JP4938728B2 - Chromatographic separation method - Google Patents

Description

  The present invention relates to a chromatographic separation method.

  Conventionally, a method of separating (chromatographic separation) a plurality of components in a liquid by a chromatographic technique has been used industrially. In the chromatographic separation, a solid adsorbent is used, and components in the raw material liquid are separated by utilizing the difference in adsorption characteristics with respect to the adsorbent. The principle of chromatographic separation is that a raw material liquid containing two or more components to be separated is supplied to a packed bed packed with an adsorbent, and this raw material liquid is caused to flow down with an eluent such as water, whereby Due to the difference in adsorptivity to the adsorbent, weakly adsorbable components (non-adsorbate components) flow down relatively fast, and strong adsorbent components (adsorbate components) flow down relatively slowly. Is separated into fractions containing.

  Examples of chromatographic separation methods include a fixed bed method, a moving bed method, and a simulated moving bed method. In the simulated moving bed method, a plurality of unit packed towers filled with an adsorbent having a selective adsorption ability for a specific component among two or more components contained in the raw material liquid are connected in series, and the most downstream By connecting the unit packed tower in the upper part and the unit packed tower in the most upstream part, the raw material liquid and the eluent are passed through the circulation system which is endless and separated. In the simulated moving bed method, a fraction containing a non-adsorbate component that moves through the circulation system (non-adsorbate fraction) and a fraction containing an adsorbate component that moves through the circulation system (adsorbate). The fractions are extracted from different positions, and the circulation direction of the circulation system is maintained while keeping the raw material supply position, eluent supply position, non-adsorbate fraction extraction position, and adsorbate fraction extraction position in a fixed positional relationship. It is well known that the apparatus realizes the processing operation of the moving layer in a pseudo manner by sequentially moving to the downstream side.

  In such a simulated moving bed type chromatographic separation apparatus, the raw material liquid and eluent are always supplied, and the circulation system is circulated while extracting the non-adsorbate fraction and the adsorbate fraction. Requires a pump, eluent supply pump, non-adsorbate fraction extraction pump, adsorbate fraction extraction pump and one or more circulation pumps. In particular, since the circulation pump is fixed between fixed unit packed towers, the discharge flow rate changes periodically. Moreover, since the density | concentration and composition of the fluid which flow in into a circulation pump also change sequentially, physical properties, such as specific gravity of a fluid, and viscosity, also change with it. As the difference between the set value and the actual flow rate increases, the separation performance decreases. In order to automatically control the flow rate of the circulation pump in accordance with such a change in the flow rate setting value or a change in physical properties, an expensive control device is required.

In response to the above-mentioned problem of the circulation pump, a chromatograph in which the unit packed tower is shifted one by one so that the circulation pump is always at a fixed position so that the change in physical properties of the fluid introduced into the circulation pump is reduced. A separation method is disclosed (for example, Patent Document 1).
Further, instead of installing a circulation pump, a raw material liquid and / or an eluent is extracted from one place of the circulation system, and a raw material liquid and / or an eluent (hereinafter sometimes referred to as a circulating fluid) extracted from the circulation system is used. A chromatographic separation method is disclosed in which circulation is performed by flowing into an eluent pump or a raw material liquid supply pump and supplying a circulating fluid to the circulation system again (for example, Patent Document 2). In the method described in Patent Document 2, the number of packed columns is small, the chromatographic separation apparatus can be controlled by two units of the raw material liquid supply pump and the eluent supply pump, the pump control is simple, and the apparatus is simple and inexpensive as a whole. In addition, a high and stable separation performance can be obtained.
Japanese Patent Publication No. 6-95090 JP 2005-288411 A

  However, in the chromatographic separation method described in Patent Document 1, the non-adsorbate mixed in the circulating fluid immediately before switching of the raw material liquid supply position, the eluent supply position, the non-adsorbate fraction extraction position, and the adsorbate fraction extraction position. Ingredients may remain in the eluent supply line. As the eluent is supplied in the next step, the non-adsorbate component is supplied to the circulation system, which disturbs the adsorption zone.

  Moreover, in the chromatographic separation method described in Patent Document 2, an eluent supply pipe and a raw material supply pipe are used to circulate the circulating fluid in the circulation system. In addition, the circulation (circulation process) of the circulating fluid to the circulation system is performed by stopping the supply of the raw material liquid and the eluent to the circulation system and the extraction of the fraction from the circulation system. At this time, the concentration of the component at the extraction location changes at the start and end of the circulation process. For this reason, the fluid of the target component density | concentration cannot be extracted through a circulation process. As a result, there is a problem that a fluid having an inappropriate component concentration is supplied at the start of the circulation process, and the liquid remaining in the eluent supply pipe and the raw material supply pipe at the end of the circulation process There is a problem of being mixed into the raw material liquid. In particular, if the components in the raw material liquid remain in the eluent supply pipe, these components are supplied into the circulation system together with the eluent when the next eluent is supplied, disturbing the adsorption zone and separating. It causes the performance to decrease.

  Further, when the extraction position at the end of the circulation process is set to be a position where the component concentration is low, the extraction position at the start of the circulation process is a position where the component concentration is relatively high. In this case, the eluent is supplied from the eluent supply pipe, the concentration distribution is divided, and the separation performance is deteriorated.

  As a method of preventing the components contained in the raw material liquid from remaining in the eluent supply pipe, a method of reducing the flow rate of the circulating fluid is conceivable. However, in this method, a liquid containing almost no non-adsorbate fraction comes out immediately after the start of the extraction step of the non-adsorbate fraction, which is performed after the circulation step. As a result, the amount of eluent used increases.

  Accordingly, the present invention focuses on various problems in the conventional apparatus as described above, and an object of the present invention is a chromatographic separation method capable of obtaining a target component in a raw material solution with high purity.

  The chromatographic separation method of the present invention comprises a circulation system in which a plurality of unit packed towers packed with an adsorbent are connected in series, and the raw material liquid and the eluent passed through the unit packed tower can be circulated, A raw material liquid containing two or more components is passed through the circulation system to form an adsorption zone in accordance with the affinity for the adsorbent, and the raw material liquid and / or the eluent is supplied to an arbitrary unit packed column. A unit in which the fraction in the optional component is extracted from the unit packed column enriched with the above, and the liquid in the circulating system is extracted from the optional unit packed column constituting the circulating system. Supply the extracted liquid to the unit packed tower downstream in the circulation direction of the packed tower, circulate in the circulating system, and then extract the liquid in the circulating system and the extracted liquid in the circulating system. A unit packed column to be fed downstream of the circulation system in the circulation direction And having a second step for circulating switch to packing bed units, a.

  The first step and the second step are repeated, and in the second step, the portion enriched with an arbitrary component by circulation of the liquid in the circulation system is transferred to the unit packed tower on the downstream side in the circulation direction. Corresponding to the movement, the first step includes a unit packed column for supplying a raw material liquid and / or an eluent and a unit packed column for extracting a fraction of the arbitrary component on the downstream side of the circulation system. The unit packed tower of the present invention has a transition operation for sequentially transferring the unit packed tower for extracting the liquid in the circulation system in the second step, and the unit packed tower for supplying the liquid in the extracted circulation system. May be performed after switching to the unit packed column on the downstream side in the circulation direction of the circulation system, and the second step includes supplying the raw material liquid and the eluent to the circulation system, and separating from the circulation system. You can stop the extraction of the minute, and extract the liquid in the circulatory system. After switching the unit packed column to the unit packed column on the downstream side in the circulation direction of the circulation system, the unit packed column for supplying the liquid in the extracted circulation system is switched to the unit packed column on the downstream side in the circulation direction of the circulation system. Also good.

  According to the chromatographic separation method of the present invention, the target component in the raw material liquid can be obtained with high purity.

  The chromatographic separation method of the present invention will be described below with examples, but the present invention is not limited to this.

