JPH1010103A - Automatic sample injector in liquid chromatography device and washing method for the injector - Google Patents

Abstract

PROBLEM TO BE SOLVED: To make it possible to perform washing safely without adverse effects on a device even when an organism body is a sample to be measured in the automatic sample injector in a liquid chromatography device having the structure, wherein the sample to be measured is sucked and injected into a sample supply pipeline system connected to a detecting means by using a sample injecting pipe. SOLUTION: A sample to be measured is injected into an introducing valve 14x by using a sample injecting pipe 22. The sample injecting pipe 22 is inserted into a port 21b for washing liquid, wherein the washing liquid having the washing capability for washing the sample to be measured is stored, and the washing liquid is sucked. The sample injecting pipe 22 under the sucked state of the washing liquid is connected to the introducing valve 14x. The injection and the sucking of the washing liquid into and out of the introducing valve 14x are repeated, and the washing is performed. After the washing is finished, the sample injecting pipe 22 is connected to a drain port 38, and the washing liquid after the washing is discharged.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an automatic sample injection device and a method for cleaning the automatic sample injection device in a liquid chromatography apparatus, and more particularly to a method for inhaling a sample to be measured using a sample injection tube and connecting the sample to a detection means. The present invention relates to an automatic sample injector and a method for cleaning the automatic sample injector in a liquid chromatography apparatus configured to inject into a sample supply piping system.

[0002]

2. Description of the Related Art Liquid chromatography apparatuses are widely used as chemical analysis means for separating and quantifying chemical substances.
In particular, a liquid chromatography apparatus using a semi-micro column having an inner diameter of 1 to 2 mm has an advantage of providing high sensitivity, high resolution, and high precision analysis results, and has been actively studied.

FIG. 7 shows an example of a conventional liquid chromatography apparatus. The liquid chromatography apparatus shown in FIG. 1 includes a concentration column 3 for performing primary separation between an autosampler 1 and a 6-way switching valve (hereinafter, referred to as a switching valve) 2 configured as a 6-way valve. I have. The concentration column 3 is filled with a column packing material.
In, the sample to be measured supplied from the autosampler 4 is concentrated.

In order to send out the sample to be measured to the separation column 4 through the switching valve 2 inside the auto sampler 1, an automatic sample injection device, various ports,
The components include components such as various valves and a syringe. Further, the pump 5A and the pump 5B
Constitutes a sample supply system for supplying the sample to be measured to the concentration column 3, the separation column 4 and the detector 6 in cooperation with the switching valve 2.

According to the configuration shown in FIG. 7, the sample to be measured is sent from the autosampler 1 to the concentration column 4 together with the solvent (mobile phase A) supplied by the pump 5A. Is performed. And
The sample to be measured concentrated in the concentration column 4 is supplied to the separation column 4 to perform a separation process together with the solvent (mobile phase B) supplied by the pump 5B. On this occasion,
Switching between the mobile phase A and the mobile phase B is performed by the switching valve 2 described above.

The conveyed sample to be measured is separated in the separation column 4, and subsequently introduced into the detector 6, where a predetermined analysis process is performed on the sample to be measured. The liquid chromatography apparatus having the above-described configuration includes a controller 7 configured by a computer, and the controller 7 controls the driving of the autosampler 1, the switching valve 2, and the pumps 5A and 5B.

Next, the switching operation of the switching valve 2 performed by the control process of the controller 7 will be described. The switching valve 2 switches the flow path of the sample to be measured formed in the liquid chromatography apparatus between the first state and the second state by the control processing of the controller 7.

When the switching valve 2 is in the first state, the sample to be measured, which is pumped together with the mobile phase A from the autosampler 1 by the pump pressure generated by the pump 5A, is switched after passing through the concentration column 3. The sample to be measured is supplied to the valve 2 and further proceeds to a waste liquid storage tank (not shown) along a line indicated by a solid line in the figure. At this time, only the substance to be analyzed contained in the sample to be measured is captured by the concentration column 3 and concentrated. At the same time, the mobile phase B is also supplied to the switching valve 2 by the pump pressure generated by the pump 5B, and flows toward the separation column 4 along the line shown by the solid line in FIG.

