WO1997040357A1

WO1997040357A1 - Automatic extracting equipment and automatic concentration measuring equipment for component substance in liquid sample

Info

Publication number: WO1997040357A1
Application number: PCT/JP1997/001366
Authority: WO
Inventors: Naotaka Sawada; Akiyoshi Baba; Katsuhiko Saito; Yasumasa Hosokawa; Hajime Miyoshi; Shintaro Nishimura; Masayoshi Murata
Original assignee: Dainippon Seiki Co., Ltd.; Fujisawa Yakuhin Kogyo Co., Ltd.
Priority date: 1996-04-19
Filing date: 1997-04-18
Publication date: 1997-10-30

Abstract

Equipment, which can automatically perform a series of operations in solvent extraction when subjecting component substances contained in liquid samples to solvent extraction and measuring concentrations of the component substances, attain reduction of an operator's labor and time and make effective use of space. An automatic extracting equipment comprises a table holding a plurality of sample tubes, a table holding a plurality of containers for extraction and a plurality of containers for storage, a sample dispensing unit for sucking liquid samples from the sample tubes to discharge the same into the containers for extraction, a solvent dispensing unit for discharging an organic solvent into the containers for extraction, a shaker for transferring target component substances, in liquid samples contained in the containers for extraction, into the organic solvent, and a separation liquid dispensing unit for sucking sample separation liquids separated in the containers for extraction to discharge the same into the containers for storage.

Description

Description Automatic extraction device and automatic concentration measurement device for component substances in liquid samples

The present invention automatically performs a solvent extraction operation of a specific component substance contained in a liquid sample such as a homogenate (refining) such as serum, plasma, whole blood, urine, and biological tissue, and a reaction mixture. An automatic extraction device capable of automatically extracting all the components contained in a liquid sample and measuring the concentration of the components, and an automatic concentration measurement device capable of automatically performing all operations up to the measurement of the concentration. The present invention also relates to a liquid dispensing device and a centrifugal sedimentation tube used for such a device.

Background art

For example, to measure the concentration of a drug contained in blood, an organic solvent is used. The drug component is dissolved in the organic solvent from blood and separated (solvent extraction), and the drug component is dissolved in the organic solvent. After concentrating the sample separation liquid as needed, the sample separation liquid is injected into analytical equipment such as high-performance liquid chromatography. One example of this series of measurement operations is to aspirate a blood sample from a sample tube and discharge a blood sample to a centrifuge sedimentation tube (hereinafter referred to as a “centrifuge tube”). —Dispense the organic solvent for extraction into the centrifuge tube—Attach the cap to the centrifuge tube—Shake the centrifuge tube—Centrifugation—Remove the cap from the centrifuge tube—Sample separation liquid from the centrifuge tube (organic solvent layer) Inhalation of the liquid containing the target component dissolved in the sample side) and discharge of the sample separation liquid into a test tube for concentration (dispensing of the sample separation liquid) —Concentration —The drug concentration in the blood is measured by sequentially performing the operations of inhaling the concentrated sample separated solution from the test tube and injecting the concentrated sample separated solution into the high performance liquid chromatography.

By the way, conventionally, operations such as shaking and centrifugation of a centrifuge tube have been performed using a shaker or a centrifuge, but dispensing of a blood sample, dispensing of an organic solvent, and dispensing of a sample separated solution are performed. Each operation in the note was performed manually using a micro- or hole-piped with a disposable disposable tip (hereinafter referred to as “disposal tip”) detachably attached to the tip. In addition, the operations of attaching and detaching the cap to and from the centrifuge tube, inhaling the concentrated sample separation liquid from the test tube, and injecting the liquid into the high-performance liquid mouth chromatography were also performed manually.

Various dispensing operations can be automated by a liquid dispensing device equipped with a dispensing nozzle, a syringe and its driving motor, a dispensing nozzle moving mechanism, and the like. However, in a general method in which the amount of liquid to be sucked into the dispensing nozzle is controlled by controlling the driving amount of the syringe, the specific gravity, viscosity, surface tension, etc. of the liquid to be sucked are different. In addition, due to changes in the temperature around the dispensing nozzle, it may not be possible to accurately adjust the amount of liquid to be suctioned as desired. In addition, when an organic solvent is sucked into the dispensing nozzle, particularly in the dispensing operation of the sample separation liquid, the organic solvent such as getyl ether, ethyl acetate, and chloroform is highly volatile and has a high viscosity. Low, low surface tension, and low specific gravity make it easy for liquid to drip from the lower end of the dispensing nozzle when moving the dispensing nozzle from the suction position to the dispensing position. Furthermore, after separating into an upper layer solution and a lower layer solution by centrifugation, the sample separation liquid separated into the lower layer side which is the organic solvent layer in the centrifuge tube It was very difficult to automate the operation of fractionating only _W without causing contamination. For these reasons, dispensing operations in liquid-liquid solvent extraction have conventionally been performed manually, and shakers and centrifugal separators have been used.However, the entire process of solvent extraction has been automated. There was no such device in the past.

Disclosure of the invention

A large number of samples must be analyzed and processed at once at a clinical laboratory center or a pharmaceutical company's laboratory where the above-mentioned analysis work such as measurement of drug concentration in blood is performed. In the case where a large amount of sample must be analyzed at one time, performing individual operations such as the dispensing operation described above requires a lot of labor and time. . Further, in the case where the above-mentioned operations are shared by a plurality of workers, a relatively large work space that does not hinder the work of each person is required.

The present invention has been made in view of the above circumstances, and solvent extraction of a specific component substance such as a drug contained in a liquid sample such as serum, plasma, whole blood, homogenate, and reaction mixture. Then, when measuring the concentration, a series of operations for solvent extraction can be performed automatically, and all the steps from the solvent extraction to concentration measurement can be performed automatically. In addition to reducing the labor and time required by the operator for the series of operations, it is possible to effectively use the space, and it is also possible to effectively use the space. It is an object of the present invention to provide an automatic extraction device and an automatic concentration measuring device for component substances in a liquid sample, and to provide a liquid dispensing device and a centrifugal sedimentation tube suitably used for those devices. Make it an issue. As means for solving the above technical problems, a first invention is a sample holding unit for holding a plurality of sample containers containing a liquid sample, an extraction container holding unit for holding a plurality of extraction containers, A predetermined amount of a liquid sample is taken out of the sample container taken out of the sample holding unit or held in the sample holding unit, and the sucked liquid sample is taken out of the extracting container holding unit or the extraction container is taken out. Sample dispensing means for discharging into an extraction container held by a holding unit; extraction solvent dispensing means for discharging a predetermined amount of an organic solvent for extraction into the extraction container; A component substance transfer means for transferring the target component substance in the liquid sample into the organic solvent for extraction, a container holding portion for holding a plurality of containers, and the inside of the extraction container A predetermined amount of the sample separation liquid in which the target component substance is dissolved in the organic solvent is separated and then aspirated, and the sucked sample separation liquid is taken out of the storage container holding section or transferred to the storage container holding section. A separation liquid dispensing means for discharging into a held storage container, wherein an automatic extraction device for component substances in the liquid sample is configured.

In the automatic extraction device having the above configuration, a predetermined amount of the liquid sample is sucked from the inside of the sample container by the sample dispensing means, and the liquid sample is discharged into the extraction container. Next, a predetermined amount of the organic solvent for extraction is discharged into the extraction container by the solvent dispensing means for extraction, and the target component substance in the liquid sample entering the container for extraction is discharged by the component substance transfer means. It is transferred to the organic solvent for extraction. Then, a predetermined amount of the sample separation liquid separated in the extraction container is sucked by the separation liquid dispensing means, and the sample separation liquid is discharged into the storage container. In this way, a sample separation liquid in which the target component substance is dissolved (migrated) in the organic solvent for extraction is automatically obtained. Using the automatic extraction device having the above configuration, a series of operations for solvent extraction of specific component substances such as drugs contained in liquid samples such as serum, plasma, whole blood, homogenate, and reaction mixture can be performed. Since the operations can be performed automatically, it is possible to reduce the labor and the time required for the operator for the series of operations conventionally, and to effectively use the space.

In addition, the automatic extraction device having the above configuration can be provided with an evaporation / drying means for evaporating the sample separation liquid contained in the storage container and drying the sample separation liquid, whereby an automatic extraction device can be configured. A dissolving solvent dispensing means for discharging a predetermined amount of the dissolving organic solvent into the receiving vessel; and a dissolving means for dissolving the dried residue in the dissolving organic solvent in the receiving vessel. A motion extraction device can be configured. As the dissolving means, a shaker for shaking the container may be provided. Further, it is preferable that the evaporating and drying means comprises a heater for ripening the storage container from the periphery thereof, and a nitrogen gas supply means for blowing nitrogen gas into the storage container through an upper opening thereof.

