JP5772313B2 - Liquid chromatography apparatus and packing method for packing material - Google Patents

Description

  The present invention relates to a liquid chromatography apparatus, and more particularly to a liquid chromatography apparatus configured to pressurize a packing material or a slurry thereof in a chromatography column with a movable stopper. The present invention also relates to a method for forming a packed bed of packing material in the chromatographic column.

  Liquid chromatography is widely used as a means for separating and purifying target substances in natural products, fermentation products, cultured products by gene recombination and the like, and synthesis reaction products at a target purity and purification rate.

  When filling a chromatographic column (liquid chromatography column) with a filler, after supplying the filler slurry into the chromatographic column, the movable stopper is driven by a driving means to pressurize the slurry, and the chromatograph having a porous plate is provided. The solvent in the slurry is pushed out of the chromatographic column through the column lid or the movable stopper to form a packed bed of packing material. After filling is completed, pressurization of the packed bed by the movable stopper is maintained (Patent Documents 1 and 2).

  In Patent Documents 1 and 2, when supplying the filler slurry into the chromatographic column, the movable stopper is detached from the chromatographic column, and after supplying the filler slurry, the movable stopper is fitted to the chromatographic column, I try to move it forward. In this way, when the movable stopper is positioned outside the chromatographic column and the filler slurry is supplied, there is a high possibility that foreign matter will enter the chromatographic column from the atmosphere.

  As a method of filling the chromatographic column apparatus with the packing material without removing the movable stopper from the column, Patent Document 3 discloses that the movable stopper is moved to a predetermined position of the column and then the valve (or nozzle) at the central part of the movable stopper. ) And injecting filler slurry and pressing with pump discharge pressure, or closing the valve (or nozzle) at the center of the movable stopper and lowering the movable stopper to press the filler. .

  In Patent Document 3, a valve (or nozzle) is provided at the central part of the movable stopper, and the liquid to be treated cannot flow through the movable stopper area corresponding to the valve (or nozzle). Further, if there is a portion where the liquid does not flow on the surface where the liquid to be processed flows into or out of the column, the flow of liquid in the column is unevenly distributed, and the separation and purification efficiency is lowered. In addition, when filling the filler slurry only with the discharge pressure of the pump, it is not easy to make the filling conditions constant because the discharge pressure of the pump varies depending on the state of the pump, for example, the temperature and the degree of deterioration of the drive unit.

JP 2000-346833 A JP-A-2005-332102 JP 4-98156

  An object of the present invention is to provide a liquid chromatography apparatus capable of filling a column with a packing material without providing a valve or nozzle for packing the packing material on a movable stopper or a flange, and a packing method for the packing material. To do.

A liquid chromatography apparatus according to a first aspect of the present invention is a liquid having a chromatographic column arranged with the column axial direction being the vertical direction, and a movable stopper for pressurizing the packing material or slurry supplied to the chromatographic column. In the chromatography apparatus, the movable stopper is provided with an upper movable stopper and a lower movable stopper, and liquid passing means are provided in the upper movable stopper and the lower movable stopper, respectively, at the lower part of the side surface of the chromatography column. A filler slurry inlet is provided , and the lower movable stopper is disposed below the filler slurry inlet, and the upper movable stopper is moved upward to fill from the filler slurry inlet. The agent slurry is configured to be drawn into the chromatographic column .

  The packing method of the packing material of the second invention is a method of filling the liquid chromatography device of the first invention with a packing material, wherein the lower movable stopper and the upper movable stopper are located at the lower part of the chromatography column, At this time, the lower movable stopper is disposed below the slurry inlet, the upper movable stopper is moved upward to supply the filler slurry into the chromatographic column from the slurry inlet, After the lower movable plug is positioned higher than the slurry introduction port, the upper movable plug is lowered or the lower movable plug is raised to form the packed bed.

