JPH10332660A - Liquid chromatograph and method using the same - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a liquid chromatograph by which a sample containing a plurality of components can be separated selectively by a method wherein the sample is introduced into a separating column, components to be separated are collected into respective component containers during the time corresponding to the eluting time of components which are required to be sampled and the components are resampled so as to be introduced into the column. SOLUTION: A sample is filled into a flow passage system, for sample introduction, from a nozzle 7 by using a syringe 2, the nozzle 7 is moved to a sample injection chamber 11, and the sample is introduced into a metering pipe 13 by using the syringe 2. A six-way valve 12 is changed over, and the sample in the metering pipe 12 is introduced into an eluting liquid flow from a liquid feed pump 14 so as to be sent to a separating column 15. A component which is separated by the separating column 15 is detected by a detector 16, a control circuit 18 controls various valves and the nozzle 7 on the basis of information on it, and the separated component is collected into a primary-separated-component container 9 from the nozzle 7. Then, the nozzle 7 is moved to a rinsing port 5, a three-way valve 3 is changed over, and the nozzle 7 and the like are rinsed by a rinsing liquid. Then, the collected component is sucked again from the primary-separated-component container 9, and the component which is separated by the separating column 15 is collected by a secondary-separated- component container 10.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a liquid chromatograph, and more particularly to a liquid chromatograph suitable for individually collecting components in a mixture sample.

[0002]

2. Description of the Related Art Conventionally, a liquid chromatograph for separating components from a sample in which multiple components are mixed has been conventionally used, as disclosed in, for example, JP-A-62-75261. When peaks are detected overlapping,
The peak of the analyte is once held in a sample loop, re-separated by repeatedly passing through the column by a flow switching operation of an eluent by a six-way valve, and collected by a fraction collector.

[0003]

In the above prior art, when the separation of components is insufficient, the unseparated component regions are separated by repeatedly passing through a separation column. In the case of individual pieces, there was a problem that other components could not be collected during that time.

It is an object of the present invention not only to be able to individually collect a plurality of components in a mixture sample, but also to selectively and automatically re-separate any of the collected components. .

[0005]

In order to achieve the above object, according to the present invention, a sample containing two or more components is introduced into a separation column, and a single sample or a plurality of samples which need to be collected for the sample introduction are introduced. The components separated by the column at the time corresponding to the elution time of the components are collected in each component container, one or each of the collected separated components is collected again, introduced into the column, and separated. And configured to collect.

In addition, a sample containing two or more components is introduced into a separation column, and the sample separated by the column at a time corresponding to the elution time of the first component with respect to the sample introduction is placed in a first container. Before the introduction of the next sample and after the elution of the first component, the sample separated by the separation column
The second sample is collected in the second container at a time corresponding to the elution time of the component, and after the second component is collected, the separated samples collected in the first container and the second container are again collected, respectively. The column was configured to be separated and collected.

Preferably, the liquid chromatograph of the present invention supplies an eluent to a separation column, introduces a mixture sample to be separated into the eluent stream, and individually captures separated components of the sample. In the collecting liquid chromatograph,
Providing a plurality of separation component containers for collecting the eluted components from the separation column, providing a component take-out channel system for individually guiding the components separated by the separation column to the separation component container, Is provided with a sample introduction channel system for collecting a collected component from any of the plurality of separated component containers in which the sample is collected and introducing the collected component upstream of the separation column. Further, preferably, the component extraction channel system and the sample introduction channel system have a common channel portion in their specific channel region, and further have a mechanism for washing the common channel portion. Have.

[0010] Preferably, a sample injection chamber is provided between the eluent liquid sending pump and the separation column, and a nozzle selectively inserted into the separation component container is provided to collect the separated components. A device for operating the nozzle so as to collect a specific component from any of the plurality of separated component containers using the nozzle and to introduce the specific component into the sample injection chamber. Preferably, a connection portion is provided for communicating the nozzle with a flow path downstream of the separation column.
In addition, it is preferable to use a valve that shuts off the flow path downstream of the separation column from the nozzle and communicates with the drainage path at the connection portion.