In addition, the automatic extraction device having the above configuration may be provided with a concentrating means for evaporating a part of the organic solvent of the sample separation solution contained in the storage container and concentrating the sample separation solution, thereby configuring the automatic extraction device. . The concentrating means may be constituted by a heater for heating the storage container from the periphery thereof, and a nitrogen gas supply means for blowing nitrogen gas into the storage container through an upper opening thereof. The separation liquid dispensing means holds a dispensing nozzle that sucks a predetermined amount of the sample separation liquid contained in the extraction container from the lower end port and discharges the sample separation liquid from the lower end port, and the dispensing nozzle. A nozzle holding means, the nozzle holding means having a lower position where the lower end of the dispensing nozzle is immersed in the sample separation liquid in the extraction container, and a lower end of the dispensing nozzle which is upward from the extraction container.

-0- A nozzle elevating means for elevating and lowering between the separated upper position, a nozzle moving means for moving the nozzle holding means between a position immediately above the extraction container and a dispensing position, and the dispensing nozzle A syringe for allowing a predetermined amount of the sample separation liquid in the extraction container to be sucked in from the lower end of the syringe, and discharging the sample separation liquid in the dispensing nozzle from the lower end of the dispensing nozzle at the dispensing position; It is preferable to provide a syringe driving means for driving and a syringe control means for controlling the syringe driving means. In such a configuration, when the lower end of the dispensing nozzle is pulled out of the sample separation liquid in the extraction container, the lower end of the dispensing nozzle is inserted into the dispensing nozzle when the lower end of the dispensing nozzle comes out of the sample separation liquid. Means to generate air bubbles inside the sample separation liquid in the dispensing nozzle and continue this state until immediately before the sample separation liquid in the dispensing nozzle is discharged. It is preferable that the separation liquid dispensing means has the following. Also, the separation liquid dispensing means is configured such that when a predetermined amount of the sample separation liquid is sucked into the dispensing nozzle having the lower end immersed in the sample separation liquid in the extraction container, the upper end of the sample separation liquid is Is provided at a height position where is located, and a liquid level sensor that photoelectrically detects whether the upper end of the sample separation liquid has reached the height position is provided, based on a detection signal of the liquid level sensor. Preferably, the syringe driving means is controlled by the syringe control means to stop driving the syringe. The nozzle raising / lowering means is constituted by a stepping motor whose driving amount is controlled by the number of pulses, and when the dispensing nozzle is lowered in order to immerse the lower end of the dispensing nozzle into the sample separation liquid in the extraction container. A nozzle detecting means for detecting whether the lower end of the dispensing nozzle has reached the reference height position is provided, and the lower end of the dispensing nozzle is set at the reference height by the nozzle detecting means. It is preferable that a signal of a fixed number of pulses is input to the stepping motor from the time when the position is detected. The dispensing nozzle may be configured using a disposable suction pipe.

Further, the automatic extraction device having the above-described configuration may be provided with a water or aqueous solution dispensing means for discharging a predetermined amount of water or an aqueous solution into the extraction container, thereby configuring the automatic extraction device.

In addition, the automatic extraction device having the above configuration can be provided with a cap attaching / detaching means for attaching / detaching the cap to / from the extraction container, thereby configuring the automatic extraction device. In addition, the automatic extraction device having the above configuration may be provided with a shaker that shakes the extraction container as a component substance transfer means, to configure the automatic extraction device.

In addition, the automatic extraction device having the above configuration is provided with a centrifugal separator for centrifuging a liquid in which a target component substance has been transferred from the liquid sample into the organic solvent for extraction by the component substance transfer means, and the automatic extraction is performed. The device can be configured.

Next, as means for solving the above technical problems, a second invention is directed to an automatic extraction apparatus having the above-described various configurations, a concentration measuring means for measuring the concentration of a component substance in a liquid sample, and By injecting a component solution obtained by dissolving a target component material in an organic solvent and injecting a predetermined amount of the sucked component solution into the concentration measuring device, the component material in the liquid sample is provided. Characterized in that the automatic concentration measuring device of (1) is constituted. The liquid injecting means may have a measuring tube for holding a predetermined amount of the component solution to be injected into the concentration measuring means.

In the automatic concentration measuring apparatus according to the second aspect of the present invention, the liquid injection means (4) A component solution (sample separation solution, sample solution or concentrated sample separation solution) obtained by the automatic extraction device and obtained by dissolving the target component substance in an organic solvent is sucked from the container and the component solution is collected. Only the quantitative amount is injected into the concentration measuring means, and the concentration of the component substance in the liquid sample is measured by the concentration measuring means.

According to a third aspect of the present invention, there is provided a sample holding unit for holding a plurality of sample containers containing a liquid sample, a container holding unit for holding a plurality of containers, A predetermined amount of a liquid sample is sucked out of a sample container held in the sample holding unit or taken out of the holding unit, and the sucked liquid sample is taken out of the container holding unit or the container holding unit is sucked out. Sample dispensing means for discharging into a container held in a container, extracting solvent dispensing means for discharging a predetermined amount of an organic solvent for extraction into the container, and a target in a liquid sample contained in the container. A component substance transfer means for causing the component substances to flow into the extraction organic solvent; a concentration measuring means for measuring the concentration of the component substances in the liquid sample; and a component substance which is separated and intended in the container. A sample separation liquid dissolved in a solvent is sucked, and a predetermined amount of the sample separation liquid thus sucked is injected into the concentration measuring means. Is constituted. An automatic concentration measuring device having such a configuration is provided with a water or aqueous solution dispensing means for discharging a predetermined amount of water or an aqueous solution into a container, whereby the automatic concentration measuring device can be constituted. In addition, the automatic concentration measuring device having the above-described configuration is provided with a cap attaching / detaching means for attaching / detaching the cap to / from the container, whereby the automatic concentration measuring device can be constituted. In addition, the automatic concentration measuring device having the above-mentioned configuration is visually checked from the liquid sample by the component substance transfer means. An automatic concentration measuring device can be configured by providing a centrifuge for centrifuging a liquid in which a target component substance is transferred into an organic solvent for extraction. In the automatic concentration measuring device according to the third aspect of the present invention, a predetermined amount of the liquid sample is sucked from the sample container and discharged into the container by the sample dispensing means. Next, a predetermined amount of the organic solvent for extraction is discharged into the container by the solvent dispensing means for extraction, and the target component substance in the liquid sample contained in the container is discharged by the component substance transfer means. Moved inside. Then, a predetermined amount of the sample separation liquid separated in the container is aspirated by the separation liquid injection means, and the predetermined amount of the sample separation liquid is injected into the concentration measurement means, and the components in the liquid sample are analyzed by the concentration measurement means. The concentration of the substance is measured.

When each of the automatic concentration measuring devices according to the second and third aspects of the present invention is used, a specific component substance such as a drug contained in the liquid sample is extracted with a solvent and then the solvent is extracted. Since the entire series of operations up to the measurement of the concentration of the component substances can be performed automatically, the labor and time required by the operator for the series of operations have been reduced, while This enables efficient use of space, and allows the operator to simply set the sample container containing the sample in the sample holder, and quickly obtain accurate data on the concentration of the components contained in the sample. can get.

Next, a fourth invention provides a dispensing nozzle for sucking a predetermined amount of liquid contained in a liquid container from a lower end port and discharging the liquid from the lower end port, and a nozzle holding means for holding the dispensing nozzle. Raising and lowering the nozzle holding means between a lower position where the lower end of the dispensing nozzle is immersed in the liquid in the liquid container and an upper position where the lower end of the dispensing nozzle is separated upward from the liquid container. Let A nozzle elevating means, a nozzle moving means for moving the nozzle holding means between a position immediately above the liquid container and a dispensing position, and a liquid in the liquid container from the lower end of the dispensing nozzle. A syringe for inhaling only a fixed amount and discharging the liquid in the dispensing nozzle from the lower end of the dispensing position at the dispensing position, a syringe driving means for driving the syringe, and a syringe control for controlling the syringe driving means Means for dispensing the dispensing nozzle, when the lower end of the dispensing nozzle is pulled out of the liquid in the liquid container, when the lower end of the dispensing nozzle comes out of the liquid, the liquid is dispensed into the dispensing nozzle. Continue to inhale a small amount of air from the lower end port to continuously generate bubbles inside the liquid in the dispensing nozzle, and keep this state until just before the liquid in the dispensing nozzle is discharged. Characterized by comprising a bubble generator.