A liquid chromatography apparatus according to a third aspect of the present invention is a liquid having a chromatographic column arranged with the column axial direction being the vertical direction, and a movable stopper for pressurizing the packing material or its slurry supplied into the chromatographic column. The chromatography apparatus includes an upper movable stopper and a lower movable stopper as the movable stoppers, and liquid passing means are provided in the upper movable stopper and the lower movable stopper, respectively, and the upper part of the side surface of the chromatography column. A filler slurry introduction port is provided, a filler slurry discharge port is provided at the bottom, the upper movable plug is disposed higher than the filler slurry introduction port, and the lower movable plug is directed downward der which filler slurry from said filler slurry inlet by moving is characterized in that it is configured to be pulled into the chromatographic column to .

  The packing method of the packing material of the fourth invention is a method of filling the liquid chromatography apparatus of the third invention with a packing material, wherein the lower movable stopper and the upper movable stopper are positioned above the chromatography column, At this time, the upper movable stopper is disposed above the slurry inlet, and the lower movable stopper is moved downward to supply the filler slurry into the chromatography column from the slurry inlet, After the upper movable plug is positioned lower than the slurry inlet, the upper movable plug is lowered or the lower movable plug is raised to form a packed bed.

  The chromatographic column apparatus of the present invention includes an upper movable stopper and a lower movable stopper as movable stoppers, and a filler slurry inlet on the side of the column. According to this liquid chromatography apparatus and the method of filling the liquid chromatography apparatus with the packing material, the packing material slurry can be packed into the column without completely opening the column interior to the atmosphere outside the column. According to this liquid chromatography apparatus, the target product can be efficiently separated and purified by uniformly dispersing it in the filler layer without causing a drift in the flow of the liquid to be treated. In addition, without completely opening the column interior to the atmosphere outside the column, the once-pressed packing material can be reslurried and easily pulled out of the column.

  According to the present invention, since the valve or nozzle for filling the filler is not installed on the movable stopper or the flange, the liquid to be treated can be uniformly dispersed in the filler layer in the column, and the separation and purification rate is not impaired.

  In the present invention, a pump for introducing the filler slurry into the column is unnecessary, the apparatus system can be simplified and the space can be saved, and the equipment cost can be reduced.

It is a longitudinal section of the liquid chromatography device concerning an embodiment. It is explanatory drawing which shows the formation method of a filled layer. It is explanatory drawing which shows the formation method of a filled layer. It is explanatory drawing which shows the discharge method of a filler.

  Hereinafter, embodiments will be described with reference to the drawings. FIG. 1 shows a liquid chromatography apparatus 1 according to an embodiment. The liquid chromatography apparatus 1 includes a chromatographic column 2, an upper movable plug 3, a lower movable plug 4, and a cylinder device for moving the movable plugs 3 and 4 forward and backward (except for the piston rods 3d and 4d). Etc. The chromatographic column 2 is installed with the cylinder axis direction as the vertical direction (vertical direction in this embodiment). A filler slurry inlet 2 a is provided on the lower side surface of the chromatographic column 2. The filler slurry is a slurry in which a filler is dispersed in a solvent. The solvent may be water or an organic solvent.

  A liquid-permeable porous plate 3a that does not allow a filler to pass therethrough is provided on the lower surface of the movable plug 3, and a flow path 3b is formed above the porous plate 3a. The flow path 3b communicates with the outside of the upper movable plug 3 through a liquid passage 3c.

  On the upper surface of the lower movable stopper 4, a liquid-permeable porous plate 4a that does not allow the filler to pass through but allows the solvent to pass therethrough is provided, and a flow path 4b is formed below the porous plate 4a. The flow path 4b communicates with the outside of the lower movable plug 4 through a liquid passage 4c.

  The cylinder device includes a cylinder, a piston (not shown) in the cylinder, a piston rod 3d or 4d connected to the piston, and the like. The lower end of the piston rod 3d is connected to the upper movable plug 3, and the upper end of the piston rod 4d is connected to the lower movable plug 4.

  Next, a method of filling the filler into the liquid chromatography apparatus 1 having the above configuration will be described with reference to FIG. 2A to 2E are longitudinal sectional views showing the liquid chromatography apparatus of FIG. 1 in a simplified manner.

