JPS63300961A - Column processor - Google Patents

Abstract

PURPOSE:To process plural columns simultaneously with high efficiency by providing a column holder, a plug body to be fitted airtightly in the upper parts of respective columns, an elevating means which fits and extracts the plug body in and from each column, a tube penetrating the plug body, etc. CONSTITUTION:The columns 10 where fillers are charged are inserted into respective holding holes 22 of the column holder 20. When a specific column 10 is positioned below the plug body 33, an air cylinder 32 drives a piston rod 32a downward and the plug body 33 fitted atop is fitted airtightly in the upper end part of a column container 11. Then a pump 35 is driven to feed a sample into the container 11 through the tube 34. Further, a changeover valve is switched to feed cleaning liquid by the pump 35, so that the column 10 is cleaned. Then an eluate is fed by the pump 35 in to the column 10. Then the rod 32a is driven upward and the plug body 33 leaves the column container 11. Then an adjacent column 10 is positioned below the plug body 33.

Description

DETAILED DESCRIPTION OF THE INVENTION (Field of Industrial Application) The present invention relates to a column processing device that can efficiently and continuously process samples.

(Conventional technology) In chromatography, which separates and analyzes substances.

High performance liquid chromatography (HPLC), which performs everything from liquid delivery to analysis of eluate, is widely used because of its high performance and excellent reproducibility. However, since HPLC is expensive and needs to be carried out under high pressure, methods have also been adopted in which substances are easily separated using open columns or mini-columns.

An open column is a type of column in which an air layer exists above the packing material packed inside the column container. A mini column is a simpler version of the open column, and is generally used as a disposable disposable column.

(Problems to be Solved by the Invention) When processing a sample using such an oven column, 1. Usually, the sample is charged to a packing material.

Cleaning treatment with a cleaning solution, elution treatment with an eluent, etc. have generally been performed manually by the measurer. Although automatic charging devices have recently been developed that automatically charge samples into packing materials, washing, elution, etc. must be performed manually by the measuring person as in the past. For this reason.

Simultaneously processing a large number of samples using multiple columns requires a lot of effort and is not efficient.

When a sample is adsorbed to the packing material packed in a mini-column and a cleaning solution is injected into the column for cleaning, it takes a long time for the cleaning solution to flow through the column, and as expected,
This causes a decrease in column processing efficiency. Therefore, by sealing the column with a stopper and pressurizing the inside of the column with a pump.

The cleaning liquid is passed through at high speed.

It is difficult to seal the column with a stopper, making it impossible to improve work efficiency. In addition, when treating multiple columns at the same time with high pressure inside the columns, it is necessary to connect a pump to each column to increase the pressure inside the columns. Impairs chromatographic properties.

The present invention solves the above-mentioned conventional problems, and its purpose is to provide a column processing apparatus that can simultaneously process a plurality of columns with high efficiency.

(Means for Solving the Problems) The column processing apparatus of the present invention includes a plurality of columns filled with a filler so as to form an air layer at the top, and a column holder capable of holding each column substantially vertically. a stopper that can be airtightly fitted to the top of each column held in the column holder; means for raising and lowering the stopper to fit into and separate from each column; Thereby, the above object is achieved.

(Example) The present invention will be described below with reference to Examples.

As shown in FIGS. 1 and 2, the column processing apparatus of the present invention includes a plurality of columns 10.10. It has a column holder 20 that holds... Each column 10 held by the column holder 20 is filled with a filler 12 in a cylindrical column container 11 having an opening at the upper end so that an air layer 13 is formed above the column container 11 . The filler 1
A filter 14 is disposed on the upper surface of 2 as required. An outlet 11a projecting downward is provided on the bottom of the column container 11, and a flange portion 11b is provided at the upper end of the column container 11.

The column holder 20 has, for example, a main body portion 21 extending substantially horizontally in a predetermined direction at a predetermined height position.

The main body portion 21 is provided with a holding hole 22°2 that penetrates in the vertical direction.
2, . . . are arranged at predetermined intervals. Each holding hole 22 is large enough to fit each column 10 therein,
Each column 10 is inserted into a respective holding hole 22.

Flange portion 11b provided at the upper end of column container 11
is held substantially vertically by being engaged with the upper surface of the main body portion 21.

A sealing mechanism 30 capable of sealing the opening at the upper end of each column 1o is disposed above the column holder 20. The sealing mechanism 3o includes a substrate 31 in a substantially horizontal state, and an air cylinder 32 fixed to the substrate 31.

The air cylinder 32 has a plug 33 attached to the tip of the piston rod 32a.

The air cylinder 32 is attached to the base plate 31 so that the tip of the piston rod 32a is located on the lower side, and the direction in which the piston rod advances is downward. A stopper 33 is attached to the tip of the piston rod 32a, and as the piston rod 32a moves up and down, the stopper 33 moves up and down.

The plug body 33 can be airtightly fitted into the upper end opening of each column 10 held in each holding hole 22 in the column holder 20 when the piston rod 32a moves downward. The plug body 33 is made of a material that can be airtightly fitted to the column container 11 when the plug body 33 is fitted to the upper end portion of the column container 11. so that they can be easily mated.