[0009] Preferably, a plurality of flow paths for guiding outflow components from the separation column are provided corresponding to the separation component container, and one of the plurality of flow paths is selected to select the separation component container. And a device for introducing the components collected in the eluent stream into the eluent. Preferably, a sample valve device is provided between the eluent solution sending pump and the separation column, and the sample valve device is provided with a port through which a sample can be injected from the outside. Preferably, the plurality of flow paths include an on-off valve for selectively opening and closing the plurality of flow paths. Preferably, a downstream side of the separation column is provided with a component taking-out channel system and a connection portion selectively connected to a drainage channel.

[0010] Preferably, in the method using a liquid chromatograph of the present invention, the step of introducing a sample into a separation column, the components separated by the separation column to a plurality of separation component containers via a common flow channel. Collecting, collecting the component from a specific separation component container through the washed common channel, and introducing the component into the separation column.

[0011] Single or plural components in the sample separated by the separation column are supplied to a separation component container provided outside the channel system by a single nozzle device or a flow for guiding elution components from the separation column. It is collected by the operation of the open / close valve provided on the road. If the collected components are insufficiently separated, they can be re-injected and re-separated, and re-collected as high purity components. For this purpose, the components in the separated component container are sucked from the common channel portion, which is a specific portion of the component extraction channel system and the sample introduction channel system, by the syringe suction operation, and then the syringe discharge operation To inject into the separation column through the sample introduction channel system.

[0012]

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Hereinafter, a first embodiment of the present invention will be described with reference to FIG. A sample is placed in a sample container 8, and the sample introduction channel system 4 is moved by the suction operation of the syringe 2.
After the sample is filled in the sample through the nozzle 7, the nozzle 7 moves to the sample injection chamber 11, and the sample is introduced into the measuring tube 13 connected to the hexagonal valve 12 which is a sample valve device by the discharge operation of the syringe 2. At this time, the six-way valve is switched so that the eluent 17 does not pass through the measuring tube 13. When aspirating and discharging the sample, the component extracting flow path system 47 that connects the nozzle 7 and the downstream of the separation column 15 is connected.
A four-way valve 4 which is a connection portion between the two, and a three-way valve connected to the syringe 2 and connected to the cleaning liquid 1 and the four-way valve 4 constitute a flow path shown in FIG. Next, the six-way valve 1
By switching 2, the sample in the measuring tube 13 is introduced into the flow of the eluent sent by the solution sending pump 14 and enters the separation column 15. The components separated by the separation column 15 are detected by a detector 16, and the detected signal is sent to a control circuit 18.
Sent to The control circuit 18 controls various valves and operating devices of the nozzle 7 based on the information of this signal.
Normally, when only the eluent is flowing, the eluent is discharged through the drain 38 of the four-way valve, but by switching the four-way valve, the eluent 17 is directed to the nozzle 7. At this time, while the eluted component from the separation column 15 does not reach the nozzle 7, the eluent 17 is discharged from the nozzle 7 to the drain 48. However, when the eluted component is detected by the detector, the nozzle 7 is discharged.
From the primary separated component container 9 of the plurality of separated component containers.
The components are collected. Next, the nozzle 7 is moved to the washing port 5, the washing liquid is sucked into the syringe 2 by switching the three-way valve 3, and the three-way valve 3 is again switched to suck the washing liquid 1 and the sample introduction flow communicating with the nozzle 7 and the nozzle 7. The common channel portion of the channel system 46 and the component extracting channel system 47 is cleaned. When the separation of the components is insufficient, the nozzle 7 is moved to the position of the primary separation component container 9 again, and the collected components are sucked by the operation of the syringe 2 and introduced into the separation column 15 from the sample injection chamber 11. To separate again. The same operation as described above is repeated, and the components collected for the second time are collected in the secondary separation component container 10. Above is primary collection and secondary collection is 1
In this case, for example, a method in which the primary collection is performed a plurality of times by the number of components and then the secondary collection is performed is also possible. Further, a specific description of the operation mechanism will be given.