  In this filling method, first, as shown in FIG. 2 (a), the lower movable plug 4 is made equal to or lower than the lower edge of the slurry inlet 2a, and the upper movable plug 3 is brought into contact with the lower movable plug 4. . In this state, a slurry tank (not shown) containing a predetermined concentration of filler slurry in which a filler is dispersed in a filling solvent is connected to the introduction port 2a, or a valve of a pipe connected to the slurry tank is connected. The upper movable plug 3 is opened and moved upward (FIG. 2B). As a result, the filler slurry is drawn into the column 2. The upper movable stopper 3 is moved upward in the column until a predetermined amount of the filler slurry is filled in the column 2 (FIG. 2 (c)).

  Thereafter, the lower movable stopper 4 is moved up or the column 2 is moved downward until the lower movable stopper 4 comes above the slurry inlet 2a (FIG. 2 (d)). Thereafter, the upper movable plug 3 is moved downward by a necessary amount to compress and fix the packed bed (FIG. 2 (e)). As a result, the filler slurry can be introduced into the column without using a filler slurry transfer pump, and can be pressed with reproducibility to an arbitrary compression rate by the movable stopper, and the treated liquid can be applied to the packed layer after filling. Can be made to flow evenly, and thus efficient separation and purification is possible.

  The filler can be extracted from the filler-filled column 2 by reversing the filling operation. That is, the upper movable plug 3 is moved upward in a state in which the solvent (filler slurry liquid) is adjusted to be sucked from the liquid passage 4 c of the lower movable plug 4. As a result, the upper movable plug 3 moves upward while the solvent is sucked in through the liquid passing pipe 4c (FIG. 2D). At this time, the packed bed that has been compressed loosens and becomes a slurry. Next, the lower movable stopper 4 is moved downward from the slurry inlet 2a, or the column 2 is moved upward to communicate the slurry inlet 2a and the column 2 (FIG. 2C). Next, while moving the upper movable stopper 3 downward, the filler slurry in the column 2 is pushed out of the column through the slurry inlet 2a (FIG. 2B). The upper movable plug 3 is lowered until it comes into close contact with the lower movable plug 4 (FIG. 2A). During this time, the solvent may continue to flow, or the supply of the solvent may be temporarily stopped and the column 2 may be flushed with the solvent immediately before the upper movable plug 3 and the lower movable plug 4 come into close contact with each other.

  Another embodiment is shown in FIG. In this embodiment, a slurry introduction port 2 a is provided at the upper part of the side surface of the column 2, and a slurry discharge port 2 b is provided at the lower part of the side surface of the column 2. Other configurations are the same as those in FIGS. 1 and 2, and the same reference numerals denote the same parts.

  In FIG. 3, in order to fill the column 2 with the packing material, the upper movable plug 3 and the lower movable plug 4 are positioned above the column 2 as shown in FIG. At this time, the upper movable plug 3 is raised above the slurry introduction port 2 a, and the lower movable plug 4 is raised until it contacts the upper movable plug 3. Next, the slurry inlet 2a is connected to a slurry tank or the like, or the valve of the pipe connected to the slurry tank is opened, and the lower movable plug 4 is moved downward as shown in FIG. Is introduced into the column 2. The lower column 4 is moved downward until a predetermined amount of the filler slurry is filled into the column 2 (FIG. 3C). At this time, the lower movable plug 4 is not lowered below the slurry discharge port 2b. Thereafter, as shown in FIG. 3D, the upper movable plug 3 is moved down or the column 2 is moved up until the upper movable plug 3 comes below the slurry inlet 2a. Finally, the upper movable plug 3 is moved downward by a necessary amount to compress and fix the packed bed (FIG. 3E).

  In this way, without using a slurry transfer pump, the filler slurry can be introduced into the column and pressed to a desired compression rate with a movable stopper with good reproducibility. Can be made to flow evenly, and thus efficient separation and purification is possible.

  3A to 3E, the packing material can be extracted from the column 2 by reversing the packing operation.