Further, in the liquid dispensing apparatus having the above configuration, a control circuit for controlling the syringe driving means so as to drive the syringe by switching at a low speed is provided in the syringe control means, and the bubble generating means is configured. be able to.

Further, in the liquid dispensing apparatus having the above configuration, the low-speed syringe, low-speed syringe driving means for driving the low-speed syringe at a low speed, low-speed syringe control means for controlling the low-speed syringe driving means, Flow path switching means for selectively connecting the above-mentioned syringe for sucking a liquid into the dispensing nozzle and discharging the liquid from the lower end of the dispensing nozzle and the low-speed syringe to the dispensing nozzle; Thus, the bubble generating means can be constituted.

When performing a liquid dispensing operation using the liquid dispensing device with the above configuration, a predetermined amount of liquid is injected into the dispensing nozzle, and the lower end of the dispensing nozzle is pulled out of the liquid in the liquid container. Between the time when the lower end of the dispensing nozzle is above the liquid and the time immediately before the liquid in the dispensing nozzle is discharged at the dispensing position. The inside of the dispensing nozzle is continuously suctioned so that a minute flow of air is continuously sucked into the dispensing nozzle from the lower end port, and bubbles are continuously generated in the liquid in the dispensing nozzle. For this reason, a slight upward suction force always acts near the lower end of the dispensing nozzle, and this is a case when dispensing liquids such as hydrochloric acid and organic solvents such as getyl ether. Also, the liquid is prevented from dripping from the lower end of the dispensing nozzle.

When the liquid dispensing device with the above configuration is used, it is easy to evaporate like an organic solvent, has a low viscosity, has a low surface tension, and dispenses a liquid with a low specific gravity or a liquid with a high specific gravity such as hydrochloric acid. Even in this case, it is possible to reliably prevent liquid dripping from the lower end of the dispensing nozzle without being affected by changes in the surrounding temperature, and to prevent a decrease in dispensing accuracy.

According to a fifth aspect of the present invention, there is provided a dispensing nozzle for sucking a predetermined amount of liquid contained in a liquid container from a lower end port and discharging the liquid from the lower end port, and a nozzle holding means for holding the dispensing nozzle. The nozzle holding means is moved up and down between a lower position where the lower end of the dispensing nozzle is immersed in the liquid in the liquid container and an upper position where the lower end of the dispensing nozzle is separated upward from the liquid container. A nozzle elevating means, a nozzle moving means for moving the nozzle holding means between a position immediately above the liquid container and a dispensing position, and a liquid in the liquid container from the lower end of the dispensing nozzle. A syringe for inhaling only a fixed amount and discharging the liquid in the dispensing nozzle from the lower end of the dispensing position at the dispensing position, a syringe driving means for driving the syringe, and the syringe driving means are controlled. In a liquid dispensing apparatus including a syringe control means, when a predetermined amount of liquid is sucked into the dispensing nozzle in a state where a lower end port is immersed in the liquid in the liquid container, an upper end of the liquid is Liquid at the level where it is located A liquid level sensor for photoelectrically detecting whether the upper end of the liquid has reached the height position, and controlling the syringe driving means by the syringe control means based on a detection signal of the liquid level sensor. In this case, the driving of the syringe is stopped.

In the liquid dispensing apparatus having the above configuration, the nozzle elevating means is constituted by a stepping motor whose driving amount is controlled by the number of pulses, and the lower end of the dispensing nozzle is immersed in the liquid in the liquid container. Nozzle detecting means for detecting whether the lower end of the dispensing nozzle has reached the reference height position when lowering the dispensing nozzle is provided, and the lower end of the dispensing nozzle is set at the reference height by the nozzle detecting means. From the point in time when it is detected that the position has been reached, a signal of a fixed number of pulses can be input to the stepping motor.

Further, in the liquid dispensing device having the above configuration, it is preferable that the nozzle detecting unit is configured by a photoelectric sensor that photoelectrically detects a lower end of the dispensing nozzle. When performing a liquid dispensing operation using the liquid dispensing apparatus according to the fifth aspect of the present invention, the lower end of the dispensing nozzle is immersed in the liquid in the liquid container and the lower end of the dispensing nozzle is closed. In the process where the syringe is driven and the liquid in the liquid container is sucked into the dispensing nozzle from the lower end of the dispensing nozzle while stopped at a fixed position, the upper end of the liquid sucked into the dispensing nozzle is in a predetermined position. If it is photoelectrically detected that the height position has been reached, the drive of the syringe is stopped at that point, and the suction operation of the liquid into the dispensing nozzle is stopped. Therefore, when the operation of sucking the liquid into the dispensing nozzle is completed, the upper end position of the liquid sucked into the dispensing nozzle is always at a position higher than the lower end of the dispensing nozzle by a certain distance, and accordingly, the liquid Regardless of the type or ambient temperature, Even if there is a slight leak at the connection part of the pipe, a certain amount of liquid will always be sucked into the dispensing nozzle.

When the liquid dispensing apparatus according to the fifth aspect of the present invention is used, the dispensing operation can be performed irrespective of the type of liquid to be dispensed and without being affected by changes in the ambient temperature. Even if there is a slight leak at the nozzle connection, etc., a predetermined amount of liquid can always be drawn into the dispensing nozzle accurately and without variation, improving dispensing accuracy. You.

According to a sixth aspect of the present invention, there is provided a centrifuge tube comprising a tubular container body having an open upper surface and a cap covering the upper surface opening of the container body, wherein the cap is provided with a through hole at the center. A tightly-sealed plug portion having an outer diameter smaller than the inner diameter of the container body, the upper end portion being connected to a through hole of the sealed plug portion, An inner tube portion having a length such that the lower end is positioned near the inner bottom surface of the container body when the stopper portion is closely fitted to the upper end portion, and a lower end of the inner tube portion is liquid-tightly closed. In addition, it is characterized by comprising a closed part which easily falls off or ruptures by a downward pressing force.

In the centrifuge tube having the above configuration, the closing portion of the cap may be a filling plug inserted into a lower end of the inner tube portion or a thin plate portion integrally formed at a lower end of the inner tube portion of the cap.

In the centrifuge tube having the above configuration, after the liquid to be separated is centrifugally injected into the container body, the inner tube portion of the cap is inserted deeply into the container body and inserted into the liquid. The sealed stopper is tightly fitted to the upper end of the container body, and the container is then centrifuged. As a result, the liquid in the centrifuge tube becomes The liquid is separated into an upper liquid and a lower liquid according to the difference between the two. At this time, the inner tube of the cap is inserted into the liquid in the container body and its lower end is located near the inner bottom surface of the container body, so that the lower end of the inner tube is liquid-tightly closed and the upper layer is closed. It is located below the boundary between the liquid and the lower liquid, and the lower end of the inner tube is inserted into the lower liquid. In order to extract only the lower layer liquid from the centrifuge tube in such a state, the lower end of the dispensing nozzle (pitta etc. in the case of manual operation. Insert the cap deeply into the inner tube through the through hole in the cap, and press the lower end of the dispensing nozzle at the lower end of the inner tube. As a result, the obstruction that obstructs the lower end of the inner pipe portion falls off or bursts, and the lower end of the dispensing nozzle is inserted into the lower liquid. After this, when the liquid is sucked into the dispensing nozzle by driving the syringe, etc., only the lower layer liquid is sucked into the dispensing nozzle because the lower end of the dispensing nozzle is inserted into the lower layer liquid. . Since the lower end of the inner tube, and thus the lower end of the dispensing nozzle, is located just below the boundary between the upper liquid and the lower liquid, a part of the upper liquid mixes with the lower liquid and contaminates the lower liquid. There is no worry about mining.

