JP2012159462A - Liquid chromatography device and packing method of packing agent - Google Patents

Abstract

[PROBLEMS] To prevent foreign substances from entering when a filler slurry is supplied to a chromatographic column to form a packed bed of the filler, and to smoothly introduce the filler slurry into the chromatographic column without using a slurry pump. A possible liquid chromatography apparatus and a packing method for a packing material are provided.
In order to fill the packing material, the movable stopper 3 is lowered to the lower limit facing the bottom lid 6 in the chromatographic column 2. The valves 9a and 20 are closed. Next, the valve 24 is opened, the valve rod 17 is pushed up, the valve body 16 is raised, and the supply port 14 is opened. Then, the movable stopper 3 is raised and the slurry S is sucked into the chromatographic column 2. Next, the valve stem 17 is pulled down to seat the valve body 16 on the flange 12a, and the supply port 14 is closed. Next, the piston rod 36 of the cylinder device 4 is projected to lower the movable plug 3 and pressurize the filler.
[Selection] Figure 1

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).

JP 2000-346833 A JP-A-2005-332102

  In Patent Document 1, 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 and moved forward. I am doing so. 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.

  In Patent Document 2, the filler slurry supply pipe is advanced and retracted into the chromatographic column, and a drive mechanism for this purpose is required.

  In both Patent Documents 1 and 2, a pump for supplying the filler slurry into the chromatographic column is required. When a slurry pump is used, the equipment cost increases accordingly. Further, when the filler slurry passes through the slurry pump, the filler may be deformed or damaged.

  The present invention prevents the introduction of foreign matters when the filler slurry is supplied to the chromatographic column to form the packed bed of the filler, and allows the filler slurry to smoothly enter the chromatographic column without using a slurry pump. It is an object of the present invention to provide a liquid chromatography apparatus that can be introduced and a packing method for a packing material.

  The liquid chromatography apparatus according to claim 1, wherein the liquid chromatography apparatus includes a chromatographic column provided with a lid on one end side, and a movable stopper movable within the chromatography column. One side is provided with a slurry supply path for allowing the filler slurry to flow into the chromatographic column, and is seated from the inside of the chromatographic column to the supply port at the end of the slurry supply path facing the chromatographic column, and the supply port is closed. A possible valve body is provided.

  A liquid chromatography apparatus according to a second aspect is the liquid chromatography apparatus according to the first aspect, wherein a cleaning fluid supply path for supplying a cleaning fluid to the vicinity of the supply port of the slurry supply path and cleaning the slurry supply path is provided. It is characterized by.

  The liquid chromatography device according to claim 3 is the liquid chromatography device according to claim 2, wherein the valve body is provided at a distal end of a tubular valve stem, and the cleaning fluid supply path is washed from the distal end portion of the valve stem through the valve stem. It is provided so that a fluid may be supplied to the slurry supply path.

  A method for packing a packing material into a chromatography column according to claim 4 is a method for packing a packing material into the chromatography column of the liquid chromatography device according to any one of claims 1 to 3, wherein the packing material slurry is chromatographed. When introducing into the column, a slurry tank is communicated with the slurry supply path, the movable stopper is disposed on the one end side of the chromatographic column, and the movable stopper is connected to the other end in a state where the valve body is separated. The filler slurry is sucked and introduced into the chromatographic column, and then the valve element is seated and the movable stopper is advanced.

  According to a fifth aspect of the present invention, there is provided the method for filling the chromatographic column with the packing material according to the fourth aspect, wherein after the valve body is seated, a cleaning fluid is introduced in the vicinity of the supply port of the slurry supply path to wash the slurry supply path. It is characterized by doing.

  In the liquid chromatography apparatus and packing method of the present invention, when the packing slurry is introduced into the chromatography column, the slurry tank and the slurry supply path are communicated with each other, and the movable stopper is connected to the lid portion on one end side of the chromatography column. Keep it in the opposite position. The movable stopper is moved toward the other end while the valve body is separated and the supply port is opened, and the slurry in the slurry tank is sucked into the chromatographic column from the slurry supply path. Thereafter, the valve body is seated, the supply port is closed, the movable stopper is advanced, the liquid content in the slurry is squeezed out, and the filler is pressurized to form a packed layer of the filler. Thus, the filler slurry can be introduced into the chromatography column without using a slurry pump. In addition, since the movable stopper is located in the chromatographic column during the period from the introduction of the filler slurry to the formation of the packed bed, foreign matters are prevented from entering the chromatographic column.

