JPH02134562A - Manufacture of separation column for liquid chromatograph - Google Patents

Abstract

PURPOSE:To make it possible to fill a filling column having the different diameter and to make it possible to make the filling column long to a large extent by driving a movable plug in the up and down direction with a driving mechanism, and using the same moving column or movable plug. CONSTITUTION:A speed reducing mechanism 4 which is rotated and driven with a motor 5 is attached on a fixed stage 1. Threaded rotary shafts 3 which are rotated at a low speed with said speed reducing mechanism 4 are attached so that the shafts are made to extend upward vertically with respect to the fixed stage 1. A movable plug attaching stage 6 is screwed to the rotary shafts 3. The upper end of a movable plug shaft 7 is attached to the movable plug attaching stage 6. A part of the shaft 7 is engaged with a supporting post 18 which is in parallel with said rotary shafts 3 and whose base plate is fixed to the fixed stage 1 in order to prevent the turning with the rotary shafts 3 so that said part can be moved freely only in the up and down direction. In order to fill a filling column 14 having the same diameter with a filler by using said apparatus, a movable plug 9 is lifted, and the insides of a moving column 12 and an adapter 13 for a pipe having the different diameter are cleaned. Thereafter, the filling column 14 is attached. The same processure is repeated. When the filling column 14 having the different diameter is filled with the filler, the adapter 13 for the pipe having the different diameter and the filling column 14 are replaced.

Description

DETAILED DESCRIPTION OF THE INVENTION [Field of Industrial Application] The present invention relates to a method for manufacturing a separation column, which is a main part of a liquid chromatograph.

[Conventional technology]

The column, which is the main component of a liquid chromatograph, is mostly a backed column (separation column) in which a packing material is packed in advance, and the type of packing material, diameter, etc.
Several types of backed columns with different lengths are prepared and used depending on the purpose.

Generally, to manufacture the above-mentioned backed column, an aerodynamic ram called a backer is connected on top of the separation aerodynamic ram (packed column), a slurry of packing material is injected into it, the lid is closed, and then the filling liquid is pumped into the backer. A slurry method has been used in which the liquid is fed at a high flow rate and the packing material is transferred to a separation column and packed tightly. However, recently, separation and purification technology using liquid chromatography has been used to separate large amounts of biochemical substances and pharmaceuticals.
Separation columns, which are equipped with a column and a packing material, have come to be used to provide optimal functionality for separating target substances. In order to improve the separation efficiency of the above-mentioned separation columns, separation columns for liquid chromatography are manufactured using the pressure packing method, but at this time, the packing material is uniformly packed into the column in the circumferential and axial directions of the column. It is necessary that there be no Further, since the pressure to be applied differs depending on the type of filler, it is necessary to select an appropriate pressure for each type of filler before filling.

In the pressurized filling method, the filler is normally filled under pressure using the pressure of a fluid such as oil or gas, but the present inventors have previously disclosed Japanese Patent Application Laid-Open No. 61-7466 and No. 61-61. No. 28864, No. 6:3-62506, and No. 63-134
Publication No. 952 proposed a method in which a movable stopper is driven using an electric motor as a power source and a mechanical transmission mechanism, and a filler filled in a column is compressed under pressure by the movable stopper.

[Problem to be solved by the invention]

However, in the above-mentioned method of pressurizing the packing material using liquid pressure, in order to pack the packing material densely, the liquid must be delivered at a high flow rate and the packing state must be stabilized, so the volume of the separation column must be reduced. It is necessary to flow several times the flow rate. Therefore, as the volume of the separation column increases, the flow rate increases.
A pump with a high discharge pressure and a large amount of filling liquid were required, making it difficult to manufacture a large-volume, high-performance separation column, and causing problems such as high filling costs.

In addition, the method of pressurizing using a movable stopper proposed earlier by the present inventors uses a packed column connected to a moving column of the same diameter that is longer than the packed column, so when producing a long separation column, The equipment must be more than twice the height, the length of the packed column used is restricted, and each time the diameter of the packed column changes, the moving column and movable stopper must be replaced, etc. There were drawbacks.

In view of the above circumstances, the present invention uses the same moving column and movable stopper when the movable stopper is driven in the vertical direction by the drive mechanism and the filler is pressurized and filled into the packed column.
It is an object of the present invention to provide a method for manufacturing a separation column for liquid chromatography, which enables packing into packed columns of different diameters and allows the length of the packed column to be packed to be significantly increased.