When the centrifugal tube having the above configuration is used, the lower layer can be formed without any contamination with the upper layer liquid from the two or more liquid phase systems separated into the upper layer liquid and the lower layer liquid by centrifugal force. Only liquid can be extracted. When the extraction operation is performed manually, almost no skill is required. On the other hand, when the extraction operation is performed automatically, the automation is easy.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 shows an embodiment of the present invention, in which an automatic detection of component substances in a liquid sample is performed. FIG. 2 is a perspective view showing the entire configuration of the concentration measuring device. FIG. 2 is a plan layout view of the automatic concentration measuring device shown in FIG. FIG. 3 is a diagram for explaining an example of a series of operation steps for measuring the concentration of a drug contained in a blood sample, and FIG. 4 is a flowchart of the operation steps shown in FIG. FIG. 5 is a diagram for explaining another example of a series of operation steps for measuring the concentration of a drug contained in a blood sample, and FIG. 6 is a diagram illustrating a series of steps for measuring the concentration of a drug contained in a blood sample. FIG. 9 is a view for explaining still another example of the operation step of FIG. Fig. 7 is a front view of the dispensing head of the sample dispensing unit, which is one of the components of the automatic concentration measuring device shown in Figs. 1 and 2, and Fig. 8 is shown in Fig. 7. FIG. 9 is a left side view of the dispensing head, and FIG. 9 is a diagram for explaining the configuration and operation of a cap chuck unit provided in the dispensing head shown in FIG. FIG. 10 is a diagram showing a configuration of a solvent dispensing unit which is one of the components of the automatic concentration measuring device shown in FIGS. 1 and 2, and FIG. 11 is a diagram showing a configuration of the solvent dispensing unit shown in FIG. It is a cross-sectional view of the dispensing arm of the dispensing unit. FIG. 12 is a side view of a dispensing head of a separation solution dispensing unit which is one of the components of the automatic concentration measuring device shown in FIGS. 1 and 2. Fig. 13 is a diagram for explaining the configuration and operation of the cap removal unit provided on the separation liquid suction stage, which is one of the components of the automatic concentration measurement device shown in Figs. 1 and 2. FIG. 14 is a front view partially showing the configuration of a centrifuge tube fixing unit provided on the separation liquid suction stage. Fig. 15 is a schematic diagram showing the configuration of the main part of the separation liquid dispensing unit, which is one of the components of the automatic concentration measuring device shown in Figs. 1 and 2. FIG. 6 is a longitudinal sectional view for explaining a method of performing a dispensing operation of a sample separation liquid in a centrifuge tube using the separation liquid dispensing unit shown in FIG. Figure 17

-is- 2 is a schematic block diagram of a separation solution dispensing unit, which is one of the components of the automatic concentration measuring apparatus shown in FIG. 2 and FIG. 2, and FIG. 18 uses the separation solution dispensing unit shown in FIG. Fig. 19 is a longitudinal sectional view for explaining a method of dispensing a sample separation liquid in a centrifuge tube by using the method shown in Fig. 19. It is a longitudinal cross-sectional view for explaining. Fig. 20 shows the centrifuge tube and cap used when a sample separation liquid separated to the lower layer side in the centrifuge tube is sucked into the disposable tip of the dispensing nozzle, and the cap is removed from the centrifuge tube. FIG. 2 is a perspective view, and FIG.

Fig. 1 is a longitudinal sectional view showing a state in which a cap is attached to the centrifuge tube. Figure

Method 2 uses a centrifuge tube with the cap shown in Figure 20 and Figure 21 to inhale only the sample separation liquid separated into the lower layer of the liquid separated into the upper liquid and the lower liquid. FIG. 4 is a view for explaining the above, and is a view showing a part in a longitudinal section. FIG. 23 is a longitudinal sectional view showing a configuration example of a centrifuge tube cap different from those shown in FIGS. 20 and 21. Fig. 24 is a front vertical sectional view showing the configuration of the concentration stage, which is one of the components of the automatic concentration measurement device shown in Figs. 1 and 2, and Fig. 25 is shown in Fig. 24. FIG. 4 is a side vertical sectional view of the concentrated stage. Fig. 26 is a partially cutaway side view showing the structure of the main part of the engine unit, which is one of the components of the automatic concentration measuring device shown in Figs. 1 and 2. Fig. 27 shows the flow path configuration of the injection unit shown in Fig. 26, and the injection unit allows the dispensing of the solvent into the test tube, the inhalation of the sample solution from the test tube, and the HPLC. FIG. 5 is a schematic diagram for explaining a method of performing an operation up to injection of a sample solution into a column. Fig. 28 and Fig. 29 are included in frozen serum by the automatic concentration measurement device shown in Fig. 1 and Fig. 2. 5 is a flowchart showing an example of a series of operations for automatically measuring the concentration of a specific component substance.

BEST MODE FOR CARRYING OUT THE INVENTION

First, three examples of a series of operation steps for measuring the concentration of a drug contained in a liquid sample, for example, blood, will be described with reference to FIGS.

First, as shown in Fig. 3 (a), for example, the blood serum collected by administering a drug to an animal is centrifuged and the frozen serum (specimen) stored in a sample tube 10 with a lid is thawed. After homogenization, remove the cap 12 of the sample tube 10 and put the sample liquid (melt) from the sample tube 10 into the disposable tip (not shown) attached to the tip of the dispensing nozzle of the liquid dispenser. 0.1 ml of serum) (Fig. 3 (b)), discharge the sucked sample solution into the centrifuge tube 14, and further add an organic solvent such as ethyl acetate, for example 4 ml. And 0.5 ml of pH buffer and 0.5 ml of methanol are dispensed into a centrifuge tube 14 ((c) in FIG. 3). Next, after attaching the cap 16 to the centrifuge tube 14 ((d) in FIG. 3), the centrifuge tube 1 is shaken by a shaker ((e) in FIG. 3). Make sure that the target component (drug) in the sample solution is sufficiently transferred into the organic solvent. Subsequently, the centrifuge tube 14 containing the sample solution and the organic solvent is set in the centrifuge 18 and the solution is centrifuged ((f) in Fig. 3). By this centrifugation, the liquid in the centrifuge tube 14 is separated into the upper layer A and the lower layer B, and the sample separation liquid in which the target component substance is dissolved in the organic solvent forms the upper layer A ( Figure 3 (g)). Therefore, after removing the cap 16 of the centrifuge tube 14, the disposable tip 22 (see (h) in Fig. 3) attached to the tip of the dispensing nozzle (not shown) of the liquid dispensing device. Centrifuge tube 14 to upper layer A For example, 3 ml of the sample separated liquid is inhaled, and the sucked sample separated liquid is discharged into the test tube 24 ((h) in FIG. 3). Next, the test tube 24 is heated from the surroundings, and nitrogen gas is blown from the gas supply nozzle 26 into the inside of the test tube 24 through the upper opening thereof, so that the organic liquid of the sample separation liquid in the test tube 24 is removed. Evaporate the solvent to dry the sample separation liquid ((i) in Fig. 3). Finally, an organic solvent, for example, methanol (〇.lml) is dispensed into the test tube 24 and stirred to dissolve the residue. Then, the sample solution is sucked from the test tube 24 with the nozzle 28. ((J) in Fig. 3) Then, the inhaled sample solution is injected into an analytical device such as high-performance liquid chromatography by, for example, 20 to 30 µl, and the concentration of the component substance is measured. Figure 4 shows a flowchart of this series of operations.

Next, in the example shown in Fig. 5, when the liquid in the centrifuge tube 14 is separated into the upper layer Α and the lower layer Β by centrifugation, the target component substance is dissolved in the organic solvent. This is an operation example in which the liquid forms the lower layer B, the sample separated liquid in the lower layer B is sucked from the centrifuge tube 14, and the sucked sample separated liquid is discharged into the test tube 24. The operation other than dispensing the sample separation liquid in the lower part B from the centrifuge tube 14 is the same as the example shown in FIG. In the operation example shown in FIG. 4, the sample separation liquid in the lower layer B is sucked without removing the cap 30 from the centrifuge tube 14. In this case, a cap 30 having a special structure is used as a cap to be attached to the centrifuge tube 14, which will be described later in detail.

FIG. 6 shows an example of a blood analysis operation using the direct protein removal method. First, as shown in Fig. 6 (a), the frozen serum (sample) contained in the sample tube 10 with a lid is thawed and homogenized, and then the sample tube 1 Remove the cap 12 of the sample dispenser and aspirate the sample solution, for example 0.1 m1, from the sample tube 10 into a disposable tip (not shown) attached to the tip of the dispensing nozzle of the liquid dispenser. ((B) of FIG. 6), the sucked sample liquid is discharged into a centrifuge tube 32 with a lid, and a predetermined amount of an organic solvent, for example, 0.2 ml of methanol or 0.5 ml of methanol is added. Dispense the acetonitrile into the centrifuge tube 32 (Fig. 6, (c)). Next, after attaching the cap 34 to the centrifuge tube 32 (FIG. 6 (d)), the centrifuge tube 32 is shaken with a shaker (FIG. 6 (e)). Next, a centrifuge tube containing the sample solution and organic solvent

Set 32 in the centrifuge 18 and centrifuge the solution (Fig. 6, (f)). By this centrifugation, the liquid in the centrifuge tube 32 is separated into a liquid layer and a sedimentation part. Then, since the target component substance is dissolved in the liquid layer, after removing the cap 34 of the centrifuge tube 32, the sample is separated from the liquid layer in the centrifuge tube 32 by the nozzle 28. The liquid is inhaled ((g) in Fig. 6), and the inhaled sample separation liquid is injected into an analytical device such as high-performance liquid chromatography by, for example, 20 to 50 µl, to obtain the concentration of the component substances. Is measured.

