintaining their relative positional relationship in step (B), and then, FIG. 16 shows a flowchart in the case where the substeps (A1-6ex), (A2-6ex) and (A3-6ex) are sequentially performed. The unit packed columns arranged in the respective sections shown in fig. 15 are shifted one by one to the downstream side in fig. 16. In this case, the procedure starts with step (a) shown in fig. 15, and then step (B) is performed as 1 group, and the procedure is performed as 5 groups, thereby returning to the method shown in fig. 15 again.
Embodiment 7-
Embodiment 7 uses a circulation system having 5 or more unit packed columns. The circulation system is divided into 5 sections 1 to 5 continuous in an annular shape from the upstream side to the downstream side so that each section has at least 1 unit packed column. In addition, as the eluent, using the desorption force different 4 kinds of eluent d-I ~ d-IV.
In embodiment 7, as the step (a), the following substeps (a1-7), (a2-7) and (A3-7) are carried out in this order.
< substep (A1-7) >)
An eluent D-I is supplied from the eluent supply port D-I with the upstream end of the zone 1 being an eluent supply port D-I, a strongly adsorbable component is extracted from the extraction port C with the downstream end of the zone 1 being a strongly adsorbable component extraction port C, a stock solution is supplied from the stock solution supply port F with the upstream end of the zone 3 being a stock solution supply port F, an eluent D-III is supplied from the eluent supply port D-III with the upstream end of the zone 4 being an eluent supply port D-III, and a weakly adsorbable component is extracted from the extraction port A with the downstream end of the zone 5 being the extraction port A,
the desorption force of the eluent flowing in the section 1 is the strongest,
the desorption force of the eluent flowing through the section 3 is weaker than that of the eluent flowing through the section 1,
the desorption force of the eluent flowing through the sections 4 and 5 is weaker than that of the eluent flowing through the section 3.
< substep (A2-7) >)
Supplying the eluent D-I from the eluent supply port D-I, extracting the strongly adsorptive component from the extraction port C, supplying the eluent D-II from the eluent supply port D-II with the upstream end of the section 2 as the eluent supply port D-II, supplying the eluent D-III from the eluent supply port D-III, extracting the weakly adsorptive component from the extraction port A,
the desorption force of the eluent flowing in the section 1 is the strongest,
the desorption force of the eluent flowing through the sections 2 and 3 is weaker than that of the eluent flowing through the section 1,
the desorption force of the eluent flowing through the sections 4 and 5 is weaker than that of the eluent flowing through the sections 2 and 3.
< substep (A3-7) >)
Supplying the eluent D-I from the eluent supply port D-I, extracting a strongly adsorptive component from the extraction port C, supplying the eluent D-II from the eluent supply port D-II, extracting a moderately adsorptive component from the extraction port B with the downstream end of the zone 4 being the moderately adsorptive component extraction port B, supplying the eluent D-IV from the eluent supply port D-IV with the upstream end of the zone 5 being the eluent supply port D-IV, extracting a weakly adsorptive component from the extraction port A,
the desorption force of the eluent flowing in the section 1 is the strongest,
the desorption force of the eluent flowing through the zone 2, the zone 3 and the zone 4 is weaker than that of the eluent flowing through the zone 1,
the desorption force of the eluent flowing through the zone 5 is made weaker than the desorption force of the eluent flowing through the zones 2, 3 and 4.
As an example of the above sub-step (A1-7), a sub-step of carrying out the following sub-step (A1-7ex) can be cited, but the above sub-step (A1-7) is not limited to the sub-step (A1-7 ex).
< substep (A1-7ex) >)
The upstream end of the zone 1 was set as an eluent supply port D1, and among the 4 eluents, the eluent D1 having the highest desorption power was supplied from the eluent supply port D1,
the downstream end of the section 1 is set as a strongly adsorbable component extraction port C, from which the strongly adsorbable component is extracted,
the upstream end of the block 3 is used as a stock solution supply port F, the stock solution is supplied from the stock solution supply port F,
the upstream end of the zone 4 was set as an eluent supply port D3, and the eluent D3 having the weakest desorption force among the 4 kinds of eluents was supplied from the eluent supply port D3,
the downstream end of the zone 5 is set as a weakly adsorptive component extraction port a, and the weakly adsorptive component is extracted from the extraction port a.
As an example of the above sub-step (A2-7), a sub-step of carrying out the following sub-step (A2-7ex) can be cited, but the above sub-step (A2-7) is not limited to the sub-step (A2-7 ex).
< substep (A2-7ex) >)
The eluent D1 was supplied from the eluent supply port D1,
a strongly adsorbable component is drawn out from the strongly adsorbable component draw-out port C,
the eluent D3 was supplied from the eluent supply port D3,
and extracting the weakly adsorptive component from the weakly adsorptive component extraction port A.
As an example of the above sub-step (A3-7), a sub-step of carrying out the following sub-step (A3-7ex) can be cited, but the above sub-step (A3-7) is not limited to the sub-step (A3-7 ex).
< substep (A3-7ex) >)
The eluent D1 was supplied from the eluent supply port D1,
a strongly adsorbable component is drawn out from the strongly adsorbable component draw-out port C,
the eluent D2 was supplied from the eluent supply port D2,
the downstream end of the zone 4 was set as a middle-adsorbable component extraction port B, from which the middle-adsorbable component was extracted,
the upstream end of the zone 5 was set as an eluent supply port D4, and the third strongest eluent D4 among the 4 eluents was supplied from the eluent supply port D4,
and extracting the weakly adsorptive component from the weakly adsorptive component extraction port A.
Taking a case where each section has 1 unit packed column as an example, a flowchart in the case where the above-described step (a) is performed in the order of the above-described substeps (a1-7ex), (a2-7ex), and (A3-7ex) is shown in fig. 17. In fig. 17, the enclosing line of a square indicates 1 unit packed tower, and the number in the enclosure indicates the number of the unit packed tower.
After completion of step (A) in which the above substeps (A1-7ex), (A2-7ex) and (A3-7ex) are sequentially performed, the flow chart in the case where the raw liquid supply port F, the eluent supply port D, the weakly adsorptive component extraction port A, the intermediate adsorptive component extraction port B and the strongly adsorptive component extraction port C are transferred to the downstream side while maintaining their relative positional relationship in step (B), and the substeps (A1-7ex), (A2-7ex) and (A3-7ex) are sequentially performed is shown in FIG. 18. The unit packed towers arranged in the respective sections shown in fig. 17 are moved downstream one by one in fig. 18. In this case, the procedure starts with step (a) shown in fig. 17, and then step (B) is performed as 1 group, and the procedure is performed as 5 groups, thereby returning to the mode shown in fig. 17 again.