  In the present specification, “downstream in the circulation direction” means that it is downstream in the circulation direction of the circulation system relative to the base point. The range of “the downstream side in the circulation direction” is not particularly limited, and the range of “the downstream side in the circulation direction” can be determined according to the type of the adsorbent, the component to be separated, and the like. For example, the range of “downstream side in the circulation direction” is the downstream side in the circulation direction of the base point, and is preferably a range including 50% of the unit packed tower of the unit packed tower constituting the circulation system. More preferably, the unit packed column is included. Further, the “upstream side in the circulation direction” means that it is upstream in the circulation direction of the circulation system relative to the base point. The range of “upstream side in the circulation direction” is not particularly limited, and the range of “upstream side in the circulation direction” can be determined according to the type of the adsorbent, the component to be separated, and the like. For example, the range of “the upstream side in the circulation direction” is preferably the range including the unit packed tower of 50% of the unit packed towers constituting the circulation system, which is the upstream side in the circulating direction of the base point. More preferably, the unit packed column is included.

(First embodiment)
A first embodiment of the present invention will be described with reference to FIG. FIG. 1 is a schematic diagram of a chromatographic separation apparatus 8 according to the present embodiment. As shown in FIG. 1, the chromatographic separation device 8 includes eight unit packed towers 11 to 18 (hereinafter, sometimes collectively referred to as unit packed towers) packed with an adsorbent connected in series by pipes 111 to 118, This is a simulated moving bed type chromatographic separation apparatus in which a circulation system 10 is formed.

  The unit packed tower 11 is connected to the unit packed tower 12 by a pipe 111 via a valve R1, and the unit packed tower 12 is connected to the unit packed tower 13 by a pipe 112 via a valve R2. The unit packed tower 13 is connected to the unit packed tower 14 by a pipe 113 via a valve R3, and the unit packed tower 14 is connected to the unit packed tower 15 by a pipe 114 via a valve R4. The unit packed tower 15 is connected to the unit packed tower 16 via a valve 115 via a pipe 115, and the unit packed tower 16 is connected to the unit packed tower 17 via a valve R6 via a pipe 116. The unit packed tower 17 is connected to the unit packed tower 18 through a valve 117 through a pipe 117, and the unit packed tower 18 is connected to the unit packed tower 11 through a valve 118 through a valve R8. A check valve 101 is provided in the pipes 111 to 118. In this way, the circulation system 10 is configured by the unit packed towers 11 to 18, the pipes 111 to 118, the valves R1 to R8, and the check valve 101.

  Pipes 34 and 36 are connected to the raw material liquid storage tank 32, and the pipes 34 and 36 are connected to a pipe 38 at a branch 39. The pipe 34 is provided with a pump PF, and the pipe 36 is provided with a valve F0. Branch pipes 51 to 58 are connected to the pipe 38, and valves F 1 to F 8 (hereinafter sometimes collectively referred to as raw material liquid supply valves F) are provided on the branch pipes 51 to 58, respectively. The branch pipe 51 is a pipe 118, the branch pipe 52 is a pipe 111, the branch pipe 53 is a pipe 112, the branch pipe 54 is a pipe 113, the branch pipe 55 is a pipe 114, and the branch pipe 56 is a pipe 115. The pipe 57 is connected to the pipe 116, and the branch pipe 58 is connected to the pipe 117. The pump PF, the pipes 34 and 38, the branch pipes 51 to 58, and the valves F1 to F8 constitute raw material supply means. A raw material liquid circulation line 31 is constituted by the raw material liquid storage tank 32, the pump PF, the pipes 34 and 36, and the valve F0.

  A pipe 44 is connected to the eluent storage tank 42, and a pump PD is provided in the pipe 44. Branch pipes 61 to 68 are connected to the pipe 44, and valves D1 to D8 (hereinafter sometimes collectively referred to as an eluent supply valve D) are provided on the branch pipes 61 to 68, respectively. The branch pipe 61 is a pipe 118, the branch pipe 62 is a pipe 111, the branch pipe 63 is a pipe 112, the branch pipe 64 is a pipe 113, the branch pipe 65 is a pipe 114, the branch pipe 66 is a pipe 115, and a branch pipe. 67 is connected to the pipe 116, and the branch pipe 68 is connected to the pipe 117. The pipe 46 is connected to the pipe 44 at the branch 45 on the primary side of the pump PD. The pump PD, the pipe 44, the branch pipes 61 to 68, and the valves D1 to D8 constitute eluent supply means.

  Branch pipes 71 to 78 (hereinafter sometimes collectively referred to as branch pipe A) and non-adsorbate fraction outflow pipe 79 are connected to pipe 70 through which the non-adsorbate fraction that has flowed down flows. Yes. The branch pipes 71 to 78 are provided with valves A1 to A8 (hereinafter, sometimes collectively referred to as extraction valves A), respectively. The branch pipe 71 is the pipe 111, the branch pipe 72 is the pipe 112, the branch pipe 73 is the pipe 113, the branch pipe 74 is the pipe 114, the branch pipe 75 is the pipe 115, the branch pipe 76 is the pipe 116, and the branch pipe. 77 is connected to the pipe 117, and the branch pipe 78 is connected to the pipe 118. The non-adsorbate fraction outflow pipe 79 is provided with a pump PA.

  Branch pipes 81 to 88 (hereinafter, sometimes collectively referred to as branch pipe C) and an adsorbate fraction outflow pipe 89 are connected to the pipe 80 through which the adsorbate fraction flowed down flows. The branch pipes 81 to 88 are provided with valves C1 to C8 (hereinafter, sometimes collectively referred to as extraction valves C), respectively. The branch pipe 81 is the pipe 111, the branch pipe 82 is the pipe 112, the branch pipe 83 is the pipe 113, the branch pipe 84 is the pipe 114, the branch pipe 85 is the pipe 115, the branch pipe 86 is the pipe 116, and the branch pipe. Reference numeral 87 denotes a pipe 117 and branch pipe 88 is connected to a pipe 118. The adsorbate fraction outflow pipe 89 is provided with a pump PC.

  A pipe 90 through which a fluid circulating in the circulation system 10 (circulating fluid) flows is connected to branch pipes 91 to 98 (hereinafter sometimes referred to as a branch pipe Z as a whole). The branch pipes 91 to 98 are provided with valves Z1 to Z8 (hereinafter, sometimes collectively referred to as extraction valves Z). The branch pipe 91 is the pipe 111, the branch pipe 92 is the pipe 112, the branch pipe 93 is the pipe 113, the branch pipe 94 is the pipe 114, the branch pipe 95 is the pipe 115, the branch pipe 96 is the pipe 116, and the branch pipe. Reference numeral 97 denotes a pipe 117 and branch pipe 98 is connected to a pipe 118. Further, the pipe 90 is connected to the pipe 46.

  The unit packed towers 11 to 18 are filled with an adsorbent. The adsorbent is not particularly limited as long as it has a selective adsorption ability with respect to two or more components contained in the raw material liquid. For example, ion exchange resin, silica gel, ODS (octadecyl group-bonded silica gel; long chain) And silica gel in which an aliphatic octadecyl group is introduced) and synthetic adsorbents. These may be used alone or in combination of two or more. Further, the unit packed towers 11 to 18 may be filled with the same adsorbent or may be filled with different adsorbents. The type and combination of the adsorbent can be determined according to the component to be separated.

  A chromatographic separation method using the chromatographic separation apparatus 8 will be described. In the chromatographic separation method in the present embodiment, a raw material liquid containing two or more components is passed through the circulation system 10 to form an adsorption zone, and the raw material liquid and the eluent are supplied to an arbitrary unit packed tower, And a first step of extracting the adsorbate fraction. In addition, a circulating fluid extracted from the circulation system 10 is supplied to the circulation system 10 for circulation, and then a unit packed tower for extracting the liquid in the circulation system 10 and a unit for supplying the extracted liquid in the circulation system 10 A second step of switching the packed tower to a unit packed tower on the downstream side in the circulation direction of the circulation system 10 for circulation.