For example, it has a hemispherical shape. In this way, if the lower part of the stopper 33 is hemispherical, even if the stopper 33 is located at a position slightly shifted from above the opening in the column 10, it will easily fit into the opening by descending. obtain.

The end of one tube 34, for example, passes through the plug 33, and its end surface is flush with the lower surface of the plug 33.

The tube 34 is connected to a pump 35 for supplying a predetermined gas or solution. The pump 35 is connected to the pump 35 through two, for example, one switching valve (not shown).

A solution such as a cleaning solution or a gas is passed through the tube 34, and the pump 35 supplies a predetermined solution or gas to the tube 34 by switching the switching valve.

The column holder 20 has a plug body 33 of a plugging mechanism 30.

The column holder 20 is moved in the longitudinal direction by a predetermined driving means such as a motor or a hydraulic cylinder so that it can be positioned above each column 10 held by the column holder 20.

The column processing apparatus of the present invention has such a configuration.

Used as follows: First, each column 10 filled with the filler 12 in the column container 11 is inserted into each holding hole 22 in the column holder 20 . As a result, each column 10 is held substantially vertically with the flange portion 11b at the upper end thereof being engaged with the upper surface of the main body portion 21 of the column holder 20.

In this state, the column holder 20 is moved, and a predetermined column 10 held on the column stand 20 is
It is located below the stopper 33. When a predetermined column 10 is positioned below the stopper 33, the air cylinder 32
The piston rod 32a is driven to advance downward, and as shown in FIG.
It is hermetically fitted to the upper end of the column container 11 at 0.

In this state, the pump 35 is driven and the tube 3
4 through the column vessel 11, e.g.

A predetermined sample is delivered. As a result, a predetermined sample is charged to the packing material 12 of the column 10. Next, by switching a switching valve (not shown) connected to the pump 35, the pump 35 supplies a cleaning liquid into the column 10, and the column 10, which is plugged with the plug body 33, is cleaned with the cleaning liquid. . moreover.

Thereafter, an eluent is fed into the column 10 by the pump 35, and the column 10 is subjected to an elution process.

The above washing process and elution process are performed by supplying a predetermined solution from the pump 35 into the column that is sealed with the stopper 33, so the inside of the column is pressurized and the time required for the two liquid passages is shortened. .

In this way, when charging the sample in one column, washing process, and elution process are completed.

The air cylinder 32 in the sealing mechanism 30 is driven to move its piston rod 32a upward;
A plug 33 attached to the tip of the piston rod 32a
is raised. As a result, the plug body 33.

The column is separated from the column container 11. Next, the column holder 20 is moved in its longitudinal direction, and the column holder 20 is moved below the stopper 33.
A column 10 adjacent to the column 10 for which the predetermined process has been completed as described above is located. below.

The same operations as those described above are repeated, and the column 10 located below the stopper 33 is charged with a sample, washed, and
and elution treatment is performed. When the elution process for this column 10 is completed, the column holder 20 is moved, the stopper 33 is fitted into the column 10 adjacent to the column IO, and a predetermined process is performed.

Thereafter, the same operation is repeated sequentially, and each column held in the column holder 20 is sequentially charged with the sample,
Washing treatment and elution treatment are performed.

In addition, in the above embodiment, the plug body 33 in the plug mechanism 30
Although the column holder 20 is moved so as to be positioned above each column 10 held by the column holder 20, the entire sealing mechanism 30 is limited to such a structure. 10 may be moved in the direction of arrangement.

Further, in the above embodiment, the closure mechanism 30 in which one closure body 33 is disposed has been described, but the closure mechanism 30 is not limited to such an embodiment.

As shown in FIG. 3, a plurality of plugs 33, 33. . . , a plurality of air cylinders 32° for raising and lowering each plug body 33, a plurality of substrates 31 for supporting each air cylinder 32, etc. may be arranged. In this case, predetermined processing can be performed independently for each column 10.

Furthermore, as shown in FIG. 4, one air cylinder 32 can have multiple plugs (two plugs in FIG. 4).
33 may be configured to be integrally driven up and down. In this case, since the plugs 33 can be fitted into and removed from a plurality of columns 10 at the same time, a plurality of columns 10 can be processed simultaneously.

As shown in FIGS. 3 and 4, in a column processing apparatus in which a plurality of plugs 33 are arranged, the type of solution or gas fed from the tube 34 attached to each plug 33 is limited. If a stopper equipped with a tube capable of supplying a specified solution or gas is fitted to a column that requires the supply of a specified solution or gas, column processing can be performed with complete division of labor. Moreover, the composition of the liquid flowing through the tube is not changed, and the sample can be processed with high reproducibility and high efficiency.

Further, in the above embodiment, sample charging, washing treatment, and elution treatment were sequentially performed once for one column 10, but instead of this, for example, one column 10
When the sample charge of 0 is completed.

The sealing mechanism 30 is moved to charge a sample into a column 10 adjacent to the column 10, and the same process is repeated.

