JP6546808B2 - Chromatographic separation method and chromatographic separation system - Google Patents

Description

  The present invention relates to a chromatographic separation method and a chromatographic separation system. More specifically, the present invention relates to a chromatographic separation method and a chromatographic separation system using a simulated moving bed system.

  Chromatographic separation methods are roughly classified into fixed bed methods and moving bed methods. In the fixed bed method, a multicomponent sample (hereinafter, also referred to as "stock solution") is injected into a column filled with an adsorbent, and the eluent is allowed to flow in one direction in the column, thereby removing the adsorbent. The target component in the stock solution is separated and purified from other components based on the difference in adsorption power. In this fixed bed system, the target component can be separated and purified only by flowing the eluent while fixing the adsorbent. However, if there is no significant difference in the adsorptive power to the adsorbent between the target component to be purified (hereinafter, also simply referred to as "component to be purified") and the other components, the fixed bed system is good. Separation and purification can not be realized. Furthermore, in the fixed bed method, it is not possible to separate the target component while continuously injecting the sample, which limits the industrial application.

On the other hand, in the moving bed method, the adsorbent is moved in the reverse direction with respect to the flowing direction of the eluting solution while flowing the eluting solution in one direction in the column. In the moving bed method, the component to be purified in the undiluted solution is moved in the direction opposite to the moving direction of the other components based on the inlet of the undiluted solution by adjusting the flow rate of the eluant and the moving velocity of the adsorbent. It can be done. Therefore, for example, when the component to be purified in the stock solution is a stronger adsorptive component than the other components, the component to be purified is upstream with respect to the inlet of the stock solution (reverse direction to the fluid flow direction) In addition, it is possible to construct a system in which other components move to the downstream side (flow direction of fluid). By extracting the purification target component on the upstream side and extracting the other components on the downstream side, the purification target component can be separated and purified continuously with higher purity while continuously injecting the stock solution.
However, industrial application of the moving bed system is not easy. In the chromatographic separation system using a large column used industrially, moving the adsorbent uniformly has high technical hurdles. Also, even if the adsorbent can be moved uniformly, the adsorbent is heavily loaded during this movement. As a result, the adsorbent is likely to be broken (fractured), and the constructed chromatographic separation system is likely to be less practical in durability and the like.

  In order to solve the problems of the moving bed system, chromatographic separation techniques using a simulated moving bed system have been proposed (for example, Patent Documents 1 and 2). The chromatographic separation by the simulated moving bed method is performed using a circulatory system in which a plurality of unit packed columns (columns) packed with adsorbents are connected in series and endlessly via piping. In this circulation system, the pipe has a stock solution supply port for supplying a stock solution containing the component to be purified, an outlet for the weakly adsorptive component, an eluent supply port, and an outlet for the strongly adsorptive component. Between the stock solution supply port and the weakly adsorptive component outlet, between the weakly adsorptive component outlet and the eluent supply port, the eluent supply port and the strongly adsorptive component outlet. At least one unit packing tower is disposed between the outlet and between the strong adsorptive component outlet and the stock solution inlet. The undiluted solution supply port, the weakly adsorptive component outlet, the eluent feed port, and the strongly adsorptive component outlet are intermittently directed in the fluid flow direction while maintaining their relative positional relationship. By moving it in the opposite direction, although the adsorbent is fixed, it is possible to obtain the same effect as moving the adsorbent in the reverse direction with respect to the flow direction of the fluid.

Japanese Patent Publication No. 60-55162 JP, 2009-36536, A

In the chromatographic separation, in addition to the stock solution containing the component to be purified, the eluent for flushing the stock solution is supplied. Therefore, the target fraction containing the purification target component (hereinafter, also referred to as "purification target fraction") is in a state where the purification target component is diluted to a certain extent by the eluent. Therefore, although the purification target fraction obtained by the chromatographic separation is usually shipped via a concentration operation, this concentration operation contributes to an increase in the purification cost.
In order to reduce the cost, it is necessary to increase the concentration of the purification target component in the purification target fraction. However, if the amount of eluent used in the chromatographic separation is reduced to increase the concentration of the component to be purified, the separation performance generally decreases, and the purity of the component to be purified in the fraction to be purified (the fraction to be purified The ratio of the component to be purified to the remainder excluding the solvent and the solvent in the undiluted solution decreases. Furthermore, when the amount of eluent used is reduced, the recovery rate of the component to be purified contained in the stock solution into the fraction to be purified tends to decrease.

The present invention is a chromatographic separation method using a simulated moving bed method, which can greatly reduce the amount of eluent used, and the recovery rate of the component to be purified into the fraction to be purified obtained, and the purification target It is an object of the present invention to provide a chromatographic separation method capable of highly enhancing any of the purity of the component to be purified in the fraction.
Another object of the present invention is to provide a chromatographic separation system suitable for carrying out the above-mentioned chromatographic separation method.

As a result of intensive investigations in view of the above problems, the inventors of the present invention have found that, in a chromatographic separation method using a simulated moving bed method, a liquid supply operation selected from a stock solution supply operation and an eluent supply operation and a strongly adsorptive fraction extraction. The step of performing a specific combination of the liquid extraction operation selected from the extraction operation and the weakly adsorptive fraction extraction operation in a specific order, the fluid in the system without performing any liquid supply operation and liquid extraction operation By combining the method with the circulation step of circulating the mixture, thereby significantly reducing the amount of the eluent used, while at the same time the purity of the component to be purified in the fraction to be purified and the recovery of the component to be purified into the fraction to be purified It has been found that either can be raised to the desired level. That is, it was found that the component to be purified can be separated and purified at high concentration, high purity, and high recovery rate in the purification target fraction.
The present invention has been further studied based on these findings and has been completed.