In the method of the present invention, the supply of the target liquid to the target site or the extraction of the target liquid from the target site can be performed by appropriately adjusting the operation of the pump provided at each point of the circulation system and the opening and closing of the valve at each point. That is, a method of supplying a target fluid or extracting a target component in a circulation system is known per se. The supply flow rate and the extraction flow rate of each liquid may be appropriately set according to the purpose such as the treatment efficiency.
In the method of the present invention, the component to be purified may be any of a strongly adsorbable component, a medium adsorbable component and a weakly adsorbable component, and among these, the method is suitable for purifying the medium adsorbable component. The method of the present invention can be suitably used for purification of protein. The method of the present invention is a suitable method for obtaining a target protein from a raw liquid containing a target protein and its degradation product and aggregates with high purity because the intermediate adsorptive component can be obtained with high purity.
The protein is not particularly limited, and for example, an antibody may be used as a component to be purified. In the present invention, the "antibody" may be a naturally occurring antibody, a chimeric antibody, or an antibody fragmented by an enzyme or the like (for example, F (ab')2Fragment, Fab' fragment, Fab fragment). In addition, alsoIncluding single chain antibodies or 2-mers (diabodies) or 3-mers (triabodies) or minibodies thereof. Alternatively, the antibody may be a single domain antibody. Further, these are examples, and all of the proteins having a specific binding ability to an antigen or derivatives thereof are included in the concept of the antibody of the present invention.
The antibody highly purified by the method of the present invention can also be used as an antibody drug. That is, the method of the present invention can be applied to separate an antibody contained in a stock solution, thereby providing a method for producing an antibody drug. More specifically, according to the method of the present invention, an antibody drug can be obtained by separating an antibody contained in a culture solution of antibody-producing cells and/or an extract solution of antibody-producing cells as a stock solution. In the present invention, the "culture solution of antibody-producing cells" and the "extract solution of antibody-producing cells" include a culture solution of antibody-producing cells or an extract solution of antibody-producing cells that are subjected to various treatments such as centrifugation or chromatography to form a certain fraction or a state of purification.
In the method of the present invention, the adsorbent to be packed in the unit packed column is appropriately selected depending on the component to be purified, and various adsorbents can be used. For example, strong acid cation exchange resins, weak acid cation exchange resins, strong base anion exchange resins, weak base anion exchange resins, synthetic adsorbents, zeolites, silica gels, and silica gels modified with functional groups (preferably octadecylsilyl-modified silica gels), and other gel filtration chromatography materials and affinity adsorbents can be used as the adsorbents.
When the component to be purified is a protein, the adsorbent is preferably an ion exchange resin. Among them, cation exchange resins can be suitably used.
The simulated moving bed mode chromatographic separation system of the present invention is a system for carrying out the method of the present invention. That is, the simulated moving bed chromatography system of the present invention has the configuration of the above-described circulation system, which is a system capable of repeating the operation of the above-described step (a) and the operation of the step (B) in this order.
[ examples ] A method for producing a compound
The present invention will be described in further detail below with reference to examples, but the present invention is not limited to the following examples.
[ preparation of stock solution ]
Cells producing human immunoglobulin G2(IgG2) were cultured, and the supernatant of the culture solution was desalted by dialysis, and then NaCl was added to adjust the salt concentration to prepare a stock solution. The contents of the antibody, its fragments and aggregates contained in the supernatant were as follows. In the following table, fragment 1 is a protein contained in a fraction having a molecular weight of less than 25000 having a peak molecular weight of around 5000, and fragment 2 is a protein contained in a fraction having a molecular weight of not less than 25000 but less than 50000. The antibody is a protein contained in a fraction having a molecular weight of 50000 or more and less than 300000 and having a peak molecular weight of about 150000. The aggregate is a protein contained in a component having a molecular weight of 300000 or more. The following composition was determined by High Performance Liquid Chromatography (HPLC) using an analytical column (Tosoh TSKgel G3000 SWXL).
[ TABLE 1]
Composition of culture supernatant Concentration (g/L)
Fragment 1 0.02
Fragment 2 0.326
Antibodies 1.564
Aggregate 0.090
[ adsorbent for Unit-packed column (column) ]
As the adsorbent, cation exchange resin (trade name: Fractogel (registered trademark)) EMD SO was used3 -(M, manufactured by Merck).
[ eluent ]
The following solutions A and B were used to prepare phosphate buffers of various NaCl concentrations and used as eluents.
< solution A >
20mM phosphate buffer pH6.0
< solution B >
20mM phosphate buffer pH6.0 containing NaCl at a concentration of 0.3M (17.53g/L)
Comparative example 1 Single column discontinuous gradient
< column >
Diameter 10mm x length 100mm 1
< stock solution >
The NaCl concentration in the stock solution was set to 2.05 g/L.
< eluent >
The following eluents were used.
[ TABLE 2]
Kind of eluent NaCl concentration (g/L) of the eluate
D1 2.39
D2 2.66
D3 17.53
< operating conditions >
The following steps 1 to 6 are sequentially performed. FIG. 10 shows a flowchart of steps 1 to 6.
The following operating conditions were set to such conditions that the recovery rate for fragments 1 and 2 of the weakly adsorptive component was 98% or more, the recovery rate for the antibody of the moderately adsorptive component was 98% or more, and the recovery rate for the aggregate of the strongly adsorptive component was 98% or more. This is the same for each comparative example and example described later.
[ TABLE 3]
Procedure (ii) Time (minutes) Flow rate (mL/min)
1 5.03 9.11
2 8.50 9.11
3 0.71 9.11
4 39.46 9.11
5 0.66 9.11
6 3.60 9.11
< results >
Recovery rate-
The following table shows the recovery rates of fragments 1 and 2 of the weakly adsorptive component, the recovery rate of the antibody of the moderately adsorptive component, and the recovery rate of the aggregate of the strongly adsorptive component. The recovery rate was calculated by 100 × [ mass in component ]/[ mass in stock solution ].
[ TABLE 4]
Figure BDA0002953809260000541
Efficiency of separation treatment
The treatment amount of the raw liquid per unit volume (unit: "L (liter) -R", R being an abbreviation of Resin) and per unit time (unit: "h (hour)", was used as the separation treatment efficiency.
The separation efficiency in comparative example 1 was 6.04 (L-dope)/(L-R). h.
Comparative example 2 Single column discontinuous gradient
< column >
Diameter 10mm x length 400mm 1
< stock solution >
The NaCl concentration in the stock solution was set to 2.05 g/L.
< eluent >
The following eluents were used.
[ TABLE 5]
Kind of eluent NaCl concentration (g/L) of the eluate
D1 2.35
D2 2.62
D3 17.53
< operating conditions >
The following steps 1 to 6 are sequentially performed. The flow charts of the steps 1 to 6 are shown in FIG. 10.
