Preparative liquid chromatograph

Abstract

A liquid chromatograph of the present application can continuously perform separation and fractionation of a plurality of samples and includes a separation flow path provided with a liquid feed pump, a separation column, and a detector; a plurality of liquid handlers each including an injection section that performs an injection operation when introduced into the separation flow path and a fractionation section that performs a fractionation operation when introduced into the separation flow path; a selector that selectively switches the liquid handlers introduced into the separation flow path; and a controller that controls the operations of the liquid handlers and the selector and introduces the plurality of liquid handlers into the separation flow path in a prescribed order and causes each liquid handler to perform the injection operation and the fractionation operation, sequentially performing separation and fractionation of a plurality of samples assigned to each of the liquid handlers, the controller causing each of the liquid handlers to perform a preparation operation for performing the injection operation during introduction of the other liquid handler into the separation flow path and to perform the injection operation immediately after being introduced into the separation flow path.

Description

TECHNICAL FIELD

[0001] The present application relates to a preparative liquid chromatograph. BACKGROUND

[0002] A preparative liquid chromatograph is an apparatus that separates components in a sample injected into a mobile phase by a syringe and detects peaks of the components, and fractionates and collects portions of eluent corresponding to the peaks into different containers. As elements constituting the preparative liquid chromatograph, there are a pump for feeding the mobile phase, a syringe for injecting the sample into the mobile phase, a separation column for separating the components in the sample, a detector for detecting peaks of the components separated by the separation column, and a fraction collector for collecting the peaks detected by the detector into different collection containers.

[0003] In addition, in the preparative liquid chromatograph, a liquid handler having a function of both the syringe and the fraction collector is sometimes used (see Patent Literature 1). If the liquid handler having the function of both the syringe and the fraction collector is used, it is possible to automatically re-inject the components fractionated and collected in the fraction collector and perform separation and collection.

[0004] PRIOR ART DOCUMENTS

[0005] PATENT LITERATURE

[0006] Patent Literature 1: Japanese Patent Laid-Open No. 2000-162217 SUMMARY

[0007] PROBLEMS TO BE SOLVED BY THE INVENTION

[0008] The liquid handler includes a needle for performing a suction and injection operation of the sample and a probe for performing a collection operation of the peaks of the separated components, and the needle and the probe cannot be operated completely independently in many cases, and the operation of the needle in the collection operation of the peaks using the probe is limited in many cases. Therefore, in a case where separation and collection of a plurality of samples are performed, after the collection operation using the probe is completed, it is necessary to perform a preparation operation such as suction of the next sample, and it is not possible to continuously perform separation and collection of a plurality of samples.

[0009] The present application has been made in view of the above problems, and an object thereof is to enable continuous performance of separation and collection of a plurality of samples in a preparative liquid chromatograph using a liquid handler.

[0010] MEANS OF SOLVING THE PROBLEMS

[0011] The preparative liquid chromatograph of the present application includes: a separation flow path provided with a liquid feed pump that feeds a mobile phase, a separation column that separates components in a sample, and a detector that detects a wave peak of a component separated by the separation column; a plurality of liquid handlers each including an injection portion that functions as an injector that performs an injection operation of injecting a sample into a mobile phase flowing in the separation flow path upstream of the separation column in the separation flow path when introduced into the separation flow path, and a fraction collection portion that functions as a fraction collector that performs a fraction collection operation of a wave peak of a component separated in the separation column when introduced into the separation flow path; a selector that selectively switches a liquid handler introduced into the separation flow path among the plurality of liquid handlers; and a controller that controls operations of the plurality of liquid handlers and the selector, and is configured to introduce the plurality of liquid handlers into the separation flow path in a prescribed order and cause each liquid handler to perform the injection operation and the fraction collection operation, sequentially perform separation and fraction collection of a plurality of samples each assigned to the plurality of liquid handlers, and cause each of the plurality of liquid handlers to perform a preparation operation for the injection operation during introduction of another liquid handler into the separation flow path and immediately after introduction into the separation flow path.

[0012] Effects of the Invention

[0013] In the preparative liquid chromatograph of the present application, since the plurality of liquid handlers are introduced into the separation flow path in the prescribed order and each liquid handler is caused to perform the injection operation and the fraction collection operation, separation and fraction collection of the plurality of samples each assigned to the plurality of liquid handlers are sequentially performed, and each of the plurality of liquid handlers is caused to perform the preparation operation for the injection operation during introduction of another liquid handler into the separation flow path and immediately after introduction into the separation flow path, so separation and fraction collection of the plurality of samples can be continuously performed. BRIEF DESCRIPTION OF DRAWINGS

[0014] Figure 1 is a flow path structure diagram showing an embodiment of a preparative liquid chromatograph.