[First step]
In the first step, the raw material liquid is passed through a circulation system to form an adsorption zone, the raw material liquid and / or the eluent are supplied to an arbitrary unit packed column, and the raw material liquid is separated from the unit packed column enriched with arbitrary components. This is a step of extracting fractions (non-adsorbate fraction and adsorbate fraction).
In the first step, the raw material liquid 30 is caused to flow through the pipes 34 and 38 by the pump PF, one of the raw material liquid supply valves F is opened, and the raw material liquid 30 is supplied to the corresponding unit packed tower. At the same time, the eluent 40 is circulated through the pipe 44 by the pump PD, one of the eluent supply valves D is opened, and the eluent 40 is supplied to the corresponding unit packed tower. Then, the extraction valve A located immediately below the unit packed column enriched with the non-adsorbed fraction is opened, and the extraction valve C located directly below the unit packed column enriched with the adsorbate fraction is opened. . Thus, the non-adsorbate fraction is passed through the open extraction valve A, branch pipe A, pipe 70, and pump PA by passing the raw material liquid 30 and the eluent 40 through the circulation system 10. It flows out from the adsorbate fraction outflow pipe 79. Further, the adsorbate fraction flows out from the adsorbate fraction outflow pipe 89 through the open extraction valve C, branch pipe C, pipe 80, and pump PC.

[Second step]
In the second step, the circulating fluid is extracted from an arbitrary unit packed tower constituting the circulating system, the extracted circulating fluid is supplied to the circulating system and circulated in the circulating system (first treatment), and then the circulating In this step, the unit packed tower for extracting the fluid and the unit packed tower for supplying the extracted circulating fluid are switched to the unit packed tower on the downstream side in the circulation direction of the circulation system (second processing).
The second step of this embodiment is performed in parallel with the first step described above. In the second step, any one of valves R1 to R8 (hereinafter sometimes collectively referred to as shut-off valve R), and a unit packed column for supplying the eluent 40 in the first step, and circulation of the unit packed column The shutoff valve R included in the pipe connecting the unit packed tower located upstream in the direction is closed. Further, the extraction valve Z included in the branch pipe Z connecting the pipe having the shutoff valve R and the pipe 90 is opened. Then, the circulating fluid is extracted from the extraction valve Z corresponding to the unit packed column located upstream of the unit packed column supplying the eluent 40 in the circulation direction. The extracted circulating fluid flows from the pipe 90 through the pipe 46, joins the eluent 40 at the branch 45, flows to the pipe 44 by the pump PD, and is supplied again to the unit packed tower together with the eluent 40. Thus, the circulating fluid extracted from the circulation system 10 is supplied to the circulation system 10 by the eluent supply means and circulates (first treatment).

  After the first processing is performed for an arbitrary period, the raw material liquid supply valve F, the extraction valve A, and the extraction valve C are opened while the eluent supply valve D and the extraction valve C are opened while maintaining the state of the first processing described above. The valve Z and the shutoff valve R to be closed are switched to the eluent supply valve D, the extraction valve Z, and the shutoff valve R on the downstream side in the circulation direction of the circulation system 10. Then, the circulating fluid is extracted from the extraction valve Z corresponding to the unit packed column located on the upstream side in the circulation direction of the shutoff valve R after switching. The extracted circulating fluid flows from the pipe 90 through the pipe 46, joins the eluent 40 at the branch 45, flows to the pipe 44 by the pump PD, and is supplied again to the unit packed tower together with the eluent 40. Thus, the unit packed tower for extracting the circulating fluid and the unit packed tower for supplying the circulating fluid are switched to the unit packed tower on the downstream side in the circulation direction of the circulation system, and the circulating fluid is circulated in the circulation system (second treatment). .

  After the second treatment is performed for an arbitrary period, the extraction valve Z and the shutoff valve R are opened while maintaining the state of the second treatment described above, the raw material liquid supply valve F, the extraction valve A, and the extraction valve D. Are switched to the raw material liquid supply valve F, the extraction valve A, and the extraction valve D on the downstream side in the circulation direction of the circulation system 10. In this way, the unit packed column for supplying the raw material liquid 30 and the eluent 40 and the unit packed column for extracting the non-adsorbate fraction and the adsorbate fraction are used as the packed column on the downstream side of the circulation system 10. Transition. Then, the raw material liquid and the eluent are passed through the circulation system and continuously separated into a non-adsorbate fraction and an adsorbate fraction. In the present embodiment, the operation of switching the raw material liquid supply valve F, the extraction valve A, and the extraction valve D to be opened to the raw material liquid supply valve F, the extraction valve A, and the extraction valve D downstream in the circulation direction of the circulation system 10 is performed. Corresponds to a “migration operation”.

  An example of the operation of each valve and each pump in the first step and the second step described above will be described below using Table 1. In Table 1, the numbers described in the F, D, A, C, and Z sections of the valves indicate valves that are “open”, and “−” indicates that all valves are “closed”. . For example, when “1” is described in the item F, only the valve F1 of the raw material liquid supply valves F is “open”. In Table 1, the number described in the R section of the valve indicates a valve that is “closed”. For example, when “1” is written in the R term, only the valve R1 of the shutoff valves R is “closed”. In the pump section, “◯” indicates the operating state.

[Operation 1-1]
As shown in Table 1, in operation 1-1, the valves F1, D5, A2, C6, and Z4 are opened, and the valve R4 is closed. Next, the raw material liquid 30 is supplied to the unit packed tower 11, and the eluent 40 is supplied to the unit packed tower 15. During this time, the non-adsorbate fraction enriched in the unit packed tower 12 flows down, flows in the order of the pipe 112, the branch pipe 71, and the pipe 70 and flows out from the non-adsorbate fraction outflow pipe 79. Further, the adsorbate fraction enriched in the unit packed column 16 flows down, flows in the order of the pipe 116, the branch pipe 86, and the pipe 80, and flows out from the adsorbate fraction outflow pipe 89. Therefore, in this operation, the raw material liquid is passed through the circulation system to form an adsorption zone, the raw material liquid and / or the eluent is supplied to an arbitrary unit packed column, and the unit packed column enriched with arbitrary components is used. This corresponds to the first step of extracting fractions (non-adsorbate fraction and adsorbate fraction).

  At the same time, the circulating fluid extracted from the unit packed tower 14 flows in the order of the pipe 114, the branch pipe 94, the pipe 90, and the pipe 46, and reaches the pipe 44 at the branch 45. Then, the circulating fluid merges with the eluent 40, flows through the pipe 44 and the branch pipe 65 in this order by the pump PD, and is supplied to the unit packed column 15. The supplied circulating fluid is unit packed tower 15, pipe 115, unit packed tower 16, pipe 116, unit packed tower 17, pipe 117, unit packed tower 18, pipe 118, unit packed tower 11, pipe 111, unit packed tower. 12, the pipe 112, the unit packed tower 13, the pipe 113, and the unit packed tower 14 are passed in this order and circulate in the circulation system 10. Therefore, this operation corresponds to the first process of the second step in which the circulating fluid is extracted from an arbitrary unit packed tower constituting the circulating system, and the extracted circulating fluid is supplied to the circulating system and circulated in the circulating system. .