On the other hand, when the switching valve 2 is brought into the second state by the control processing of the controller 7, the sample to be measured concentrated in the concentration column 3 is analyzed along a line shown by a broken line in FIG. It is supplied to the flow separation column 4. On the other hand, the mobile phase B supplied from the pump 5B is configured to flow out to a waste liquid storage tank (not shown) along a line shown by a broken line in the figure.

Here, FIG. 8 shows a main part of an automatic sample injection device incorporated in the autosampler 1. In the figure, reference numeral 7 denotes a disk-shaped sample holding unit, and a plurality of sample ports 7a for loading a plurality of types of samples to be measured.
Are formed. The sample holder 7 is configured to be rotatable as indicated by an arrow in the figure.

Reference numeral 8 denotes a sample injection tube, which is a robot 9
Is movable. Further, a suction / discharge device is connected to the sample injection tube 8, and the sample injection tube 8
It is configured such that a liquid can be sucked into the inside and the sucked liquid can be discharged. Accordingly, the sample injection tube 8 is moved by the robot 9 and inserted into the port 7a for the sample to be measured of the sample holding unit 7, and then a suction process is performed by the suction / discharge device. More sample injection tube 8
The sample to be measured can be taken out.

In the figure, reference numeral 10 denotes an introduction valve, which is connected to a pipe connected to the above-mentioned concentration column 3. Therefore, the sample injection tube 8 from which the sample to be measured has been sucked as described above is moved by the robot 9 and inserted into the introduction valve 10, and the sample to be measured in the sample injection tube 8 is transferred to the introduction valve 10 by the suction / discharge device. By injecting (discharging)
A predetermined sample to be measured can be supplied to the concentration column 3.

[0013]

When the sample to be measured is injected into the introduction valve 10 using the sample injection tube 8 as described above, a certain amount of the sample to be measured is placed in the sample injection tube 8 and the introduction valve 10. Adhere to. Therefore, the sample injection tube 8
When the analysis process of another sample to be measured is subsequently performed in a state where the sample to be measured adheres to the introduction valve 10, the previous sample to be measured affects the analysis of the next sample to be measured, and the analysis after the next time is performed. It is conceivable that the accuracy of the processing is reduced.

For this reason, it is necessary to wash the sample injection pipe 8 and the introduction valve 10 every time the analysis processing is completed. Conventionally, the mobile phase A is used as a washing liquid, and this mobile phase A is used as the sample injection pipe. The sample injection tube 8 and the introduction valve 10 can be introduced into the sample injection tube 8 and the introduction valve 10.
Cleaning was performed.

On the other hand, in recent years, the range of use of the liquid chromatography apparatus has been widened, and analysis using a biological substance such as a protein as a sample to be measured has been performed. Therefore, even in the analysis using a biological material as a sample to be measured, it is necessary to wash the biological material as a sample to be measured after completion of the analysis processing.

However, biological materials such as proteins are difficult to wash, and biological materials cannot be washed with a solvent generally used as the mobile phase A. Therefore, the conventional configuration has a problem that a biological substance cannot be used as a sample to be measured.

It is known that biological substances such as proteins are washed with an alkaline solution and a surfactant solution. However, these washing liquids are dangerous if they adhere to the skin, or they constitute a liquid chromatography apparatus. It is a troublesome substance to handle, for example, if it adheres to a metal part or the like that is deteriorated. For this reason, an alkaline solution and a surfactant solution cannot be used in place of the mobile phase A from the viewpoint of maintaining the safety and reliability of the liquid chromatography device.

The present invention has been made in view of the above points, and is a liquid capable of performing a washing treatment safely and without adversely affecting the apparatus even when a biological substance such as a protein is used as a sample to be measured. It is an object of the present invention to provide an automatic sample injection device in a chromatography device and a method for cleaning the automatic sample injection device.

[0019]

The above object can be attained by taking the following means. 2. The automatic sample injection device in the liquid chromatography apparatus according to claim 1, wherein the port for the sample to be measured in which the sample to be measured is stored, and the port for the cleaning solution in which a cleaning solution having a cleaning capability for washing the sample to be measured is stored. And a drain port from which the cleaning liquid is discharged, and a detection unit for performing an analysis process on the sample to be measured, and an introduction valve for introducing the sample to be measured, and a structure movable by a sample injection tube moving unit. And a sample injection tube selectively connected to the port for the sample to be measured, the port for the washing solution, the drain port, or the introduction valve, and suctioning or sucking the sample to be measured or the cleaning solution into or from the sample injection tube. Suction / discharge means for discharging processing, and the sample injection pipe in a state where the cleaning liquid is sucked by driving the sample injection pipe moving means. While connected to the inlet valve, the suction /
A cleaning control unit that repeatedly executes a process of injecting and suctioning the cleaning liquid into the introduction valve by driving a discharge unit.