For each column 10 after all columns 10 held in the column holder 20 have been charged with sample.

In the same manner, with the stopper 33 fitted in the column, cleaning liquid was charged, and then the stopper 33 was pulled up.

The application to the next column is repeated in sequence, the cleaning solution is charged, and the cleaning process is performed without sealing each column 10 with the stopper 33, and then thereafter.

For each column 10, eluent may be sequentially charged and elution processing may be performed in the same manner.

Movement control of column holder 20 or sealing mechanism 30,
Control of the air cylinder 32 in the sealing mechanism 30;
Furthermore, the control of the pump 35, the control of the switching valve, etc. can also be performed automatically using a microcomputer.

Next, using the column processing apparatus of the present invention, glycated hemoglobin Lc, which is a good indicator of diabetes, was quantified, and the details thereof will be explained.

A minicolumn filled with an affinity gel in which 1rR1 m-aminophenylboronic acid was immobilized on agarose beads was used as the column. and.

This packing material is then equilibrated with a washing solution into a minicolumn.

100 μl of a sample obtained by hemolyzing EDTA anticoagulated blood with 20 times the amount of water was charged. In this way, 10 mini-columns charged with samples from 10 people.

Washing and elution treatments were carried out using the column processing apparatus of the present invention shown in FIG. The column processing apparatus was configured to be automatically controlled by a microcomputer.

Automatic control by a microcomputer first involves fitting a stopper airtightly into one column, and applying 25 mL of washing liquid to the column.
0mM ammonium acetate (pl+ 8.0) -50m
20m of M magnesium chloride solution! The column was charged and washed under pressure. Next, the column was charged with 200 mM sorbitol-10On + M Tris/HCl buffer (pH 8,5) for 5 d as an eluent.
Elution treatment was performed under pressure. The amount of glycated hemoglobin Alc in the blood was determined from the absorbance at 414 nm of the solution eluted with the eluate.

Thereafter, the column treatment device was automatically operated by a microcomputer, and the same treatment was sequentially performed once on other columns held in the column holder. For each column, it took 15 minutes for washing and 3 minutes for elution, 2 for a total of 18 minutes.

Denaturation of "Devitrification" Ten mini-columns charged with samples in the same manner as in Example 1 were held in one column holder in the column processing apparatus of the present invention. Next, in order to sequentially charge each column with 10 ml of the same washing solution as in Example 1, the column processing device of the present invention was automatically operated by a microcomputer, and the stopper was fitted into the column. The washing solution was charged in the same state, and then the stopper was pulled up and applied to the next column, and this process was repeated in sequence. Each column has
The cleaning liquid was charged in 10 seconds. When each column was left unsealed with a stopper for 10 minutes, the washing solution flowed out of the column. After that, apply the cleaning solution again for 10 minutes.
ml was charged to each column sequentially.

Each column was charged with wash solution in 10 seconds. Each column was then left for 10 minutes to allow the washing solution to flow out from each column.

Next, 5 d each of the same solution as in Example 1 was added.

Two column processing apparatuses of the present invention were automatically operated by a microcomputer to sequentially charge each column.

Each column was charged with eluent in approximately 5 seconds. Then, the eluate eluted from each column was collected into an acceptor container. It took 5 minutes for each column eluent to elute from the column.

In any of the above embodiments, it is much more efficient than manually processing each column, and by controlling the column processing apparatus of the present invention using a microcomputer, each column can be processed manually. This is because the processing conditions for the sample in the column are approximately constant.

Reproducibility was good. Furthermore, in Example 1, the time required for the liquid passage was shortened because the liquid passage treatment was performed under pressure.

(Effects of the Invention) As described above, the column processing apparatus of the present invention can efficiently process a plurality of columns with good reproducibility. Furthermore, when charging a processing liquid such as a cleaning liquid into the column, it can be carried out with the stopper fitted.

The processing liquid can be reliably charged. moreover,
When carrying out liquid passage treatment under pressure, the time required for one liquid passage can be shortened.

Figure 1 is a perspective view showing an example of the column processing apparatus of the present invention;
FIG. 2 is a sectional view for explaining its operation.

FIG. 3 is a sectional view showing another example of the column processing apparatus of the present invention, and FIG. 4 is a sectional view showing still another example of the column processing apparatus of the invention.

10... Column, 11... Column container, 12...
Filling agent, 20... Column holder, 30... Sealing mechanism, 31... Substrate, 32... Air cylinder, 3
3... Plug body, 34... Tube, 35... Pump.

C) Thief decrease

Claims (1)

[Claims] 1. A plurality of columns filled with a filler so as to form an air layer at the top, a column holder capable of holding each column substantially vertically, and each column held by the column holder. A column treatment comprising: a plug that can be airtightly fitted to the top of a column; means for raising and lowering the plug to fit into and remove from each column; and a tube passing through the plug. Device. 2. The column processing apparatus according to claim 1, wherein the plug body is moved relative to the column holder to a position where it can fit into each column held by the column holder. 3. The column processing apparatus according to claim 1, wherein the plurality of plugs are integrally raised and lowered so as to fit into the plurality of columns held by the column holder.