The above problems of the present invention are solved by the following means.
[1]
A chromatographic separation method of separating and purifying components in a stock solution by a simulated moving bed method using a circulating system in which a plurality of unit packed towers filled with an adsorbent are connected in series and endlessly via a pipe,
The circulation system includes the undiluted solution feed port F, the weakly adsorptive fraction outlet A, the eluent feed port D, and the strongly adsorptive fraction outlet C in the pipe in this order in the fluid flow direction. Between the undiluted solution feed port F and the weakly adsorptive fraction outlet A, between the weakly adsorptive fraction outlet A and the eluent feed port D, and the eluent feed port D and At least one unit packing tower is disposed between the strongly adsorptive fraction outlet C and between the strongly adsorptive fraction outlet C and the stock solution feed port F,
Repeat the following steps (a) and (b) in sequence:
(A) performing the following sub-steps (i) to (iv) in this order;
(I) A substep of supplying a stock solution from the stock solution supply port F and extracting a strongly adsorptive fraction from the strongly adsorptive fraction outlet C;
(Ii) a substep of supplying a stock solution from the stock solution supply port F and extracting a weakly adsorptive fraction from the weakly adsorptive fraction outlet A;
(Iii) a substep of supplying an eluent from the eluent supply port D and extracting a weakly adsorptive fraction from the weakly adsorptive fraction outlet A;
(Iv) A substep of circulating the fluid in the circulatory system without supplying the stock solution and the eluent and without extracting the strongly adsorptive fraction and the weakly adsorptive fraction,
(B) After the step (a) is completed, the undiluted solution supply port F, the weakly adsorptive fraction outlet A, the eluent feed port D, and the strong adsorptive fraction outlet C are relative to one another. Step of moving in the fluid flow direction while maintaining the positional relationship.
[2]
The chromatographic separation method according to [1], wherein in the step (a), the total amount of the strong adsorptive fraction extracted from the outlet C is smaller than the total amount of the undiluted solution supplied from the supply port F. .
[3]
The chromatographic separation method according to [1] or [2], wherein the stock solution contains glucose and fructose, and the fructose is separated and purified in the strongly adsorptive fraction.
[4]
The chromatographic separation method according to any one of [1] to [3], wherein the circulating system has at least four unit packed towers.
[5]
In the step (a), the ratio of the total supply amount of the eluent to the total supply amount of the stock solution satisfies, in a volume ratio, [total supply amount of the eluent] / [total supply amount of the stock solution] <1.2 The chromatographic separation method according to any one of [1] to [4].
[6]
A chromatographic separation system for separating and purifying components in a stock solution by a simulated moving bed method using a circulating system in which a plurality of unit packed towers packed with adsorbents are connected in series and endlessly via a pipe,
The circulation system includes the undiluted solution feed port F, the weakly adsorptive fraction outlet A, the eluent feed port D, and the strongly adsorptive fraction outlet C in the pipe in this order in the fluid flow direction. Between the undiluted solution feed port F and the weakly adsorptive fraction outlet A, between the weakly adsorptive fraction outlet A and the eluent feed port D, and the eluent feed port D and At least one unit packing tower is disposed between the strongly adsorptive fraction outlet C and between the strongly adsorptive fraction outlet C and the stock solution feed port F,
System which repeats the following steps (a) and (b) in order:
(A) performing the following sub-steps (i) to (iv) in this order;
(I) A substep of supplying a stock solution from the stock solution supply port F and extracting a strongly adsorptive fraction from the strongly adsorptive fraction outlet C;
(Ii) a substep of supplying a stock solution from the stock solution supply port F and extracting a weakly adsorptive fraction from the weakly adsorptive fraction outlet A;
(Iii) a substep of supplying an eluent from the eluent supply port D and extracting a weakly adsorptive fraction from the weakly adsorptive fraction outlet A;
(Iv) A substep of circulating the fluid in the circulatory system without supplying the stock solution and the eluent and without extracting the strongly adsorptive fraction and the weakly adsorptive fraction,
(B) After the step (a) is completed, the undiluted solution supply port F, the weakly adsorptive fraction outlet A, the eluent feed port D, and the strong adsorptive fraction outlet C are relative to one another. Step of moving in the fluid flow direction while maintaining the positional relationship.

As used herein, the terms "upstream" and "downstream" are used with respect to the flow direction of fluid in the circulatory system. That is, "upstream" with respect to a part of the circulatory system means the side through which the fluid flows toward the site, and "downstream" means the side from which the fluid flows out from the site means.
In the present specification, the "strongly adsorptive component" means a component having a strong adsorptive power to the adsorbent among a plurality of components contained in the stock solution, and the "weakly adsorptive component" means the above-mentioned strongly adsorptive component It means a component that is less adsorptive to the adsorbent. In other words, the terms "strongly adsorptive" and "weakly adsorptive" refer to the degree of strength of the adsorptive power of the components contained in the stock solution when the adsorptive powers of the components are relatively compared. It is a thing.
The “strongly adsorptive component” and the “weakly adsorptive component” may each be composed of a single component or may be composed of a plurality of components having different adsorptive powers. Since the component to be purified is often a single component, when the strongly adsorptive component is the component to be purified, the strongly adsorptive component is usually the component having the strongest adsorptive power to the adsorbent in the stock solution. However, the present invention is not limited to this embodiment. The weakly adsorptive component is one or two or more components that are less adsorptive to the adsorbent than the strong adsorptive component. Similarly, when the weakly adsorptive component is a component to be purified, the weakly adsorptive component is usually the component having the weakest adsorptive power to the adsorbent in the undiluted solution, but the present invention is limited to this aspect It is not a thing. The strongly adsorptive component is one or two or more components that are more adsorptive to the adsorbent than the weakly adsorptive component.

  According to the chromatographic separation method of the present invention, the components to be purified in the stock solution can be separated and purified at high concentration, high purity, and high recovery rate. Further, the chromatographic separation system of the present invention can be used to carry out the above-mentioned chromatographic separation method of the present invention.

FIG. 1 is a system diagram showing an embodiment of the chromatographic separation system of the present invention. FIG. 2 is an explanatory view for explaining the basic concept of the chromatographic separation by the simulated moving bed method.

  A preferred embodiment of the chromatographic separation method of the present invention (hereinafter, also simply referred to as "the present method") will be described.

The method of the present invention is carried out using a circulation system in which a plurality of unit packed towers packed with adsorbent are connected in series and endlessly via piping. The circulatory system itself used in the simulated moving bed system is known, and, for example, JP-A-2009-36536 and JP-B-7-46097 can be referred to.
Although the said circulatory system is demonstrated below using drawing, this invention is not limited to these aspects.
Note that the drawings referred to below are explanatory diagrams for facilitating the understanding of the present invention, and the sizes of relative configurations and relative magnitude relationships may be different in magnitude for convenience of explanation, and actual relationships It does not show as it is. Further, other than the matters specified in the present invention, the present invention is not limited to the shapes, relative positional relationships, etc. shown in these drawings.

A preferred embodiment of the circulatory system used in the method of the present invention is shown in FIG. The circulatory system 100 shown in FIG. 1 includes four unit packed towers (unit packed towers 10a, 10b, 10c, 10d) packed with the adsorbent Ab, and the outlet of each unit packed column is an adjacent unit packed tower The unit packing towers are connected in series as a whole.
And, the outlet of the last unit packed tower (for example, unit packed tower 10d) is connected to the inlet of the foremost unit packed tower (for example, unit packed tower 10a) through piping 1, and all unit packed towers are endless. Are connected in a ring shape. With this configuration, it is possible to circulate the fluid in the circulatory system 100. It is preferable that unit packing towers 10a to 10d have the same internal shape, size, and adsorbent loading amount (preferably the same).

