JPH03115832A - Sampling column - Google Patents

Abstract

PURPOSE:To effectively utilize a separation agent by filling a column container having a taper formed thereto so that the cross-sectional area thereof is monotonously reduced from an inflow port to an outflow port with the separation agent. CONSTITUTION:When a mobile phase supply apparatus is connected to the inflow port 1a of a column container 1 to inject a definite amount of a sample in the column container 1, the injected sample is distributed in a definite volume and moves through a sample separation agent 2 toward the outflow port 1b by the flow of a mobile phase and the concn. of the sample per unit length is diluted. As mentioned above, since the concn. per unit length of the sample is lowered with the advance of separation, treatment quantity becomes little toward the outflow port 1b and, therefore, even when the inner diameter of the column container 1 becomes fine toward the outflow port 1b and the separation gent 2 per unit length is little, separation treatment can be furnished and the separation agent 2 can be used at the limit of its capacity.

Description

DETAILED DESCRIPTION OF THE INVENTION (Industrial Application Field) The present invention is directed to the structure of a preparative column suitable for extracting a specific target component from a sample.

(Prior Art) When separating a target component from a sample, a conventional analytical column is used to separate the sample into components.

(Problems to be Solved by the Invention) By the way, the amount of sample that can be processed by a column, so-called processing capacity, is determined by the cross-sectional area of the column inlet, while for analytical columns, it is necessary to identify components based on retention time. First, the axial cross-sectional area of the column is uniform, and the components are diluted as their separation progresses, so from the perspective of preparative separation, the column usage efficiency is low. there were.

The present invention has been made in view of these problems, and its purpose is to provide a novel preparative column that can effectively utilize a separating agent.

(Means for solving the problem) In order to solve such a problem, in the present invention,
A separating agent was filled in a column container that was tapered so that the cross-sectional area monotonically decreased from the inlet to the outlet.

(Function) As the target component is separated from the sample and the target component becomes diluted, the cross-sectional area of the column also decreases, so the cross-sectional area of the column for the target component is always at the very edge of the limit that satisfies the processing capacity at each position. Therefore, processing capacity can be improved with the same amount of filler.

(Embodiment) Fig. 1 shows an embodiment of the present invention, in which reference numeral 1 indicates a container constituting a column, from one end serving as an inlet port 1a to the other end serving as an outlet port 1b. It is formed so that the inner diameter monotonically decreases toward the end, and the separating agent 2 is filled inside.

In this example, when a mobile phase supply bag M is connected to the inlet 1a and a fixed amount A of sample is injected, the injected sample has a fixed volume determined by the volume near the inlet 1a of the column and the solvent of the sample. distributed in

The flow of the mobile phase causes the sample to flow through the separation agent 2 to the outlet 1b.
When moving to the side, the sample is separated into components by interaction with the separation agent 2 and spread in the axial direction, so that the concentration of the sample per unit length is diluted.

Incidentally, the inner diameter of the container 1 constituting the column becomes narrower as it goes toward the outlet 1b, but as mentioned above, as the separation progresses, the concentration per unit length decreases. As the amount of treatment decreases toward the outlet 1b, the amount of separating agent per unit length can be reduced.
It can cover layer separation processing.

Each component thus separated is discharged from the outlet 1b after a period of time determined by the column shape and separation agent.

EXAMPLE When a sample of jlA (micrograms) is injected into a column of uniform internal diameter, for example an analytical column, the sample is distributed within the volume at the inlet of the column. This distribution volume is determined by the solvent that makes up the sample,
Although it depends on the amount of sample, the upper limit is determined by the total volume of the column1.For example, a column with an inner diameter of 6 mm and a length of 15 cm with the relationship between the injection volume and the number of theoretical plates N as shown in the M2 diagram. Taking an analytical column as an example, if we find its upper limit (the upper limit usually refers to the injection volume at which the number of plates decreases to 90% when the injection volume is infinitesimal) for sample solvent C, , approximately 5 microliters (note that depending on the type of solvent that makes up the sample, concentration may occur at the column inlet during injection, so even if the sample is injected with an apparent size of 200 to 1000 microliters, it will actually be Distributed below 5 microliters,
Therefore, there may be cases where the number of stages does not decrease).

As mentioned above, the upper limit of the distribution volume at the column inlet is 5
Assuming microliters, this amount is equivalent to the column volume [πX (6/2)X (6/2)x 150]
=4239 (microliter), which is approximately 1/800 of the column volume.

On the other hand, if the porosity of the filler is α and the total volume of the voids is VO (microliter), the concentration of sacsiple at the time of injection is A+ [(VO/(1)/800] = 800(!A/
VO (L19/L11) ...(1).

On the other hand, the sample is diluted at the outlet of the column, so when calculating this dilution rate by focusing on the peak in the distribution ratio, if the half-width of the peak is W+/2h, the concentration at the top of the peak is A. Since it is expressed by /W+/2h, A/Wl/2h=A/ rJ stone 54/NX (1+K)
xVO] (u9/ui)--(2) becomes (
However, N represents the number of theoretical plates in the column).

Therefore, the dilution rate is calculated from the above formulas (1) and (2) as follows: [800aA/VO] / [, /”-Stone 54/NX (
1+K) be able to handle the case.

Therefore, when this condition is applied and calculated from equation (3), the dilution rate at the outlet becomes 3760α/ff.

From this, inflow side inner diameter D1: outflow side inner diameter D2 = 1:
By selecting the internal diameters of the inlet and outlet so that α/F is equal to If a separating agent is used, the absolute amount to be treated will increase.

In this embodiment, the cross-section of the column is circular, but it is clear that the same effect can be obtained even if the column has another shape, such as an ellipse.

(Effects of the Invention) As explained above, in the present invention, the separation agent is filled in one column container which is tapered so that the cross-sectional area decreases monotonically from the inlet to the outlet, so that the separation progresses. According to the dilution of the sample that occurs over time, the amount of separation agent required in each separation step can be reduced, making it possible to effectively utilize the separation agent in the entire column and obtain a column with high throughput using less separation agent. can.

[Brief explanation of drawings]

FIG. 1 is a block diagram of an apparatus showing an embodiment of the present invention, and FIG. 2 is a diagram showing the relationship between the sample injection volume I of the column and the number N of theoretical plates.

Claims (1)

[Claims] A preparative column is made by filling a separation agent into a column container that is tapered so that the cross-sectional area decreases monotonically from the inlet to the outlet.