According to a second aspect of the present invention, in the automatic sample injector of the liquid chromatography apparatus according to the first aspect, a biological substance is used as the sample to be measured, and an alkaline solution or a surfactant solution is used as the cleaning solution. Is used.

According to the third aspect of the present invention, the sample to be measured is inhaled from the port for the sample to be measured in which the sample to be measured is stored using a sample injection tube, and the sample injection tube is used to analyze the sample to be measured. A cleaning method for an automatic sample injection device that performs a cleaning process on an automatic sample injection device that is connected to an introduction valve connected to a detection unit that performs processing, and that automatically injects the measured sample into the introduction valve, After injecting the sample to be measured into the introduction valve by using the sample injection tube, the sample injection tube is inserted into a cleaning solution port storing a cleaning solution having a cleaning ability for washing the sample to be measured, and the cleaning solution is inserted. Then, the sample injection pipe in a state where the cleaning liquid is sucked is connected to the introduction valve, and the cleaning processing is performed by repeatedly injecting and suctioning the cleaning liquid into the introduction valve. , After it said washing process is completed, and is characterized in that for discharging the cleaning liquid after the cleaning process by connecting the sample injection tube to the drain port.

According to a fourth aspect of the present invention, in the cleaning method of the automatic sample injector according to the third aspect, a biological substance is used as the sample to be measured, and an alkaline solution or a surfactant solution is used as the cleaning solution. It is characterized by using.

The first and third aspects of the present invention operate as follows. That is, the cleaning liquid having the cleaning ability for the sample to be measured is stored in the cleaning liquid port.
Further, the cleaning liquid in the cleaning liquid port is sucked into the sample injection tube and connected to the introduction valve only when the cleaning processing is performed by the cleaning control means.

In a state where the sample injection tube and the introduction valve are connected, the washing control means drives the suction / discharge means to inject the cleaning liquid into the introduction valve, and from the introduction valve to the sample injection tube again. The suction process is repeatedly performed. By repeatedly performing the injection and suction processing, the sample to be measured attached to the sample injection tube and the introduction valve is washed.

The used cleaning liquid is discharged to the drain port. Therefore, the washing liquid is not introduced into any part other than the washing liquid port, the sample injection tube, and the introduction valve. Thereby, even if it is difficult to use the cleaning liquid, for example, an alkaline liquid and a surfactant solution, etc., it is possible to prevent the components of the liquid chromatography apparatus from deteriorating and to prevent the cleaning liquid from adhering to the skin or the like carelessly. be able to.

The most problematic of the sample to be measured is the sample injection tube and the introduction valve. That is, the undiluted sample to be measured is directly introduced into the sample injection tube and the introduction valve, but the sample to be measured in a diluted state after the addition of the solvent is introduced to other portions. For this reason, the portions other than the sample injection tube and the introduction valve can be sufficiently cleaned by using a solvent serving as a mobile phase as a cleaning solution as in the related art.

[0027]

Embodiments of the present invention will now be described with reference to the drawings. FIG. 2 shows a schematic configuration of a liquid chromatography apparatus to which an automatic sample injection device and a washing method according to an embodiment of the present invention are applied.

As shown in FIG. 2, the liquid chromatography apparatus has a sample holding section 21 in which a plurality of ports 21a for a sample to be measured are formed. Each of the test sample ports 21a has a capacity of, for example, about 200 μl, and different test samples are stored in the respective test sample ports 21a. In the present embodiment, it is assumed that the measurement sample stored in each measurement sample port 21a contains a biological substance such as a protein.

In this embodiment, one of the plurality of ports 21a for the sample to be measured formed in the sample holder 21 is filled with a cleaning liquid (hereinafter, the port for the sample to be measured filled with this cleaning liquid). Is called a washing liquid port 21b). The washing solution port 21b stores an alkali solution or a surfactant solution as a washing solution for the biological substance such as the protein described above.