[ TABLE 6]
Procedure (ii) Time (minutes) Flow rate (mL/min)
1 5.99 43.3
2 6.35 43.3
3 0.63 43.3
4 22.48 43.3
5 0.58 43.3
6 3.55 43.3
< results >
Recovery rate-
The following table shows the recovery rates of fragments 1 and 2 of the weakly adsorptive component, the recovery rate of the antibody of the moderately adsorptive component, and the recovery rate of the aggregate of the strongly adsorptive component.
[ TABLE 7]
Figure BDA0002953809260000551
Efficiency of separation treatment
The separation efficiency in comparative example 2 was 12.51 (L-dope)/(L-R). h.
Comparative example 3 Multi-column gradient simulated moving bed System
< column >
Diameter 10mm x length 100mm 4
< stock solution >
The NaCl concentration in the stock solution was 2.23 g/L.
< eluent >
The following eluents were used.
[ TABLE 8 ]
Kind of eluent NaCl concentration (g/L) of the eluate
D1 17.53
D2 2.28
D3 2.23
< operating conditions >
Fig. 11 shows a flowchart of the operation of comparative example 3. The 1 st to 4 th steps shown in FIG. 11 were performed for 10 cycles as 1 cycle. Between the steps, the following steps were performed in which 1 column was transferred to the downstream side while maintaining the relative positional relationship among the raw liquid supply port F, the eluent supply ports D (D1 to D3), the weakly adsorptive component extraction port a, the intermediate adsorptive component extraction port B, and the strongly adsorptive component extraction port C.
The 1 st to 4 th steps shown in fig. 11 correspond to the step (a) of the present invention, but are not configured by a plurality of substeps (in other words, 1 substep is configured by 1 substep) unlike the step (a). In fig. 11, "D1", "D2", and "D3" are eluent supply ports, and eluents D1, D2, and D3 are supplied, respectively. "C" in FIG. 11 is a strongly adsorbable component withdrawal port for withdrawing the strongly adsorbable component. Similarly, "B" is a medium-adsorbable component withdrawal port from which a medium-adsorbable component is withdrawn, and "A" is a weak-adsorbable component withdrawal port from which a weak-adsorbable component is withdrawn.
The supply flow rates of the eluents (D1 to D3) and the stock solution (F) in the respective steps shown in fig. 11 are as follows. In addition, the flow rate of the liquid withdrawn is not described in the following table, but the flow rate of the strongly adsorbable component withdrawn from the strongly adsorbable component withdrawal port C is the same as the flow rate of the eluent D1 supplied thereto. In addition, the flow rate of the middle adsorptive component withdrawn from the middle adsorptive component withdrawing port B was the same as the flow rate of the feed eluent D2. The flow rate of the weakly adsorptive component extracted from the weakly adsorptive component extraction port a is the sum of the flow rate of the eluent D3 supplied and the flow rate of the raw liquid supplied from the raw liquid supply port F. That is, the supply flow rate and the extraction flow rate are always the same, and this is the same in the following comparative examples and examples.
[ TABLE 9 ]
Figure BDA0002953809260000571
< results >
Recovery rate-
The following table shows the recovery rates of fragments 1 and 2 of the weakly adsorptive component, the recovery rate of the antibody of the moderately adsorptive component, and the recovery rate of the aggregate of the strongly adsorptive component.
[ TABLE 10 ]
Figure BDA0002953809260000572
Efficiency of separation treatment
The separation efficiency in comparative example 3 was 7.11 (L-dope)/(L-R). h.
Comparative example 4 Multi-column gradient simulated moving bed System
< column >
Diameter 10mm x length 100mm 4
< stock solution >
The NaCl concentration in the stock solution was set to 2.24 g/L.
< eluent >
The following eluents were used.
[ TABLE 11 ]
Kind of eluent NaCl concentration (g/L) of the eluate
D1 17.53
D2 2.26
D3 2.24
< operating conditions >
Fig. 12 shows a flowchart of the operation of comparative example 4. The 1 st to 4 th steps shown in FIG. 12 were performed for 10 cycles as 1 cycle. Each step shown in fig. 12 is composed of 2 substeps, i.e., the 1 st substep and the 2 nd substep, and the 2 nd substep is a step of circulating the fluid in the circulation system without performing any of the supply and extraction of the liquid.
The supply flow rates of the eluents (D1 to D3) and the stock solution (F) in the respective steps shown in fig. 12 are as follows.
[ TABLE 12 ]
Figure BDA0002953809260000581
< results >
Recovery rate-
The following table shows the recovery rates of fragments 1 and 2 of the weakly adsorptive component, the recovery rate of the antibody of the medium adsorptive component, and the recovery rate of the aggregate of the strongly adsorptive component.
[ TABLE 13 ]
Figure BDA0002953809260000591
Efficiency of separation treatment
The separation efficiency in comparative example 4 was 7.19 (L-dope)/(L-R). h.
Example 1 Multi-column gradient simulated moving bed System
< column >
Diameter 10mm x length 100mm 4
< stock solution >
The NaCl concentration in the stock solution was 1.93 g/L.
< eluent >
The following eluents were used.
[ TABLE 14 ]
Kind of eluent NaCl concentration (g/L) of the eluate
D1 17.53
D2 3.59
D3 1.93
D4 2.21
< operating conditions >
Step (a) is constituted by a combination of sub-steps shown in fig. 2. This step (a) and the subsequent step (B) were set as 1 group, and 4 groups were performed to set 1 cycle, and 10 cycles were performed. The flow rates of the eluents (D1 to D4) and the stock solution (F) supplied in each step (a) are as follows.
[ TABLE 15 ]
Figure BDA0002953809260000601
< results >
Recovery rate-
The following table shows the recovery rates of fragments 1 and 2 of the weakly adsorptive component, the recovery rate of the antibody of the medium adsorptive component, and the recovery rate of the aggregate of the strongly adsorptive component.
[ TABLE 16 ]
Figure BDA0002953809260000602
Efficiency of separation treatment
The separation efficiency in example 1 was 19.696 (L-dope)/(L-R). h.
Example 2 Multi-column gradient simulated moving bed System
< column >
Diameter 10mm x length 100mm 4
< stock solution >
The NaCl concentration in the stock solution was set to 2.02 g/L.
< eluent >
The following eluents were used.
[ TABLE 17 ]
Kind of eluent NaCl concentration (g/L) of the eluate
D1 17.53
D2 3.57
D3 2.02
D4 2.21
< operating conditions >
Step (a) is constituted by a combination of sub-steps shown in fig. 4. This step (a) and the subsequent step (B) were set as 1 group, and 4 groups were performed to set 1 cycle, and 10 cycles were performed. The flow rates of the eluents (D1 to D4) and the stock solution (F) supplied in each step (a) are as follows.