  In the circulation system 100, a circulation pump P1 for circulating a fluid in the arrow direction is disposed. The circulation pump P1 is preferably a metering pump. Further, in the circulation system 100, the piping 1 between two adjacent unit packing towers is provided with shutoff valves R1, R2, R3, and R4 capable of blocking the flow of fluid to the unit packing towers on the downstream side thereof. It is done.

  A fraction containing a large amount of a weakly adsorptive component to the adsorbent Ab between each of the shutoff valves R1 to R4 and the outlets of the unit packed towers 10a to 10d located upstream thereof (herein, “the fraction The weakly adsorptive fraction extraction lines 2a, 2b, 2c and 2d for extracting the weakly adsorptive fraction "or simply the" weakly adsorptive fraction "to the adsorbent Ab are branched. In each weakly adsorptive fraction extracting line 2a, 2b, 2c, 2d, weakly adsorptive fraction extracting valves A1, A2, A3, A4 capable of opening and closing each weakly adsorptive fraction extracting line are provided. It is provided. The weakly adsorptive fraction extracting lines 2a, 2b, 2c and 2d are merged and combined into one weakly adsorptive fraction combining pipe 3.

  Similarly, between each shutoff valve R1 to R4 and the outlet of each unit packed tower 10a to 10d located upstream thereof, a fraction containing a large amount of strongly adsorptive component to the adsorbent Ab (this specification The strongly adsorptive fraction extraction lines 4a, 4b, 4c and 4d for extracting the "strongly adsorptive fraction to the adsorbent Ab" or simply the "strongly adsorptive fraction" are branched. In each strongly adsorptive fraction extracting line 4a, 4b, 4c, 4d, strongly adsorptive fraction extracting valves C1, C2, C3, C4 capable of opening and closing each strongly adsorptive fraction extracting line are provided. It is provided. The strongly adsorptive fraction extracting lines 4a, 4b, 4c and 4d are merged and combined into one strongly adsorptive fraction merging pipe 5.

  In step (a) to be described later, any one of the weakly adsorptive fraction extraction valves A1, A2, A3 and A4 is opened. The connection site between the weakly adsorptive fraction extraction line in which the opened valve is installed and the pipe 1 is the outlet A of the weakly adsorptive fraction in the step (a). In step (a), one of the strongly adsorptive fraction extracting valves C1, C2, C3 and C4 is opened. The connection site between the pipe 1 and the strongly adsorptive fraction extraction line in which the opened extraction valve is installed, and the pipe 1 form the outlet C of the strongly adsorptive fraction in step (a).

  In order to prevent the pressure of the circulatory system 100 from rising excessively, the circulatory system 100 is preferably provided with a safety valve (or a relief valve), not shown, at an appropriate site. It is also preferable to provide check valves T1, T2, T3 and T4 for backflow prevention between two adjacent unit packing towers.

In the circulation system 100, as shown in FIG. 1, the stock solution 7 contained in the stock solution tank 6 and the eluent 9 contained in the eluent tank 8 can be supplied. The undiluted solution 7 is supplied via the undiluted solution supply line 11 by an undiluted solution supply pump P2 whose feed flow rate can be controlled. The stock solution supply pump P2 is preferably a metering pump. The stock solution supply line 11 preferably includes a relief valve U that returns the stock solution to the stock solution tank 6 when the supply pressure exceeds the set pressure while the stock solution is being supplied. The stock solution supply line 11 is branched into four stock solution supply branch lines 11a, 11b, 11c and 11d as shown in FIG. 1, and the stock solution is respectively supplied via the stock solution supply branch lines 11a, 11b, 11c and 11d. It can be supplied to the inlet of each unit packed tower 10a, 10b, 10c, 10d. Each of the stock solution supply branch lines 11a, 11b, 11c and 11d is provided with openable stock solution supply valves F1, F2, F3 and F4, and is passed through the stock solution supply branch line having an open stock solution supply valve. The stock solution is supplied to a unit packed tower connected downstream.
In step (a) to be described later, any one of the stock solution supply valves F1, F2, F3, and F4 is opened. The connection part with the undiluted | stock solution supply branch line in which the said undiluted | flowed undiluted | stock solution supply valve was installed, and the piping 1 becomes the undiluted | stock solution supply port F in step (a).

The eluent 9 is supplied via the eluent supply line 12 by the eluent supply pump P3 capable of controlling the supply flow rate. The eluent supply pump P3 is preferably a metering pump. The eluent supply line 12 preferably includes a relief valve V that returns the eluent to the eluent tank 8 when the supply pressure exceeds the set pressure during the supply of the eluent. The eluent supply line 12 is branched into four eluent supply branch lines 12a, 12b, 12c and 12d as shown in FIG. 1, and elution is performed via the respective eluent supply branch lines 12a, 12b, 12c and 12d. The liquid can be supplied to the inlet of each unit packed column 10a, 10b, 10c, 10d. Each of the eluent supply branch lines 12a, 12b, 12c and 12d is provided with an eluent supply valve D1, D2, D3, D4 which can be opened and closed, and the eluent supply branch line having an opened eluent supply valve. Eluate is fed through to the unit packed column connected downstream thereof.
In step (a) to be described later, any one of the eluent supply valves D1, D2, D3 and D4 is opened. The connection site between the pipe 1 and the eluent supply branch line where the opened eluent supply valve is installed, and the pipe 1 form the eluent supply port D in step (a).

  Subsequently, the operation of the circulatory system when the method of the present invention is carried out by the circulatory system will be described. In the method of the present invention, the following steps (a) and (b) are sequentially repeated using the above-mentioned circulatory system.

(A) performing the following sub-steps (i) to (iv) in this order:
(I) A substep of supplying the stock solution from the stock solution supply port F and extracting the strongly adsorptive fraction from the strongly adsorptive fraction outlet C.
(Ii) A substep of supplying the stock solution from the stock solution supply port F and extracting the weakly adsorptive fraction from the weakly adsorptive fraction outlet A.
(Iii) A substep of supplying the eluent from the eluent supply port D and extracting the weakly adsorptive fraction from the weakly adsorptive fraction outlet A.
(Iv) A substep of circulating the fluid in the circulatory system without supplying the stock solution and the eluent and without extracting the strongly adsorptive fraction and the weakly adsorptive fraction.
(B) After the step (a) is completed (that is, after the substep (iv) is completed), the stock solution supply port F, the weakly adsorptive fraction outlet A, the eluent supply port D and the strong adsorption Moving the sex fraction outlet C in the fluid flow direction while maintaining the relative positional relationship between them.

[Step (a)]
The step (a) is a step of sequentially executing the four sub steps (i) to (iv). Each substep will be described with reference to the circulatory system shown in FIG.