[Operation 1-2]
As shown in Table 1, in operation 1-2, the valves F1, D6, A2, C6, and Z5 are opened, and the valve R5 is closed. That is, the raw material liquid supply valve F, the extraction valve A, and the extraction valve C are closed with the eluent supply valve D and the extraction valve Z that are opened while maintaining the state in the first process of the second step described above. The shutoff valve R is switched to the eluent supply valve D, the extraction valve Z, and the shutoff valve R on the downstream side in the circulation direction of the circulation system 10. The circulating fluid extracted from the unit packed tower 15 flows in the order of the pipe 115, the branch pipe 95, the pipe 90, and the pipe 46, and reaches the pipe 44 at the branch 45. Then, the circulating fluid merges with the eluent 40, flows through the pipe 44 and the branch pipe 66 in this order by the pump PD, and is supplied to the unit packed tower 16. The supplied circulating fluid is unit packed tower 16, pipe 116, unit packed tower 17, pipe 117, unit packed tower 18, pipe 118, unit packed tower 11, pipe 111, unit packed tower 12, pipe 112, unit packed tower. 13, the pipe 113, the unit packed tower 14, the pipe 114, and the unit packed tower 15 are passed in this order, and circulate in the circulation system 10. Therefore, in this operation, after the first treatment in the second step, the unit packed tower for extracting the circulating fluid and the unit packed tower for supplying the circulating fluid are switched to the unit packed tower on the downstream side in the circulation direction of the circulation system for circulation. This corresponds to the second process of the second step.

[Operation 2-1]
As shown in Table 1, in operation 2-1, the valves F2, D6, A3, C7, and Z5 are opened, and the valve R5 is closed. As a result, the supply position of the raw material liquid, the supply position of the eluent, the extraction position of the non-adsorbate fraction, and the extraction position of the adsorbate fraction are moved to the downstream side in the circulation direction of the circulation system 10, and the non-adsorbate fraction is extracted. And adsorbate fraction are extracted.

  In the series of operations 1-1 to 1-2 by switching the valves as described above, the supply position of the raw material liquid and the eluent, the extraction position of the non-adsorbate fraction and the adsorbate fraction, and the extraction position of the circulating fluid are as follows: This is performed while maintaining a specific positional relationship. When such a series of operations 1-1 to 1-2 is completed, each valve to be controlled is shifted by one unit packed tower downstream in the circulation direction while maintaining the above-described specific positional relationship. A series of operations 2-1 to 2-2 are performed. By performing this transition sequentially, the simulated moving bed type chromatographic separation apparatus is operated.

[Operation 2-1 and later]
As shown in Table 1, operations 2-1 to 2-2, operations 3-1 to 2-3, operations 4-1 to 4-2, and operations 5-1 to 5- 2, operation 6-1 to 6-2, operation 7-1 to 7-2, and operation 8-1 to 8-2, the raw material liquid supply valve F, the extraction valve A, the extraction valve C, and the extraction valve Z to be opened And the shutoff valve R to be closed are sequentially switched to the raw material liquid supply valve F, the extraction valve A, the extraction valve C, the extraction valve Z, and the cutoff valve R on the downstream side in the circulation direction of the circulation system, and the operation 1-1 to The same operation as 1-2 is performed. When the operations 1-1 to 8-2 are performed, one cycle of chromatographic separation is completed.

  The raw material liquid 30 is not particularly limited as long as it contains two or more components, and examples thereof include a sugar liquid during the production of sucrose, a biological extract, and a culture liquid obtained by microbial fermentation.

  The eluent 40 is not particularly limited as long as each component in the raw material liquid is allowed to flow down into the unit packed tower and the components to be separated can be appropriately separated, and can be determined according to the purpose. it can. For example, water, basic aqueous solution such as sodium hydroxide aqueous solution, acidic aqueous solution such as hydrochloric acid aqueous solution, alcohols such as methanol, ethanol, n-propanol, isopropanol, butanol, isobutanol and the like can be mentioned. As the eluent, one kind of these may be used alone, or a mixed solution obtained by mixing two or more kinds may be used as the eluent. The eluent may contain an acid, a base, a buffer, and a salt.

  The supply amount of the raw material liquid 30 to the unit packed column in the first step is not particularly limited, and is determined in consideration of the components to be separated, the type of adsorbent packed in the unit packed column, the state of the adsorption zone, and the like. It is preferable to do. The amount of eluent 40 supplied to the unit packed column in the first step is not particularly limited, and is determined in consideration of the components to be separated, the type of adsorbent packed in the unit packed column, the state of the adsorption zone, and the like. It is preferable.

  The amount of circulating fluid flowing into the unit packed column in the second step is not particularly limited, and is determined in consideration of the components to be separated, the type of adsorbent packed in the unit packed column, the state of the adsorption zone, etc. It is preferable to do.

  The unit packed tower for extracting the circulating fluid in the first treatment of the second step is a circulating fluid in which the components of the raw material liquid 30 (non-adsorbed fraction and adsorbed fraction) in the circulating fluid have a relatively low concentration. Is preferably a unit packed tower from which Moreover, the period of the 1st process of a 2nd process is not specifically limited, It can set in the range which the density | concentration of a non-adsorbate fraction and an adsorbate fraction in the extracted circulating fluid does not rise. When the extracted circulating fluid contains the component of the raw material liquid 30 at a high concentration, the adsorption zone in the circulating layer 10 is disturbed, and the non-adsorbate fraction and the adsorbate fraction obtained by the subsequent operation are disturbed. This is because the purity is lowered.

  The period of the second treatment in the second step is not particularly limited, and can be set in a range in which the concentration of the non-adsorbate fraction or the adsorbate fraction in the extracted circulating fluid does not increase.

  In the chromatographic separation method, the pump PF and the pump PD may be set to a constant flow rate discharge setting or may perform flow rate control.

  According to the chromatographic separation method of this embodiment, by performing the second treatment in the second step, the component of the raw material liquid is continuously supplied with a relatively low concentration, that is, a circulating fluid having physical properties close to those of the eluent. Can be extracted and circulated into the circulatory system. Further, since the circulating fluid supplied to the circulation system in the second treatment is extracted from the unit packed tower on the downstream side in the circulation direction of the circulation system than in the first treatment, the content of the component of the raw material liquid may be reduced. it can. In addition, after the second treatment in the second step, the raw material liquid supply valve F, the extraction valve A, and the extraction valve D to be opened are connected to the raw material liquid supply valve F, the extraction valve A, and the extraction valve on the downstream side in the circulation direction of the circulation system. A transition operation to switch to the valve D is performed, and the non-adsorbate fraction and the adsorbate fraction are extracted. For this reason, the components of the raw material liquid do not remain at a high concentration in the eluent supply means. In other words, a high concentration of components corresponding to the non-adsorbate fraction and components corresponding to the adsorbate fraction in the raw material liquid are supplied to the unit packed tower in which neither the non-adsorbate component nor the adsorbate component is enriched. Thus, it is possible to prevent the adsorption zone formed in the circulation system from being disturbed. As a result, high-purity non-adsorbate fraction and adsorbate fraction can be obtained continuously.

(Second embodiment)
A second embodiment of the present invention will be described with reference to FIG. FIG. 2 is a schematic diagram of a chromatographic separation apparatus 300 according to the present embodiment. As shown in FIG. 2, in the chromatographic separation apparatus 300, four unit packed columns 311 to 314 (hereinafter, sometimes collectively referred to as unit packed columns) packed with an adsorbent are connected in series by pipes 321 to 324, This is a simulated moving bed type chromatographic separation apparatus in which a circulation system 310 is formed.

  The unit packed tower 311 is connected to the unit packed tower 312 through the valve 321 via the pipe 321, and the unit packed tower 312 is connected to the unit packed tower 313 through the valve r 2 through the valve 322. The unit packed tower 313 is connected to the unit packed tower 314 through a valve 323 by a pipe 323, and the unit packed tower 314 is connected to the unit packed tower 311 through a valve r4 through a valve 324. A check valve 301 is provided in each of the pipes 321 to 324. Thus, the circulation system 310 is constituted by the unit packed towers 311 to 314, the pipes 321 to 324, the valves r1 to r4, and the check valve 301.