[ TABLE 18 ]
Figure BDA0002953809260000611
< results >
Recovery rate-
The following table shows the recovery rates of fragments 1 and 2 of the weakly adsorptive component, the recovery rate of the antibody of the medium adsorptive component, and the recovery rate of the aggregate of the strongly adsorptive component.
[ TABLE 19 ]
Figure BDA0002953809260000612
Efficiency of separation treatment
The separation efficiency in example 2 was 18.610 (L-dope)/(L-R). h.
EXAMPLE 3 Multi-column gradient simulated moving bed System
< column >
Diameter 10mm x length 805mm 5
< stock solution >
The NaCl concentration in the stock solution was set to 2.46 g/L.
< eluent >
The following eluents were used.
[ TABLE 20 ]
Kind of eluent NaCl concentration (g/L) of the eluate
D1 17.53
D2 2.46
D3 0.00
D4 1.98
< operating conditions >
Step (a) is constituted by a combination of sub-steps shown in fig. 6. This step (a) and the subsequent step (B) were set as 1 group, and 5 groups were performed to set 1 cycle, and 10 cycles were performed. The flow rates of the eluents (D1 to D4) and the stock solution (F) supplied in each step (a) are as follows.
[ TABLE 21 ]
Figure BDA0002953809260000621
< results >
Recovery rate-
The following table shows the recovery rates of fragments 1 and 2 of the weakly adsorptive component, the recovery rate of the antibody of the medium adsorptive component, and the recovery rate of the aggregate of the strongly adsorptive component.
[ TABLE 22 ]
Figure BDA0002953809260000631
Efficiency of separation treatment
The separation efficiency in example 3 was 16.499 (L-dope)/(L-R). h.
EXAMPLE 4 Multi-column gradient simulated moving bed System
< column >
Diameter 10mm x length 100mm 7
< stock solution >
The NaCl concentration in the stock solution was set to 2.57 g/L.
< eluent >
The following eluents were used.
[ TABLE 23 ]
Kind of eluent NaCl concentration (g/L) of the eluate
D1 17.53
D2 2.57
D3 0.00
D4 0.37
D5 1.80
< operating conditions >
Step (a) is constituted by a combination of sub-steps shown in fig. 8. This step (a) and the subsequent step (B) were set as 1 group, and 7 groups were performed to set 1 cycle, and 10 cycles were performed. The flow rates of the eluents (D1 to D5) and the stock solution (F) supplied in each step (a) are as follows.
[ TABLE 24 ]
Figure BDA0002953809260000641
< results >
Recovery rate-
The following table shows the recovery rates of fragments 1 and 2 of the weakly adsorptive component, the recovery rate of the antibody of the medium adsorptive component, and the recovery rate of the aggregate of the strongly adsorptive component.
[ TABLE 25 ]
Figure BDA0002953809260000642
Efficiency of separation treatment
The separation efficiency in example 4 was 15.225 (L-dope)/(L-R). h.
As described above, in the simulated moving bed chromatography, when 2 or more kinds of eluents are used, the positional relationship among the weakly adsorptive component extraction port a, the intermediate adsorptive component extraction port B, the strongly adsorptive component extraction port C, and the raw liquid supply port F in the circulation system is set to a specific relationship defined in the present invention, whereby the weakly adsorptive component, the intermediate adsorptive component, and the strongly adsorptive component can be separated with sufficiently high purity with a smaller amount of the adsorbent used. According to this example, it was shown that the target antibody can be obtained with high purity and high efficiency in the medium-adsorbable fraction.
EXAMPLE 5 Multi-column gradient simulated moving bed System
< column >
Diameter 10mm x length 80mm 5
< stock solution >
The NaCl concentration in the stock solution was set to 2.46 g/L.
< eluent >
The following eluents were used.
[ TABLE 26 ]
Kind of eluent NaCl concentration (g/L) of the eluate
D1 17.53
D2 2.46
D3 0.00
D4 1.98
Step (a) is constituted by a combination of sub-steps shown in fig. 13. This step (a) and the subsequent step (B) were set as 1 group, and 5 groups were performed to set 1 cycle, and 10 cycles were performed. The flow rates of the eluents (D1 to D4) and the stock solution (F) supplied in each step (a) are as follows.
[ TABLE 27 ]
Figure BDA0002953809260000651
< results >
Recovery rate-
The following table shows the recovery rates of fragments 1 and 2 of the weakly adsorptive component, the recovery rate of the antibody of the medium adsorptive component, and the recovery rate of the aggregate of the strongly adsorptive component.
[ TABLE 28 ]
Figure BDA0002953809260000661
Efficiency of separation treatment
The separation efficiency in example 5 was 16.502 (L-dope)/(L-R). h.
EXAMPLE 6 Multi-column gradient simulated moving bed System
< column >
Diameter 10mm x length 80mm 5
< stock solution >
The NaCl concentration in the stock solution was set to 2.55 g/L.
< eluent >
The following eluents were used.
[ TABLE 29 ]
Kind of eluent NaCl concentration (g/L) of the eluate
D1 17.53
D2 2.55
D3 0.00
D4 2.03
< operating conditions >
Step (a) is constituted by a combination of sub-steps shown in fig. 15. This step (a) and the subsequent step (B) were set as 1 group, and 5 groups were performed to set 1 cycle, and 10 cycles were performed. The flow rates of the eluents (D1 to D4) and the stock solution (F) supplied in each step (a) are as follows.
[ TABLE 30 ]
Figure BDA0002953809260000671
< results >
Recovery rate-
The following table shows the recovery rates of fragments 1 and 2 of the weakly adsorptive component, the recovery rate of the antibody of the medium adsorptive component, and the recovery rate of the aggregate of the strongly adsorptive component.
[ TABLE 31 ]
Figure BDA0002953809260000672
Efficiency of separation treatment
The separation efficiency in example 6 was 18.898 (L-dope)/(L-R). h.
EXAMPLE 7 Multi-column gradient simulated moving bed System
< column >
Diameter 10mm x length 80mm 5
< stock solution >
The NaCl concentration in the stock solution was set to 2.55 g/L.
< eluent >
The following eluents were used.
[ TABLE 32 ]
Kind of eluent NaCl concentration (g/L) of the eluate
D1 17.53
D2 2.55
D3 0.00
D4 2.03
< operating conditions >
Step (a) is constituted by a combination of sub-steps shown in fig. 17. This step (a) and the subsequent step (B) were set as 1 group, and 5 groups were performed to set 1 cycle, and 10 cycles were performed. The flow rates of the eluents (D1 to D4) and the stock solution (F) supplied in each step (a) are as follows.