On the other hand, reference numeral 22 in the figure denotes a sample injection tube, which is configured to be movable by a robot 22a (corresponding to the above-described injection tube moving means). Wherein each of the sample port 21a and the cleaning liquid port 21b that is inserted into the measured sample port 21a at a position P ₁ where the injection needle of the sample injection tube 22 which is movably held by the robot 22a is shown in FIG. Is done.

The sample injection tube 22 has a thin tube shape, and its material is selected, for example, from stainless steel which is resistant to corrosion. The syringe 15 is connected to the sample injection tube 22 via a resin pipe 32 made of Diflon, Teflon or the like and a joint 15a. The syringe 15 is configured to be driven by a syringe driving device 39, and the sample solution 30 is transferred from the selected sample port 21 a to the sample injection tube 22 by driving the syringe 15.
(The syringe 15 and the syringe driving device 39)
Corresponds to the suction / discharge means described above).

As described above, when the syringe driving device 39 is driven in a state where the sample injection tube 22 is inserted into the sample port 21a, the sample port 21a is inserted.
The sample solution 30 loaded therein is sucked into the sample injection tube 22, and the cleaning liquid is supplied by driving the syringe driving device 39 while the sample injection tube 22 is inserted into the cleaning liquid port 21 b. 22 can be inhaled.

Further, a drain port 38 for discharging the used washing liquid is provided at a position near the sample holding section 21 as described later. This drain port 3
The used cleaning liquid discharged to 8 is discarded in a waste liquid reservoir (not shown). To connect the sample injection tube 22 to the drain port 38, the sample injection tube 22 moved to the position shown in the drawing P ₃ by driving the robot 22a, inserting the sample injection tube 22 to the drain port 38 on its . By driving the syringe driving device 39 in this inserted state, the used cleaning liquid injected into the sample injection tube 22 can be discharged to the drain port 38.

On the other hand, in the sample storage loop 14 ₇ to work hexagonal valve 14 two cooperating, inlet valve 14x for receiving the tip of the sample injection tube 22 is formed. When the liquid chromatography apparatus is in the analysis processing mode, the introduction valve 14x is connected to the sample injection tube 2 from the port 21a for the sample to be measured.
The sample to be measured inhaled in 2 is injected.

When the liquid chromatography apparatus is in the washing mode, a sample injection pipe 22 for sucking the cleaning liquid is connected to the introduction valve 14x, as will be described later, and between the introduction valve 14x and the sample injection pipe 22. The cleaning liquid is injected and sucked in the inlet valve.
The cleaning process of the 4x and the sample injection tube 22 is performed.

As a specific injection method for connecting the sample injection tube 22 to the introduction valve 14x, a robot 22a moves the sample injection tube 22 to a position P ₂ corresponding to the introduction valve 14x.
Then, the sample injection tube 22 is inserted into the introduction valve 14x, and the syringe driving device 39 is operated in this state. This allows the sample to be measured or the cleaning liquid sucked into the sample injection tube 22 to be introduced into the introduction valve 14x.

The six-way valve 14 connected to the introduction valve 14x is connected to the detector 17 via the column 16 and the pump 11 and the waste liquid reservoir 18. The six-way valve 14 connects the introduction valve 14 x to the column 16 when the liquid chromatography apparatus is in the analysis processing mode, and first supplies the sample to be measured to the column 16 together with the solvent supplied from the container 12 by the pump 11. After performing the separation process, the sample is supplied to the detector 17 to perform the analysis process on the sample to be measured.

A second six-way valve 23 is connected to the sample outlet of the detector 17. The sample to be separated separated by the column 16 and further detected by the detector 17 is supplied to the second six-way valve 2.
3 is discharged to the waste liquid reservoir 18 by switching. Depending on the switching state of the six-way valve 23, the sample to be separated can be supplied to the sample injection pipe 22 via the joint 15a.

In the state shown in FIG. 2, the sample to be separated discharged from the detector 17 is a waste liquid reservoir 18 as indicated by an arrow.
Sent to On the other hand, when the sample to be separated is fed to the sample injection tube 22, the sample is supplied to the joint 1 provided on the syringe 15.
The sample injection tube 22 passes through the flow path 15b formed in the
Supplied to The flow path 15b is formed in a T-shape so that both the sample supplied from the syringe 15 and the sample supplied from the six-way valve 23 can be sent to the sample injection tube 22.