<Substep (i)>
In substep (i), the strong adsorptive fraction is extracted from the strongly adsorptive fraction outlet C while supplying the undiluted solution from the undiluted solution supply port F. For example, when the undiluted solution supply valve F1 is opened and the undiluted solution supply pump P2 is operated to connect the undiluted solution supply branch line 11a and the pipe 1 to the undiluted solution supply port F, the extraction valve C3 is opened. A connecting portion between the strongly adsorptive fraction extraction line 4 c and the pipe 1 is taken as an outlet C.

  In substep (i), all of the stock solution supply valves F2 to F4, the weakly adsorptive fraction extraction valves A1 to A4, the strongly adsorptive fraction extraction valves C1, C2 and C4, and the shutoff valve R3 are closed. In addition, the eluent supply valves D1 to D4 may be opened as long as the eluent supply pump P3 is stopped, but in order to carry out the supply and withdrawal of the solution with high accuracy, the eluent supply is performed. The valves D1 to D4 are preferably closed. The shutoff valves R1 and R2 may be open, and the shutoff valve R4 may be open or closed. The circulation pump P1 and the eluent feed pump P3 are normally stopped.

  In the substep (i), the amount of the stock solution supplied from the above-mentioned stock solution supply port F is not particularly limited, but per hour, per liter of adsorbent Ab packed in the unit packed tower 10a, 0.032 It is preferable to set it as -0.059 liter, and it is more preferable to set it as 0.036-0.054 liter. The total supply amount of the stock solution in sub-step (i) is substantially the same as the total amount of the strongly-adsorbable fraction extracted from the strongly-adsorbable fraction outlet C in sub-step (i).

<Sub-step (ii)>
In sub-step (ii), the weakly adsorptive fraction is extracted from the weakly adsorptive fraction outlet A while supplying the undiluted solution from the undiluted solution supply port F. That is, the strong adsorptive fraction extraction valve C3 which has been opened in the above sub-step (i) is closed, and instead the weakly adsorptive fraction extraction valve A1 is opened. That is, the connection site between the weakly adsorptive fraction extraction line 2a having the weakly adsorptive fraction extraction valve A1 and the pipe 1 is taken as the weakly adsorptive fraction extraction outlet A, and the weakly adsorptive fraction from the relevant outlet A Pull out.

  In substep (ii), all of the undiluted solution supply valves F2 to F4, the weakly adsorptive fraction extraction valves A2 to A4, the strongly adsorptive fraction extraction valves C1 to C4, and the shutoff valve R1 are closed. In addition, the eluent supply valves D1 to D4 may be opened as long as the eluent supply pump P3 is stopped, but in order to carry out the supply and withdrawal of the solution with high accuracy, the eluent supply is performed. The valves D1 to D4 are preferably closed. The shutoff valves R2 to R4 may be open or closed. Further, as in sub-step (i), the circulation pump P1 and the eluent feed pump P3 are normally stopped.

  In sub-step (ii), the amount of the stock solution supplied from the above-mentioned stock solution feed port F is not particularly limited, but per hour, per liter of adsorbent Ab packed in unit packed tower 10a, 0.020 to 20%. It is preferable to set it as 0.036 liter, and it is more preferable to set it as 0.022-0.034 liter. The total supply amount of the stock solution in sub-step (ii) is substantially the same as the total amount of the weakly adsorptive fraction withdrawn from the weakly adsorptive fraction outlet A in sub-step (ii).

<Substep (iii)>
In substep (iii), the eluent is supplied from the eluent supply port D, and the weakly adsorptive fraction is withdrawn from the weakly adsorptive fraction outlet A. That is, the supply valve F1 which has been opened in the substep (ii) is closed to stop the operation of the stock solution supply pump P2, and instead the eluent supply valve D3 is opened to operate the eluent supply pump P3. That is, the connection portion between the eluent supply branch line 12c having the supply valve D3 and the pipe 1 becomes the eluent supply port D, the eluent is supplied from the eluent supply port D, and the weakly adsorptive fraction The weakly adsorptive fraction is withdrawn from the outlet A.

  In substep (iii), the eluent supply valves D1, D2 and D4, the weakly adsorptive fraction extraction valves A2 to A4, the strongly adsorptive fraction extraction valves C1 to C4, and the shutoff valve R1 are all closed. ing. Undiluted solution supply valves F1 to F4 may be opened as long as undiluted solution pump P2 is stopped, but undiluted solution supply valves F1 to F4 may be opened to carry out liquid supply and extraction with higher accuracy. It is preferably closed. The shutoff valve R2 may be open or closed. The shutoff valves R3 and R4 are both open. Further, the circulation pump P1 may be operated or may be stopped.

  In sub-step (iii), the amount of the eluent supplied from the eluent supply port D is not particularly limited, but per liter of the adsorbent Ab packed in the unit packed tower 10a per hour, it is 0. It is preferable to set it as 051-0.095 liter, and it is more preferable to set it as 0.058-0.088 liter. The total amount of eluent supplied in substep (iii) is, in effect, substantially the same as the total amount of weakly adsorptive fraction withdrawn from weakly adsorptive fraction outlet A in substep (iii).

<Substep (iv)>
In sub-step (iv), the circulation pump P1 is operated without supplying the stock solution and the eluent and without extracting the strongly adsorptive fraction and the weakly adsorptive fraction. Simply circulate. If the undiluted solution supply pump P2 and the eluant solution supply pump P3 are stopped, the undiluted solution supply valves F1 to F4 and the eluant solution supply valves D1 to D4 may be open, but from the viewpoint of controlling the fluid with higher accuracy, The stock solution supply valves F1 to F4 and the eluent supply valves D1 to D4 are preferably closed. The extraction valves A1 to A4 and the extraction valves C1 to C4 are all closed, and the shutoff valves R1 to R4 are all open.

  In sub-step (iv), the circulating flow rate of the fluid in the system is preferably 0.133 to 0.247 liters per liter of adsorbent Ab packed in unit packed tower 10a per hour, It is more preferable to set it as 0.152-0.228 liter.

  The relationship between the supply amount of the liquid and the circulation flow rate in each substep is not particularly limited, and is appropriately adjusted in accordance with the type of the component in the stock solution. Usually, the total amount (I) of the undiluted solution supplied from the undiluted solution supply port F in the sub-step (i) and the total amount (II) of undiluted solution supplied from the undiluted solution supply port F in the sub-step (ii) In terms of volume ratio, it is preferable to set the total amount (II) / total amount (I) = 0.30 to 0.70, and more preferably 0.40 to 0.60.

  Further, in the sub-step (i), the total amount (I) of the undiluted solution supplied from the undiluted solution supply port F, and in the sub-step (iii), the total amount of eluant supplied from the eluant It is preferable to set it as total amount (III) / total amount (I) = 1.14-2.11 by a volume ratio, and, as for a relationship with III), it is more preferable to set it as 1.30-1.95.