  Pipes 334 and 336 are connected to the raw material liquid storage tank 32, and the pipes 334 and 336 are connected to the pipe 338 at a branch 339. The pipe 334 is provided with a pump Pf, and the pipe 336 is provided with a valve f0. The pipe 338 is connected to the branch pipes 351 to 354, and the branch pipes 351 to 354 are provided with valves f 1 to f 4 (hereinafter sometimes collectively referred to as a raw material liquid supply valve f), respectively. The branch pipe 351 is connected to the pipe 324, the branch pipe 352 is connected to the pipe 321, the branch pipe 353 is connected to the pipe 322, and the branch pipe 354 is connected to the pipe 323. The pump Pf, the pipes 334 and 338, the branch pipes 351 to 354, and the valves f1 to f4 constitute a raw material liquid supply unit. A raw material liquid circulation line 330 is configured by the raw material liquid storage tank 32, the pump Pf, the pipes 334 and 336, and the valve f0.

  A pipe 344 is connected to the eluent storage tank 42, and the pipe 344 is provided with a pump Pd. The pipe 344 is connected to the branch pipes 361 to 364, and the branch pipes 361 to 364 are provided with valves d1 to d4 (hereinafter sometimes collectively referred to as an eluent supply valve d), respectively. The branch pipe 361 is connected to the pipe 324, the branch pipe 362 is connected to the pipe 321, the branch pipe 363 is connected to the pipe 322, and the branch pipe 364 is connected to the pipe 323. The pipe 346 is connected to the pipe 344 at a branch 345 on the primary side of the pump Pd. In the pipe 346, a valve d0 is provided between the eluent storage tank 42 and the branch 345. The pump Pd, the pipe 344, the branch pipes 361 to 364, and the valves d0 to d4 constitute an eluent supply means.

  Branch pipes 371 to 374 (hereinafter sometimes collectively referred to as branch pipe a) and non-adsorbate fraction outflow pipe 379 are connected to pipe 370 through which the non-adsorbate fraction flowed down flows. Yes. The branch pipes 371 to 374 are provided with valves a1 to a4 (hereinafter sometimes collectively referred to as extraction valves a), respectively. The branch pipe 371 is connected to the pipe 321, the branch pipe 372 is connected to the pipe 322, the branch pipe 373 is connected to the pipe 323, and the branch pipe 374 is connected to the pipe 324.

  Branch pipes 381 to 384 (hereinafter sometimes collectively referred to as branch pipe c) and an adsorbate fraction outflow pipe 389 are connected to the pipe 380 through which the adsorbate fraction flowed down flows. The branch pipes 381 to 384 are provided with valves c1 to c4 (hereinafter sometimes collectively referred to as extraction valves c), respectively. The branch pipe 381 is connected to the pipe 321, the branch pipe 382 is connected to the pipe 322, the branch pipe 383 is connected to the pipe 323, and the branch pipe 384 is connected to the pipe 324.

  Branch pipes 391 to 394 (hereinafter sometimes collectively referred to as branch pipes z) are connected to the pipe 390 through which the circulating fluid flows. The branch pipes 391 to 394 are provided with valves z1 to z4 (hereinafter sometimes collectively referred to as extraction valves z), respectively. The branch pipe 391 is connected to the pipe 321, the branch pipe 392 is connected to the pipe 322, the branch pipe 393 is connected to the pipe 323, and the branch pipe 394 is connected to the pipe 324. Further, the pipe 390 is connected to the pipe 346.

  The adsorbent packed in the unit packed columns 311 to 314 is the same as the adsorbent packed in the unit packed columns 11 to 18 in the first embodiment. Moreover, the unit packed towers 311 to 314 may be filled with the same adsorbent, or may be filled with different adsorbents. The type and combination of the adsorbent can be determined according to the component to be separated.

  A chromatographic separation method using the chromatographic separation apparatus 300 will be described. In the chromatographic separation method according to the present embodiment, a raw material liquid containing two or more components is passed through the circulation system 310 to form an adsorption zone, and the raw material liquid and the eluent are supplied to an arbitrary unit packed tower, And a first step of extracting the adsorbate fraction. In addition, the supply of the raw material liquid and the eluent to the circulation system 310 and the extraction of the fraction from the circulation system 310 are stopped, and the circulation fluid extracted from the circulation system 310 is supplied to the circulation system 310 and circulated. Next, the unit packed tower for extracting the liquid in the circulation system 310 and the unit packed tower for supplying the extracted liquid in the circulation system 310 are switched to the unit packed tower on the downstream side in the circulation direction of the circulation system 310 for circulation. A second step.

[First step]
In the first step, the raw material liquid is passed through a circulation system to form an adsorption zone, the raw material liquid and / or the eluent are supplied to an arbitrary unit packed column, and the raw material liquid is separated from the unit packed column enriched with arbitrary components. This is a step of extracting fractions (non-adsorbate fraction and adsorbate fraction).
In the first step, the raw material liquid 30 is caused to flow through the pipes 334 and 338 by the pump Pf, one of the raw material liquid supply valves f is opened, and the raw material liquid 30 is supplied to the corresponding unit packed tower. The extraction valve a corresponding to the unit packed column corresponding to the extraction position and enriched with the non-adsorbate fraction is opened, and valves r1 to r4 (hereinafter sometimes referred to as shut-off valve r). The shut-off valve r located on the downstream side in the circulation direction of the unit packed column enriched with the non-adsorbate fraction is closed. In this way, a substantially non-adsorbed fraction equivalent to the supplied raw material liquid 30 is extracted. The extracted non-adsorbate fraction flows out from the non-adsorbate fraction outflow pipe 379 via the open extraction valve a, branch pipe a, and pipe 370.

  Next, the valve f0 is opened, the valves f1 to f4 are closed, and the supply of the raw material liquid 30 to the circulation system 310 is stopped. Thereafter, the eluent 40 is caused to flow through the pipe 344 by the pump Pd, one of the eluent supply valves d is opened, and the eluent 40 is supplied to the corresponding unit packed tower. Then, the extraction valve c located immediately below the unit packed column where the adsorbate fraction is enriched is opened. In this way, an adsorbate fraction substantially equal to the supplied eluent 40 is extracted. The extracted adsorbate fraction flows out from the adsorbate fraction outflow pipe 389 via the open extraction valve c, branch pipe c, and pipe 380.

  Next, the valve f0 is opened, the valves f1 to f4 are closed, the supply of the raw material liquid 30 to the circulation system 310 is stopped, and all the extraction valves c are closed. Thereafter, the eluent 40 is caused to flow through the pipe 344 by the pump Pd, one of the eluent supply valves d is opened, and the eluent 40 is supplied to the corresponding unit packed tower. And the extraction valve a located just under the unit packed column in which the non-adsorbate fraction is enriched is opened. In this way, a substantially non-adsorbed fraction equivalent to the supplied eluent 40 is extracted. The extracted non-adsorbate fraction flows out from the non-adsorbate fraction outflow pipe 379 via the open extraction valve a, branch pipe a, and pipe 370.

[Second step]
In the second step, the circulating fluid is extracted from an arbitrary unit packed tower constituting the circulating system, the extracted circulating fluid is supplied to the circulating system and circulated in the circulating system (first treatment), and then the circulating In this step, the unit packed tower for extracting the fluid and the unit packed tower for supplying the extracted circulating fluid are switched to the unit packed tower on the downstream side in the circulation direction of the circulation system (second processing).
The second step of the present embodiment is performed by stopping the supply of the raw material liquid 30 and the eluent 40 to the circulation system 310 and the extraction of the non-adsorbate fraction and the adsorbate fraction. In the second step, the shutoff valve r included in the pipe connecting the unit packed column for supplying the eluent 40 in the first step and the unit packed column located upstream in the circulation direction of the unit packed column is closed. To do. Further, the extraction valve z of the branch pipe z connecting the pipe having the shut-off valve r and the pipe 390 is opened. The circulating fluid is extracted from an extraction valve z corresponding to the unit packed column located upstream in the circulation direction of the unit packed column supplying the eluent 40. The extracted circulating fluid flows from the pipe 390 through the pipe 346, merges with the eluent 40 at the branch 345, flows into the pipe 344 by the pump Pd, and is supplied again to the unit packed column that supplies the eluent 40. . Thus, the circulating fluid is supplied to the circulation system 310 by the eluent supply means and circulates (first process).