Next, a configuration example of an automatic concentration measuring device for a component substance in a liquid sample will be described. FIG. 1 is a perspective view showing the entire configuration of the automatic concentration measuring device, and FIG. 2 is a plan layout diagram thereof.

This automatic concentration measurement device automatically performs a series of operations to extract a specific component substance (for example, a drug) contained in a sample solution such as blood with a solvent. Syringe pump unit for sending organic solvent, etc. to 6 40 or multiple storage containers for storing organic solvent, etc.

4 2, Supply liquid with waste liquid tank (not shown) etc. Drain section 38 Automatic solvent extraction section 36 Automatically measures the concentration of component substances extracted by solvent It consists of an analytical instrument and, on this side, high-performance liquid chromatography (hereinafter referred to as “HPLC”). The automatic solvent extraction unit 36 is disposed on the upper surface of the apparatus, and the liquid supply / drainage unit 38 and the HPLC 44 are housed in the cabinet below the automatic solvent extraction unit 36, respectively. . An operation panel 46 is provided on the front of the device. Although not shown, the automatic solvent extraction unit 36 is covered with a transparent cover that can be freely opened and closed.

It should be noted that, instead of providing an integrated analytical instrument such as HPLC as shown in the figure, a device for automatically extracting the component substances in the liquid sample with a solvent was obtained by the automatic extraction device. The liquid may be injected into an attached analytical instrument or into a separate analytical instrument.

The automatic solvent extraction unit 36 includes a circular turntable 48, a processing turntable 50, a sample dispensing unit 52 (the structure is not shown in FIG. 2), a sample suction stage 54, and a solvent dispensing unit. 56, Shaking stage 58, Centrifuge 60, Separation liquid dispensing unit 62 (Structure is not shown in Fig. 2), Separation liquid suction stage 64, Evaporation to dryness stage 66, Solvent content It is composed of a stage 68, an injection unit 70, a disposable chip rack 72, a waste port 73, and the like.

The circular turntable 48 has a large number of sample tube holders 74 that hold sample tubes (eg, 1.5 m1 microtubes) 10 with lids containing samples such as frozen serum, and a disposable tip. And a plurality of disposable chip holding portions 76 holding the disposable chips, which are rotated by a rotation drive mechanism (not shown) to control the stop position. In addition, The table 50 has a number of centrifuge tube holders 78 holding centrifuge tubes (for example, 7 cc centrifuge tubes) 14 and a number of cap holders 80 holding centrifuge tube caps 16. In addition, it has a number of test tube holders 82 that hold the glass test tubes 24 on the outer periphery, and is rotated by a rotation drive mechanism (not shown) to control the stop position. .

The sample dispensing unit 52 has an arm 84 that reciprocates in the front-rear direction (Y-axis direction) and a dispensing head that is supported by this arm 84 and moves back and forth in the left-right direction (X-axis direction). (The reciprocating drive mechanism is not shown), and the dispensing head 86 has a sample dispensing nozzle 88 and a chuck unit for cap which reciprocate in the vertical direction (Z-axis direction). 90 are provided. The dispensing nozzle 88 is connected through a tube 89 to a motor-driven syringe (not shown). As shown in the front view in Fig. 7 and the left side view in Fig. 8, the dispensing nozzle 88 has an upper and lower slide member that is supported by being engaged with the dispensing head 86 and reciprocates vertically.

9 is held in two. The dispensing nozzle 88 is driven by a forward / reverse drive motor 94 fixed to the dispensing head 88, with a pinion 96 fixed to the motor rotating shaft and a vertical slide member 92. Driving force is transmitted through a rack 98 fixed to the fin, and the cradle reciprocates up and down. In FIG. 7, reference numeral 100 denotes an ascending limit sensor for detecting the ascending limit position of the dispensing nozzle 88, and 102 detects an origin position in the upward and downward direction of the dispensing nozzle 888. The upper and lower origin sensor for measuring the dispensing nozzle 108 is a descending limit sensor for detecting the descending limit position of the dispensing nozzle 88.

Reference numeral 6 denotes a sensor detection plate, and reference numeral 108 denotes a compression coil spring. Also the figure

Reference number 1 in 8 is 1 1 0 slide bearing, 1 1 2 is vertical slide guide Reference numeral 114 denotes a chip presence / absence sensor for confirming that the disposable chip 116 is attached to the lower end of the dispensing nozzle 88.

Further, as shown in FIG. 9, the cap chuck unit 90 includes a pair of chuck jaws i 18, 1 18 and an opening / closing mechanism for opening and closing the pair of chuck jaws 1 18, 1 18. And a tubular solenoid 122 for driving the opening and closing mechanism 120. As shown in FIG. 7, the chuck unit 90 is held by an upper and lower slide member 124 that is supported by being engaged with the dispensing head 86 and that reciprocates vertically. The chuck unit 90 is fixed to a dispensing head 88 by a forward / reverse rotatable drive motor (not shown). And rack fixed to upper and lower slide members 1 2 4

The driving force is transmitted via 1 2 8, so that it reciprocates up and down. Reference numeral 13 0 in FIG. 7 denotes an ascending limit sensor for detecting the ascending limit position of the chuck unit 90, and 13 2 denotes a chuck unit 9.

Vertical origin sensor for detecting the origin position in the vertical direction of 、, 1 3

Reference numeral 4 denotes a lower limit sensor for detecting the lower limit position of the chuck unit 90, and reference numeral 1336 denotes a sensor detection plate.

The operation of attaching the cap 16 to the centrifuge tube 14 by the chuck unit 90 will be described with reference to Fig. 9. First, as shown in Fig. 9 (a), a pair of chuck claws 1 18 and 1 18 With the arm open, move the arm 84 in the Y-axis direction, move the dispensing head 88 in the X-axis direction, and move the chuck unit 90 in the Z-axis direction. 5 0 cap holder 8 0

(See Fig. 2) Cap 16 held by the pair of jaws 1 18 and 1

Move the chuck unit 90 so that it is located between 18. next, By driving the tubular solenoid 122, the pair of chuck jaws 111, 118 are closed as shown in FIG. 9 (b), and the cap is closed by the pair of chuck jaws 111, 118. 1 Hold 6. Then, after raising the chuck unit 90, the dispensing head 88 is moved in the X-axis direction and the arm 84 is moved in the Y-axis direction, and the centrifuge tube holding part of the processing turntable 50 is moved. Move the cap 16 gripped by the chuck jaws 1 18 and 1 18 to the position immediately above the centrifuge tube 14 held at 7 8 (see Fig. 2). Next, the chuck unit 90 is lowered, and as shown in FIG. 9 (c), the cap 16 held by the chuck claws 118, 118 is pushed into the upper end opening of the centrifuge tube 14. . Then, the tubular solenoid 122 is driven to open the pair of chuck claws 111, 118 as shown in FIG. 9 (d), and then the chuck unit 90 is raised. .

As shown in FIGS. 1 and 2, the sample suction stage 54 is provided with a cap attaching / detaching mechanism 1 38. With this cap attaching / detaching mechanism 13 8, the sample tube 10 with the lid containing the sample liquid is taken out from the sample tube holding portion 7 4 of the circular turntable 48, and the sample tube I 0 is placed in the sample suction stage. The cap 12 of the sample tube 10 mounted on the stage 54 and fixed on the stage 54 is removed. A cap 12 is attached to the sample tube 10 after the sample is sucked by the cap attachment / detachment mechanism 13, and the sample tube 10 is held from above the sample suction stage 54 by the sample tube of the circular turn table 48. Returned to Part 7 4.

