JP3864876B2 - Preparative device and preparative liquid chromatograph - Google Patents

Description

[0001]
BACKGROUND OF THE INVENTION
The present invention relates to a fractionation device that collects sample components based on the output of a detector, and a fractionation liquid chromatograph equipped with the fractionation device.
[0002]
[Prior art]
The preparative liquid chromatograph is composed of a separation column, a liquid feed pump for feeding the mobile phase contained in the mobile phase container to the separation column, an injector for injecting the sample into the mobile phase flow path to the separation column, and a sample from the separation column. The detector which detects a component, and the fractionation apparatus which extract | collects the elution sample component after passing a detector to a fractionation container are provided. The fractionation device includes a fraction collector, which is a mechanism part that collects the eluted sample component in a fractionation container, and a fractionation control device that controls the operation of the fraction collector based on the detection signal of the detector.
[0003]
The sorting control device has a function of automatically determining whether or not to perform fractionation according to the peak height, but the fractionation amount to be sorted into one sorting container is fixed.
In the liquid chromatograph, the sample components separated and eluted by the separation column have a concentration distribution, and a concentration gradient is generated in which the concentration decreases from the center to the front and back of the sample component in terms of elution time. When the sample components are fractionated in the sorting container, it is desirable to fractionate the portion where the concentration of the sample components is as high as possible. Therefore, for sample components with a large amount of elution, one sample component is divided into a plurality of preparative containers and fractionated, and the portion with a high peak height (high sample component concentration) corresponding to the chromatograph is effective. As an ingredient.
[0004]
[Problems to be solved by the invention]
When the sample components eluted from the separation column are collected in multiple collection containers, if the fraction is fixed, the dark and light portions of the sample components will be treated in the same way. The degree of freedom for processing after collection is small. For example, as post-treatment after fractionation, the solvent in the collection container from which the sample component is collected is volatilized and the target sample component is recovered. In that case, when one sample component is divided into a plurality of sorting containers, it is preferable that each sorting container contains the same amount of the sample component as much as possible. If fixed, such a request cannot be met.
[0005]
Therefore, the present invention provides a fractionation device adapted to meet various demands on the fractionated sample components when the sample components eluted from the separation column are collected in a plurality of fractionation containers. An object of the present invention is to provide a liquid chromatograph provided with a fractionation device.
[0006]
[Means for Solving the Problems]
The present invention includes a fraction collector that collects an elution sample component after passing through a detector of a liquid chromatograph in a sorting container, and a fractionation control device that controls the operation of the fraction collector based on a detection signal of the detector. The fractionation control device has a function capable of variably setting the fractionation capacity in the fractionation container of the fraction collector according to the magnitude of the detection signal of the sample component intended for fractionation. It is characterized by this.
[0007]
The preparative liquid chromatograph detects a separation column, a liquid feeding means for sending a mobile phase to the separation column, an injector for injecting a sample into the mobile phase flow path to the separation column, and a sample component eluted from the separation column. And a fractionation device that collects the eluted sample components after passing through the detector in a fractionation container, and the fractionation device of the present invention described above is used as the fractionation device. .
[0008]
The setting in the sorting control device is such that the volume fractionated in the sorting container decreases as the peak height of the target sample component increases.
[0009]
DETAILED DESCRIPTION OF THE INVENTION
FIG. 1 shows an embodiment of the preparative liquid chromatograph of the present invention, which is a high-performance liquid chromatograph equipped with a preparative device. The preparative liquid chromatograph includes a separation column 7, a liquid feed pump 3 that feeds the mobile phase contained in the mobile phase container 1 to the separation column 7, and an injector that injects a sample into the 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 fractionation device are provided. The fractionation device is a fraction collector 11 of a fractionation mechanism that collects the eluted sample components after passing through the detector 9 in a fractionation container, and a fraction that controls the operation of the fraction collector 11 based on the detection signal of the detector 9. A 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 be realized by a CPU or a personal computer only for the sorting device.