  After performing the first treatment for an arbitrary period, the raw material liquid supply valve f, the extraction valve a, and the extraction valve c are opened while the eluent supply valve d and the extraction valve are opened while maintaining the state of the first treatment described above. z and the shut-off valve r to be closed are switched to the eluent supply valve d, the extraction valve z, and the shut-off valve r on the downstream side of the circulation system 310 in the circulation direction. Then, the circulating fluid is extracted from the extraction valve z corresponding to the unit packed column located on the upstream side in the circulation direction of the shutoff valve r after switching. The extracted circulating fluid flows from the pipe 390 through the pipe 346, merges with the eluent 40 at the branch 345, flows into the pipe 344 by the pump Pd, and is supplied again to the unit packed column that supplies the eluent 40. . Thus, the unit packed tower for extracting the circulating fluid and the unit packed tower for supplying the circulating fluid are switched to the downstream side in the circulating direction of the circulating system, and the circulating fluid is circulated in the circulating system (second processing).

  After the second process is performed for an arbitrary period, the eluent supply valve d, the extraction valve z, and the shut-off valve r are opened with the raw material liquid supply valve f being opened while maintaining the state of the second process described above. The valve a and the extraction valve c are switched to the raw material liquid supply valve f, the extraction valve a, and the extraction valve c on the downstream side in the circulation direction of the circulation system 310. In this way, the unit packed column for supplying the raw material liquid 30 and the eluent 40 and the unit packed column for extracting the non-adsorbate fraction and the adsorbate fraction are packed in the packed column on the downstream side of the circulation system 310. Transition. Then, the first step is performed again. In this embodiment, the raw material liquid supply valve f, the extraction valve a, and the extraction valve c to be opened are switched to the raw material liquid supply valve f, the extraction valve a, and the extraction valve c on the downstream side of the circulation system 310 in the circulation direction. The operation to perform corresponds to the “migration operation”.

  An example of the operation of each valve and each pump in the first step and the second step described above will be described below using Table 2. In Table 2, the numbers described in the f, d, a, c, and z sections of the valves indicate valves that are “open”, and “−” indicates that all valves are “closed”. . For example, when “1” is described in the item f, only the valve f1 of the raw material liquid supply valve f is “open”. In Table 2, the number described in the section of the valve r indicates a valve that is “closed”. For example, when “1” is described in the item r, only the valve r1 of the shutoff valves R is “closed”. In the pump section, “◯” indicates the operating state, and “−” indicates the stopped state.

[Operation I-1]
As shown in Table 2, in operation I-1, the valves f1 and a1 are opened and the valve r1 is closed. The pump Pf is in an operating state, and the pump Pd is in a stopped state. The raw material liquid 30 is supplied to the unit packed tower 311 through the pipe 334, the pipe 338, and the branch pipe 351 in this order by the pump Pf. The raw material liquid 30 supplied to the unit packed column 311 causes the non-adsorbate fraction enriched in the unit packed column 311 to flow down. The flowed non-adsorbate fraction flows in the order of the pipe 321, the branch pipe 371, and the pipe 370 and flows out from the non-adsorbate fraction outflow pipe 379. Therefore, in this operation, the raw material liquid is passed through a circulation system to form an adsorption zone, the raw material liquid is supplied to an arbitrary unit packed column, and a fraction (non-adsorbed) is supplied from the unit packed column enriched with arbitrary components. This corresponds to the first step of extracting the quality fraction).

[Operation I-2]
As shown in Table 2, in the operation I-2, the valves f0, d0, d3, and c3 are opened, the valve r3 is closed, and the pumps Pf and Pd are in the operating state. The raw material liquid 30 is circulated by the pump Pf from the pipe 334 through the pipe 336 and returned to the raw material liquid storage tank 32. The eluent 40 flows through the pipe 344 and the branch pipe 363 by the pump Pd and is supplied to the unit packed tower 313. The eluent 40 supplied to the unit packed column 313 causes the adsorbate fraction enriched in the unit packed column 313 to flow down. The adsorbate fraction that has flowed down flows in the order of the pipe 323, the branch pipe 383, and the pipe 380, and flows out from the adsorbate fraction outflow pipe 389. Therefore, this operation corresponds to the first step of supplying an eluent to an arbitrary unit packed column and extracting a fraction (adsorbate fraction) from the unit packed column enriched with an arbitrary component.

[Operation I-3]
As shown in Table 2, in the operation I-3, the valves f0, d0, d3, a1 are opened, the valve r1 is closed, and the pumps Pf, Pd are put into operation. The raw material liquid 30 is circulated by the pump Pf from the pipe 334 through the pipe 336 and returned to the raw material liquid storage tank 32. The eluent 40 flows through the pipe 344 and the branch pipe 363 by the pump Pd and is supplied to the unit packed tower 313. The eluent 40 supplied to the unit packed tower 313 flows through the pipe 323, the unit packed tower 314, the pipe 324, and the unit packed tower 311 in this order. Then, the eluent 40 allows the non-adsorbent fraction enriched in the unit packed column 311 to flow down. The flowed non-adsorbate fraction flows in the order of the pipe 321, the branch pipe 371, and the pipe 370 and flows out from the non-adsorbate fraction outflow pipe 379. Therefore, this operation corresponds to the first step of supplying the eluent to an arbitrary unit packed column and extracting a fraction (non-adsorbed fraction) from the unit packed column enriched with an arbitrary component.

[Operation I-4]
As shown in Table 2, in the operation I-4, the valves f0, d3, and z2 are opened, the valve r2 is closed, and the pumps Pf and Pd are set in an operating state. The raw material liquid 30 is circulated by the pump Pf from the pipe 334 through the pipe 336 and returned to the raw material liquid storage tank 32. The eluent 40 is closed from the eluent storage tank 32 to the circulation system 310 by closing the valve d0. Moreover, all the extraction valves a and all the extraction valves c are closed, and extraction of the non-adsorbate fraction and the adsorbate fraction is stopped. The circulating fluid flows from the unit packed tower 312 through the pipe 322, the branch pipe 392, the pipe 390, and the pipe 346, and flows into the pipe 344 at the branch 345. The circulating fluid flows through the pipe 344, the branch pipe 363, and the pipe 322 by the pump Pd, and is supplied to the unit packed tower 313. The supplied circulating fluid flows in the order of the unit packed tower 313, the pipe 323, the unit packed tower 314, the pipe 324, the unit packed tower 311, the pipe 321, and the unit packed tower 312, and circulates in the circulation system 310. Therefore, this operation stops the supply of the raw material liquid and the eluent to the circulation system, and the extraction of the fraction from the circulation system, and the circulation fluid is extracted from an arbitrary unit packed column constituting the circulation system, This corresponds to the first process of the second step in which the extracted circulating fluid is supplied to the circulation system and circulated in the circulation system.

[Operation I-5]
As shown in Table 2, in the operation I-5, the valves f0, d4, and z3 are opened, the valve r3 is closed, and the pumps Pf and Pd are put into operation. That is, the eluent supply valve d, the extraction valve z, and the shutoff valve r to be opened are switched to the downstream side in the circulation direction of the circulation system 310 while maintaining the state in which the valve f0 of the operation I-4 is opened. The raw material liquid 30 is circulated by the pump Pf from the pipe 334 through the pipe 336 and returned to the raw material liquid storage tank 32. The eluent 40 is closed from the eluent storage tank 42 to the circulation system 310 by closing the valve d0. Moreover, all the extraction valves a and all the extraction valves c are closed, and extraction of the non-adsorbate fraction and the adsorbate fraction is stopped.