  Further, in the sub-step (i), the total amount (I) of the undiluted solution supplied from the undiluted solution supply port F, and in the sub-step (ii), the total amount (II) of undiluted solution supplied from the undiluted solution supply port F The relationship between the total amount (I + II) and the total amount (III) of the eluent supplied from the eluent supply port D in the above sub-step (iii) is the total volume (III) / total amount by volume ratio It is preferable to set it as (I + II) = 0.5-1.5, and it is more preferable to set it as 0.7-1.2.

  Further, in the sub-step (i), the relationship between the total amount (I) of the undiluted solution supplied from the undiluted solution supply port F and the total circulating flow rate in the sub-step (iv) The total amount (I) is preferably 3.0 to 5.0, and more preferably 3.5 to 4.5.

In the circulating system used in the present invention, the loading amount of the adsorbent charged in one unit packing tower is not particularly limited and may be appropriately selected according to the purpose, but it is usually 10 mL to 150 m ³ , preferably Is 150 mL to ³⁰ m ³ , more preferably 300 mL to 15 m ³ .
Further, the relationship between the total volume of the adsorbent Ab filled in all unit packed towers in the circulation system and the total volume of the pipe 1 (total volume in the cavity of the pipe 1) is the volume ratio [Pipe 1 The total volume of [(total volume of adsorbent Ab)] = [0.01 to 0.2] is preferable.

  The ratio of the total feed of the eluent to the total feed of the stock solution in step (a) (that is, in the above substeps (i) to (iv)) (corresponding to the total (III) / total (I + II) above) It is more preferable that [total supply amount of eluent] / [total supply amount of stock solution] <1.2, and [total supply amount of eluent] / [total supply amount of stock solution] ≦ 1.1. It is further preferable to satisfy. There is no particular limitation on the lower limit value of the ratio, and it is more practical to set [total supply amount of eluent] / [total supply amount of stock solution] 原 0.5. The ratio is more preferably 0.5 ≦ [total supply amount of eluent] / [total supply amount of stock solution] ≦ 1.1, and 0.8 ≦ [total supply amount of eluent] / [total stock amount] It is also preferable to set the total supply amount] ≦ 1.1.

The temperature at which the method of the present invention is carried out is not particularly limited as long as the fluid in the circulation system is liquid, and may be appropriately selected depending on the purpose. It is usually carried out at 40-80 ° C.
In the method of the present invention, the flow rate of the supplied liquid or the flow rate of the liquid circulating in the circulatory system may be constant or may be varied during each substep, but is usually constant. . In addition, the flow rate of the supplied liquid or the flow rate of the liquid circulating in the circulatory system may be constant or may be varied between the substeps, but is usually constant. That is, in sub-steps (i) to (iii), the flow rate of the supplied liquid is preferably constant, and in sub-step (iv), the flow rate of the liquid circulating in the circulatory system is also sub-step (i) to It is preferable to make it the same as the flow rate of the liquid supplied in (iii).

[Step (b)]
After the above step (a) is completed (after the completion of sub step (iv)), step (b) is performed. The step (b) maintains the relative positional relationship among the undiluted solution supply port F, the weakly adsorptive fraction outlet A, the eluent supply port D, and the strong adsorptive fraction outlet C. This is a step of shifting the flow direction of the fluid as it is.
In the present specification, “the undiluted solution feed port F, the weakly adsorptive fraction outlet A, the eluent feed port D, and the strongly adsorptive fraction outlet C are maintained in their relative positional relationship, and the fluid flow direction is maintained. In the previous step (a), the stock solution supply port F, the weakly adsorptive fraction extraction port A, the eluent supply port D, and the strongly adsorptive property are arranged in order in the flow direction of the fluid. It means that the line of the fraction outlet C is switched to the line of the strong adsorptive fraction outlet C, the stock solution feed port F, the weakly adsorptive fraction outlet A, and the eluent feed port D, respectively.
In other words, the positions of the undiluted solution feed port F, the weakly adsorptive fraction outlet A, the eluent feed port D and the strongly adsorptive fraction outlet C with all the unit packing towers fixed in position By switching
1) A unit packed tower between the undiluted solution feed port F and the weakly adsorptive fraction outlet A in the immediately preceding step (a), between the strongly adsorptive fraction outlet C and the undiluted solution feed port F,
2) A unit packed tower between the weakly adsorptive fraction outlet A and the eluent feed port D in the immediately preceding step (a), between the stock solution feed port F and the weakly adsorptive fraction outlet A,
3) The unit packed tower between the eluent supply port D and the strong adsorptive fraction outlet C in the step (a) immediately before step A, between the weakly adsorptive fraction outlet A and the eluent supply port D ,
4) The unit packed tower between the strongly adsorptive fraction outlet C and the stock solution feed port F in the previous step (a), and between the eluent feed port D and the strongly adsorptive fraction outlet C It means to be in a placed state.
In the form of the circulatory system of FIG. 1, this step (b) is not a step of physically changing the structure of the circulatory system, but a preparatory step for performing step (a) following this step (b) It is.

The step (b) will be specifically described with reference to FIG.
In the step (a) immediately before the step (b), it is assumed that the supply valve F1 is opened to supply the stock solution. In this case, in the step (a), the stock solution supply port F is a connection portion between the stock solution supply branch line 11 a and the pipe 1,
The weakly adsorptive fraction outlet A is a connection site between the weakly adsorptive fraction withdrawal line 2 a and the pipe 1,
The eluent supply port D is a connection portion between the eluent supply branch line 12 c and the pipe 1,
The strongly adsorptive fraction extraction outlet C is a connection site between the strongly adsorptive fraction extraction line 4 c and the pipe 1.
The respective ports F, A, D and C in the step (a) are switched as follows by the step (b). That is,
The undiluted solution supply port F is a connection portion between the undiluted solution supply branch line 11 b and the pipe 1,
The weakly adsorptive fraction extraction outlet A is a connection site between the weakly adsorptive fraction extraction line 2 b and the pipe 1,
The eluent supply port D is a connection site between the eluent supply branch line 12 d and the pipe 1,
The strongly adsorptive fraction outlet C is a connection site between the strongly adsorptive fraction withdrawal line 4 d and the pipe 1.
That is, in the step (a) following immediately after the step (b), the supply of the stock solution in sub-steps (i) and (ii) is performed through the stock solution supply branch line 11b, and the eluent in the sub-step (iii) Feeding is performed through eluent feed branch line 12d, withdrawal of the strongly adsorbed fraction in sub-step (i) is carried out through strongly adsorbed fraction withdrawal line 4d, weak in sub-steps (ii) and (iii) Extraction of the adsorptive fraction will be carried out through the weakly adsorptive fraction extraction line 2b.