[Means to solve the problem]

In order to achieve the above object, in the method of the present invention,
One opening of the cylindrical moving column and one opening of the cylindrical packed column having an inner diameter smaller than the inner diameter of the moving column are made to coincide with their respective central axes using a removable cone-shaped different diameter pipe adapter. connected in series,
The other opening of the connected packed columns is closed by a removable closing plate having a filter on the inner surface and a capillary tube communicating with the inner surface on the outer surface, and the central axis of the connected columns is vertical. The moving column is attached to a fixed base so that the filler slurry is introduced from the other opening of the moving column, and a filter is provided on the inner surface, a movable stopper shaft is provided in the center of the outer surface, and the filler slurry is introduced into the inner surface through the above-mentioned filter. A piston-shaped movable stopper, to which a capillary tube communicating with the piston is attached, is fitted into the movable column, and the movable stopper is lowered via the movable stopper shaft by a drive mechanism, so that the packing material in the packed column is at a predetermined pressure. After pressing it so that It is closed by a removable closing plate provided with.

[Effect]

Since the present invention has the above-mentioned configuration, the same moving force ram can be used to accommodate packed columns of various diameters by exchanging different diameter adapters. Furthermore, since the moving column has a larger diameter than the packed column, the length is shorter and the capacity can be increased.

Therefore, not only can a long packed column be filled without increasing the height of the packing equipment, but also the slurry concentration range of the usable packing material is wide (and by selecting the slurry concentration conditions, high performance can be achieved. It becomes possible to manufacture packed columns of

The different diameter pipe adapters are cone-shaped and connected so that they share the same center axis as the upper and lower columns, so the pressurizing force from the movable plug in the moving column is easily transmitted to the packed column, and the movable plug Since the applied pressure can be freely selected from a relatively low pressure to a high pressure range, the filling pressure in the packed column can be freely adjusted.

〔Example〕

FIG. 1 shows an example of an apparatus for carrying out the method of the present invention, in which reference numeral 1 denotes a horizontal fixed base having a column attachment part 2 in the center. On the upper surface of the fixed base 1, two rotary shafts 3 with threads cut around the periphery are vertically provided with the column mounting portion 2 in between and are rotatably supported at both ends. A deceleration mechanism 4 is provided at the base of these rotating shafts 3 to rotate them synchronously. These speed reduction mechanisms 4 are driven by an electric motor 5.

Both ends of a movable stopper mounting stand 6 are screwed onto the rotating shaft 3, and the rotating shaft 3 moves up and down while maintaining a horizontal state. In this movable stopper installation, the upper end of the movable stopper shaft 7 is attached to the center of the lower surface of the stand 6, and the lower end (
A disk-shaped movable stopper 9 having a filter 8 made of sintered metal or the like provided on the inner surface thereof is attached, and is formed in the shape of a piston.

One end of a capillary tube 1o is connected to the upper surface (outer surface) side of the movable stopper 9 and communicates with the inner surface via the filter 8. The other end is pulled out from the side hole ll.

To fill a packed column with packing material using the above device,
By bolting the flanges of the stainless steel cylindrical moving column 12 that has connection flanges at both ends, the cone-shaped different diameter pipe adapter 13, and the packed column 14 having a smaller diameter than the moving column 12, The central axes of each are aligned and connected in a liquid-tight manner. A filter 15a is attached to the inner surface of the lower end opening of the connected packed column 14, and the filter 15a is attached to the outer surface.
It is closed by a blockage flliE15 provided with a bovine epithelial tube 15b that communicates with the inner surface via a.

The upper end of the moving column 12 of the connecting column to which the closing plate 15 is attached is fixed to the column mounting section 2 of the fixing base I. Next, a slurry filler adjusted to a predetermined concentration is injected from the upper opening of the moving column 12, and the electric motor 5
The movable stopper mounting table 6 is lowered by driving the movable stopper 9, and the movable stopper 9 is placed into the moving column 12. By further lowering the liquid, the liquid flows out from the capillary tube 15b, the filler is pressed by the movable stopper 9, and the filler 16 is filled into the packed column 14 under pressure.

After pressurized filling is completed, the packed column 14 is connected to the different diameter pipe adapter 1.
Remove from 3. Next, as shown in FIG. 2, a filter 17a made of sintered metal or the like with a porous inner surface, similar to the closing plate 15, is attached to the upper end of the packed column 14. Separation column A is obtained by bolting the closing plate 17 to which the capillary tube 17b for liquid discharge communicating with the outer surface side is attached in a liquid-tight manner. In this case, it is necessary that the filler is sufficiently filled up to the upper end of the packing column 14 so that no gap is created between the closing plate 17 and the filler 16.