  The present invention does not require a filler slurry supply pipe that advances and retreats in the chromatographic column and its advance and retreat drive mechanism, and the equipment cost is low.

  In the present invention, since the valve body is seated from the inside of the chromatographic column with respect to the slurry supply port, the valve body is pressed so as to be pressed against the movable plug from the slurry or filler when the movable plug is advanced, and the supply port is sealed. Is done.

  In the present invention, after the filler slurry is introduced into the chromatographic column, the valve body is seated to close the supply port, and then the cleaning fluid is supplied to the vicinity of the slurry supply port to clean the slurry supply path. . For example, the filler slurry remaining in the slurry supply path can be pushed out to the slurry tank side.

  In one embodiment of the present invention, the valve stem is tubular, and the cleaning fluid supply path is provided in the valve stem. As a result, the valve stem is also used as the conduit portion of the fluid supply passage, and the device configuration is simplified.

It is sectional drawing of the liquid chromatography apparatus which concerns on embodiment. FIG. 2 is an enlarged view of a part of the liquid chromatography apparatus of FIG. 1. 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.

  Hereinafter, embodiments will be described with reference to the drawings. 1 to 3 show a liquid chromatography apparatus 1 according to the first embodiment, in which a chromatographic column 2 is installed with the cylinder axis direction as the vertical direction (vertical direction in this embodiment). Yes.

  The liquid chromatography apparatus 1 includes a chromatographic column 2, a movable plug 3 that can move up and down in the chromatographic column 2, and a cylinder device 4 for moving the movable plug 3 back and forth (down and up) (FIG. 3). ) Etc. The chromatographic column 2 includes a cylindrical column main body 5 and a bottom lid 6 as a lid attached to the lower end of the column main body 5 in a detachable manner. A liquid-permeable porous plate 7 that does not allow the filler to pass therethrough is provided on the upper surface of the bottom lid 6, and a flow path 8 formed on the lower side of the porous plate 7 leads to the liquid outlet 9. The liquid is taken out from the liquid outlet 9 through the valve 9a and the extraction line 9b. The bottom lid 6 is connected to the column body 5 by bolts 10.

  A short tube 11 protrudes downward from the bottom lid 6. As clearly shown in FIG. 2, a female screw is engraved on the upper inner peripheral surface of the tube body 11, and a male screw on the lower outer peripheral surface of the joint tube 12 is screwed into the female screw. The joint pipe 12 has a pipe axis direction as the vertical direction, and extends to the upper side of the porous plate 7 through an opening 13 provided in the porous plate 7. A flange 12 a is provided at the upper end of the joint pipe 12, and the flange 12 a overlaps the upper surface of the edge of the opening 13 of the perforated plate 7 via a seal packing (not shown). The upper end portion of the joint pipe 12 is a slurry supply port 14. A seal ring 15 surrounding the joint pipe 12 is interposed between the perforated plate 7 and the bottom surface of the flow path 8.

  A valve body 16 is disposed on the upper side of the porous plate 7 so as to be seated on the upper surface of the flange 12a. A sealing member (not shown) such as an O-ring concentric with the valve body 16 is provided on the lower surface of the valve body 16 so as to surround the slurry supply port 14.

  The valve body 16 is attached to the upper end of the valve rod 17. The valve rod 17 extends through the joint pipe 12 and the axial center of the pipe body 11 to the lower side of the pipe body 11. The tube 11 is tubular, and a solvent outlet 17a is provided at the upper end thereof.

  As shown in FIG. 1, a solvent supply hose 19 is connected to the lower end of the valve rod 17 via a joint 18. The hose 19 can be supplied with a solvent from a solvent tank (not shown) via a valve 20.