  Further, the filler can be extracted from the column 2 as shown in FIG. 4 from the filler-packed column 2 in the state of FIG. 4A, the upper movable plug 3 is moved upward as shown in FIG. 4B and stopped below the slurry inlet 2a. At this time, it is adjusted so that the solvent can be sucked from the liquid passage 4c of the lower movable stopper 4, and is sucked into the column 2 by the upward movement of the upper movable stopper 3, thereby slurrying the packed bed (see FIG. 4 (b)). Next, as shown in FIG. 4 (c), the lower movable stopper 4 is moved downward, or the column 2 is moved upward so that the lower movable stopper 4 is positioned below the slurry discharge port 2b. The slurry discharge port 2b and the column 2 are communicated. Next, as shown in FIG. 4D, the filler slurry in the column 2 is pushed out of the column 2 through the slurry discharge port 2b while moving the upper movable plug 3 downward. As shown in FIG. 4E, the entire amount of slurry in the column 2 is pushed out of the column 2 by lowering the upper movable plug 3 until it comes into contact with the lower movable plug 4. During this time, the solvent may continue to flow, or the solvent supply may be temporarily stopped and flushed immediately before the upper movable plug 3 and the lower movable plug 4 come into contact with each other.

  In the above embodiment, the chromatographic column 2 is illustrated as being installed vertically, but the chromatographic column 2 may be slightly inclined (for example, about several degrees to several tens of degrees with respect to the vertical line).

  In the above embodiment, the upper movable plug is lowered when pressurizing the packed bed, but the lower movable plug may be raised. Further, the upper movable stopper may be lowered and the lower movable stopper may be raised.

DESCRIPTION OF SYMBOLS 1 Liquid chromatography apparatus 2 Chromatography column 3 Upper movable stopper 4 Lower movable stopper

Claims (4)

In a liquid chromatography apparatus having a chromatographic column arranged with the column axial direction being the vertical direction, and a movable stopper for pressurizing the packing material or slurry supplied in the chromatographic column,
The movable stopper includes an upper movable stopper and a lower movable stopper,
The upper movable stopper and the lower movable stopper are respectively provided with liquid passing means,
An inlet for the filler slurry is provided at the bottom of the side surface of the chromatographic column ,
The lower movable stopper is arranged below the filler slurry inlet, and the upper slurry stopper is moved upward to draw the filler slurry into the chromatographic column from the filler slurry inlet. The liquid chromatography apparatus characterized by the above-mentioned. A liquid chromatography apparatus according to claim 1, wherein the liquid chromatography apparatus is filled with a filler.
The lower movable stopper and the upper movable stopper are positioned at the lower part of the chromatographic column. At this time, the lower movable stopper is disposed below the slurry inlet, and the upper movable stopper is moved upward to move the upper movable stopper. The filler slurry is supplied into the chromatographic column from the slurry inlet, and then the lower movable stopper is positioned higher than the slurry inlet, and then the upper movable stopper is lowered or the lower movable stopper is raised. And a filling layer is formed. In a liquid chromatography apparatus having a chromatographic column arranged with the column axial direction being the vertical direction, and a movable stopper for pressurizing the packing material or slurry supplied in the chromatographic column,
The movable stopper includes an upper movable stopper and a lower movable stopper,
The upper movable stopper and the lower movable stopper are respectively provided with liquid passing means,
An inlet for the filler slurry is provided at the upper part of the side surface of the chromatographic column, and an outlet for the filler slurry is provided at the lower part .
The upper movable stopper is arranged above the filler slurry inlet, and the lower movable stopper is moved downward so that the filler slurry is drawn into the chromatographic column from the filler slurry inlet. liquid chromatography apparatus characterized by being configured. A method for filling a liquid chromatography apparatus according to claim 3 with a filler,
The lower movable stopper and the upper movable stopper are positioned at the upper part of the chromatographic column. At this time, the upper movable stopper is disposed higher than the slurry inlet, and the lower movable stopper is moved downward to move the lower movable stopper. After the filler slurry is supplied into the chromatographic column from the slurry inlet, the upper movable stopper is positioned below the slurry inlet, and then the upper movable stopper is lowered or the lower movable stopper is raised. A filling method of a filler characterized by forming a packed bed.

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