FIG. 2 shows a mechanical system of a main part in the embodiment of FIG. The same parts as those in FIG. 1 are given the same numbers.
The nozzle 7 is driven up and down by a vertical drive mechanism 30. When the sample is moved deeply downward when a sample or a component collected for the first time is sucked from the container, when the nozzle 7 is inserted into the sample injection chamber 11 and the sample is introduced into the measuring tube 13, the washing port is used. The case where the nozzle 7 is inserted into the container 5 and the case where the diluting liquid is sucked from the container containing the diluting liquid, if necessary, for example, when the collected components cannot be suctioned from the container due to the minute amount. . The forward / backward moving mechanism 31 moves a mechanism system including the nozzle 7, the arm 6, and the vertical drive mechanism 30 back and forth by the action of the pulse motor 33. The left / right driving mechanism 32 moves the front / rear moving system right and left together with the nozzle 7, the arm 6 and the vertical driving mechanism 30 integrated therewith by the action of the pulse motor 34. The six-way valve 12 is driven by a pulse motor 35, the four-way valve 4 is driven by a pulse motor 43,
The three-way valve 3 is driven by a pulse motor 36. The syringe 2 performs a suction operation or a discharge operation by the action of the syringe drive mechanism 44 driven by the pulse motor 37. Each pulse motor is controlled by the control circuit 18. A sample container 8, a primary separation component container 9, and a secondary separation component container 10 are set in the container storage rack 40, and the component in the sample is collected as described with reference to FIG. If the component to be collected is two components,
In the primary collection, each component is collected in the primary separation component container 9 and the primary separation component container 41 for the component 2, and in the secondary collection, the secondary separation component container 10 and the secondary separation component for the component 2 are collected. Container 4
The operation of collecting each of the components is performed in 2. If there are three or more components to be collected, three or more primary separation component containers,
A similar collection operation is performed using three or more secondary separation component containers.

In the present system, it is necessary to discharge components and eluents that do not need to be collected, cleaning liquid after cleaning the nozzle 7 and the common flow path, etc., and discharge the liquid directly from the nozzle 7 or the drain 38. And collected in a waste bin 39.

In addition, although not shown in the figure, there are a liquid feed pump 14, a separation column 15, and a detector 16. The liquid sending pump 14 and the separation column 15 are connected to the six-way valve 12, and the detector 16 is connected to the four-way valve 4.

FIG. 5 is a time chart for explaining the operation of each section in the embodiment of FIG. 1, and the operation will be described in relation to the operation of the mechanical section.

When the nozzle 7 descends into the sample container 8,
The three-way valve 3 is in a state where a flow path from the syringe 2 to the four-way valve 4 is opened. That is, the state is as shown in FIG. By operating the syringe 2 in the suction direction in this state, the sample in the sample container 8 is sucked into the sample introduction channel system 46 through the nozzle 7. During this time, the six-way valve 12 is in the connected state shown in FIG. 1, and the eluent is sent to the separation column 15 by the liquid sending pump 14.

In the time chart of FIG.
Is in the injection state, that is, when the nozzle 7 is inserted into the sample injection chamber 11, the six-way valve 12 rotates to the sample injection side. FIG. 7 shows the connection state of the six-way valve 12 on the sample injection side. The injected sample is held in the measuring tube 13. When returning to the liquid sending side from this state, the connection state of the six-way valve 12 shown in FIG.
The sample therein is introduced into the separation column 15.

The time chart of FIG. 5 shows that the four-way valve 4 is always in a conductive state between the nozzle 7 and the syringe 2 during the suction and injection of the sample as described above. Therefore, the liquid that has passed through the separation column 15 and the detector 16 is discharged to the drain 38. When the four-way valve 4 becomes conductive between the nozzle 7 and the detector 16 from this state, the liquid that has passed through the separation column 15 and the detector 16 is
It is sent to the nozzle 7 and discharged from the nozzle 7. FIG. 8 shows the state of the four-way valve 4 at this time. In this state, the liquid is first discharged to the drain 38. When the detector 16 detects the rise of the component peak 1, the nozzle 7 moves onto the primary separation component container 9 at a timing considering a slight time lag in which the component moves from the detector 16 to the nozzle 7, and the component 1 is captured. Perform a collection. When the detector 16 detects the fall of the component peak 1, the nozzle 7 moves from the primary separation component container 9 after the same time lag, and the collection of the component indicated as collection 1 in FIG. 5 is completed. . Subsequently, the collection 2 for the component peak 2 based on the information from the detector 16 is performed, but the separated component container is a container different from the collection 1.
When there is no more component peak to be collected, the nozzle 7 moves to the drain 48.