  The valve rod 17 and the inner peripheral surface of the tube body 11 are sealed in a watertight manner by a sliding packing 21. The sliding packing 21 can slide along the inner peripheral surface of the tube body 11.

  The pipe 11 has a branch pipe 23 branched from a position higher than the sliding packing 21, and the branch pipe 23 is connected to the bottom of the slurry tank 26 via a valve 24 and a pipe 25 as shown in FIG. 3. It is connected to the.

  The slurry tank 26 is provided with a stirrer 27 for stirring the slurry S. The slurry tank 26 is preferably arranged at substantially the same level as the chromatographic column 2.

  A liquid-permeable porous plate 30 that does not allow the filler to pass therethrough is provided on the lower surface of the movable plug 3, and a flow path 31 is formed above the porous plate 30. The flow path 31 communicates with the outside of the movable plug 3 via the liquid flow path 32.

  The cylinder device 4 includes a cylinder, a piston in the cylinder, a piston rod 36 connected to the piston, and the like. The lower end of the piston rod 36 is detachably connected to the movable plug 3 via a connector 37.

  Next, a filling method of the filler into the liquid chromatography apparatus 1 having the above configuration will be described with reference to FIG. FIG. 3 is a schematic cross-sectional view showing the filling method of the filler in the order of the steps.

  In this packing method, first, as shown in FIG. 3 (a), the movable stopper 3 is lowered to the lower limit facing the bottom lid 6 in the chromatographic column 2. The valves 9a and 20 are closed. Next, the valve 24 is opened, the valve rod 17 is pushed up manually or the like to raise the valve body 16 as shown in FIG. 2A, and the slurry supply port 14 is opened. And the movable stopper 3 is raised. Thereby, as shown in FIG. 3B, the slurry S in the slurry tank 26 is sucked into the chromatographic column 2 and introduced. After raising the movable plug 3 to a specified height, as shown in FIG. 3C, the valve rod 17 is lowered to seat the valve body 16 on the flange 12a, and the slurry supply port 14 is closed.

  Next, the valve 9a is opened, the piston rod 36 of the cylinder device 4 is projected, the movable stopper 3 is lowered, and the filling layer of the filler is pressurized. At this time, piping (not shown in FIG. 3) is connected to the liquid flow path 32, and the slurry solvent squeezed from the packed bed to the movable plug 3 side is extracted. The solvent squeezed out to the bottom lid 6 side flows out from the liquid outlet 9. The packed bed is formed by pressurizing the packed bed to a predetermined pressure by the movable stopper 3.

  After the valve 16 is closed, the valve 20 is opened as shown in FIG. 3 (c), and the solvent is allowed to flow from the valve rod 17 and the outlet 17a into the tube 11, the branch pipe 23, the valve 24, and the pipe 25. The remaining filler slurry is pushed back into the slurry tank 26. Thereafter, the valves 20 and 24 are closed. In this case, the same solvent as that of the slurry S is used as the solvent.

  The sample solution and the developing solution are passed through the packed bed from the movable stopper 3 side through the liquid flow path 32 to the chromatographic column 2 on which the packed bed is formed, and the fractionated fraction is separated from the liquid outlet 9. To take.

  According to this embodiment, the slurry in the slurry tank 26 can be introduced into the chromatographic column 2 without using a slurry pump, and the slurry pump becomes unnecessary and the equipment cost is reduced. Further, deformation and damage of the filler particles due to passing through the slurry pump are prevented.

  In this embodiment, after the slurry is introduced, the slurry supply path (the tube 11, the branch pipe 23, the valve 24, and the pipe 25) is washed with a solvent, so that the slurry supply path is not blocked and the filler particles are prevented from sticking. The In this embodiment, since the cleaning fluid (solvent) flows out from the outlet 17a immediately below the valve body 16, the entire slurry supply path including the lower surface of the valve body 16 is cleaned with the solvent.

