JPS5866845A - Separating method for sample by column chromatograph - Google Patents

Abstract

PURPOSE:To accurate by perform an analysis at a constant flow rate and under constant conditions, by placing a sample on a carrier packed an upstanding column and keeping the liquid face of a solvent higher than the face of the sample and then, equalizing the quantity of liquid to be sent from the column and the quantity of the solvent to be supplemented so that compressed air having a constant, pressure is hermetically sealed on the liquid face. CONSTITUTION:A glass wool 21 and sea sand 22 are laid on the bottom part of the main body 7 of an upstanding column 1 and carriers 23 are packed on it and hereafter, a cover 8 is united with a flange. Next, a solvent 24 is put in up to the top of the carrier and bubbles or the excess solvent are discharged from a feeding and discharging pipe 15 and then, a sample 25 is placed on the carrier 23 from the pipe 15 through a pipe 20 and a hand cock 18 from an adapter 19. The solvent 24 is sent to the column 1 by closing the cock 18 and sending from a solvent vessel 3 through a control valve and a pipe 5 by two pumps and then, keeping the pressure constant by a relief valve 6 and a constant-pressure vacant part is formed on the upper part of column 1. Further, an eluate is sent to a collecting vessel 26 through a detector 11 by opening a manual opening and closing valve 9 and a solenoid valve 10, and the solvent 24 of the quantity equal to that of sent-out liquid is sent from the vessel 3 and then, pressurized air is kept at a constant pressure always. In this manner, an accurate analysis is performed by eluting the components in the sample with constant pressure and flow rate.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention is directed to a method for separating samples using column chromatography.

A column is installed vertically, a carrier made of particles is loaded, a sample is placed on the carrier, and the sample is eluted into a solvent and flows out from the bottom of the column to analyze, isolate, purify and synthesize the sample. For separation and purification of samples consisting of There are ways to get it out. This so-called flash chromatography has an excellent separation ability that enables separation in a short time compared to conventional sample separation methods using column chromatography. ?
! ! A small amount of strategy is enough. Suitable for unstable substances. It is difficult to set the amount of text IE, which has the following advantages. Since it is a continuous outflow, it is difficult to achieve 7 rakunyeon. Commercially available columns have problems such as the inability to load a large amount of sample at one time due to the container.

The present invention provides a sample separation method that does not involve the above-mentioned problems by improving a method in which a column is loaded with a carrier, a sample, and a solvent, and the solvent that has eluted the sample flows out from the bottom of the column by the action of gas pressure. The purpose is to maintain the solvent level in the column above the sample and to supply the solvent almost continuously from above along the inner wall of the column while sealing air above the liquid level. The method is characterized in that the pressure generated by compressing the sealed air causes the solvent that has eluted the sample to be sent out from the bottom of the column.

Embodiments of the invention will be described below with reference to the drawings.

FIG. 1 is a flow sheet of an example of carrying out the present invention, where l indicates a column, 2 a pump, and 3 a solvent tank, where two reciprocating pumps 2.2 alternately discharge the solvent. enters the supply pipe 5 without being sucked into the pump during the suction stroke by the control valve 4, and the pressure above a certain amount is released by the safety valve 6 and sent to the upper part of the column ◎The column 1 is connected to the main body 7 and the lid 8. It consists of 9. At the center of the bottom of the main body 7 is a manual on-off valve 9.
A delivery pipe 12 having a solenoid valve 10 and a detector 11 is connected.

The ts2 diagram shows an example of column 1, with a flat-bottomed cylindrical body 7
W8 is removably flange connected to the upper end, and a joint 13 connecting the supply pipe 5 to the side surface is screwed and fixed to the upper part of the stirrup 8, and the joint 13 and the center hole 1 of the lid 8 are fixed.
A supply/discharge pipe 15 is loosely passed through the main body 7.
It has reached the upper part of 0. The center hole 14 is isolated from the outside air by an O-ring 17 held down by an outer 116 screwed into the joint 13, and the supply/discharge pipe Is is connected to an auxiliary supply/discharge pipe zoKil having a manual cock 18 and an adapter 1g.

The method of sample separation using this column 1 is as follows.

That is, first of all, glass wool 21 and an appropriate amount of sea sand 22 are spread on the bottom of the main body 7 to prevent the carrier from flowing out, and then a carrier made of particles such as Kurikagel is loaded to a height of 23 feet on top of the glass wool 21, and then the lid 8 is placed. (Figure 3). Second, open the on-off valve 9 and the cock 18, and inject the solvent 24 from the auxiliary supply/discharge pipe 2G and the supply/discharge pipe 15 until the liquid level is above the carrier 23 (Fig. 4). 3 and discharges the air in the gap, and excess solvent is suctioned and discharged from the supply/discharge pipe 15. Thirdly, the auxiliary supply/discharge pipe 20. A predetermined amount of sample 25 is placed on the center of the carrier 23 through the supply/discharge pipe Is (FIG. 5). Fourth, the cock 1B is closed and the pump 2 is operated to send the solvent to the center hole 14 of the solvent fitting 13t IF 8 of the tank 3. The solvent 24 is supplied by flowing down along the inner wall surface of the lid 8, and the sample 25 is immersed in the solvent 24, so that the liquid level gradually rises (FIG. 6).