[0015] Figure 2 is a block diagram showing a control system of the embodiment.

[0016] Figure 3 is a flowchart for explaining an example of an operation of the embodiment.

[0017] Figure 4 is a timing chart for explaining an example of an operation of each liquid handler in the embodiment. Detailed Implementation

[0018] The following is a reference to the appendix. Figure 1 An embodiment of the preparative liquid chromatograph of the present invention will be described below.

[0019] like Figure 1 As shown, the preparative liquid chromatograph of the embodiment includes a separation flow path 1 for separating and separating samples. From the upstream side, a delivery pump 2, a switching valve 10, a switching valve 12, a separation column 4, a detector 6, and a switching valve 14 are arranged on the separation flow path 1. The preparative liquid chromatograph also includes two or more liquid processors 8-1 to 8-n, each having an injection section 16 and a separation section 18. Switching valves 10, 12, and 14 are all multi-port valves and are interconnected to form a selector that selectively introduces any one of the liquid processors 8-1 to 8-n into the separation flow path 1. Regarding the introduction of the liquid processor into the separation flow path 1, taking liquid processor 8-1 as an example, the injection section 16 of liquid processor 8-1 exists between the delivery pump 2 and the separation column 4, and the separation section 18 of liquid processor 8-1 is connected downstream of the detector 6.

[0020] When the liquid processors 8-1 to 8-n are introduced into the separation flow path 1, each injection unit 16 functions as an injection syringe that performs an injection operation to inject a sample into the mobile phase flowing in the separation flow path. When the liquid processors 8-1 to 8-n are introduced into the separation flow path 1, each fractionation unit 18 functions as a fraction collector that performs a capture operation to collect the peaks of the sample separated in the separation column 4.

[0021] Each injection unit 16 of liquid processors 8-1 to 8-n includes a needle 20, a high-pressure valve 22, a syringe 24, a moving mechanism 26, and an injection port 28. One port of the high-pressure valve 22 of each injection unit 16 is connected to the port of the switching valve 10 via an inlet flow path 30, and the other port is connected to the switching valve 12 via an outlet flow path 32. Thus, the switching valves 10 and 12 can selectively introduce any one of the injection units 16 of liquid processors 8-1 to 8-n into the space between the delivery pump 2 and the separation column 4.

[0022] A sampling flow path 21, a needle cylinder 24, a drain, and an injection port 28 are connected to the other port of the high-pressure valve 22 of each injection section 16, which communicates with the needle 20. The high-pressure valve 22 is a 2-position valve that switches between an injection mode and a loading mode. In the injection mode, the port connected to the inlet flow path 30 communicates with the port connected to the injection port, and the port connected to the outlet flow path 32 communicates with the port connected to the sampling flow path 21. In the loading mode, the port connected to the inlet flow path 30 communicates with the port connected to the outlet flow path 32, the port connected to the needle cylinder 24 communicates with the port connected to the sampling flow path 21, and the port connected to the injection port 28 communicates with the port leading to the drain.

[0023] The needle 20 of the injection section 16 is three-dimensionally movable by a moving mechanism 26. The moving mechanism 26 is, for example, an arm that can move three-dimensionally. The injection section 16 can pull out the needle 20 from the injection port 28 and perform a preparation operation such as washing the needle 20 or sucking a sample from a sample container when the high-pressure valve 22 is in the loading mode. After sucking a sample from the tip of the needle 20, the needle 20 is inserted into the injection port 28 and connected, and the high-pressure valve 22 is switched to the injection mode, whereby the sample is injected into the mobile phase flowing in the separation flow path 1. In the embodiment, each injection section 16 is configured to function as a syringe of a full-volume injection method when introduced into the separation flow path 1, but the present application is not limited thereto, and each injection section 16 can be configured to function as a syringe of a loop injection method when introduced into the separation flow path 1.

