JPH04221759A - High performance liquid chromatograph - Google Patents

Abstract

PURPOSE:To analyze a plurality of samples, needing pretreatment in prior to analysis, in a short time. CONSTITUTION:At least three columns 1, 2, and 3 for pretreatment are connected, in order, to a column 9 for analysis through a switching valve 4 in a manner that the columns are capable of switching. The columns 2 and 3 except the column 1 for pretreatment used in a process to feed a sample to the column 9 for analysis are transferred to pretreatment and conditioning of an analyzing sample, and simultaneous progressive different treatment is effected on the column for analysis. This constitution orderly analyzes a sample, pretreatment of which is completed, during a time in which the sample is separated at the column for analysis.

Description

[Detailed description of the invention]

0001

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a high performance liquid chromatograph for analyzing multiple samples requiring pretreatment in a short time.

0002

[Prior Art] When analyzing a sample containing many components such as blood, a sample is prepared from which components that interfere with the analysis, such as proteins, are removed and used for analysis. ing. Since such preprocessing is usually performed manually, there is a problem in that variations in processing conditions occur and analysis errors occur.

[0003] In order to solve this problem, a high-pressure channel switching means is provided in the analysis channel, and after the interfering components in the sample are discharged from the system, only the components to be analyzed are injected into the analytical column. High performance liquid chromatography has also been proposed (Analytical Chemistry vol. 35, no. 3, 1986
Year).

0004

[Problems to be Solved by the Invention] According to such an apparatus, the reliability of the analysis results is improved because pre-treatment can be carried out automatically, but on the other hand, the reliability of the analysis results is improved. The problem is that it requires time to regenerate the pretreatment column, resulting in low analysis throughput. The present invention was made in view of such problems, and
The objective is to provide a new high performance liquid chromatograph that can analyze multiple samples in succession in a short period of time.

[0005]

[Means for Solving the Problems] In order to solve this problem, in the present invention, a plurality of pretreatment columns are connected between the sample injection mechanism and the analysis column via a flow path switching means. The pretreatment column is selectively connected to the analysis column by the flow path switching means, and the pretreatment column that has been processed is transferred to the conditioning step.

[0006]

[Operation] In parallel with the process of supplying the analysis target component from which impurities have been removed by one pretreatment column to the analytical column, the other pretreatment column that has been pretreated is subjected to a conditioning process. The pretreatment column, which has undergone a conditioning step, is used for sample pretreatment. Thereby, the analysis time in the analytical column can be used to simultaneously perform sample pretreatment and conditioning for regeneration of the pretreatment column.

[0007]

DESCRIPTION OF THE PREFERRED EMBODIMENTS The details of the present invention will be explained below based on illustrated embodiments. FIG. 1 shows an embodiment of the present invention, and reference numerals 1, 2, and 3 in the figure are pretreatment columns, for example, Shim-pac SP manufactured by Shimadzu Corporation.
C-RP1 (product name), high pressure switching valve 4, which will be described later.
In this embodiment, one end communicates with the first mobile phase liquid pump 6 via the sample injection mechanism 5 and the other end communicates with the first waste port 7 by switching the six-way valve; 2
The other end is connected to an analytical column 9, for example Shim-pac CLC-ODS (trade name) manufactured by Shimadzu Corporation, to the mobile phase liquid pump 8, and one end is connected to a third mobile phase liquid pump 10. , and the process of connecting the other end to the second waste port 11.

The analytical column 9 is connected at its outlet to a waste port 13 via a detector 12 for high performance liquid chromatography, such as an ultraviolet spectrophotometer. In addition, the symbols 14, 15, and 16 in the figure each indicate a tank containing a mobile phase.

FIG. 2 shows an embodiment of the above-mentioned high pressure valve, which is composed of a rotating part 20 and fixed parts 21 and 22, and the rotating part 20 has a shaft passing through the fixed parts 21 and 22. The fixed parts 21, 2 are connected to the motor 24 via 23.
It is rotatable relative to 2. Further, the fixing portion 21 is fixed to the base via an attachment member 25. The rotating part 20 and the fixed parts 21, 22 have through holes 20a, 20a, 20a, which communicate with each other at a certain angle on the same circumference, at intervals of 60 degrees in this embodiment.
21a, 21a, 21a..., 22a, 22a, 22a
... are bored, and the through holes 20a, 20a on the rotating part side
, 20a... are connected to the inlets and outlets of the pretreatment columns 1, 2, and 3 by pipes, and on the fixed part side are a sample injection port, second and third liquid pumps 6, 8, 10 discharge ports, an inlet of the analytical column 9, and first and second waste ports 7
, 11 are connected.