[ TABLE 33 ]
Figure BDA0002953809260000681
< results >
Recovery rate-
The following table shows the recovery rates of fragments 1 and 2 of the weakly adsorptive component, the recovery rate of the antibody of the medium adsorptive component, and the recovery rate of the aggregate of the strongly adsorptive component.
[ TABLE 34 ]
Figure BDA0002953809260000682
Efficiency of separation treatment
The separation efficiency in example 7 was 16.502 (L-dope)/(L-R). h.
Although the present invention has been described in connection with other embodiments, there is no intention to limit the invention in any detail of the description unless specifically stated, but rather should be construed broadly within its spirit and scope as indicated in the appended claims.
The present application claims priority based on japanese patent application nos. 2018-215950, 2018, 16, and 2019-088523, 2019, 8, which are filed in japan, and the contents of which are incorporated herein by reference as part of the description of the present specification.
(description of reference numerals)
100 circulation system
10a, 10b, 10c, 10d unit packed column (column)
Ab adsorbent
R1, R2, R3 and R4 stop valves
2a, 2b, 2c, 2d weak adsorptive component extraction line
A1, A2, A3 and A4 weak adsorptive component extraction valves
3a, 3b, 3c, 3d, and a line for drawing out the adsorptive component
Sucking valve for adsorptive component in B1, B2, B3 and B4
4a, 4b, 4c, 4d strong adsorptive component extraction line
C1, C2, C3 and C4 strong adsorptive component extraction valve
T1, T2, T3, T4 check valves
1 piping
2J weak-adsorbability component flow combining pipe
3J middle adsorption component flow combining pipe
4J strong adsorptive component flow combining pipe
6 stock solution pot
7 stock solution
8a, 8b, 8c, 8d eluent tank
9a, 9b, 9c, 9d eluent
11 stock solution supply line
11a, 11b, 11c, 11d stock solution supply branch line
F1, F2, F3 and F4 stock solution supply valves
12. 13, 14, 15 eluent supply lines
12a, 12b, 12c, 12d eluent supply branch line
13a, 13b, 13c, 13d eluent supply branch line
14a, 14b, 14c, 14d eluent supply branch line
15a, 15b, 15c, 15d eluent supply branch line
E1a, E2a, E3a, E4a eluent supply valve
E1b, E2b, E3b, E4b eluent supply valve
E1c, E2c, E3c, E4c eluent supply valve
E1d, E2d, E3d, E4d eluent supply valve
P1 circulating pump
P2 stock solution supply pump
P3, P4, P5, P6 eluent supply pumps.

Claims (13)

1. A simulated moving bed mode chromatographic separation method comprises the following steps: separating weakly adsorptive components, strongly adsorptive components and intermediate adsorptive components contained in a raw liquid with respect to an adsorbent by using 2 or more kinds of eluents, the adsorbability of the intermediate adsorptive component being between that of the weakly adsorptive component and that of the strongly adsorptive component, with a circulation system in which a plurality of unit packed columns packed with the adsorbent are connected in series via pipes and connected in a ring shape,
the piping of the circulation system is provided with a raw liquid supply port F, 2 or more eluent supply ports D corresponding to the 2 or more eluents, a drawing port a for weakly adsorbing components containing the weakly adsorbing components, a drawing port B for moderately adsorbing components containing the moderately adsorbing components, and a drawing port C for strongly adsorbing components containing the strongly adsorbing components, and the positions of the raw liquid supply port F, the drawing port a, the drawing port B, and the drawing port C are set as follows (a) to (C):
(a) the extraction port B is provided downstream of the raw liquid supply port F with at least 1 unit packed column interposed therebetween;
(b) the extraction port C is provided in a pipe having the raw liquid supply port F, or the extraction port C is provided on the upstream side of the raw liquid supply port F with at least 1 unit packed column interposed therebetween;
(c) the extraction port A is provided in a pipe having the extraction port B, or the extraction port A is provided downstream of the extraction port B with at least 1 unit packed column interposed therebetween,
the chromatographic separation method comprises the following steps of sequentially repeating the step A and the step B,
the step A is a step of simultaneously or separately supplying a raw liquid and 2 or more kinds of eluents from the raw liquid supply port F and the 2 or more kinds of eluent supply ports D, and simultaneously or separately extracting a weakly adsorptive component, a medium adsorptive component and a strongly adsorptive component from the extraction port A, the extraction port B and the extraction port C,
and a step B of transferring the raw liquid supply port F, the eluent supply port D, the extraction port a, the extraction port B, and the extraction port C to a downstream side while maintaining a relative positional relationship after the step a is completed.
2. A simulated moving bed mode chromatographic separation process according to claim 1, wherein,
the step a is composed of a plurality of substeps including a substep of supplying the stock solution and a substep of not supplying the stock solution.
3. A simulated moving bed mode chromatographic separation process according to claim 1 or 2, wherein,
the extraction port C is provided on the downstream side of an eluent supply port D1 through which an eluent D1 having the strongest desorption force among 2 or more kinds of eluents is supplied, at least 1 unit packed column is arranged between the eluent supply port D1 and the extraction port C, and in the step a, a strongly adsorbable component in the same amount as the supply amount of the eluent D1 is extracted from the extraction port C while the eluent D1 is supplied.
4. A simulated moving bed-based chromatographic separation process according to any one of claims 1 to 3, wherein,
the extraction port B is provided on the downstream side of the eluent supply port D2 to which the eluent D2 having the second strongest desorption force among 2 or more kinds of eluents is supplied, at least 1 unit packed column is arranged from the eluent supply port D2 to the extraction port B, and in the step a, while the eluent D2 is supplied, a period of time for extracting the middle adsorptive component in the same amount as the supply amount of the eluent D2 from the extraction port B is set.
5. A simulated moving bed-based chromatographic separation process according to any one of claims 1 to 4, wherein,
4 to 6 kinds of eluents having different desorption forces are used.