In the sample separation/preparation operation, the sample is injected through the capillary tube 15b, and the liquid whose components have been separated in the separation column A is caused to flow out from the capillary tube 17b.

In order to fill the packed column I4 of the same diameter with the packing material using the above device, the movable stopper 9 is raised, the inside of the moving column 12 and the different diameter pipe adapter 13 are cleaned, and then the packed column 14 is attached. repeats the same operation. Furthermore, when filling the packing columns 14 with different diameters,
The different diameter pipe adapter 13 and packed column 14 may be replaced.

Furthermore, FIG. 3 shows another example of an apparatus for carrying out the method of the present invention, in which there is only one rotating shaft 3, and the same parts as in FIG. omitted.

A speed reduction mechanism 4 rotatably driven by an electric motor 5 is attached to the fixed base l, and a threaded rotating shaft 3 rotated at a low speed is extended upward perpendicularly to the fixed base 1 by the speed reduction mechanism 4. It is installed out of the way. This rotating shaft 3 has
For mounting the movable stopper, a stand 6 is screwed together, and the upper end of the movable stopper shaft 7 is attached to the stand 6, and a part of the moveable stopper shaft 7 is attached to the stand 6 to prevent co-rotation with the rotating shaft 3. Above rotating shaft 3
A support column 18 whose base is parallel to and fixed to the fixed base 1,
They are engaged for free movement only in the vertical direction.

In this device as well, by driving the motor 5, the movable stopper mounting table 6 moves up and down, and the movable stopper 9 moves the filler 1
6 is pressurized.

The above explanation is not limited to the case where two or one rotating shaft 3 is rotated by an electric motor to drive the movable stopper to apply pressure, but the movable stopper 9 is moved downward via the movable stopper shaft 7. There are no restrictions on the driving method, as long as it presses the movable stopper shaft 7, for example, a rod of a hydraulic cylinder or a pneumatic cylinder may be used as the movable stopper shaft 7. Further, the filler is usually pressed at a predetermined pressure by using a strain gauge, a load cell, a hydraulic pressure measurement, or the like to keep the pressure constant.

〔Effect of the invention〕

As described above, the method of the present invention can fill a long packed column with filler without increasing the height of the packing device, and also has a wide adjustment range for the slurry concentration to be injected.
It is an excellent method that can accommodate a variety of conditions and allows the creation of various separation columns that can be selectively used for the separation and fractionation of each component without replacing the movable stopper or moving column.

[Brief explanation of the drawing]

FIG. 1 is a schematic diagram showing an example of an apparatus for implementing the method of the present invention, FIG. 2 is a longitudinal sectional view of a separation column produced by the method of the present invention, and FIG. 3 is an apparatus for implementing the method of the present invention. It is a schematic diagram showing another example. Fig. 1 1...Fixed stand, 2...Column mounting part, 3...Rotating shaft, 4...Deceleration mechanism, 5...Electric motor, 6...Movable stopper mounting stand, 7 Movable Plug shaft, 8...Filter, 9...
Movable stopper, IO...capillary tube, 11...side hole, 1
2... Moving column, 13... Different diameter pipe adapter, 14
... Packed column, 15 ... Closure plate, 15a ... Filter, 15b ... Capillary tube, 16 ... Filler,
17... Closure plate, 17a... Filter, 17b... Capillary tube, 18... Support column, A... Separation column.

Claims (1)

[Claims] 1. Connect one opening of the cylindrical moving column and one opening of the cylindrical packed column with an inner diameter smaller than the inner diameter of the moving column to each other using a removable cone-shaped different diameter pipe adapter. The other opening of the connected packed columns is closed by a removable closing plate having a filter on the inner surface and a capillary tube communicating with the inner surface on the outer surface. The movable column is mounted on a fixed base so that the center axis of the column is vertical, and the filler slurry is introduced from the other opening of the movable column.A filter is provided on the inner surface, and a movable stopper shaft is provided at the center of the outer surface. A piston-shaped movable stopper to which a capillary tube communicating with the inner surface through the filter is attached is fitted into the movable column, and the movable stopper is lowered via the movable stopper shaft by the drive mechanism to fill the above-mentioned filling material. After pressing the packing material in the column to a predetermined pressure, the different diameter pipe adapter and the packed column are separated, and one opening of the separated packed column is provided with a filter on the inner surface side, and a filter is provided on the outer surface side. A method for producing a separation column for liquid chromatography, characterized in that the column is closed by a removable closing plate provided with a capillary tube communicating with the inner surface.