In this embodiment, the valve 4 is operated to a position where one end of the first pretreatment column 1 communicates with the sample injection port 5 and the other end communicates with the first waste port 7, and the first liquid supply is started. pump 6
When the first mobile phase, e.g. 0.2 molar phosphate buffer (pH 6.8) is activated, the pump 6
It is delivered at a rate of milliliter/minute, flows into the first pretreatment column through the sample injection mechanism 5, and is discharged from the first waste port 7.

In this state, when a sample is injected into the sample injection mechanism 5, the sample flows into the first pretreatment column 1 by the first mobile phase, and is processed under conditions determined by the pretreatment column 1 and the mobile phase. separated. Among the components separated by the first pretreatment column 1, undesired components that pose an obstacle to analysis are transferred out of the system through the first waste port 7. When the high-pressure switching valve 4 is rotated to the second position when the miscellaneous substances are discharged from the system, the inlet of the first pretreatment column 1 is connected to the second mobile phase liquid feeding pump 8. The outflow port is connected to the inflow port of the analytical column 9. This allows a second mobile phase, for example 0.1 molar phosphate buffer (pH 2.
A mixture of 1) and ethanol in a ratio of 10:9 is supplied to the first pretreatment column 1 and the analysis column 9 at a flow rate of 0.5 ml/min. The target component eluted from the pretreatment column 1 with the second mobile phase is transferred to the analytical column 9.
It is further separated there and detected by a detector 12.

On the other hand, when the first pretreatment column 1 is connected to the analysis column 9, the second pretreatment column 2 is connected between the sample injection mechanism 5 and the first waste port 7. connected and will be supplied with the first mobile phase. Therefore, the sample to be analyzed next is injected into the second pretreatment column 2 in such a way as to match the timing when the analysis column 9 becomes ready for the next analysis. As a result, the next sample is pretreated in the second pretreatment column 2 in parallel with the analysis in the analysis column 9.

When the analysis of the components held in the first pretreatment column 1 is completed, the high pressure switching valve 4 is rotated to the third position, and the first pretreatment column 1 is The inlet is connected to the discharge port of the third liquid feeding pump 10, and the outflow port is connected to the second waste port 11, and the third mobile phase,
For example, cleaning is performed by supplying a solution containing a 1:1 mixture of 0.1% triethylamine solution and acetonitrile at a flow rate of 0.5 ml/min.

While the first pretreatment column 1 is being washed, the third pretreatment column 3 is connected to the sample injection mechanism 5.
Since it is connected to the first waste port 7, the next sample can be pretreated in the same manner as described above, and the second pretreatment column 2 is connected to the second liquid pump 2. It is connected to the analytical column 9, so that the pretreated sample can be discharged to the analytical column 9.

[0015] By repeating the above process, the sample is pretreated by effectively utilizing the processing time in the analytical column.

[0016] In this embodiment, an example has been explained in which three pre-treatment columns are provided, but if three or more pre-treatment columns are provided, the pre-treatment capacity will not only be improved; More complex pre-treatments and conditioning treatments can be performed.

[0017]

[Effects of the Invention] As explained above, in the present invention,
A plurality of pretreatment columns are connected between the sample injection mechanism and the analysis column via a flow path switching means, and the pretreatment columns are selectively connected to the analysis column by the flow path switching means, so that the processing is performed. Since the completed pretreatment column is transferred to the conditioning process, the processing time in the analytical column can be used to pretreat the next sample to be analyzed, and to wash and condition the column after pretreatment. This allows the processing capacity per unit time to be improved.

[Brief explanation of the drawing]

FIG. 1 is a configuration diagram of an apparatus showing an embodiment of the present invention.

FIG. 2 is an exploded perspective view showing an embodiment of a high-pressure switching valve used in the above device.

[Explanation of symbols]

1, 2, 3 Pretreatment column 4 High pressure switching valve 5 Sample injection mechanism 6, 8, 10 Liquid pump 9 Column for analysis

Claims (1)

[Claims] 1. A plurality of pretreatment columns are connected between a sample injection mechanism and an analytical column via a flow path switching means, and the pretreatment column is selectively connected to the analytical column by the flow path switching means. A high-performance liquid chromatograph that is connected to a column and transfers the pretreatment column to the conditioning process after processing.