The solvent dispensing unit 56 has a dispensing arm 140 that rotates in a horizontal plane around one end, and as shown in FIG. 10, is attached to the tip of the dispensing arm 140. A nozzle section 142 is provided. Nozzle 1 4 2 has multiple Several, in this example, three liquid transfer tubes 144, 146, 148 are fixed at the tips, as shown in the cross-sectional view of dispensing arm 140 in Figure 11 . The three feeding tubes 1 4 4, 1 4 6, and 1 4 8 are for the liquid supply / drainage section 3 8 Syringe pump unit 40 Methanol supply syringe 150, ethyl acetate (organic solvent) The supply syringe 15 2 and the pH buffer supply syringe 15 4 (see Fig. 1) are connected to the respective flow paths via switching valves. The channels 154 are respectively connected to the respective storage containers (not shown) storing the required liquids.

As shown in FIG. 10, the dispensing arm 140 has one end lower surface fixed to the arm support shaft 156. The arm support shaft 156 is connected to the ball spline shaft 158, and is supported so as to be rotatable around a vertical axis and to be movable up and down along the vertical axis. Then, the arm support shaft 156 is reciprocated in the vertical direction by the lifting / lowering drive mechanism, and is rotated by the rotary drive mechanism, whereby the dispensing fixed to the arm support shaft 156 is performed. The arm 140 moves up and down and rotates. The lifting drive mechanism includes a drive motor 16 2 fixed to the upper mounting plate 160, a timing pulley 16 4 fixed to the rotating shaft of the drive motor 16 2, an upper mounting plate 16 0, and a lower mounting. A screw shaft 1688 whose upper and lower ends are rotatably supported on a plate 166, respectively, and a timing tool 170 near the upper end of this screw shaft 168, both timings The timing belt 172, which is stretched between the nipples 1 6 4 and 1 70, the change nut 1 7 4 screwed to the screw shaft 1 6 8, and this change nut 1 7 4 An elevating member 176 engaged with the arm support shaft 156 so as to allow its rotation and move integrally in the vertical direction, and an upper mounting plate 160 The upper and lower ends are fixed to the lower and lower mounting plates 166, and the guide bars 180 are slidably engaged with the elevating members 176 via the bearings 1-8. . Also, although not shown, an upward limit sensor and a downward limit sensor for detecting the upper limit position and the lower limit position of the dispensing arm 140, respectively, and the vertical direction of the dispensing arm 140. An upper and lower origin sensor for detecting the origin position at, and a sensor detection plate are provided. The rotary drive mechanism includes a drive motor 18 2 fixed to the lower mounting plate 16 6, a timing pulley 18 2 fixed to the rotating shaft of the drive motor 18 2, and a lower mounting plate 16 6 The support block 18 4 is fixed to the support block 18 4, and is rotatably supported by the support block 18 4. The key groove allows the ball spline shaft 15 8 to move up and down and rotates integrally therewith. Rotating member 1 8 6 having a boss hole formed with it, this rotating member 1 8 6 and a timing pulley 1 8 8 that rotates physically, and a tie bridged between both timing pulleys 1 8 2 and 1 8 8 And a positioning sensor 192 for detecting the rotation angle position of the dispensing arm 140.

In the solvent dispensing unit 56 configured as described above, the dispensing arm 140 is rotated by the rotary drive mechanism, and as shown by the solid line in FIG. Move the nozzle part 14 2 at the tip to the position immediately above the centrifuge tube 14 held in the centrifuge tube holding part 78 (see Fig. 2) of the processing turntable 50, and then move up and down The dispensing arm 140 is lowered by the drive mechanism, and as shown by the two-dot chain line in Fig. 10, the J-sleeve 144 at the tip of the dispensing arm 140 is placed in the centrifuge tube 14. Insert into Then, each syringe 150, 152, 154 (see Fig. 1) of the syringe pump unit 40 is driven, and each liquid feeding tube is turned on. —Methanol, ethyl acetate (organic solvent) and PH buffer should be discharged into the centrifuge tube 14 from the discharge ports at the tips of the tubes I 44, 146, and 148. The shaking stage 58 is provided with a shaker 194. The centrifugal separator 60 is installed inside the annular processing turntable 50 in order to use the space effectively and compact the apparatus. The shaker 194 and the centrifuge 60 are conventionally used, and the detailed structure and illustration thereof are omitted. As a means for transferring the target component substance in the sample liquid into the organic solvent in the centrifuge tube 14, an ultrasonic vibrator or a stirrer may be used instead of the shaker. Also, when the centrifuge tube 14 into which the sample solution and the organic solvent are dispensed is shaken, and the centrifuge tube 14 is allowed to stand still, the liquid in the centrifuge tube 14 is rapidly separated into layers. The centrifuge 60 may not be particularly provided.

The separated liquid dispensing unit 62 has an arm 196 that reciprocates in the front-rear direction (Y-axis direction) and an arm 196 that reciprocates in the left-right direction (X-axis direction) supported by the arm 196. It has an injection head 198 (the illustration of each reciprocating drive mechanism is omitted), and the dispensing head 198 has a vertical direction as shown in FIG.

A separation liquid dispensing nozzle 200 reciprocating (in the Z-axis direction) is provided, and the dispensing nozzle 200 is driven by a motor (not shown) via a tube 201. (See Figure 15). The nozzle raising / lowering mechanism for reciprocating the dispensing nozzle 200 in the vertical direction includes a drive motor (stepping motor) 204 fixed to the mounting plate 202 fixed to the dispensing head 198, The timing plate 206 fixed to the rotating shaft of the drive motor 204 and the upper mounting plate 208 fixed to the dispensing head 198 and the upper mounting plate 210 fixed to the lower mounting plate 210 And the lower end can be rotated freely. Supported screw shaft 2 1 2, Timing pulley 2 14 fixed to the lower end of this screw shaft 2 1 2, Timing belt 2 stretched between both timing pulleys 2 0 6 and 2 14 16 The change nut 2 18 that is screwed into the screw shaft 2 12 and reciprocates up and down with the forward and reverse rotation of the screw shaft 2 1 2, and the change nut 2 18 It is composed of an elevating member 220 that is connected and engages with the dispensing nozzle 200 to move in the vertical direction integrally with the change nut 210 and the dispensing nozzle 200. Although not shown, an ascending limit sensor and a descending limit sensor for detecting the ascending limit position and descending limit position of the dispensing nozzle 200, respectively, and the dispensing nozzle 200 An upper / lower origin sensor for detecting the origin position in the vertical direction and a sensor detection plate are provided, and a disposable tip 222 is attached to the lower end of the dispensing nozzle 200. A chip presence / absence sensor is provided to confirm this. The disposable tip 222 attached to the lower end of the dispensing nozzle 200 is held by a large number of disposable tip holding parts 222 of the disposable tip rack 72.

Although not shown, the dispensing head 198 has a cap chuck unit 90 provided on the dispensing head 86 of the sample dispensing unit 52 (see FIGS. 7 and 8). A check unit for transferring the centrifuge tubes, which has the same configuration as that described in Section 9), is provided.

At the separator suction stage 64, a cap removal unit 22 6 as shown in Fig. 13 and a centrifuge tube fixing unit 2 28 as shown in Fig. 14 are fixed to the mounting base 230. Have been. The cap removal unit 2 26 has an arm 2 32 that rotates in a horizontal plane with one end as the center.The tip of the arm 2 32 is horizontal to the centrifuge tube cap 16. Approach from the direction An engaging claw 234 for engaging with the cap 16 is provided. The arm 232 has one end fixed to the upper end of the ball spline shaft 236 and is supported in a horizontal posture. Although not shown, the ball spline shaft 2 36 is reciprocated vertically by a lifting / lowering drive mechanism, and is rotated by a rotary drive mechanism. The structure is such that the fixed arm 2 32 moves up and down and rotates.

The operation of removing the cap 16 from the centrifuge tube 14 by the cap removal unit 2 26 will be described. When the arm 2 32 is in the lowered position, the arm 2 32 is rotated to set the centrifuge tube fixing unit. Engage the engaging claw 23 of the arm 23 with the cap 16 attached to the centrifuge tube 14 (see Fig. 14) fixed to the rack 2 28. Next, as shown by a two-dot chain line in FIG. 13, the arm 2 32 is raised, and the cap 16 is pulled out from the upper end opening of the centrifuge tube 14. Then, the arm 2 32 is turned by 180 ° and then lowered, and the arm 2 32 is stopped at the position shown by the solid line in FIG.