The air sealed in the upper part of the carrier 23 is compressed and applied to the solvent 24 under pressure, and the solvent that has eluted the specific component of the sample or the specific substance in the mixture sample passes through the gap of the carrier 23 under the air pressure. It is pumped to the bottom of the main body 7 and sent out from the delivery pipe 12.

After the elution is recorded by the detector 11, it enters the container 26 of the fraction collector.

FIG. 7 shows another example of the column 1, in which a supply pipe 5 and a nozzle pipe 28 are fixed in communication with each other at the center of a joint 27 which is screwed and fixed to the column 8.

The tip of the nozzle pipe 2s reaches the top of the main body 7 and has a plurality of nozzle holes 2g that open in the radial direction.

The method of sample separation using this column lik is as follows.

That is, first of all, the glass school 21. Sea sand 22. Loading the carrier 23 (Fig. 8),
Second, open the on-off valve 9 so that the liquid level is on the carrier 23 from above the main body 7.
Inject the solvent 241 until it reaches the upper part of the carrier 23 to wet the carrier 23 and exhaust the air in the gap (Figure 9) .

Fourth, the lid 8 is attached and the pump 2 is operated to send the solvent in the tank 3 from the supply pipe 5 to the nozzle pipe 28 and spray it in the radial direction from the nozzle hole 29. The solvent is injected toward the inner wall surface of the main body 7 or l [8 and flows down along the inner wall surface, and the solvent 24 immerses the sample 25 and the liquid level gradually rises (Fig. 11). The air sealed in is compressed into solvent 2
4, the solvent that has eluted the specific component of the sample or the specific substance of the mixture sample enters the container 26 of the fraction collector after recording the elution curve.

In the above two embodiments, the solenoid valve lO is linked to the fraction collector to open and close the container and stop the outflow of the solvent. Also.

Pump 2 operates continuously to supply solvent little by little almost continuously to column 1, and after the sealed air at the top of column 1 reaches a certain pressure, it is pumped out from the bottom in an amount equal to the supplied amount. .

As described above, in the present invention, a column arranged vertically is loaded with a carrier, a sample is placed on the column, the liquid level of the solvent is kept above the sample, and the solvent is released in a state where the air above the liquid level is sealed. Since it is supplied almost continuously, once the amount of solvent in the column increases and the sealed air is compressed to a constant pressure, the solvent that eluted the sample from the bottom of the column is sent out almost continuously at this pressure. I can do it.

In addition, since an amount of solvent equal to the amount supplied is pushed out with a constant air pressure, the sample can always be immersed in a constant amount of solvent and eluted under constant conditions.

Also,? Since the I medium is constantly replenished, not only can a large amount of sample be loaded and the column can be made smaller, but since the solvent is supplied from above along the inner wall of the column, the liquid level is hardly disturbed, so the sample is not absorbed into the solvent. Since there is no diffusion, the elution conditions do not change, and there is no inconvenience that would adversely affect analytical performance. Since the sealed air is delivered at a constant pressure, it is easier to set the supply amount to a predetermined value than the flow rate membrane, and since the sealed air has a cushioning property, there is no problem even if the delivery is temporarily interrupted, and the fraction can be maintained. It makes it easy.

[Brief explanation of drawings]

FIG. 1 is a diagram showing an example of a flow sheet for carrying out the present invention, FIG. 2 is a longitudinal sectional view of an example of a column, FIGS. 3, 4,
FIGS. 5 and 6 are schematic vertical cross-sectional views for explaining the sequence of sample separation operations, FIG. 7 is a vertical cross-sectional view of different examples of columns, and FIG. 8. FIG. 9, FIG. 1θ, and FIG. 11 are schematic vertical cross-sectional views illustrating the operational sequence of sample separation. l...Column, 2...--Pump, 5...
...Feed pipe, 7...Main body, 8...
Lid, 9... Open/close valve. 1G...-Solenoid valve, 12... Delivery pipe, 2
3...Carrier. 24...Solvent, 25...Sample. Agent Mutsuaki Nozawa 11 Figure 2

Claims (1)

[Claims] A carrier is loaded into a column installed vertically, a sample is placed on it, and the solvent is placed on the inner wall of the column while keeping the liquid level of the solvent Km above the sample and sealing air above the liquid level. A sample separation method using column chromatography, characterized in that the solvent is supplied almost continuously from above along the column, and the solvent eluted from the sample fk is sent out from the bottom of the column by the pressure generated by compressing the sealed air.