Further, in the liquid chromatography apparatus according to the present embodiment, a container 2 filled with a washing liquid for washing the column 16 and a piping system (sample supply piping system) cooperating with the column 16 is provided.
The cleaning liquid is supplied from the container 24 to the pump 25.
Is supplied to the above-mentioned six-way valve 23. In the state shown in FIG. 2, the cleaning liquid is sent to the sample injection tube 22 via the joint 15a and cleans it. When cleaning the sample injection tube 22,
Robot 22a is connected to the sample injection tube 22 to the drain port 38 is located P _3.

However, as shown in FIG. 2, since the washing liquid filled in the container 24 moves widely inside the liquid chromatography apparatus, the washing solution for the biological substance is an alkaline solution or an interface solution. Activator solutions cannot be used. This is because, when an alkaline solution or a surfactant solution is used as a washing solution, the pump 25 and the piping 3
2, because the joint 15a and the like may be deteriorated. For this reason, a liquid agent used as a mobile phase is selected as the washing liquid.

The sample holder 21, the robot 22a, the syringe driving device 39, and the six-way valves 14, 2
3 and the like are connected to a controller 50 (constituting an inhalation / injection control means), and the driving thereof is controlled by the controller 50. Controller 5
Reference numeral 0 denotes, for example, a microcomputer, and each of the devices 1 according to a preset program.
4, 21, 22a, 23, and 39 are driven.

FIG. 3 shows a six-way valve 2 in the apparatus shown in FIG.
3 shows a rotated state. In the state shown in FIG.
The sample to be separated discharged from the detector 17 passes through the six-way valve 23 along the arrow, and is sent to the sample injection pipe 22 via the flow path 15b in the joint 15a. Further, the washing liquid from the container 24 is blocked by the six-way valve 23.

As a result, the sample injection tube 22 is moved to a position corresponding to the appropriate sample port 21a on the sample holder 21 by the robot 22a, so that the sample to be separated discharged from the detector 17 is moved. It becomes possible to collect the sample into the predetermined sample port 21a.

In particular, when a semi-micro column having an inner diameter of 1.0 to 2.0 mm is used as the column 16 for liquid chromatography, since the total flow rate of the solvent is very small at 50 to 200 μl / min, the sample is diluted by the solvent and the sample is diluted. The increase in the volume of the separation sample does not matter, and the sample to be separated can be sufficiently collected by the measurement sample port 21a having a capacity of several hundred μl.

FIGS. 4A and 4B are diagrams illustrating the switching of the flow path of the sample to be separated discharged from the detector 17 by the six-way valve 23. FIG. Referring to FIG. 4 (A), the hexagonal valve 23 has a rotatable valve element 23R, the valve body 23R straight tubular through channel 23 _{1-23 6,} connecting channel connecting these 23a~ 23c are formed. Here, connecting channel 23a is connected to the ₁ and 23 ₆ straight pipe through channel 23,
Connection channel 23b is connected to the straight pipe-shaped through channel 23 ₂ and 23 _3, further connecting channel 23c to connect the straight pipe-shaped through channel 23 ₅ and 23 _6.

In the state shown in FIG.
Channel 23₁Is the sample injection pipe 2 through the syringe joint 15a.
2 and the straight tubular through flow path 23
_TwoMatches the port D connected to the outlet of the detector 17,
Straight tubular through channel 23_ThreeIs the port connected to the waste liquid reservoir 18.
Match with W. On the other hand, the straight tubular through channel 23₆Is the wash
Align with the port C connected to the holding container 24, and
Straight through-flow channel 23 _FiveAnd 23₆Is any port
Also not connected.

In the above configuration, in the state shown in FIG. 4A, the sample to be separated discharged from the detector 17 is discarded from the port D through the connection channel 23b and the port W to the waste liquid reservoir 18. Further, the washing liquid from the container 24 is supplied to the sample injection tube 22 through the connection channel 23a.

The state where the valve element 23R is rotated in the direction of the arrow is shown.
It is shown in FIG. In the state shown in FIG.
I is a straight tubular through channel 23_Two, 23b and 23_ThreeThrough
To the detector 17 and the port C is a straight tubular through-flow
Road 23₁, 23a and 23 ₆Connected to. Scold
The straight tubular through flow path 23₆Is not connected to any port
Therefore, the washing liquid supplied from the container 24 is a straight tubular through flow path.
23₆Is stopped at. Furthermore, it is connected to the waste liquid reservoir 18.
In the state of FIG. 4B, the connected port W is connected to the connection flow path 23c.
Through the straight tubular through channel 23_FiveConnected to, but straight tubular
Through channel 23_FiveBecause there is no port connected to
No discharge of the sample into the waste reservoir 18 occurs.