  By repeating the above steps (a) and (b) in order, the method of the present invention according to the pseudo movement method is implemented.

By repeating the above steps (a) and (b) sequentially, the amount of eluent used can be greatly reduced as compared with the conventional chromatographic separation by the simulated moving bed method, and adsorption in the strongly adsorptive fraction and weak adsorption can be achieved. The proportion of eluent in the sex fraction can be effectively reduced (ie, the concentration of components other than the eluent in the strongly adsorbed fraction and in the weakly adsorbed fraction can be increased). Moreover, the method of the present invention can also enhance the separation performance of the component to be purified and other components. Therefore, when the component to be purified is a strongly adsorptive component, the strongly adsorbable component can be obtained with high purity and high recovery rate in the strongly adsorptive fraction, and the component to be purified is weakly adsorbed. In the case of the sex component, the weakly adsorptive component can be obtained with high purity and high recovery rate in the weakly adsorptive fraction. That is, according to the method of the present invention, the target component to be purified can be obtained at high concentration, in high purity, and in high recovery rate, in the fraction selected from the strongly adsorptive fraction and the weakly adsorptive fraction. .
In the method of the present invention, the component to be purified is preferably a strong adsorptive component.

  Here, the concept of the simulated moving bed method shown in FIG. 2 is applied to the general conditions of liquid supply and extraction which should be satisfied in order to realize good separation and purification in the chromatographic separation by the simulated moving bed method. Description will be made with reference to the drawings. FIG. 2 shows that the section between the eluent supply port D and the strong adsorptive component outlet C is section 1, the section between the strong adsorptive component outlet C and the stock solution feed port F is the section 2, the stock solution inlet F and weak adsorption The section to the sex component outlet A is shown as section 4. Also, V1 to V4 respectively mean the flow rates of the sections 1 to 4 per one cycle (during which each port D, C, F, A is moved once). Arrows in FIG. 2 indicate the flow direction of the fluid

Focusing on sections 2 and 3, in order to realize the simulated moving bed method, the weakly adsorptive component moves over the length of section 2 at the flow rate V2, and the movement of the strongly adsorptive component is the distance of section 2 It is necessary to keep the amount less than this, and at the flow rate V3, the weakly adsorptive component moves over the length of the section 3 and the movement of the strongly adsorptive component needs to be suppressed less than the distance of the section 3. By doing this, the weakly adsorptive component is further downstream, and the strong adsorptive component is upstream when the respective ports D, C, F, A are moved downstream based on the position of each port D, C, F, A It can be moved relatively (pseudo). That is, the weakly adsorptive component can be extracted from the outlet A, and the strong adsorptive component can be extracted from the outlet C.
On the other hand, focusing on the section 1, if the moving distance of the strong adsorptive component is less than the length of the section 1 at the flow rate V1, the strong adsorptive component (passed to the upstream side without being extracted from the outlet C The strong adsorptive component) can not be extracted from the outlet C. Therefore, in section 1, it is said that the moving distance of the strong adsorptive component needs to move over the length of section 1 at flow rate V1.
Also, focusing on the section 4, when the moving distance of the weakly adsorptive component exceeds the length of the section 4 at the flow rate V4, the weakly adsorptive component (weakly adsorbed component which has passed without being extracted from the outlet A) Component can not be extracted from the outlet A. Therefore, in section 4, it is said that the moving distance of the weakly adsorptive component needs to be less than the length of section 4 at flow rate V4.

Focusing on the relationship between section 3 and section 1 above, it can be understood that it is necessary to set flow rates V3 and V1 to be V3 <V1 (section 1 moves the strong adsorptive component farther) Need to).
Here, V3 = V2 + [supply amount of stock solution], and V1 = V2 + [extraction amount of strongly adsorbed fraction], so
V3 <V1 is V2 + [supply amount of stock solution] <V2 + [extraction amount of strongly adsorbed fraction], that is, [supply amount of stock solution] <[extraction amount of strongly adsorbed fraction].
Therefore, in the simulated moving bed type chromatographic separation, it is usual to make the supply amount of the stock solution smaller than the removal amount of the strongly adsorptive fraction.

On the other hand, in the method of the present invention, it is preferable to make the supply amount of the stock solution larger than the removal amount of the strongly adsorptive fraction. In the step (a), by carrying out the above-mentioned respective sub-steps so that the supply amount of the stock solution is larger than the withdrawal amount of the strongly adsorptive fraction, the component to be purified is further purified It can be obtained in high concentration, in higher purity, and in higher recovery.
In the method of the present invention, the ratio of the supply amount (total supply amount) of the stock solution in step (a) to the extraction amount (total extraction amount) of the strongly adsorptive fraction in step (a) is the volume ratio [Supply amount of stock solution] / [extraction amount of strongly adsorptive fraction] = 1.0 to 2.0 is preferable, and [supply amount of stock solution] / [extraction amount of strongly adsorptive fraction] = 1.1. 2-1.8 are more preferable.

The method of the present invention includes various modifications in addition to the modes specifically described above. For example, between the undiluted solution feed port F and the weakly adsorptive fraction outlet A, between the weakly adsorptive fraction outlet A and the eluent feed port D, the eluent feed port D and the strongly adsorptive fraction outlet A unit packing tower may be disposed one by one between C and between the strongly adsorptive fraction outlet C and the stock solution feed port F, or two or more may be disposed. May be That is, the circulation system preferably has at least four unit packed columns, and more preferably four unit packed columns.
In addition, substeps other than the above substeps (i) to (iv) may be inserted for a short time during the above step (a), as long as the effects of the present invention are not substantially impaired. Such forms are also included in the method of the present invention. For example, when the adsorbent has deteriorated due to long-term use, etc., in substep (ii), the stock solution is supplied from the stock solution supply port F and the weakly adsorptive fraction is extracted from the weakly adsorptive fraction outlet A Separation is improved by incorporating a process (DC process) for supplying the eluent from the eluent supply port D and extracting the strongly adsorptive fraction from the strongly adsorptive fraction outlet C in addition to the process (FA process) It can. When incorporating a DC process into step (ii), it is preferable to carry out the DC process simultaneously with the FA process in step (ii). However, from the viewpoint of effectively reducing the amount of eluent, it is preferable to carry out only the FA step in sub-step (ii).