The centrifuge tube fixing unit 228 has a tube holder 238 into which the centrifuge tube 14 is inserted from above. The lower end of the tube holder 238 is fixed to the upper surface of the mounting substrate 230. Further, on the mounting substrate 230, a pair of columns 240 and 240 are erected on both left and right sides of the tube holder 238. A portion of the peripheral surface of the pipe holder 238 near the upper end is fitted to an arc-shaped receiving plate 244 attached to a bracket 242 fixed to one of the columns 240. The other side receives a resilient pressing force from a bracket 2464 fixed to the other support column 240 via a compression coil spring 248. Then, the centrifuge tube 14 inserted into the tube holder 2 38 from above is transferred to the tube holder 2 38. Therefore, it is held and fixed resiliently.

Further, a sensor mounting plate 250 is fixedly provided at the upper end of the pair of columns 240 and 240, respectively. A tip tip detecting photoelectric sensor for detecting the tip (lower end) of the disposable tip 222 attached to the tip of the dispensing nozzle 200 is mounted on the pair of sensor mounting plates 250 and 250. Sensors 25 2 a, 25 2 b, and two sets of liquid level detection photoelectric sensors for detecting the upper end of the sample separation liquid sucked into the disposable tip 22 from the centrifuge tube 14 254a, 254b; 258a, 256b force; respectively, the light projecting part and the light receiving part oppose each other across the movement path of the dispensing tip 222 of the dispensing nozzle 200 It is attached to be. Of the two sets of photoelectric sensors for liquid level detection, the upper photoelectric sensor 2554a and 2554b are used to suck the sample separated liquid separated into the upper layer in the centrifuge tube 14, and The photoelectric sensors 255 a and 255 b on the side are used when a sample separated liquid separated to the lower layer side in the centrifuge tube 14 is sucked.

Furthermore, even when dispensing a sample separation liquid (organic solvent) that is easy to evaporate, has low viscosity, low surface tension, and low specific gravity, the separation liquid dispensing unit 62 has a low Without being affected by the change, it is possible to add a means for reliably preventing the liquid from dripping from the lower end of the separated liquid dispensing nozzle 200 and preventing a decrease in dispensing accuracy. This means is used when the lower end of the dispensing nozzle 200 is pulled out of the sample separation liquid in the centrifuge tube 14 and when the lower end of the dispensing nozzle 200 comes out above the sample separation liquid. Continue to inhale a small amount of air into the sample 200 from its lower end, and generate air bubbles inside the sample separation liquid in the dispensing nozzle 200. To continue until just before the sample separation liquid is discharged. It is a foam generating means. The configuration will be described with reference to FIG. 15 and FIG.

FIG. 15 is a schematic diagram showing a configuration of a main part of the separation liquid dispensing unit 62. In the dispensing unit 62, the dispensing nozzle 200 is lowered, and the distal end of the disposable tip 222 attached to the distal end of the dispensing nozzle 200 is accommodated in the centrifuge tube 14. Then, the liquid is sucked into the disposable chip 222 from the lower end port. Then, the dispensing nozzle 200 is raised and then moved to a position directly above the test tube 24 (dispensing position), and the liquid sucked into the day spot tip 222 is dispensed under the disposable tip 222. Discharge into the test tube 24 from the end opening.

The dispensing nozzle 200 is connected by a tube 201 to a flow path connected to a syringe 258 installed in a liquid supply / drainage section 38 (see FIG. 1). The syringe 258 is driven by a motor 260 and is provided with a controller 262 for controlling the driving of the motor 260. Then, by controlling the drive of the motor 260 by the controller 26 2, a predetermined amount of liquid is sucked from the centrifuge tube 14 into the disposable tip 222 of the dispensing nozzle 200, and the dispensing position is adjusted. At, the liquid in the disposable tip 222 is discharged from the lower end port thereof into the test tube 24. In addition, the dispensing unit is configured so that the controller 262 can control the driving of the motor 260 to switch the syringe 258 to a low speed.

A method of dispensing the sample separation liquid in the centrifuge tube 14 using the dispensing unit having the above configuration will be described with reference to FIG. In this example, the sample separation liquid 264 separated into the upper layer side in the centrifuge tube 14 is sucked into the disposable tip 222 of the dispensing nozzle 200. The dispensing nozzle 200 is moved to a position directly above the centrifuge tube 14 and, as shown in FIG. 16 (A) (only the disposable tip 222 of the dispensing nozzle 200 is shown in FIG. 16). ), The dispensing nozzle 200 is lowered. Then, as shown in FIG. 16 (B), the lower end of the disposable tip 222 is immersed in the sample separation liquid 264 stored in the centrifuge tube 14, and then the syringe 258 Is driven at a normal speed, and the sample separation solution 264 in the centrifuge tube 14 is sucked into the disposable tip 222 from the lower end locator. The suction speed at this time depends on the diameter of the lower end of the disposable tip 222 and the viscosity of the sample separation liquid 264.For example, when the lower end of the disposable tip 222 is 1 mm, the sample separation liquid 26 When the organic solvent in (4) is ethyl acetate, getyl ether or a substance having properties similar to those, it is 0.2 to 0.3 cc / sec.

When a predetermined amount of the sample separation liquid 264 is sucked into the disposable tip 222, the syringe 2β8 is temporarily stopped, and the dispensing nozzle 200 is raised. In the process of pulling the lower end of the disposable tip 220 of the dispensing nozzle 200 out of the sample separation liquid 26 4 in the centrifuge tube 14, as shown in (C) of FIG. When the lower end of 2 is above the liquid level, the syringe 258 is switched to low speed and driven. As a result, a small amount of air continues to be sucked into the disposable chip 222 through its lower end, and the air becomes fine bubbles 266 as shown in FIG. 16 (D). The sample floats in the sample separation solution 264 in the step 222 toward the liquid surface and flows to the gas portion above the sample separation solution 264 in the disposable tip 222. Under the same conditions as described above, the suction speed at this time is suitably, for example, a force of 0.04 to 0.2 cc / sec and about 0.1 cc Z sec. The suction of the minute flow rate into the disposable tip 222 is performed until the dispensing nozzle 200 moves to the position above the centrifuge tube 14 and then to the position above the test tube 24 at the dispensing position, or Further, the dispensing nozzle 200 descends and continues until the lower end of the disc tip 222 is inserted into the test tube 24. During this time, an upward suction force is always applied to the vicinity of the lower end of the disposable tip 2 2 2, so that even if the sample separation liquid 2 64 There is no drip of sample separation solution 2 64 from the lower end of 22. When the lower end of the disposable tip 222 is inserted into the test tube 24, the syringe 258 is switched to a normal speed and driven, and the syringe 258 is dispensed through the tube 201 from the nozzle 250. Air is fed into the sample tube 0, and the sample separation liquid 2 64 in the disposable tip 222 is pushed down and discharged into the test tube 24 from its lower end. In the above description, when the sample separation solution 264 is sucked into the disposable tip 222, the syringe 258 is driven at a normal speed, and the dispensing nozzle 200 is raised in the process of rising. When the lower end of the tip 222 comes above the liquid level of the sample separation liquid 2664 in the centrifuge tube 14, the force that switches the syringe 258 to a low speed is used. A low-speed syringe and a low-speed syringe driven at a low speed may be provided, and a switching valve may be used to selectively connect the normal-speed syringe and the low-speed syringe to the dispensing nozzle 200 through the flow path. Good. In that case, a vacuum pump may be used instead of the low-speed syringe.

In addition, regardless of the type of sample separation liquid to be dispensed, the separation liquid dispensing unit 62 is not affected by the ambient temperature of the separation liquid dispensing nozzle 200, Also, the connection part of disposable chip 2 2 2 etc. Even if there is a slight leak, a means to improve the dispensing accuracy by always inhaling a predetermined amount of the sample separation liquid into the dispensing nozzle 200 accurately and without causing dispersion. can do. This means is to connect the above photoelectric sensors 252a, 252b; 254a, 2554b; 256a, and 256b provided on the centrifuge tube fixing unit 228. It is used to control the drive of the syringe 258. The configuration will be described with reference to FIG. 17 to FIG.