FIG. 5 is a detailed view of the above-described automatic sample injection device. As shown in the figure, the robot 22a constituting the automatic sample injection device has a base 40 and an arm 44. The base 40 is supported by horizontal shafts 41a and 41b, and is configured to be movable in an arrow X direction in the figure. The arm 44 is supported by vertical shafts 45a and 45b implanted in the base 40, and is indicated by an arrow Z in the drawing.
It is configured to be movable in the direction.

A horizontal movement motor 42 composed of a stepping motor is attached to a chassis (not shown). The horizontal movement motor 42 and the base 40 are connected by a timing belt 43. Therefore, the base 40 moves in the arrow X direction by driving the horizontal movement motor 42.

On the other hand, an elevating motor 46 composed of a stepping motor is attached to the base 40, and the elevating motor 46 and the arm 44 are connected by a timing belt 47. Therefore, the arm 44 moves up and down in the arrow Z direction by driving the up / down motor 46.

The sample injection tube 22 described above is fixed to the arm 44 by a stopper 49. Further, the base 40 is provided with a guide member 48 for protecting the sample injection tube 22 extending downward from the arm 44, particularly near the distal end. Robot 22 configured above
a, each motor 42,
The sample injection tube 22 is moved in the directions indicated by arrows X and Z in the figure by controlling the driving of the sample injection tube 46, and the sample injection tube 22 is moved along with the moving operation to the sample valve provided in the introduction valve 14 x and the sample holder 21. Selectively into or out of the cleaning port 21a, the cleaning liquid port 21b, and the drain port 38. By operating the syringe driving device 39 in accordance with the movement of the sample injection tube 22, the sample to be tested or the cleaning liquid can be sucked into the sample injection tube 22 and injected into the introduction valve 14x.

Subsequently, the sample injection tube 2 which is the main part of the present invention
2 and the cleaning process of the introduction valve 14x will be described.
The cleaning process of the sample injection tube 22 and the introduction valve 14x is executed according to the cleaning process program executed by the controller 50 described above. FIG. 6 shows a cleaning processing program executed by the controller 50, and FIG. 1 shows an operation of the sample injection tube 22 performed according to the cleaning processing program.

In the embodiments shown in FIGS. 2 to 5, the cleaning liquid port 21b is provided in the sample holding section 21, but the cleaning liquid port 21b is not necessarily provided in the sample holding section 21.
It is not necessary to provide the sample holding unit 21, and a configuration provided separately from the sample holding unit 21 may be adopted. Therefore, for convenience of illustration, FIG. 1 shows an example in which the cleaning liquid port 21b is provided separately from the sample holding unit 21.

The specific operation of the cleaning process will be described below with reference to FIGS. When the cleaning processing program shown in FIG. 6 is started, first, in step 10, it is determined whether or not a test sample of a biological substance has been injected into the sample injection tube 22 before the cleaning processing program is started.

If a negative determination is made in step 10, that is, the test sample of the biological substance is
If it is determined that the cleaning liquid is not injected into the cleaning liquid port 2, cleaning processing using the cleaning liquid (alkaline solution or surfactant solution) filled in the cleaning liquid port 21b performed in step 12 and subsequent steps is unnecessary. The cleaning processing program ends without performing the processing of.

On the other hand, if an affirmative determination is made in step 10, that is, if it is determined that the test sample of the biological substance has been injected into the sample injection tube 22, the liquid chromatography apparatus switches to the cleaning mode and the automatic sample injection apparatus is cleaned. Processing is started. First, in step 12, the controller 50 drives the robot 22a to move the sample injection tube 22 to the cleaning liquid port 21b and insert the sample injection tube 22 into the cleaning liquid port 21b. In the subsequent step 14, the controller 50 operates the syringe driving device 39, and uses the syringe 15 to suck a predetermined amount of a cleaning solution such as an alkali solution or a surfactant solution into the sample injection tube 22 from the cleaning solution port 21b. FIG.
(A) shows the operation of step 12 and step 14.