6. A simulated moving bed-based chromatographic separation process according to any one of claims 1 to 5, wherein,
the circulation system has 4 or more unit packed columns, is divided into 4 sections 1 to 4 sections, which are continuous in an annular shape, from the upstream side to the downstream side so that each section has at least 1 unit packed column, and performs the following substep A1-1, substep A2-1, and substep A3-1 in step A using the 2 or more eluents,
in the substep A1-1, the eluent D-I is supplied from the eluent supply port D-I with the upstream end of the section 1 being the eluent supply port D-I, the strongly adsorptive component is extracted from the extraction port C with the downstream end of the section 1 being the extraction port C, the eluent D-II is supplied from the eluent supply port D-II with the upstream end of the section 2 being the eluent supply port D2, the raw liquid is supplied from the raw liquid supply port F with the upstream end of the section 3 being the raw liquid supply port F, and the weakly adsorptive component is extracted from the extraction port A with the downstream end of the section 4 being the extraction port A,
the desorption force of the eluent flowing in the section 1 is the strongest,
the desorption force of the eluent flowing through the section 2 is weaker than that of the eluent flowing through the section 1,
making the desorption force of the eluent flowing in the section 3 and the section 4 weaker than that of the eluent flowing in the section 2;
in the substep A2-1, the eluent D-I is supplied from the eluent supply port D-I, the strongly adsorptive component is extracted from the extraction port C, the eluent D-II is supplied from the eluent supply port D-II, the eluent D-III is supplied from the eluent supply port D-III at the upstream end of the section 3 as the eluent supply port D-III, and the weakly adsorptive component is extracted from the extraction port A,
the desorption force of the eluent flowing in the section 1 is the strongest,
the desorption force of the eluent flowing through the section 2 is weaker than that of the eluent flowing through the section 1,
making the desorption force of the eluent flowing in the section 3 and the section 4 weaker than that of the eluent flowing in the section 2;
in the substep A3-1, the eluent D-I is supplied from the eluent supply port D-I, the strongly adsorptive component is extracted from the extraction port C, the eluent D-II is supplied from the eluent supply port D-II, the downstream end of the zone 3 is taken as the extraction port B, the intermediate adsorptive component is extracted from the extraction port B, the upstream end of the zone 4 is taken as the eluent supply port D-IV, the eluent D-IV is supplied from the eluent supply port D-IV, and the weakly adsorptive component is extracted from the extraction port A,
the desorption force of the eluent flowing in the section 1 is the strongest,
the desorption force of the eluent flowing through the sections 2 and 3 is made weaker than that of the eluent flowing through the section 1,
the desorption force of the eluent flowing through the zone 4 is made weaker than the desorption force of the eluents flowing through the zones 2 and 3.
7. A simulated moving bed-based chromatographic separation process according to any one of claims 1 to 5, wherein,
the circulation system has 4 or more unit packed columns, is divided into 4 sections 1 to 4 sections, which are continuous in an annular shape, from the upstream side to the downstream side so that each section has at least 1 unit packed column, and performs the following substep a1-2, substep a2-2, and substep A3-2 in step a using the 2 or more eluents:
in the substep A1-2, the eluent D-II is supplied from the eluent supply port D-II with the upstream end of the section 1 being the eluent supply port D-II, the raw liquid is supplied from the raw liquid supply port F with the upstream end of the section 3 being the raw liquid supply port F, and the weakly adsorptive component is extracted from the extraction port A with the downstream end of the section 4 being the extraction port A,
the desorption force of the eluent flowing through the zone 1 and the zone 2 is maximized,
making the desorption force of the eluent flowing in the section 3 and the section 4 weaker than that of the eluent flowing in the section 1 and the section 2;
in the substep A2-2, the eluent D-I is supplied from the eluent supply port D-I with the upstream end of the section 1 being the eluent supply port D-I, the strongly adsorptive component is extracted from the extraction port C with the downstream end of the section 1 being the extraction port C, the eluent D-II is supplied from the eluent supply port D-II with the upstream end of the section 2 being the eluent supply port D-II, the eluent D-III is supplied from the eluent supply port D-III with the upstream end of the section 3 being the eluent supply port D-III, and the weakly adsorptive component is extracted from the extraction port A,
the desorption force of the eluent flowing in the section 1 is the strongest,
the desorption force of the eluent flowing through the section 2 is weaker than that of the eluent flowing through the section 1,
making the desorption force of the eluent flowing in the section 3 and the section 4 weaker than that of the eluent flowing in the section 2;
in the substep A3-2, the eluent D-I is supplied from the eluent supply port D-I, a strongly adsorbable component is extracted from the extraction port C, the eluent D-II is supplied from the eluent supply port D-II in the substep A2-2, a moderately adsorbable component is extracted from the extraction port B with the downstream end of the zone 3 being the extraction port B, an eluent D-IV is supplied from the eluent supply port D-IV with the upstream end of the zone 4 being the eluent supply port D-IV, and a weakly adsorbable component is extracted from the extraction port A,
the desorption force of the eluent flowing in the section 1 is the strongest,
the desorption force of the eluent flowing through the sections 2 and 3 is made weaker than that of the eluent flowing through the section 1,
the desorption force of the eluent flowing through the zone 4 is made weaker than the desorption force of the eluents flowing through the zones 2 and 3.
8. A simulated moving bed-based chromatographic separation process according to any one of claims 1 to 5, wherein,
the circulation system has 5 or more unit packed columns, is divided into 5 sections 1 to 5 sections, which are continuous in an annular shape, from the upstream side to the downstream side so that each section has at least 1 unit packed column, and performs the following substep a1-3, substep a2-3, and substep A3-3 in step a using the 2 or more eluents:
in the substep A1-3, the eluent D-II is supplied from the eluent supply port D-II with the upstream end of the segment 1 being the eluent supply port D-II, the raw liquid is supplied from the raw liquid supply port F with the upstream end of the segment 3 being the raw liquid supply port F, the eluent D-III is supplied from the eluent supply port D-III with the upstream end of the segment 4 being the eluent supply port D-III, and the weakly adsorptive component is extracted from the extraction port A with the downstream end of the segment 5 being the extraction port A,
the desorption force of the eluent flowing through the zone 1 and the zone 2 is maximized,
the desorption force of the eluent flowing through the zone 3 is made equal to or weaker than the desorption force of the eluent flowing through the zones 1 and 2,
making the desorption force of the eluent flowing in the section 4 and the section 5 weaker than that of the eluent flowing in the section 3;
in the substep A2-3, an eluent D-I is supplied from the eluent supply port D-I with the upstream end of the section 1 being the eluent supply port D-I, a strongly adsorptive component is extracted from the extraction port C with the downstream end of the section 1 being the extraction port C, an eluent D-II is supplied from the eluent supply port D-II with the upstream end of the section 2 being the eluent supply port D-II, the eluent D-III is supplied from the eluent supply port D-III, and a weakly adsorptive component is extracted from the extraction port A,
the desorption force of the eluent flowing in the section 1 is the strongest,
the desorption force of the eluent flowing through the sections 2 and 3 is made weaker than that of the eluent flowing through the section 1,
making the desorption force of the eluent flowing in the section 4 and the section 5 weaker than that of the eluent flowing in the section 2 and the section 3;
in the substep A3-3, the eluent D-I is supplied from the eluent supply port D-I, a strongly adsorbable component is extracted from the extraction port C, the eluent D-II is supplied from the eluent supply port D-II in the substep A2-3, a moderately adsorbable component is extracted from the extraction port B with the downstream end of the zone 4 being the extraction port B, an eluent D-IV is supplied from the eluent supply port D-IV with the upstream end of the zone 5 being the eluent supply port D-IV, and a weakly adsorbable component is extracted from the extraction port A,
the desorption force of the eluent flowing in the section 1 is the strongest,
the desorption force of the eluent flowing through the zone 2, the zone 3 and the zone 4 is weaker than that of the eluent flowing through the zone 1,
the desorption force of the eluent flowing through the zone 5 is made weaker than the desorption force of the eluent flowing through the zones 2, 3 and 4.