In the method of the present invention, the adsorbent packed in the unit packed tower is appropriately selected according to the component to be purified, and various adsorbents can be adopted. For example, strong acid cation exchange resin, weak acid cation exchange resin, strong base anion exchange resin, weak base anion exchange resin, synthetic adsorbent, zeolite, and silica gel (preferably octadecylsilyl modified silica gel) are adsorbed It can be used as an agent.
Among them, the method of the present invention is a method suitable for separating and purifying a specific monosaccharide from a stock solution containing a plurality of monosaccharides using a strongly acidic cation exchange resin as an adsorbent. In particular, it is a method suitable as a method for separating and purifying fructose from a stock solution containing glucose and fructose. The strongly acidic cation exchange resin is preferably in the form of calcium, silver, lead or strontium, and more preferably in the form of calcium in consideration of economy and safety. As this calcium type strong acid cation exchange resin, for example, Amberlite (registered trademark) CR-1310Ca, CR-1320Ca (all are manufactured by Organo), DOWEX (registered trademark) Monosphere 99Ca / 310, Monosphere 99Ca / 320 ( In all, Dow Chemical Co., Ltd., Diaion (registered trademark) UBK 535, UBK 555 (all manufactured by Mitsubishi Chemical Co., Ltd.), PCR 642Ca (Purolite Co., Ltd.), CS11 GC, CS 16 GC (all manufactured by Finex Co.) can be mentioned.

  The chromatographic separation system of the present invention is a system for carrying out the method of the present invention. That is, the chromatography separation system of the present invention is a system having the configuration of the circulatory system described above, and the circulatory system sequentially repeats the operation of the step (a) and the operation of the step (b) described above.

  Hereinafter, the present invention will be described in more detail based on examples, but the present invention is not limited to the following examples. Moreover, in the following examples, “%” representing the component composition is on a mass basis unless otherwise noted.

[Preparation of stock solution]
53.49 parts by mass of glucose, 41.81 parts by mass of fructose and 4.70 parts by mass of oligosaccharide were dissolved in deionized water to prepare an aqueous solution with a total sugar concentration of 60%, and used as a stock solution.

[Preparation of eluent]
Ion exchange water was used as the eluent.

Chromatographic separation system
Using the circulating system shown in FIG. 1, fructose was separated and purified from the stock solution. A cylindrical packed tower with an inner diameter of 22 mm and a height of 1.5 m was used as the unit packed towers 10a to 10d. In addition, in the unit packed towers 10a to 10d, 0.57 L of a strongly acidic cation exchange resin (trade name: Monoshere 99Ca / 320, manufactured by Dow Chemical Company) in the form of calcium was packed as an adsorbent. The inside of each unit packed column was adjusted to 60 ° C.

[Example]
The above circulatory system was operated according to the operation steps shown in the following Table 1-1, and the chromatographic separation was performed according to the simulated moving bed system. The time taken for each substep, and the amount of weakly adsorbed fraction (hereinafter referred to as "A fraction") extracted in each substep and the strongly adsorbed fraction (hereinafter referred to as "C fraction") The amount of is shown in Table 1-2 below.
As the conditions shown in Table 1-2, the operation steps shown in Table 1-1 were adopted, and the conditions where the supply amount of the eluent was reduced as much as possible under the condition that the purity of fructose became 90% or more in the C fraction It is.
As shown in Table 1-1 below, when the cycle consisting of the combination of step (a) and the subsequent step (b) is repeated four times from one cycle to four cycles, the positions of the respective supply valves and extraction valves become , Return to the position of step (a) of the first cycle.
Moreover, the numbers described in the column of the valve switching table in Table 1-1 correspond to FIG. For example, the fact that 1, 2, 3 and 4 are described in the column of F means that the stock solution supply valves F1, F2, F3 and F4 are opened, and the numbers are not described. When (empty), it means that the stock solution supply valve is closed. The same applies to the columns of the eluent supply valve D, the weakly adsorptive fraction extraction valve A, the strongly adsorptive fraction extraction valve C and the shutoff valve R.
Moreover, "(circle)" of the column of the circulation pump of Table 1-1 means operating the circulation pump, and when blank, it means that the circulation pump is not working.

  In this chromatographic separation, fructose is used as a component to be purified (strongly adsorptive component), and glucose and an oligosaccharide are combined as a weakly adsorptive component.

The sugar composition of the above-mentioned A fraction and C fraction was analyzed using Toso HPLC 8020 series. The analysis conditions were as follows. The results are shown in Table 1-3 below.
<Analytical conditions>
Dual pump DP-8020
Differential refraction system RI-8020
Auto sampler AS-8020
Column: Tosoh SCX-Ca 7.8 mmφ × 300 mm
Eluent: deionized water Flow rate: 0.8 ml / min
Column temperature: 70 ° C
Sample injection volume: 10 μL

  As shown in the above Table 1-3, in the chromatographic separation of the example, fructose can be recovered as high as 91.2% in the C fraction, and the sugar concentration in the C fraction is It was as high as 40.8%. That is, it was found that fructose in the C fraction can be recovered with high purity and high concentration by the chromatographic separation of the example.

  Subsequently, the recovery rate of the oligosaccharides recovered in the A fraction (the ratio of the oligosaccharides recovered in the A fraction of the oligosaccharides in the stock solution, the same applies hereinafter), in the A fraction Recovery rate of recovered glucose (proportion of glucose recovered in fraction A of glucose in the stock solution, and so on), and recovery rate of fructose recovered in the above-mentioned C fraction (in stock solution) The ratio of fructose recovered in the C fraction among the fructose, the same applies hereinafter) is shown in Table 1-4 below.

  As shown in Table 1-4, in the chromatographic separation of the example, 92.7% of fructose contained in the stock solution could be recovered in the C fraction.

Comparative Example 1
In the examples, except that the circulatory system was operated according to the operation steps shown in Table 2-1 and Table 2-2 below, the chromatographic separation by the simulated moving bed system was performed in the same manner as the examples. This comparative example corresponds to the method described in Example 3 of Japanese Patent Publication No. 60-55162.
The conditions shown in Table 2-2 adopt the operation process shown in Table 2-1, and under the conditions that the purity of fructose is 90% or more in the C fraction, the condition that the supply of the eluent is minimized is there.

  The sugar composition of the A fraction and the C fraction was analyzed in the same manner as the analysis in Example 1 above. The results are shown in Table 2-3 below.

  As shown in the above Table 2-3, in the chromatographic separation of Comparative Example 1, when fructose is recovered with a purity of 90.2% in the C fraction, the sugar concentration in the C fraction is 38.1%. The That is, it was found that fructose in the C fraction had a low purity and the fructose concentration in the C fraction was significantly reduced as compared with the examples.

  Next, the recovery rate of the oligosaccharide recovered in the A fraction, the recovery rate of glucose recovered in the A fraction, and the recovery rate of fructose recovered in the C fraction are shown in Table 2 below. -4.

