JP2004101198A - Preparative device and preparative liquid chromatograph - Google Patents

Abstract

An object of the present invention is to respond to various requests for a sample component that has been collected.
A chromatogram is obtained from a detection signal obtained from a detector, and a peak to be fractionated is set according to the magnitude (peak height) of the detection signal from the detector. The level of the magnitude of the detection signal for determining the fraction peak is L1, and the fraction volume to be collected in a test tube for collecting the target sample component by the fraction collector is set according to the level of the peak height. . When the peak height is between the levels L1 and L2, the fractionation volume is set to AmL, and when the peak height exceeds the level L2, the fractionation volume is set to CmL, which is smaller than that. Thus, the setting is made so that the fractionation capacity is changed in two stages according to the peak height.
[Selection] Fig. 2

Description

[0001]
TECHNICAL FIELD OF THE INVENTION
The present invention relates to a preparative apparatus for collecting a sample component based on the output of a detector, and a preparative liquid chromatograph provided with the preparative apparatus.
[0002]
[Prior art]
The preparative liquid chromatograph is composed of a separation column, a liquid sending pump that sends the mobile phase contained in a mobile phase container to the separation column, an injector that injects a sample into the mobile phase flow path to the separation column, and a sample from the separation column. It is provided with a detector for detecting the components, and a separation device for collecting the eluted sample components after passing through the detector into a collection container. The fractionation device includes a fraction collector, which is a mechanism for collecting the eluted sample components into a fractionation container, and a fractionation control device that controls the operation of the fraction collector based on a detection signal of a detector.
[0003]
The fractionation control device has a function of automatically determining whether to fractionate or not according to the peak height, but the fractionation amount to be fractionated into one fractionation container is fixed.
In a liquid chromatograph, a sample component separated and eluted by a separation column has a concentration distribution, and a concentration gradient occurs in which the concentration decreases from the center to the front and rear of the sample component in terms of elution time. When fractionating a sample component into a fractionation container, it is desirable to fractionate a portion where the concentration of the sample component is as high as possible. Therefore, for a sample component with a large amount of elution, one sample component is divided into a plurality of preparative containers and fractionated, and a portion having a high peak height (a sample component concentration is high) corresponding to a chromatograph is effective. As a component.
[0004]
[Problems to be solved by the invention]
When the sample components eluted from the separation column are collected in multiple preparative containers, if the fractionation volume is fixed, the high and low sample component concentrations will be treated the same. In addition, the degree of freedom for post-sampling processing is small. For example, as post-processing after fractionation, the solvent in the collection container from which the sample components were collected is volatilized to recover the target sample components. In such a case, when one sample component is fractionated into a plurality of preparative containers, it is preferable that each preparative container contain the same amount of the sample component as much as possible. If they are fixed, they cannot meet such demands.
[0005]
Accordingly, the present invention provides a separation apparatus that can meet various requests for sampled sample components when the sample components eluted from the separation column are collected in a plurality of separation containers and are collected. It is an object of the present invention to provide a liquid chromatograph equipped with a fractionating device.
[0006]
[Means for Solving the Problems]
The present invention includes a fraction collector that collects an eluted sample component after passing through a detector of a liquid chromatograph into a collection container, and a fractionation control device that controls the operation of the fraction collector based on a detection signal of the detector. A fractionation device, wherein the fractionation control device has a function of variably setting the volume of fractionation in the fractionation vessel of the fraction collector according to the magnitude of a detection signal of a sample component intended for fractionation. It is characterized by the following.
[0007]
Further, the preparative liquid chromatograph includes a separation column, a liquid sending means for sending a mobile phase to the separation column, an injector for injecting a sample into a mobile phase flow path to the separation column, and detecting a sample component eluted from the separation column. And a fractionation device for collecting the eluted sample components after passing through the detector into a fractionation container, wherein the fractionation device of the present invention described above is used as the fractionation device. .
[0008]
A preferable example of the setting in the preparative control device is that the higher the peak height of the target sample component, the smaller the volume fractionated in the preparative container.
A further preferable example of the setting in the fractionation control device is to make the difference in the amount of the target sample component collected in each fractionation container small.
[0009]
BEST MODE FOR CARRYING OUT THE INVENTION
FIG. 1 shows an embodiment of a preparative liquid chromatograph according to the present invention, in which a high-performance liquid chromatograph is provided with a preparative apparatus. The preparative liquid chromatograph includes a separation column 7, a liquid sending pump 3 for sending a mobile phase contained in a mobile phase container 1 to the separation column 7, and an injector for injecting a sample into a mobile phase flow path to the separation column 7. 5, a detector 9 for detecting a sample component from the separation column 7 and a fractionating device. The fractionation device includes a fraction collector 11 of a fractionation mechanism that collects the eluted sample component after passing through the detector 9 into a fractionation container, and a fractioner that controls the operation of the fraction collector 11 based on a detection signal of the detector 9. A pick-up control device 13 is provided.
[0010]
The preparative control device 13 can be realized as being incorporated in a control device that performs operations and data processing of the preparative liquid chromatograph. In addition to the control device, it can also be realized by a CPU or a personal computer for only the sorting device.