  In the above embodiment, the slurry supply path is provided on the bottom lid 6 side. However, as shown in FIG. 4, the slurry supply path may be provided on the movable plug 3 side. In this case, the movable plug 3 is provided with a slurry supply path that is upside down in FIG. That is, although detailed illustration is omitted, the joint pipe 12 is provided through the perforated plate 30 of the movable plug 3, and the tube body 11 protrudes upward from the upper surface of the movable plug 30. This tubular body 11 is connected to a slurry tank 26 via a branch pipe 23, a valve 24 and a pipe 25. The seal packing 15 surrounding the joint pipe 12 is disposed between the upper surface of the perforated plate 30 and the upper surface of the flow path 31.

  The slurry filling procedure and the subsequent washing procedure with the solvent in the slurry supply path are the same as in FIG. That is, as shown in FIG. 4 (a), the valves 9a and 20 are closed and the movable plug 3 is lowered to the lower limit. As shown in FIG. 4B, the movable plug 3 is raised with the valve 24 opened, the valve body 16 separated, and the slurry supply port 14 opened. Thereby, the slurry S in the slurry tank 26 is sucked and introduced into the chromatographic column 2.

  After raising the movable plug 3 to a specified height, as shown in FIG. 4 (c), the valve body 16 is seated on the flange 12a, and the slurry supply port 14 is closed. Next, the valve 9a is opened, the piston rod 36 of the cylinder device 4 is projected, the movable stopper 3 is lowered, and the filling layer of the filler is pressurized. At this time, piping (not shown in FIG. 4) is connected to the liquid flow path 32, and the slurry solvent squeezed from the packed bed to the movable plug 3 side is extracted. The solvent squeezed out to the bottom lid 6 side flows out from the liquid outlet 9. The packed bed is formed by pressurizing the packed bed to a predetermined pressure by the movable stopper 3.

  After the valve 16 is closed, the valve 20 is opened as shown in FIG. 4C, and the solvent is allowed to flow from the valve rod 17 and the outlet 17a into the tube 11, the branch pipe 23, the valve 24, and the pipe 25. The remaining filler slurry is pushed back into the slurry tank 26. Thereafter, the valves 20 and 24 are closed.

  The above embodiment is an example of the present invention, and the present invention may have a configuration other than the above. For example, in the above embodiment, the movable stopper 3 is on the upper side and the bottom lid 6 is on the lower side, but conversely, the movable stopper 3 may be on the lower side and the lid may be provided on the top of the column. In addition, the cleaning drainage of the slurry supply path may be discharged out of the system. In this case, the cleaning fluid may be different from the slurry solvent.

DESCRIPTION OF SYMBOLS 1 Liquid chromatography apparatus 2 Chromatography column 3 Movable stopper 5 Column main body 11 Tubing body 12 Joint pipe 14 Supply port 16 Valve body 17 Valve rod 17a Outflow port

Claims (5)

In a liquid chromatography apparatus having a chromatography column provided with a lid on one end side and a movable stopper movable within the chromatography column,
A slurry supply path for allowing the filler slurry to flow into the chromatographic column is provided on one of the lid and the movable stopper,
A liquid chromatography apparatus comprising a valve body that is seated from the inside of a chromatographic column and can close the supply port at a supply port at the end of the slurry supply path facing the chromatographic column.   2. The liquid chromatography apparatus according to claim 1, wherein a cleaning fluid supply path for supplying a cleaning fluid to the vicinity of the supply port of the slurry supply path to clean the slurry supply path is provided. In claim 2, the valve body is provided at the tip of a tubular valve stem,
The liquid chromatography apparatus according to claim 1, wherein the cleaning fluid supply path is provided so as to supply a cleaning fluid to the slurry supply path from the tip of the valve rod through the valve rod. A method for filling a chromatographic column of a liquid chromatography apparatus according to any one of claims 1 to 3, with a filler,
In introducing the filler slurry into the chromatography column, a slurry tank is connected to the slurry supply path,
The movable stopper is arranged on the one end side of the chromatography column,
With the valve body in a separated state, the movable stopper is retracted to the other end side to suck the filler slurry and introduce it into the chromatographic column,
After that, the valve body is seated and the movable stopper is advanced, and the method for filling the chromatographic column with the packing material is provided.   5. The method of filling a chromatographic column with a packing material according to claim 4, wherein after the valve body is seated, a cleaning fluid is introduced in the vicinity of the supply port of the slurry supply path to wash the slurry supply path. .

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