As shown in Table 2-4, in the chromatographic separation of Comparative Example 1, 92.3% of fructose contained in the stock solution could be recovered in the C fraction. This recovery rate is slightly lower than the example.
Moreover, in Comparative Example 1, the purity of fructose could not be made 90% or more unless the eluent was used more than in the Examples (see “Dv / Fv” in Tables 1-2 and 2-2). See the column of

Comparative Example 2
Comparative Example 1 except that the time spent on each substep, and the amount of the A fraction and the amount of the C fraction extracted in each substep in Comparative Example 1 were changed as shown in Table 3-1 below. In the same manner as in the above, chromatographic separation by the simulated moving bed system was performed.
Under the conditions shown in Table 3-1, the total supply amount of stock solution in sub-steps (ic) to (iiic) is the same as the total supply amount of stock solution in sub-steps (i) to (iii) of the embodiment, and In the steps (ic) to (iiic), the total supplied amount of eluent is the same as the total supplied amount of eluent in sub-steps (i) to (iii) of the example, and the fructose purity in the C fraction is as much as possible. It is a condition set to increase.

  The sugar composition of the A fraction and the C fraction was analyzed in the same manner as the analysis in Example 1 above. The results are shown in Table 3-2 below.

  As shown in the above Table 3-2, in the chromatographic separation of Comparative Example 2, the purity of fructose in the C fraction could not be 90% or more. The sugar concentration in the C fraction was 39.5%. That is, it was found that the purity of fructose in the C fraction was greatly reduced and the fructose concentration in the C fraction was also reduced as compared with the examples.

  Next, the recovery rate of the oligosaccharide recovered in the A fraction, the recovery rate of glucose recovered in the A fraction, and the recovery rate of fructose recovered in the C fraction are shown in Table 3 below. -3.

  As shown in Table 3-3, in the chromatographic separation of Comparative Example 2, only 85.7% of fructose contained in the stock solution could be recovered in the C fraction.

100 Circulating system 10a, 10b, 10c, 10d unit packed tower Ab Adsorbent R1, R2, R3, R4 Shutoff valve 2a, 2b, 2c, 2d weak adsorptive fraction extraction line A1, A2, A3, A4 weak adsorptive Fractional withdrawal valve 4a, 4b, 4c, 4d Strongly adsorptive fractionated line C1, C2, C3, C4 Strongly adsorptive fractionated withdrawal valve T1, T2, T3, T4 Check valve 1 Piping 3 Weak adsorption Extractable fraction combining pipe 5 strong adsorptive fraction combining pipe 6 stock solution tank 7 stock solution 8 eluent liquid tank 9 eluent 11 stock solution supply line 11a, 11b, 11c, 11d stock solution supply branch line F1, F2, F3, F4 stock solution supply valve 12 Eluent Supply Line 12a, 12b, 12c, 12d Eluent Supply Branch Line D1, D2, D3, D4 Eluent Supply Valve P1 Circulation Pump P2 Undiluted Liquid Supply Pump P3 Eluent Supply Pump U, V Leaf valve

Claims (6)

A chromatographic separation method of separating and purifying components in a stock solution by a simulated moving bed method using a circulating system in which a plurality of unit packed towers filled with an adsorbent are connected in series and endlessly via a pipe,
The circulation system includes the undiluted solution feed port F, the weakly adsorptive fraction outlet A, the eluent feed port D, and the strongly adsorptive fraction outlet C in the pipe in this order in the fluid flow direction. Between the undiluted solution feed port F and the weakly adsorptive fraction outlet A, between the weakly adsorptive fraction outlet A and the eluent feed port D, and the eluent feed port D and At least one unit packing tower is disposed between the strongly adsorptive fraction outlet C and between the strongly adsorptive fraction outlet C and the stock solution feed port F,
Repeat the following steps (a) and (b) in sequence:
(A) performing the following sub-steps (i) to (iv) in this order;
(I) A substep of supplying a stock solution from the stock solution supply port F and extracting a strongly adsorptive fraction from the strongly adsorptive fraction outlet C;
(Ii) a substep of supplying a stock solution from the stock solution supply port F and extracting a weakly adsorptive fraction from the weakly adsorptive fraction outlet A;
(Iii) a substep of supplying an eluent from the eluent supply port D and extracting a weakly adsorptive fraction from the weakly adsorptive fraction outlet A;
(Iv) A substep of circulating the fluid in the circulatory system without supplying the stock solution and the eluent and without extracting the strongly adsorptive fraction and the weakly adsorptive fraction,
(B) After the step (a) is completed, the undiluted solution supply port F, the weakly adsorptive fraction outlet A, the eluent feed port D, and the strong adsorptive fraction outlet C are relative to one another. Step of moving in the fluid flow direction while maintaining the positional relationship.   The chromatography separation method according to claim 1, wherein the total amount of the strong adsorptive fraction extracted from the outlet C in the step (a) is smaller than the total amount of the stock solution supplied from the supply port F. .   The chromatographic separation method according to claim 1 or 2, wherein the stock solution contains glucose and fructose, and the fructose is separated and purified in the strongly adsorptive fraction.   The chromatographic separation method according to any one of claims 1 to 3, wherein the circulating system has at least four unit packed towers.   In the step (a), the ratio of the total supply amount of the eluent to the total supply amount of the stock solution satisfies, in a volume ratio, [total supply amount of the eluent] / [total supply amount of the stock solution] <1.2 The chromatographic separation method according to any one of claims 1 to 4. A chromatographic separation system for separating and purifying components in a stock solution by a simulated moving bed method using a circulating system in which a plurality of unit packed towers packed with adsorbents are connected in series and endlessly via a pipe,
The circulation system includes the undiluted solution feed port F, the weakly adsorptive fraction outlet A, the eluent feed port D, and the strongly adsorptive fraction outlet C in the pipe in this order in the fluid flow direction. Between the undiluted solution feed port F and the weakly adsorptive fraction outlet A, between the weakly adsorptive fraction outlet A and the eluent feed port D, and the eluent feed port D and At least one unit packing tower is disposed between the strongly adsorptive fraction outlet C and between the strongly adsorptive fraction outlet C and the stock solution feed port F,
System which repeats the following steps (a) and (b) in order:
(A) performing the following sub-steps (i) to (iv) in this order;
(I) A substep of supplying a stock solution from the stock solution supply port F and extracting a strongly adsorptive fraction from the strongly adsorptive fraction outlet C;
(Ii) a substep of supplying a stock solution from the stock solution supply port F and extracting a weakly adsorptive fraction from the weakly adsorptive fraction outlet A;
(Iii) a substep of supplying an eluent from the eluent supply port D and extracting a weakly adsorptive fraction from the weakly adsorptive fraction outlet A;
(Iv) A substep of circulating the fluid in the circulatory system without supplying the stock solution and the eluent and without extracting the strongly adsorptive fraction and the weakly adsorptive fraction,
(B) After the step (a) is completed, the undiluted solution supply port F, the weakly adsorptive fraction outlet A, the eluent feed port D, and the strong adsorptive fraction outlet C are relative to one another. Step of moving in the fluid flow direction while maintaining the positional relationship.

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