The fraction collector 11 is provided with a switching valve and a test tube as a sorting container.
[0011]
The sample is injected from the injector 5 into the channel, sent to the separation column 7 by the mobile phase, separated and eluted. When the detector 9 is an optical detector, for example, a chromatogram based on absorbance or fluorescence at a specific wavelength is monitored, and the fractionation control device 13 performs the operation based on the detection signal of the detector 9 at the specific wavelength. Valve switching control of the fraction collector 11 is performed.
[0012]
The operation of the sorting control device 13 will be described.
A chromatogram is obtained from the detection signal obtained from the detector 3, and the peak to be fractionated is set according to the magnitude (peak height) of the detection signal from the detector 9. Now, for example, as shown in FIG. 2, the level of the magnitude of the detection signal for determining the fractionation peak is L1. The fraction volume to be collected in a test tube (collection container) for collecting the target sample component by the fraction collector is set according to the level of the peak height.
[0013]
In the example of FIG. 2, the fractionation volume is set to AmL (milliliter) when the peak height is between levels L1 and L2, and the fractionation volume is set to CmL which is smaller at the portion where the peak height exceeds level L2. Keep it. Thus, it is assumed that the fractionation volume is set to be changed in two stages according to the peak height.
[0014]
The operation will be described based on the flowchart of FIG.
Fractionation is not performed until the peak signal, which is a detection signal of the detector, exceeds the level L1, and when the detection signal of the detector exceeds the peak height L1, fractionation of the sample component is started (step S1). → S2). Then, the fractionation is continued as it is until the fractionation volume V into one test tube, which is a fractionation container, exceeds AmL (steps S3 → S5 → S8 → S7 → S3).
[0015]
When the fractionation capacity in one test tube exceeds AmL, the process moves to the next test tube and fractionation is continued (steps S3 → S4 → S5 → S8 → S7 → S3).
Eventually, when the detection signal exceeds the level L2, the fraction volume V is changed to CmL (steps S5 → S6 → S7 → S3). In the range where the detection signal exceeds the level L2, if the fractionation capacity for one test tube exceeds CmL, the process moves to the next test tube and continues fractionation (steps S3 → S4 → S5 → S8 → S7 → S3).
[0016]
Eventually, the detection signal of the detector starts to decrease after passing the peak apex, and when it falls below the level L2, the fractionation capacity is returned to the original AmL (step S8 → S9). In this state, the fractionation is continued while changing the test tube with the fractionation volume AmL (step S7 → S3 → S4 → S5 → S8 → S7).
[0017]
When the detection signal level further decreases and falls below the level L1, the fractionation operation for the sample component ends (step S7).
When the peak of the next sample component is detected, the fractionation is started in the same manner, and the fractionation volume is changed in two stages according to the detection signal level.
[0018]
The setting of the change of the fraction volume is two steps in the above embodiment, but may be set to be changed to more steps.
Further, the relationship between the fractionation volume and the detection signal level can be set in multiple stages so that the amount of the target sample component fractionated in each test tube is substantially uniform.
[0019]
【The invention's effect】
In the preparative liquid chromatograph of the present invention, the volume of fractionation in the fraction collection container can be variably set according to the magnitude of the detection signal of the sample component to be fractionated. It becomes possible to respond to various requests for the taken sample components.
Also, for example, even when one sample component is fractionated into a plurality of fractionation containers in preparation for fractionation errors or abnormalities, only the necessary portion can be fractionated finely, compared to fine fractionation over the entire region. Thus, the number of sorting containers required for fractionation can be reduced.
It is set so that the higher the peak height of the target sample component is, the smaller the volume fractionated in the preparative container is, and further, the difference in the amount of the target sample component collected in each preparative container is reduced. In this case, the uniformity of the amount of the sample to be fractionated in each of the preparative containers increases.
[Brief description of the drawings]
FIG. 1 is a schematic configuration diagram showing a preparative liquid chromatograph of one embodiment.
FIG. 2 is a diagram for explaining a fractionation operation of one embodiment, and is a peak waveform diagram of one sample component of a chromatogram.
FIG. 3 is a flowchart illustrating the operation of one embodiment.
[Explanation of symbols]
DESCRIPTION OF SYMBOLS 1 Mobile phase container 3 Liquid sending pump 5 Injector 7 Separation column 9 Detector 11 Fraction collector 13 Preparative control device

Claims (4)

A fraction collector comprising: a fraction collector for collecting an eluted sample component after passing through a detector of a liquid chromatograph into a fractionation container; and a fractionation control device for controlling an operation of the fraction collector based on a detection signal of the detector. In the device,
The fractionation control device is characterized in that the fractionation control device has a function of variably setting a volume of fractionation in the fractionation container according to the magnitude of a detection signal of a sample component intended for fractionation. Sorting device. The fractionation device according to claim 1, wherein the fractionation control device is set such that the higher the peak height of the target sample component, the smaller the volume fractionated in the fractionation container. The sorting device according to claim 2, wherein the sorting control device is set so that a difference between the amounts of the target sample components collected in each sorting container is reduced. A separation column, a liquid sending means for sending a mobile phase to the separation column, an injector for injecting a sample into a mobile phase flow path to the separation column, a detector for detecting sample components eluted from the separation column, passing through the detector In a preparative liquid chromatograph equipped with a preparative device for collecting the subsequent eluted sample components in a preparative container,
4. A preparative liquid chromatograph, wherein the preparative apparatus is one according to any one of claims 1 to 3.

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