9. A simulated moving bed-based chromatographic separation process according to any one of claims 1 to 5, wherein,
the circulation system has 7 or more unit packed columns, is divided into 5 sections, namely, sections 1 to 5, which are continuous in an annular shape, from the upstream side to the downstream side so that each section has at least 1 unit packed column, and performs the following substep a1-4, substep a2-4, and substep A3-4 in step a using the 2 or more eluents:
in the substep A1-4, the eluent D-II is supplied from the eluent supply port D-II with the upstream end of the section 1 being the eluent supply port D-II, the raw liquid is supplied from the raw liquid supply port F with the upstream end of the section 3 being the raw liquid supply port F, the eluent D-III is supplied from the eluent supply port D-III with the upstream end of the section 4 being the eluent supply port D-III, and the weakly adsorptive component is extracted from the extraction port A with the downstream end of the section 5 being the extraction port A,
the desorption force of the eluent flowing through the zone 1 and the zone 2 is maximized,
the desorption force of the eluent flowing through the zone 3 is made equal to or weaker than the desorption force of the eluent flowing through the zones 1 and 2,
making the desorption force of the eluent flowing in the section 4 and the section 5 weaker than that of the eluent flowing in the section 3;
in the substep A2-4, the upstream end of the section 1 is used as an eluent supply port D-I, and an eluent D-I is supplied from the eluent supply port D-I, the downstream end of the section 1 is used as the extraction port C, and a strongly adsorptive component is extracted from the extraction port C, the upstream end of the section 2 is used as an eluent supply port D-II, and an eluent D-II is supplied from the eluent supply port D-II, the upstream end of the section 4 is used as an eluent supply port D-IV, and an eluent D-IV is supplied from the eluent supply port D-IV, and a weakly adsorptive component is extracted from the extraction port A,
the desorption force of the eluent flowing in the section 1 is the strongest,
the desorption force of the eluent flowing through the sections 2 and 3 is made weaker than that of the eluent flowing through the section 1,
making the desorption force of the eluent flowing in the section 4 and the section 5 weaker than that of the eluent flowing in the section 2 and the section 3;
in the substep A3-4, the eluent D-I is supplied from the eluent supply port D-I, the strongly adsorptive component is extracted from the extraction port C, the eluent D2 is supplied from the eluent supply port D2 in the substep A2-4, the intermediate adsorptive component is extracted from the extraction port B with the downstream end of the block 4 being the extraction port B, the eluent D-V is supplied from the eluent supply port D-V with the upstream end of the block 5 being the eluent supply port D-V, and the weakly adsorptive component is extracted from the extraction port A,
the desorption force of the eluent flowing in the section 1 is the strongest,
the desorption force of the eluent flowing through the zone 2, the zone 3 and the zone 4 is weaker than that of the eluent flowing through the zone 1,
the desorption force of the eluent flowing through the zone 5 is made weaker than the desorption force of the eluent flowing through the zones 2, 3 and 4.
10. A simulated moving bed-based chromatographic separation process according to any one of claims 1 to 5, wherein,
the circulation system has 5 or more unit packed columns, is divided into 5 sections 1 to 5 sections, which are continuous in an annular shape, from the upstream side to the downstream side so that each section has at least 1 unit packed column, and performs the following substep a1-5, substep a2-5, and substep A3-5 in step a using the 2 or more eluents:
in the substep A1-5, the raw liquid is supplied from the raw liquid supply port F with the upstream end of the zone 3 being the raw liquid supply port F, the eluent D-III is supplied from the eluent supply port D-III with the upstream end of the zone 4 being the eluent supply port D-III, and the weakly adsorptive component is extracted from the extraction port A with the downstream end of the zone 5 being the extraction port A,
the desorption force of the eluent flowing in the section 3 is the strongest,
making the desorption force of the eluent flowing in the section 4 and the section 5 weaker than that of the eluent flowing in the section 3;
in the substep A2-5, the upstream end of the section 1 is used as an eluent supply port D-I, an eluent D-I is supplied from the eluent supply port D-I, the downstream end of the section 1 is used as the extraction port C, and a strongly adsorptive component is extracted from the extraction port C, the upstream end of the section 2 is used as an eluent supply port D-II, an eluent D-II is supplied from the eluent supply port D-II, the eluent D-III is supplied from the eluent supply port D-III, and a weakly adsorptive component is extracted from the extraction port A,
the desorption force of the eluent flowing in the section 1 is the strongest,
the desorption force of the eluent flowing through the sections 2 and 3 is made weaker than that of the eluent flowing through the section 1,
making the desorption force of the eluent flowing in the section 4 and the section 5 weaker than that of the eluent flowing in the section 2 and the section 3;
in the substep A3-5, the eluent D-I is supplied from the eluent supply port D-I, the strongly adsorptive component is extracted from the extraction port C, the eluent D-II is supplied from the eluent supply port D-II, the downstream end of the section 4 is taken as the extraction port B, the intermediate adsorptive component is extracted from the extraction port B, the upstream end of the section 5 is taken as the eluent supply port D-IV, the eluent D-IV is supplied from the eluent supply port D-IV, and the weakly adsorptive component is extracted from the extraction port A,
the desorption force of the eluent flowing in the section 1 is the strongest,
the desorption force of the eluent flowing through the zone 2, the zone 3 and the zone 4 is weaker than that of the eluent flowing through the zone 1,
the desorption force of the eluent flowing through the zone 5 is made weaker than the desorption force of the eluent flowing through the zones 2, 3 and 4.