When the cleaning liquid is sucked into the sample injection tube 22 as described above, the processing proceeds to step 16 where the controller 5
0 moves the sample injection tube 22 to the introduction valve 14x and connects the sample injection tube 22 to the introduction valve 14x by driving the robot 22a. Next step 18
Then, the controller 50 operates the syringe driving device 39 to discharge the syringe 15 to inject the cleaning liquid from the sample injection tube 22 into the introduction valve 14x.

In step 20, the controller 5
In the case of 0, the syringe driving device 39 is operated so that the syringe 15 performs the suction operation, and the cleaning liquid introduced into the introduction valve 14x is again sucked into the sample injection tube 22. This step 1
Steps 8 and 20 are performed a predetermined number of times (X times) in steps 22 and 24.

As described above, the controller 50 drives the syringe driving device 39 and the syringe 15 in a state where the sample injection tube 22 and the introduction valve 14x are connected, and injects the cleaning liquid into the introduction valve 14x. 14
By repeatedly performing the process of sucking the sample from the x into the sample injection tube 22 again, the sample injection tube 22 and the introduction valve 14
x cleaning process is performed.

By the way, as described above, the sample injection tube 22
And the introduction valve 14x is a portion into which the undiluted sample to be measured is directly introduced, so that the adhesion of the sample to be measured, which is a biological substance (protein, etc.), is most problematic.
The sample injection tube 22 and the introduction valve 14x. Therefore, as described above, the cleaning liquid is introduced into the introduction valve 14x and the sample injection pipe 22.
By repeatedly performing the injection / suction process between the sample injection tube 22 and the sample injection tube 22 where the adhesion of the sample to be measured is most problematic.
In addition, the inlet valve 14x can be intensively cleaned. FIG. 1B shows the operation of the above-described steps 16 to 24.

On the other hand, if an affirmative determination is made in step 22, that is, if the injection / suction process is performed a sufficient number of times (X times) to wash the sample injection tube 22 and the introduction valve 14x, step 26 is executed. After the counter n used in steps 22 and 24 is cleared, the process proceeds to step 28.

In step 28, the controller 50 drives the robot 22 a to move the sample injection pipe 22 to the drain boat 38 and to move the sample injection pipe 22
Is inserted into the drain boat 38. Next step 30
Then, the controller 50 operates the syringe driving device 39 to drain the cleaning liquid (ie, the used cleaning liquid after the cleaning processing) sucked in the processing of the X-th step 20 using the syringe 15 from the sample injection tube 22. It is discharged to the boat 38.

When the discharging process in step 30 is completed, the process proceeds to step 32, in which the controller 50
4, 21, 22a, 23, and 39 are set in the state shown in FIG. 2, a normal cleaning process is performed using the cleaning solution loaded in the container 24, and the cleaning process is completed. FIG.
(C) shows the operations of step 28 and step 32.

According to the cleaning process shown in FIG. 6 carried out by the controller 50, even if a cleaning solution having a strong erodability of an alkaline solution or a surfactant solution is used, the contamination of the biological fluid is the most problematic with this cleaning solution. The cleaning liquid which is introduced only between the sample injection tube 22 and the introduction valve 14x and which has been used is discharged to the drain port 38. Therefore, even if a strong erosive cleaning liquid is used as described above, it is possible to reliably prevent the components of the liquid chromatography apparatus from deteriorating, and the cleaning liquid may be inadvertently attached to the tester's skin or the like. Can be prevented.

In the liquid chromatography apparatus other than the sample injection tube 22 and the introduction valve 14x,
Since the sample to be measured diluted by the addition of the solvent is introduced, the sample injection pipe 22 and the introduction valve 1
The cleaning of the portion other than 4x can sufficiently wash the sample to be measured by the normal cleaning process performed in step 32.

It should be noted that the cleaning control means described in claim 1 corresponds to steps 18 to 18 performed by the controller 50.
This corresponds to the processing of step 24. In the above-described embodiment, a biological substance such as a protein is used as the sample to be measured.
In addition, although an example in which an alkaline solution or a surfactant solution capable of washing biological substances is used as the washing liquid has been described, the sample to be measured and the washing liquid are not limited to the above substances, and other substances may be used. Of course, you can.