[0024] A tapping flow path 38 leading to each tapping section 18 of the liquid handler 8-1 to the liquid handler 8-n is connected to each port of the switching valve 14. The switching valve 14 can selectively connect any one of the tapping sections 18 of the liquid handlers 8-1 to 8-n to the downstream of the detector 6. Each tapping section 18 of the liquid handlers 8-1 to 8-n includes a probe 34 and a solenoid valve 36. The solenoid valve 36 of each tapping section 18 selectively connects the tapping flow path 38 to either the probe 34 or the drain. The probe 34 is three-dimensionally movable by the moving mechanism 26 shared with the needle 20 of the injection section 16.

[0025] As Figure 2As shown, the operations of the liquid handlers 8-1 to 8-n, the switching valve 10, the switching valve 12, and the switching valve 14 can be controlled by the controller 40. The controller 40 is configured to introduce the liquid handlers 8-1 to 8-n into the separation flow path 1 in a prescribed order while switching the switching valve 10, the switching valve 12, and the switching valve 14 in accordance with a program set in advance, and perform separation and fractionation of a plurality of samples assigned to each of the liquid handlers 8-1 to 8-n. In the fractionation of the samples, the peaks of components in the samples are detected from a chromatogram obtained based on the output signal of the detector 6, and the eluent portions corresponding to each of the peaks are fractionally captured into different capture containers. The controller 40 can be realized by a computer circuit including a Central Processing Unit (CPU) or the like.

[0026] With Figure 1 and Figure 2 together Figure 3 The operation of the preparative liquid chromatograph of the embodiment will be described with reference to a flowchart.

[0027] A plurality of samples to be separated and fractionated are set by a user in the liquid handlers 8-1 to 8-n in a dispersed manner. When an instruction to start separation and fractionation is input to the controller 40, the controller 40 introduces the liquid handler in which a sample to be separated and fractionated is initially set into the separation flow path 1 (step 101), and injects the sample into the injection portion 16 of the liquid handler (step 102). Thereby, the fractionation operation of the components in the sample is started. The preparation operation in the initial liquid handler, that is, the operation of cleaning the needle 20 or the operation of sucking the sample from the sample container can be performed before the initial liquid handler is introduced into the separation flow path 1, or can be performed after being introduced into the separation flow path 1.

[0028] In the case where there is a sample (next sample) for which separation and fractionation should be performed next (step 103), the controller 40 causes the liquid handler (next liquid handler) to which the next sample is assigned (provided) to perform a preparation action for performing sample injection (step 104). The preparation action includes at least an action of aspirating the sample from the tip 20. When the fractionation action of the sample being performed is completed (step 105: Yes), the controller 40 switches the switching valve 10, the switching valve 12, and the switching valve 14 to introduce the next liquid handler into the separation flow path 1 (step 106), and immediately injects the sample that has been previously aspirated from the tip 20 by the preparation action into the mobile phase (step 102). Thereafter, steps 102 to 106 are repeated for all the samples, and the process ends when the fractionation action of the last sample is completed (step 107). Further, whether the fractionation action of the sample has been completed can be determined by whether a prescribed time has elapsed from the last detected peak or whether a prescribed time has elapsed from the injection of the sample into the mobile phase.

[0029] By the action, continuous separation and fractionation of a plurality of samples as shown in Figure 4 Fig. 6 can be achieved. That is, during the performance of the separation and fractionation of the first sample using the liquid handler (LH1) to which the first sample is assigned, the preparation action of the liquid handler (LH2) to which the second sample is assigned is completed, and the separation and fractionation of the second sample using the liquid handler (LH2) is started at the same time as the completion of the separation and fractionation of the sample using the liquid handler (LH1). Then, during the performance of the separation and fractionation of the second sample using the liquid handler (LH2), the preparation action of the liquid handler (LH3) to which the third sample is assigned is completed, and the separation and fractionation of the third sample using the liquid handler (LH3) is started at the same time as the completion of the separation and fractionation of the second sample using the liquid handler (LH2).

[0030] Further, in the example, the next liquid handler is caused to start the preparation action triggered by the injection of the previous sample into the mobile phase and the start of separation and fractionation, but the present application is not limited thereto. In general, as long as the preparation action of the predetermined liquid handler that performs the injection action or the fractionation action of the sample is completed before the liquid handler is introduced into the separation flow path 1, the present application is not limited thereto.

[0031] The above-described embodiment merely shows one example of the preparative liquid chromatograph according to the present application. The preparative liquid chromatograph according to the present application is as shown below.