11. A simulated moving bed-based chromatographic separation process according to any one of claims 1 to 5, wherein,
the circulation system has 5 or more unit packed columns, is divided into 5 sections 1 to 5 sections, which are continuous in an annular shape, from the upstream side to the downstream side so that each section has at least 1 unit packed column, and performs the following substeps a1-6, substeps a2-6, and substeps A3-6 in step a using the 2 or more eluents:
in the substep A1-6, an eluent D-II is supplied from the eluent supply port D-II with the upstream end of the section 1 being the eluent supply port D-II, the mid-adsorptive component is extracted from the extraction port B with the downstream end of the section 3 being the extraction port B, an eluent D-IV is supplied from the eluent supply port D-IV with the upstream end of the section 4 being the eluent supply port D-IV, and the weakly adsorptive component is extracted from the extraction port A with the downstream end of the section 5 being the extraction port A,
the desorption force of the eluent flowing through the zone 1, the zone 2 and the zone 3 is the strongest,
making the desorption force of the eluent flowing in the section 3 and the section 5 weaker than that of the eluent flowing in the section 1, the section 2 and the section 3;
in the substep A2-6, an eluent D-I is supplied from the eluent supply port D-I with the upstream end of the section 1 being the eluent supply port D-I, a strongly adsorptive component is extracted from the extraction port C with the downstream end of the section 1 being the extraction port C, a stock solution is supplied from the stock solution supply port F with the upstream end of the section 3 being the stock solution supply port F, an eluent D-III is supplied from the eluent supply port D-III with the upstream end of the section 4 being the eluent supply port D-III, and a weakly adsorptive component is extracted from the extraction port A,
the desorption force of the eluent flowing in the section 1 is the strongest,
the desorption force of the eluent flowing through the section 3 is weaker than that of the eluent flowing through the section 1,
making the desorption force of the eluent flowing in the section 4 and the section 5 weaker than that of the eluent flowing in the section 3;
in the substep A3-6, the eluent D-I is supplied from the eluent supply port D-I, the strongly adsorptive component is extracted from the extraction port C, the eluent D-II is supplied from the eluent supply port D-II with the upstream end of the section 2 being the eluent supply port D-II, the eluent D-III is supplied from the eluent supply port D-III, and the weakly adsorptive component is extracted from the extraction port A,
the desorption force of the eluent flowing in the section 1 is the strongest,
the desorption force of the eluent flowing through the sections 2 and 3 is made weaker than that of the eluent flowing through the section 1,
the desorption force of the eluent flowing through the zone 4 and the zone 5 is made weaker than the desorption force of the eluent flowing through the zone 2 and the zone 3.
12. A simulated moving bed-based chromatographic separation process according to any one of claims 1 to 5, wherein,
the circulation system has 5 or more unit packed columns, is divided into 5 sections 1 to 5 sections, which are continuous in an annular shape, from the upstream side to the downstream side so that each section has at least 1 unit packed column, and performs the following substeps a1-7, substeps a2-7, and substeps A3-7 in step a using the 2 or more eluents:
in the substep A1-7, the eluent D-I is supplied from the eluent supply port D-I with the upstream end of the section 1 being the eluent supply port D-I, the strongly adsorptive component is extracted from the extraction port C with the downstream end of the section 1 being the extraction port C, the stock solution is supplied from the stock solution supply port F with the upstream end of the section 3 being the stock solution supply port F, the eluent D-III is supplied from the eluent supply port D-III with the upstream end of the section 4 being the eluent supply port D-III, and the weakly adsorptive component is extracted from the extraction port A with the downstream end of the section 5 being the extraction port A,
the desorption force of the eluent flowing in the section 1 is the strongest,
the desorption force of the eluent flowing through the section 3 is weaker than that of the eluent flowing through the section 1,
making the desorption force of the eluent flowing in the section 4 and the section 5 weaker than that of the eluent flowing in the section 3;
in the substep A2-7, the eluent D-I is supplied from the eluent supply port D-I, the strongly adsorptive component is extracted from the extraction port C, the eluent D-II is supplied from the eluent supply port D-II with the upstream end of the section 2 being the eluent supply port D-II, the eluent D-III is supplied from the eluent supply port D-III, and the weakly adsorptive component is extracted from the extraction port A,
the desorption force of the eluent flowing in the section 1 is the strongest,
the desorption force of the eluent flowing through the sections 2 and 3 is made weaker than that of the eluent flowing through the section 1,
making the desorption force of the eluent flowing in the section 4 and the section 5 weaker than that of the eluent flowing in the section 2 and the section 3;
in the substep A3-7, the eluent D-I is supplied from the eluent supply port D-I, the strongly adsorptive component is extracted from the extraction port C, the eluent D-II is supplied from the eluent supply port D-II, the downstream end of the section 4 is taken as the extraction port B, the intermediate adsorptive component is extracted from the extraction port B, the upstream end of the section 5 is taken as the eluent supply port D-IV, the eluent D-IV is supplied from the eluent supply port D-IV, and the weakly adsorptive component is extracted from the extraction port A,
the desorption force of the eluent flowing in the section 1 is the strongest,
the desorption force of the eluent flowing through the zone 2, the zone 3 and the zone 4 is weaker than that of the eluent flowing through the zone 1,
the desorption force of the eluent flowing through the zone 5 is made weaker than the desorption force of the eluent flowing through the zones 2, 3 and 4.
13. A simulated moving bed type chromatographic separation system wherein a plurality of unit packed columns packed with an adsorbent are connected in series via a pipe and connected in a ring-like manner in a circulating system, wherein weakly adsorptive components, strongly adsorptive components and intermediate adsorptive components are separated from the adsorbent, the components being contained in a raw liquid, using 2 or more kinds of eluents, the adsorbability of the intermediate adsorptive component being between the adsorbability of the weakly adsorptive component and the adsorbability of the strongly adsorptive component,
the piping of the circulation system is provided with a raw liquid supply port F, 2 or more eluent supply ports D corresponding to the 2 or more eluents, a drawing port a for weakly adsorbing components containing the weakly adsorbing components, a drawing port B for moderately adsorbing components containing the moderately adsorbing components, and a drawing port C for strongly adsorbing components containing the strongly adsorbing components, and the positions of the raw liquid supply port F, the drawing port a, the drawing port B, and the drawing port C are set as follows (a) to (C):
(a) the extraction port B is provided downstream of the raw liquid supply port F with at least 1 unit packed column interposed therebetween;
(b) the extraction port C is provided in a pipe having the raw liquid supply port F, or the extraction port C is provided on the upstream side of the raw liquid supply port F with at least 1 unit packed column interposed therebetween;
(c) the extraction port A is provided in a pipe having the extraction port B, or the extraction port A is provided downstream of the extraction port B with at least 1 unit packed column interposed therebetween,
the chromatographic separation system has a unit for repeating the following steps A and B in this order:
the step A is a step of simultaneously or separately supplying a raw liquid and 2 or more kinds of eluents from the raw liquid supply port F and the 2 or more kinds of eluent supply ports D, and simultaneously or separately extracting a weakly adsorptive component, a medium adsorptive component and a strongly adsorptive component from the extraction port A, the extraction port B and the extraction port C,
step B is a step of shifting the raw liquid supply port F, the eluent supply port D, the extraction port a, the extraction port B, and the extraction port C to the downstream side while maintaining a relative positional relationship after the end of step a.
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