  The circulating fluid flows from the unit packed tower 313 through the pipe 323, the branch pipe 393, the pipe 390, and the pipe 346, and flows into the pipe 344 at the branch 345. The circulating fluid flows through the pipe 344, the branch pipe 364, and the pipe 323 by the pump Pd, and is supplied to the unit packed tower 314. The supplied circulating fluid flows in the order of the unit packed tower 314, the pipe 324, the unit packed tower 311, the pipe 321, the unit packed tower 312, the pipe 322, and the unit packed tower 313, and circulates in the circulation system 310. Therefore, in this operation, after the first treatment in the second step, the unit packed tower for extracting the circulating fluid and the unit packed tower for supplying the circulating fluid are switched to the unit packed tower on the downstream side in the circulation direction of the circulation system for circulation. This corresponds to the second process of the second step.

  In the series of operations I-1 to I-5 by switching the valves as described above, the supply position of the raw material liquid and the eluent, the extraction position of the non-adsorbate fraction and the adsorbate fraction, and the extraction position of the circulating fluid are as follows: This is performed while maintaining a specific positional relationship. When such a series of operations I-1 to I-5 is completed, each valve to be controlled is shifted by one unit packed tower downstream in the circulation direction while maintaining the specific positional relationship described above. A series of operations II-1 to II-5 are performed. By performing this transition sequentially, the simulated moving bed type chromatographic separation apparatus is operated.

  As shown in Table 2, in the operations II-1 to II-5, the operations III-1 to III-5, and the operations IV-1 to IV-5 following the operations I-1 to I-5, the raw material liquid supply to be opened is used. The valve f, the extraction valve a, the extraction valve c, the extraction valve z, and the shut-off valve r to be closed are sequentially supplied to the raw material liquid supply valve f, the extraction valve a, the extraction valve c, and the extraction valve on the downstream side in the circulation direction of the circulation system. Switching to z and shut-off valve r, the same operation as operations I-1 to I-5 is performed. When operations I-1 to IV-5 are executed, one cycle of chromatographic separation is completed.

  In the first step, the flow rate of the raw material liquid 30 to the unit packed column is particularly limited as long as the total amount of the non-adsorbed fraction can be extracted from the unit packed column enriched with the non-adsorbed fraction. Not. In the first step, the flow rate of the raw material liquid 40 to the unit packed column is determined from the non-adsorbed fraction or the adsorbed fraction from the unit packed column enriched with the non-adsorbed fraction or the adsorbed fraction. The amount is not particularly limited as long as the entire amount can be extracted. It is preferable to determine the flow rates of the raw material liquid 30 and the eluent 40 in the first step in consideration of the components to be separated and the type of adsorbent packed in the unit packed column.

  The amount of circulating fluid flowing through the unit packed tower in the second step is not particularly limited, and is preferably determined in consideration of the component to be separated, the type of adsorbent packed in the unit packed tower, and the like.

  In the unit packed tower for extracting the circulating fluid in the first treatment of the second step, the circulation of the components of the raw material liquid 30 (non-adsorbed fraction and adsorbed fraction) in the circulating fluid is at the relatively lowest concentration. A unit packed column from which a fluid is obtained is preferable. Moreover, the period of the 1st process of a 2nd process is not specifically limited, It can set in the range which the density | concentration of a non-adsorbate fraction and an adsorbate fraction in the extracted circulating fluid does not rise. If the component of the raw material liquid 30 is contained in the extracted circulating fluid at a high concentration, the adsorption zone in the circulation system 310 is disturbed, and the non-adsorbate fraction and the adsorbate fraction obtained by the subsequent operation are disturbed. This is because the purity is lowered.

  The period of the second treatment in the second step is not particularly limited, and can be set in a range in which the concentration of the non-adsorbate fraction or the adsorbate fraction in the extracted circulating fluid does not increase.

  In the chromatographic separation method, the pump Pf and the pump Pd may be set to a constant flow rate discharge setting or may perform flow rate control.

  In the chromatographic separation method of this embodiment, in the first step, the entire amount of non-adsorbate fraction or adsorbate fraction to be extracted is extracted. For this reason, it is possible to focus on the fraction to be extracted and to separate and extract, and to separate and extract the non-adsorbate fraction and the adsorbate fraction at the same time, the number of unit packed towers is smaller. Separation with high accuracy is possible by number. In addition, since the entire amount of one target fraction is extracted, there is no need to consider the balance of the extraction amount of the non-adsorbate fraction and the adsorbate fraction as in the case of extracting two fractions at the same time. No flow rate control means or back pressure control means is required. As a result, the entire chromatographic separation apparatus is greatly simplified by a small number of unit packed columns and simplification on the extraction side.

  Further, in the chromatographic separation method of the present embodiment, in the second step, the raw material liquid and the eluent are supplied, and the non-adsorbate fraction and the adsorbate fraction are not extracted, and the separation is performed in the circulation system. The components are developed to form an adsorption zone. For this reason, the adsorption zone of a desired state can be formed accurately and easily, and higher-purity non-adsorbate fraction and adsorbate fraction can be obtained.

  The present invention is not limited to the embodiment described above. In the second step of the first and second embodiments, the circulating fluid extracted from the circulating system using the eluent supplying means is supplied to the circulating system, but the circulating fluid supply means is not limited to this, A raw material liquid supply unit may be used, or a circulation unit installed separately may be used to supply the circulation system.

  In the first and second embodiments, the second treatment in the second step is to switch the unit packed tower for supplying the circulating fluid to the unit packed tower on the downstream side in the circulating direction and to circulate the unit packed tower for extracting the circulating fluid. Switching to the unit packed tower on the downstream side is performed at the same time. However, after the unit packed column for extracting the circulating fluid is switched to the unit packed column on the downstream side in the circulation direction, the unit packed column for supplying the circulating fluid may be switched to the unit packed column on the downstream side in the circulation direction. By adopting such a switching system, the component packed in the raw material liquid remaining in the eluent supply means is not packed with either the non-adsorbate fraction or the adsorbate fraction in the unit packed column on the downstream side in the circulation direction. To be supplied to. As a result, it is possible to prevent a new adsorption zone from being formed in the adsorption zone of the circulation system and the disturbance of the adsorption zone.

  In the first embodiment, the number of unit packed towers is eight, and in the second embodiment, the number of unit packed towers is four. However, the number of unit packed towers is not limited as long as the number is four or more.

  In the second step of the second embodiment, the raw material liquid 30 is circulated through the raw material liquid circulation line 330. However, even if the pump Pf is stopped and the supply of the raw material liquid 30 to the circulation system 310 is stopped. good. However, it is preferable to circulate the raw material liquid 30 in the raw material liquid circulation line 330 in order to maintain the viscosity of the raw material liquid 30. Also in the first embodiment, the raw material liquid 30 may be circulated through the raw material liquid circulation line 31 as necessary.

  An eluent circulation line similar to the raw material liquid circulation line in the first and second embodiments may be provided.

  In the second embodiment, the valve d0 provided between the eluent storage tank 42 and the branch 345 is closed and the second step is performed. However, in order to smoothly circulate the liquid in the circulation system 310, the valve You may perform a 2nd process, adjusting the opening degree of d0.

  In the second embodiment, the first step and the second step are alternately repeated, but the present invention is not limited to this, and the second step may be performed between the first steps. For example, the order of operations I-1 to I-5 in the second embodiment may be operation I-1 → operation I-3 → operation I-4 → operation I-5 → operation I-2. Further, for example, the operation I-1, the operation I-3, the operation I-4, the operation I-2, the operation I-4, and the operation I-5 may be performed. The order of such operations can be appropriately combined in consideration of the types of components contained in the raw material liquid and the size and number of unit packed towers.

  EXAMPLES Hereinafter, although an Example is given and this invention is demonstrated concretely, it is not limited to an Example.

Production Example 1 Production of Chromatographic Separation Apparatus A cylindrical stainless steel column with an inner diameter of 20 mm and a height of 0.9 m and a calcium-type strongly acidic cation exchange resin (Amberlite (trade name) CR1220, ROHM as an adsorbent) (Manufactured by And Haas Co.) 1.13 L was packed to prepare four unit packed towers. A chromatographic separation apparatus A similar to the chromatographic separation apparatus 300 shown in FIG. 2 was manufactured using the prepared unit packed tower. The jacket of the unit packed tower was connected to a constant temperature water tank.