Next, the four-way valve 4 returns to a conductive state between the nozzle 7 and the syringe 2 and the liquid flowing out of the detector 16 is
The water is discharged from the four-way valve 4 to the drain 38. In this state, the three-way valve 3 conducts to the cleaning liquid side, and at the same time, the syringe 2
Operates in the suction direction to suck the cleaning liquid 1. Next, the three-way valve 3 is switched, the nozzle 7 and the three-way valve 3 are electrically connected via the four-way valve 4, and the syringe 2 starts the discharging operation. At this time, the nozzle 7 is inserted into the cleaning port 5, and the cleaning liquid 1 discharged from the nozzle 7 cleans the nozzle 7 and the common flow path communicating with the nozzle 7.

A component that improves the purity of the component is selected from the components collected at the first time, and the second collection operation is subsequently performed. Since the sequence at this time is almost the same as that described above, a description thereof will be omitted. However, since a component that has already been collected once is separated again, only one component peak appears unless the amount of impurity components is extremely large. Component 1 re-collection shown in FIG. After completing the collection of the separated components again as needed among the collected components, the above sequence is started for the next new sample.

FIG. 3 shows the structure of a flow channel of a liquid chromatograph according to a second embodiment of the present invention. A plurality of sample containers 49 and a plurality of separation component containers 50 to 52 are arranged in the container storage rack 19. Each of these containers is provided with a plurality of flow passages 53 communicating with the component extraction flow passage system 46, and the flow passages 53 are provided with opening / closing valves 23a to 23e. The sample valve device 12 provided with a sample injection port 54 connected to the sample introduction channel system 47 between the liquid feed pump 14 and the channel of the separation column 15 is provided. Further, a three-way valve 22 is provided at a connection portion between the downstream of the detector 16 and the flow path system 46 for removing components.

FIG. 3 shows a second embodiment of the present invention.
A specific operation will be described with reference to FIG. 6 showing a time chart of the second embodiment.

First, the open / close valve 23a provided in the channel inserted into the sample container 49 is opened.
At this time, the stop valve 21 is closed and the stop valve 20 is open. Also, the drain valve 23e
Is electrically connected to the drain 55 side, that is, between the detector 16 and the drain. In this state, when the syringe is operated for suction, the sample is sucked, and the sample holder 45 is filled with the sample. Next, the stop valve 20 is closed and the stop valve 21 is closed.
Is opened, and the syringe 2 performs a discharging operation. At this time, the six-way valve 12 rotates toward the sample introduction side, and the sample holding section 4
The sample filled with 5 is introduced into the measuring tube 13 from a sample injection port 54 provided in the six-way valve 12. The sample is sent to the separation column 15 by the liquid sending pump 14 by making the flow path of the six-way valve 12 conductive to the liquid sending side.

Next, when the three-way valve 22 is connected to the downstream of the detector 16 and the flow path 46 for removing components, the effluent from the detector 16 flows from the separation column 16 to the separation component containers 50 to 52. Go to Initially, the on-off valve 23e of the drain 55 is open, but when the detector 16 detects the component peak 1, the on-off valve 23b corresponding to the separated component container 50 that collects the component 1 is opened, and the 1 is collected. Similarly, when the component peak is detected, the open / close valve 23c corresponding to the separated component container 50 that collects the component 2 is opened, and the component 2 is collected.
When the collection of the components is completed, the open / close valve of the drain 55 is opened again, and the eluent 17 is discharged. Next, the three-way valve 22 connects the drain 56 with the downstream of the detector 16 to discharge the eluent 17 from the detector 16 from the drain 56. Next, when the three-way valve 3 connected to the syringe 2 communicates with the cleaning liquid 1, the syringe 2 performs a suction operation, and sucks the cleaning liquid 1 into the syringe 2. When the flow path of the three-way valve 3 is switched again, the stop valve 20 is opened and the stop valve 21 is closed, and the sample introduction flow path system 47 and the component extraction flow path system 46 are shared by the syringe discharge operation. The channel 57 and the sample holder 45 are cleaned.