[0069]

As described above, according to the present invention, in a state where the sample injection tube and the introduction valve are connected, the cleaning control means drives the suction / discharge means to inject the cleaning liquid into the introduction valve. Since the process of sucking the sample into the sample injection tube again from the introduction valve is repeatedly performed, the sample injection tube and the introduction valve, in which the contamination of the sample to be measured is particularly problematic, can be reliably washed.

The washing liquid is not introduced into any part other than the sample injection tube and the introduction valve, and the used washing liquid is discharged to the drain port. Even if it is used, it is possible to prevent the components of the liquid chromatography apparatus from deteriorating and to prevent the cleaning liquid from adhering to the skin or the like carelessly.

[Brief description of the drawings]

FIG. 1 is a diagram showing a state in which a sample solution is sucked into an injector by a sample injection method and an injection apparatus according to one embodiment of the present invention.

FIG. 2 is a diagram showing a configuration of a liquid chromatography apparatus to which a sample injection method and an injection apparatus according to one embodiment of the present invention are applied.

FIG. 3 is a diagram for explaining the operation of the liquid chromatography apparatus of FIG.

FIGS. 4A and 4B are diagrams showing the configuration and operation of flow path control means used for collecting a sample to be separated in the apparatus of FIG.

FIG. 5 is an enlarged view showing a configuration near an injector provided in the apparatus of FIG. 3;

FIG. 6 is a flowchart showing the operation of the injection device.

FIG. 7 is a diagram showing an outline of a conventional liquid chromatography apparatus.

FIG. 8 is a schematic diagram of an automatic sample injection device provided in a conventional liquid chromatography device.

[Explanation of symbols]

 11, 25 Pump 14 First six-way valve 14x Introducing valve 15 Syringe 15a Joint 16 Column 17 Detector 18 Waste liquid reservoir 21 Sample holder 21a Port for sample to be measured 21b Port for cleaning solution 22 Sample injection pipe 22a Robot 23 Second hexagon Valve 24 Cleaning liquid container 38 Drain port 39 Syringe driving device 40 Base 42 Horizontal movement motor 44 Arm 46 Elevating motor 48 Guide member 50 Controller

Claims (4)

[Claims] 1. A port for a sample to be measured in which a sample to be measured is stored; a port for a cleaning solution in which a cleaning solution having a cleaning ability for washing the sample to be stored is stored; a drain port from which the cleaning solution is discharged; It is connected to detection means for performing analysis processing on the sample to be measured, and is configured to be movable by an introduction valve for introducing the sample to be measured and a sample injection tube moving means. A sample injection tube selectively connected to a port for use, a drain port, or an introduction valve; suction / discharge means for suctioning or discharging the sample to be measured or the cleaning solution to or from the sample injection tube; By driving the moving means, the sample injection tube in which the washing liquid is sucked is connected to the introduction valve, and the suction / discharge means is driven. An automatic sample injection device in the liquid chromatography apparatus, further comprising: a washing control means for repeatedly performing an injection process and a suction process of the cleaning liquid into the introduction valve. 2. The liquid sample injection device according to claim 1, wherein a biological substance is used as the sample to be measured, and an alkaline solution or a surfactant solution is used as the cleaning solution. Automatic sample injection device in chromatography equipment. 3. The sample to be measured is sucked in from a port for the sample to be measured in which the sample to be measured is stored using a sample injection tube, and the sample injection tube is connected to a detecting means for analyzing the sample to be measured. A cleaning method for an automatic sample injection device that performs a cleaning process on an automatic sample injection device that is connected to the introduction valve, and that automatically injects the sample to be measured into the introduction valve. After injecting the sample to be measured into the introduction valve, the sample injection tube is inserted into a cleaning solution port storing a cleaning solution having a cleaning ability for washing the sample to be measured, and the cleaning solution is sucked in. The sample injection tube in the state in which the cleaning liquid has been connected is connected to the introduction valve, and the cleaning liquid is repeatedly injected into the introduction valve and a suction process is performed to perform a cleaning process. After the cleaning process is completed, Cleaning method in an automatic sample injection apparatus characterized by discharging the cleaning liquid after the cleaning process by connecting the sample injection tube to the drain port. 4. The cleaning method according to claim 3, wherein a biological substance is used as the sample to be measured, and an alkaline solution or a surfactant solution is used as the cleaning solution. Cleaning method in the injection device.