The fraction collector 11 is provided with a switching valve and a test tube as a sorting container.
[0011]
A sample is injected from the injector 5 into the flow path, sent to the separation column 7 by the mobile phase, and separated and eluted. When the detector 9 is an optical detector, for example, the absorbance or fluorescence chromatogram at a specific wavelength is monitored, and the sorting control device 13 is based on the detection signal of the detector 9 at that 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 9, 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 that determines the fractional peak is L1. The fraction volume to be collected in a test tube (collection container) for collecting a target sample component with a fraction collector is set according to the level of the peak height.
[0013]
In the example of FIG. 2, the fraction volume is set to AmL (milliliter) when the peak height is between levels L1 and L2, and the fraction volume is set to CmL smaller than that when the peak height exceeds level L2. Keep it. In this way, it is assumed that the fractionation capacity 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 height that is the detection signal of the detector exceeds the level L1, and fractionation of the sample component is started when the detection signal of the detector exceeds the peak height L1 (step S1). → S2). Then, the fractionation is continued as it is until the fractionation volume V to one test tube which is a fractionation container exceeds AmL (steps S3 → S5 → S8 → S7 → S3).
[0015]
When the fractionation volume to one test tube exceeds AmL, it moves to the next test tube and continues fractionation (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, when the fractionation volume to 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, when the detection signal of the detector starts to decrease past the peak apex and falls below the level L2, the fractional volume is returned to the original AmL (steps S8 → S9). In this state, the fractionation is continued while changing the test tube with the fraction volume AmL (steps 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 is terminated (step S7).
When the peak of the next sample component is detected, fractionation is started in the same manner, and the fraction volume is changed in two stages according to the detection signal level.
[0018]
The setting for changing the fraction capacity is two steps in the above embodiment, but it may be set so as to be changed to a larger number of steps.
Furthermore, the relationship between the fraction 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 can be made substantially uniform.
[0019]
【The invention's effect】
In the preparative liquid chromatograph of the present invention, the volume to be fractionated into the fractionation container of the fraction collector can be variably set according to the magnitude of the detection signal of the sample component intended for fractionation. It will be possible to meet various demands for sample components taken.
In addition, for example, even in the case of fractionating one sample component into multiple collection containers in preparation for mistakes or abnormalities in fractionation, only the necessary part can be fractionated finely, compared to fine fractionation over the entire region. Thus, the number of sorting containers required for fractionation can be reduced.
The higher the peak height of the target sample component is, the smaller the volume fractionated in the sorting container is, so that the uniformity of the amount of sample fractionated in each sorting container 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 the fractionation operation of one embodiment, and is a peak waveform diagram of one sample component of the chromatogram.
FIG. 3 is a flowchart showing the operation of one embodiment.
[Explanation of symbols]
DESCRIPTION OF SYMBOLS 1 Mobile phase container 3 Liquid feed pump 5 Injector 7 Separation column 9 Detector 11 Fraction collector 13 Preparative control device

Claims (2)

A fraction collector that collects the eluted sample components after passing through the detector of the liquid chromatograph in a sorting container, and a fractionation control device that controls the operation of the fraction collector based on the detection signal of the detector In the device
In accordance with the magnitude of the detection signal of the sample component intended for fractionation, the fractionation control device is configured such that the higher the peak height of the target sample component, the smaller the volume fractionated in the sorting container. A fractionation device having a function of variably setting a volume to be fractionated in a fractionation container of the fraction collector. Separation column, liquid feeding means for sending the mobile phase to the separation column, an injector for injecting a sample into the mobile phase flow path to the separation column, a detector for detecting a sample component eluted from the separation column, and passing through the detector In a preparative liquid chromatograph equipped with a fractionation device that collects later eluted sample components in a fractionation container,
The preparative liquid chromatograph according to claim 1, wherein the preparative device is the one according to claim 1.

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