Hereinafter, it is the same as in the case of FIG. 1 that each of the collected components can be re-separated and re-collected, for example, if the separation is insufficient, or the same operation can be performed if necessary. Detailed description is omitted.

As an effect peculiar to the present embodiment, in addition to the above-described normal mode, a mode in which, when only one kind of component is to be collected, is not collected in a container but is recycled as it is is also possible.

In the above description, the first sample sampling and the sampling for re-separating the collected components were performed from the container arranged in the container storage rack. FIG. This is an example in which the sampling is performed in the same manner as in the second embodiment shown in FIG. It is used in trial collection applications in which a specific sample is injected by manual injection, or in the case of continuously collecting extremely many types of samples. An autosampler can be used instead of the injection valve.

[0029]

According to the present invention, as described above, it is possible to collect a separated component with high accuracy.

Further, since a plurality of separated component containers are provided outside the channel system and a component extracting channel system and a sample introducing channel system are provided, single or plural components in the sample can be individually separated. Not only can a large number of samples be collected,
The components once collected can be introduced into the separation column when necessary, and any components can be repeatedly introduced.

Further, since the component taking-out channel system and the sample introducing channel system are communicated with each other through the common channel section, the sample and the collected components can be sucked and injected in one channel, so that the sample introduction is performed. The structure of the system is simplified.

Further, since the sample and the collected components can be sucked and injected with a single nozzle, the structure of the sample introduction system is simplified and the cost is reduced.

Further, a plurality of flow paths are provided for each of the plurality of separation component containers, and the flow path system is circulated by providing an opening / closing valve in the flow path, so that it can be used as a normal recycled liquid chromatograph. it can.

Since the separation component container serves as both the sampler and the fraction collector, it is not necessary to prepare them separately.

[Brief description of the drawings]

FIG. 1 is a first embodiment of the present invention.

FIG. 2 is an explanatory diagram of a mechanism of a main part in the first embodiment.

FIG. 3 is a second embodiment of the present invention.

FIG. 4 is a modification of the second embodiment.

FIG. 5 is a time chart relating to the operation of the first embodiment.

FIG. 6 is a time chart of the second embodiment.

FIG. 7 is a state diagram of each unit at each timing for explaining FIG. 5 in detail;

8 is a state diagram of each unit at each timing for explaining FIG. 5 in detail.

9 is a state diagram of each unit at each timing for explaining FIG. 5 in detail.

[Explanation of symbols]

1: cleaning solution, 2: syringe, 3, 22: three-way valve, 4
... four-way valve, 5 ... washing port, 7 ... nozzle, 8,49
... sample container, 9 ... primary separation component container, 10 ... secondary separation component container, 11 ... sample injection chamber, 12 ... hexagonal valve, 13 ...
Measuring tube, 14 ... Liquid sending pump, 15 ... Separation column, 16 ...
Detector, 17 ... Eluent, 18, 24 ... Control circuit, 19,
40: container storage rack, 20, 21: stop valve,
23: open / close valve set, 23a to 23e: open / close valve, 25: injection valve, 29: three-way joint, 30: vertical drive mechanism, 31: front / rear drive mechanism, 32
... left and right drive mechanism, 41 ... primary collection container for component 2, 42 ...
Secondary collection container for component 2, 44 ... Syringe drive mechanism, 45
.., A sample holding section, 46, a component taking-out channel system, 47, a sample introducing channel system, 48, a drain, 50, 51, 52, a separation component container, 53, a channel portion, 54, a sample injection port, 5
7 ... Common flow path section.

Continuing on the front page (72) Inventor Katsuo Tsukada 4-6-6 Kanda Surugadai, Chiyoda-ku, Tokyo Inside Hitachi, Ltd.