[0032] In the preparative liquid chromatograph of the present application, there are included: a separation flow path provided with a liquid feed pump for feeding a mobile phase, a separation column for separating components in a sample, and a detector for detecting a wave peak of components separated by the separation column; a plurality of liquid handlers each including an injection portion which functions as an injector performing an injection operation of injecting a sample into a mobile phase flowing in the separation flow path upstream of the separation column when introduced into the separation flow path, and a fraction collection portion which functions as a fraction collector performing a fraction collection operation of the wave peak of components separated in the separation column when introduced into the separation flow path; a selector for selectively switching a liquid handler introduced into the separation flow path among the plurality of liquid handlers; and a controller which controls operations of the plurality of liquid handlers and the selector, and which is configured to introduce the plurality of liquid handlers into the separation flow path in a prescribed order and cause each liquid handler to perform the injection operation and the fraction collection operation, sequentially perform separation and fraction collection of a plurality of samples each assigned to the plurality of liquid handlers, and cause each of the plurality of liquid handlers to perform a preparation operation for the injection operation during a period when the other liquid handler is introduced into the separation flow path and perform the injection operation immediately after being introduced into the separation flow path.

[0033] In a first aspect of the embodiment, the controller is configured to cause a predetermined liquid handler to be introduced next into the separation flow path to start the preparation operation while the liquid handler introduced into the separation flow path is performing the injection operation.

[0034] In a second aspect of the embodiment, the injection portion includes a needle for performing suction and dispensing of a sample, and the fraction collection portion includes a probe for dropping an eluate from the separation column into a recovery container, and the needle and the probe are configured to be three-dimensionally moved by a common moving mechanism. In a liquid handler in which the moving mechanisms of the needle of the injection portion and the probe of the fraction collection portion are shared, the needle and the probe cannot be operated independently of each other, and therefore, during performance of the fraction collection operation of the sample using the probe, the suction operation of the next sample and the like for preparation of injection of the next sample cannot be performed. Therefore, in a preparative liquid chromatograph including only one liquid handler, separation and fraction collection of a plurality of samples cannot be continuously performed. In the present application, even in a liquid handler in which the moving mechanisms of the needle of the injection portion and the probe of the fraction collection portion are shared, a plurality of such liquid handlers can be used in combination to continuously perform separation and fraction collection of a plurality of samples.

[0035] Explanation of symbols

[0036] 1: Separation flow path

[0037] 2: feed pump

[0038] 4: separation column

[0039] 6: detector

[0040] 8-1 to 8-n: liquid handler

[0041] 10, 12, 14: switching valve

[0042] 16: injection section

[0043] 18: fraction collection section

[0044] 20: needle

[0045] 22: high pressure valve

[0046] 24: syringe

[0047] 26: moving mechanism

[0048] 28: injection port

[0049] 30: inlet flow path

[0050] 32: outlet flow path

[0051] 34: probe

[0052] 36: solenoid valve

[0053] 38: fraction collection flow path

Claims

1. A preparative liquid chromatograph, comprising: A separation flow path provided with a pump for delivering a mobile phase, a separation column for separating components in a sample, and a detector for detecting a peak of a component separated by the separation column; A plurality of liquid handlers each including an injection portion that functions as an injector that performs an injection operation of injecting a sample into a mobile phase flowing in the separation flow path when introduced into the separation flow path, and a fraction collection portion that functions as a fraction collector that performs a fraction collection operation of a peak of a component separated in the separation column when introduced into the separation flow path; A selector for selectively switching a liquid handler among the plurality of liquid handlers that is introduced into the separation flow path; and A controller that controls operations of the plurality of liquid handlers and the selector, and is configured to sequentially perform separation and fraction collection of a plurality of samples assigned to the plurality of liquid handlers by introducing the plurality of liquid handlers into the separation flow path in a prescribed order and causing each liquid handler to perform the injection operation and the fraction collection operation, The injection portion includes a needle for performing suction and dispensing of a sample, and the fraction collection portion includes a probe for dropping an eluate from the separation column into a recovery container, and the needle and the probe are configured to be three-dimensionally moved by a common moving mechanism, The controller is configured to cause each of the plurality of liquid handlers to perform a preparation operation for performing the injection operation during a period in which another liquid handler is introduced into the separation flow path, and to perform the injection operation immediately after being introduced into the separation flow path, the preparation operation including an operation of sucking a sample from a tip of the needle. The controller is configured to cause a predetermined liquid handler to be introduced into the separation flow path next to the liquid handler that is performing the injection operation to start the preparation operation while the liquid handler that is performing the injection operation is performing the injection operation.

2. The preparative liquid chromatograph of claim 1, wherein, ​

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