Example 1
Warm water was passed through the jacket of each unit packed column of the chromatographic separation apparatus A, and the inside of the unit packed column was kept at 60 ° C. Next, chromatographic separation using the raw material solution according to the valve operation shown in Table 2 and glucose as the non-adsorbate fraction and fructose as the adsorbate fraction under the following operating conditions (described as the amount per cycle). Went. As the raw material liquid, a sugar liquid having 52% by mass of glucose, 42% by mass of fructose and 6% by mass of disaccharide and having a total sugar concentration of 60% by mass was used. Pure water was used as the eluent. The chromatographic separation was performed after 20 cycles were repeated, and it was assumed that the steady state was reached. The non-adsorbate fraction and the adsorbate fraction were collected, the sugar composition and the recovery rate were measured, and the results are shown in Table 4.

<Operating conditions>
(1) Supply amount of raw material liquid ... 0.023L
(2) Eluent supply amount ... 0.230L
(3) Extraction amount of non-adsorbate fraction accompanying supply of raw material liquid: 0.023L
(4) Amount of non-adsorbate fraction extracted with eluent supply: 0.112L
(5) Amount of adsorbate fraction extracted ... 0.118L
(6) Flow rate of circulating fluid in the second process, first treatment ... 0.380L
(7) Flow rate of circulating fluid in the second process, second treatment ... 0.380L
(8) Time per cycle ... 0.44H

(Comparative Example 1)
Warm water was passed through the jacket of each unit packed column of the chromatographic separation apparatus A, and the inside of the unit packed column was kept at 60 ° C. Next, according to the valve operation shown in Table 3, the raw material solution was set to one cycle of operations i-1 to iv-4, and glucose was not adsorbed according to the following operating conditions (described as the amount per cycle). Chromatographic separation was performed using the fraction and fructose as the adsorbate fraction. As the raw material liquid, a sugar liquid having 52% by mass of glucose, 42% by mass of fructose and 6% by mass of disaccharide and having a total sugar concentration of 60% by mass was used. Pure water was used as the eluent. Chromatographic separation was performed after 20 cycles were repeated, and it was assumed that a steady state was reached. The non-adsorbate fraction and the adsorbate fraction were collected, the sugar composition and the recovery rate were measured, and the results are shown in Table 4. .

<Operating conditions>
(1) Supply amount of raw material liquid: 0.023L
(2) Eluent supply amount: 0.230L
(3) Extraction amount of non-adsorbate fraction: 0.135L
(4) Amount of adsorbate fraction extracted ... 0.118L
(5) Flow rate of circulating fluid in the second step ... 0.760L
(6) Time per cycle ... 0.44H

(Total sugar content)
The total amount of sugar in the raw material liquid, the non-adsorbate fraction, and the adsorbate fraction was measured using a Brix meter.

(Sugar composition)
The sugar composition in the raw material liquid, the non-adsorbate fraction, and the adsorbate fraction was measured by HPLC (LC-VP series, manufactured by Shimadzu Corporation) under the following conditions.
[Measurement condition]
Column: TSKgel SCX, manufactured by Tosoh Corporation Eluent: pure water Flow rate: 0.8 mL / min
Sample volume: 10 μL
Temperature: 60 ° C
Detector: Differential refractometer (RI)

(Recovery rate)
The recovery rate was expressed as the ratio of each component in the raw material liquid contained in the non-adsorbate fraction and the adsorbate fraction.

  From the composition of the non-adsorbate fraction and the adsorbate fraction shown in Table 4, the non-adsorbate fraction of Example 1 had a composition of the target glucose in the total sugar of 87.5% by mass. Moreover, the composition of the adsorbate fraction of Example 1 in the total sugar of the target fructose was 98.0% by mass. On the other hand, the non-adsorbent fraction of Comparative Example 1 had a composition of glucose in the total sugar of 86.1% by mass. Moreover, the composition of the adsorbate fraction of Comparative Example 1 in the total sugar of fructose was 97.6% by mass. As a result, in Example 1 in which the second treatment was performed in the second step, since the glucose concentration in the non-adsorbate fraction and the fructose concentration in the adsorbate fraction were higher than those in Comparative Example 1, the target component was It was found that it can be separated with high purity.

  From the recovery rate of the non-adsorbate fraction and the adsorbate fraction shown in Table 4, in Example 1, 95.2% by mass of the added fructose was contained in the adsorbate fraction. In addition, 6.6% by mass of the added disaccharide was contained in the adsorbate fraction. On the other hand, in Comparative Example 1, 93.0% by mass of the added fructose was contained in the adsorbate fraction. In addition, 12.3% by mass of the added disaccharide was contained in the adsorbate fraction. As a result, in Example 1 in which the second treatment was performed in the second step, compared to Comparative Example 1, disaccharides separated into the adsorbate fraction can be reduced, and the purity of fructose in the adsorbate fraction can be improved. I found out.

It is a schematic diagram which shows the chromatographic separation apparatus concerning 1st embodiment of this invention. It is a schematic diagram which shows the chromatographic separation apparatus concerning 2nd embodiment of this invention.

Explanation of symbols

8,300 Chromatographic separation apparatus 10,310 Circulation system 11-18, 311-314 Unit packed tower 34, 36, 38, 44, 46, 70, 80, 90, 111-118, 321-324, 334, 336, 338 370, 380, 390 Piping 51-58, 61-68, 71-78, 81-88, 91-98, 351-354, 361-364, 371-374, 381-384, 391-394 Branch piping 101, 301 Check valve RF, RD, PA, PC, Pf, Pd pump F0-F8, f0-f4, D1-D8, d0-d4, A1-A8, a1-a4, C1-C8, c1-c4, Z1- Z8, z1-z4, R1-R8, r1-r4 valves

Claims (4)

A plurality of unit packed towers packed with an adsorbent are connected in series to form a circulation system in which the raw material liquid and the eluent passed through the unit packed tower can be circulated, and a raw material liquid containing two or more components Is passed through the circulation system to form an adsorption zone according to the affinity for the adsorbent, and the raw material liquid and / or the eluent is supplied to an arbitrary unit packed column, and the unit packed column enriched with an arbitrary component From the first step of extracting the fraction of the arbitrary component,
The liquid in the circulation system is extracted from an arbitrary unit packed column constituting the circulation system, and the extracted liquid is supplied to the unit packed column on the downstream side in the circulation direction of the unit packed column from which the liquid has been extracted. Then, the unit packed tower for extracting the liquid in the circulation system and the unit packed tower for supplying the extracted liquid in the circulation system are switched to the unit packed tower on the downstream side in the circulation direction of the circulation system for circulation. And a second step of performing a chromatographic separation method. Repeat the first step and the second step,
In the second step, the part enriched with an arbitrary component is moved to the unit packed column on the downstream side in the circulation direction by circulation of the liquid in the circulation system,
Corresponding to the movement, the first step includes unit packing tower for supplying the raw material liquid and / or eluent and unit packing tower for extracting the fraction of the arbitrary component on the downstream side of the circulation system. The tower has a transition operation that transitions sequentially,
The transfer operation includes a unit packed tower for extracting the liquid in the circulation system in the second step and a unit packed tower for supplying the extracted liquid in the circulation system to the unit packed tower on the downstream side in the circulation direction of the circulation system. The chromatographic separation method according to claim 1, wherein the method is performed after switching.   The chromatography according to claim 1 or 2, wherein the second step is performed by stopping the supply of the raw material liquid and the eluent to the circulation system and the extraction of the fraction from the circulation system. Separation method.   In the second step, the unit packed column for extracting the liquid in the circulation system is switched to the unit packed column on the downstream side in the circulation direction of the circulation system, and then the unit packed column for supplying the liquid in the extracted circulation system is circulated in the second step. The chromatographic separation method according to any one of claims 1 to 3, wherein the unit separation tower is switched to the downstream side in the circulation direction of the system.

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