Claims (15)

[Claims] 1. A liquid sending pump for sending a mobile phase, an introducing means for introducing a sample containing two or more components into the mobile phase to form a mixed solution, a separation column for separating the mixed solution, In a liquid chromatograph having detection means arranged downstream of the separation column, the components separated by the separation column at a time corresponding to the elution time of one or more components that need to be collected for the sample introduction Is collected in each component container, one or each of the collected separated components is collected again, introduced into the column, and separated and collected. . 2. A liquid sending pump for sending a mobile phase, an introducing means for introducing a sample containing two or more components into the mobile phase to form a mixed solution, a separation column for separating the mixed solution, In a liquid chromatograph having detection means arranged downstream of the separation column, a sample separated by the column is captured in a first container at a time corresponding to an elution time of a first component with respect to the sample introduction. Collecting the sample separated by the separation column before the introduction of the next sample and after elution of the first component in a second container at a time corresponding to the elution time of the second component, The liquid chromatography is characterized in that the separated samples collected in the first container and the second container after collection of the second component are again separated and collected in the column, respectively. Graph. 3. The method according to claim 1, wherein a plurality of separated component containers are provided for individually collecting the separated components of the sample, and for collecting the eluted components from the separation column, and separating the separated columns by the separation column. Providing a component take-out channel system for individually guiding the separated components to the separated component container, collecting the component from any of the plurality of separated component containers in which the separated components are collected, and performing the separation. A liquid chromatograph comprising a sample introduction channel system for introducing upstream of a column. 4. A liquid chromatograph according to claim 3, wherein said component extracting channel system and said sample introducing channel system are provided with a common channel portion in their specific channel regions. . 5. The liquid chromatograph according to claim 4, further comprising a mechanism for cleaning said common flow path. 6. A liquid chromatograph according to claim 1, wherein a plurality of separated component containers are provided in the liquid chromatograph for individually collecting the separated components of the sample, and a plurality of separated component containers are provided between the eluent liquid sending pump and the separation column. Providing a sample injection chamber, providing a nozzle that is selectively inserted into the separated component container, collecting a specific component with the nozzle from any of the plurality of separated component containers in which separated components are collected, A liquid chromatograph comprising a device for operating the nozzle so as to introduce the liquid into the sample injection chamber. 7. The liquid chromatograph according to claim 6, further comprising a connecting portion for communicating the nozzle with a flow path downstream of the separation column. 8. The liquid chromatograph according to claim 7, wherein the connection portion is a valve that shuts off a flow path downstream of the separation column from the nozzle and communicates with a drain passage. 9. A liquid chromatograph according to claim 1 or 2, wherein a plurality of separated component containers are provided in a liquid chromatograph for individually collecting separated components of the sample, and a plurality of containers are provided for guiding elution components from the separation column. A channel portion corresponding to the separation component container, and a device for selecting any one of the plurality of flow channels and introducing a component collected in the separation component container into the flow of the eluent. A liquid chromatograph characterized by the above-mentioned. 10. A sample valve according to claim 9, further comprising a sample valve device between the eluent liquid sending pump and the separation column, wherein the sample valve device is provided with a port through which a sample can be injected from the outside. Liquid chromatograph. 11. A liquid chromatograph according to claim 9, wherein said plurality of flow paths are provided with on-off valves for selectively opening and closing said flow paths. 12. The liquid chromatograph according to claim 9, further comprising a connection portion selectively connected to a component take-out channel system and a drainage channel downstream of the separation column. 13. A step of introducing a sample containing two or more components into a separation column, and separating the sample by the column at a time corresponding to the elution time of one or more components that need to be collected for the sample introduction. Collecting the collected components in each component container, collecting one or each of the collected separated components again, introducing the collected components into the column, separating and collecting the separated components. A method using a liquid chromatograph. 14. A step of introducing a sample containing two or more components into a separation column, wherein the sample separated by the column at a time corresponding to the elution time of the first component with respect to the sample introduction is subjected to the first step. Collecting the sample separated by the separation column before the introduction of the next sample and after elution of the first component, at a time corresponding to the elution time of the second component. Collecting in a container, and after collecting the second component, separating the separated samples collected in the first container and the second container by the column again and collecting the separated samples. A method using a liquid chromatograph, comprising: 15. The method according to claim 13, wherein the components separated by the separation column are collected in a plurality of separation component containers via a common flow channel, the common flow channel is washed, Collecting a component from a specific separation component container through the common channel and introducing the component into the separation column.