FIG. 17 is a schematic block diagram of the separation liquid dispensing unit 62. The controller 26 2 of the motor 260 driving the syringe 258 and the driving motor (stepping motor) 204 driving the raising / lowering member 220 holding the dispensing nozzle 200 12) are connected to the CPU 267, and the control signals from the CPU 267 control the drive of the syringe driving motor 260 and the driving of the stepping motor 204, respectively. In addition, the photoelectric sensors 25 2 a and 25 2 b for detecting the tip of the chip and the photoelectric sensors 25 24 a and 25 4 b for detecting the liquid level are connected to the CPU 2667, respectively. The predetermined operation of the stepping motor 204 is controlled based on the detection signal from the light receiving section 255 b of the photoelectric sensor for detection, and based on the detection signal from the light receiving section 255 b of the liquid level detection photoelectric sensor. The predetermined operation of the syringe driving motor 260 is controlled. FIG. 17 shows only the upper photoelectric sensor 2554a and 2δ4b of the two sets of photoelectric sensors for liquid level detection. In the following description, an operation in which the sample separated liquid separated into the upper layer in the centrifuge tube 14 is sucked into the disposable tip 222 of the dispensing nozzle 200 is also exemplified. The operation for inhaling the sample separated liquid separated to the lower side in 4 is also performed by centrifugation. The operation itself is exactly the same, only the structure of the tube cap changes.

A method of dispensing the sample separation liquid in the centrifuge tube 14 using the dispensing unit 6 shown in Fig. 17 will be described based on Figs. 18 and 19. .

The dispensing nozzle 200 is moved to a position directly above the centrifuge tube 14 and the stepping motor 204 is driven, as shown in FIG. 18 (a) (dispensing in FIGS. 18 and 19). Only the disposable tip 222 of the nozzle 200 is shown), and the dispensing nozzle 200 is lowered. At this time, the light emitted from the light emitting portion 25a of the photoelectric sensor for detecting the tip of the chip is directly incident on the light receiving portion 252b, and a predetermined output signal is output from the light receiving portion 252b of the photoelectric sensor. Sent to CPU 26 7 Then, as shown in FIG. 18 (b), when the lower end of the disposable chip 22 reaches the standard height position where the photoelectric sensors 25a and 25b are arranged, The light emitted from the light emitting part 25 2 a of the sensor is blocked by the lower end of the disposable chip 22 2, and the amount of light incident on the light receiving part 25 2 b decreases. The output from b changes and the output signal is sent to the CPU 267 to detect that the lower end of the disposable chip 222 has reached the reference height position. Even after the lower end of the disposable tip 222 reaches the reference height position, the stepping motor 204 continues to be driven and the dispensing nozzle 200 descends; The number of pulses output from the pulse generation circuit (not shown) is counted from the time when the lower end of the disposable tip 222 reaches the reference height position, and the stepping is performed until the predetermined number of pulses is counted. Motor 204 power; driven. Then, when a predetermined number of pulses are counted in the CPU 267, the driving of the stepping motor 204 is stopped. As shown in (c), the lowering operation of the dispensing nozzle 200 stops. At this time, the lower end of the disposable tip 222 is located at a distance L corresponding to a predetermined number of pulses from the installed reference height position. As a result, the lower end of the disposable chip 222 is immersed in the sample separation liquid 264 in the centrifuge tube 14. In this way, when the lower end of the disposable tip 2 22 descends from the reference height position where the photoelectric sensors 25 2 a and 25 2 b are installed by the distance L corresponding to the predetermined number of pulses, the minute Since the injection nozzle 200 stops, the lower end position of the disposable tip 222 is always a fixed position. The lower end of the disposable chip 222 is located below the liquid level even after a predetermined amount of the sample separation liquid 264 is inhaled.

When the dispensing nozzle 200 stops descending and the lower end of the disposable tip 222 is immersed in the sample separation liquid 2664 in the centrifuge tube 14, the syringe drive motor 260 is activated. The syringe 255 is actuated, and the sample separation solution 26 4 in the centrifuge tube 14 is sucked into the disposable tip 22 2 from its lower end as shown in Fig. 19 (d). Is done. At the time of this suction operation, the light emitted from the light projecting portion 254a of the liquid level detecting photoelectric sensor passes through the disc tip 222, enters the light receiving portion 254b, and enters the light receiving portion 254b. A signal of a predetermined output is sent from the light receiving section 254 b to the CPU 267. Then, as shown in (e) of FIG. 19, the upper end of the sample separation liquid 264 sucked into the disposable tip 222 is provided with the photoelectric sensors 254a and 254b. When the light reaches the predetermined height, the light emitted from the light-emitting unit 25 4 a of the photoelectric sensor is blocked by the sample separation liquid 26 4 in the disposable tip 22 2, and the light-receiving unit 25 4 b When the amount of light incident on the sensor decreases, the output from the light-receiving unit 254b of the photoelectric sensor changes, and the output signal is sent to the CPU 267. It is detected that the upper end of the sample separation solution 264 in the disposable tip 222 has reached a predetermined height. Based on this detection signal, a control signal from the CPU 267 is sent to the controller 262, and the drive of the syringe drive motor 260 is stopped by a signal from the controller 262. As a result, the syringe 258 stops, and the suction operation of the liquid into the disposable tip 222 stops. As described above, when the upper end of the sample separation liquid 26 4 sucked into the disposable tip 22 2 reaches the predetermined height position where the photoelectric sensors 25 4 a and 25 4 b are installed, Since the suction operation of the sample separation solution 2 64 into the disposable tip 2 222 stops, the lower end of the disposable tip 2 222 is located at the position where the photoelectric sensors 25 4 a and 25 4 b are installed. The sample separation liquid 264 is sucked up to the corresponding height, and the suction amount of the sample separation liquid 264 into the disposable tip 222 is always constant.

When a predetermined amount of the sample separation solution 264 is sucked into the disposable tip 222, the stepping 204 is activated, and the dispensing nozzle 200 is raised as shown in (f) of Fig. 19. Then, the lower end of the disposable tip 2 2 2 is pulled up from the sample separation solution 2 6 4 in the centrifuge tube 14. Then, the dispensing nozzle 200 is moved to a position above the centrifuge tube 14 and then to the position above the test tube 24 at the dispensing position, and then the dispensing nozzle 200 is lowered to disposable tip. The lower end of 222 is inserted into the test tube 24, and then the syringe 258 is driven to discharge the liquid in the disposable tip 222 from the lower end into the test tube 24.

In the above description, the photoelectric sensor is used to detect when the lower end of the disposable tip 222 reaches the reference height position.The detection is performed using a mechanical contact sensor or the like. May be performed. Also, When a photoelectric sensor is used, a photoelectric sensor that detects when the upper end of the liquid reaches a predetermined height position when the liquid is sucked into the disposable tip 222 is disposed at a lower end of the disposable tip 222 as a reference height. It may be shared to detect when the position is reached. Stepping motor 20

If the lower end position of the disposable tip 2 2 2 can be adjusted to always be exactly the same position by precisely controlling the

It is not necessary to provide a sensor for detecting that the lower end of 222 has reached the reference height position.

The above explanation is for the case where the sample separated liquid separated into the upper layer side in the centrifuge tube 14 is sucked into the disposable tip 220 of the dispensing nozzle 200. When the sample separated liquid separated into the lower layer side is drawn into the dispensing tip 222 of the dispensing nozzle 200, only the sample separated liquid on the lower layer side can be sucked in without causing any contamination. A preferred centrifuge tube for this purpose will be described with reference to FIGS. 20 to 23.

FIG. 20 is a perspective view showing a state in which the cap 30 is extracted from the centrifuge tube 14, and FIG. 21 is a longitudinal sectional view showing a state in which the cap 30 is attached to the centrifuge tube 14. The cap 30 for closing the upper surface opening of the centrifuge tube 14 in a liquid-tight manner is composed of a sealing portion 268, an inner tube portion 270, and a closing portion 272. The hermetic plug portion 268 is inserted into the upper end of the centrifuge tube 14 so that the outer peripheral surface is closely fitted, and a through hole 274 is formed in the center. The inner tube portion 270 has an outer diameter smaller than the inner diameter of the centrifuge tube 14, and has a tubular shape in which the lower portion is gradually made smaller in diameter, and the upper end portion of the inner tube portion 270 penetrates the plug portion 268. It is fixed to the inner peripheral part of the hole 274 and is integrated with the sealing part 268. Also, the inner pipe part 270 When the hermetic plug part 2688 is closely fitted to the upper end of the centrifuge tube 14, the lower end is formed to have a length such that it is located near the inner bottom surface of the centrifuge tube 14. The closing part 272 is formed by a filling plug inserted upward into the lower end of the inner pipe part 270, and closes the lower end of the inner pipe part 270 in a liquid-tight manner. The closing portion 272 made of the filling plug is easily dropped by a downward pressing force, that is, a force pressed downward by the lower end of the disposable tip 222 of the dispensing nozzle 200.

Next, using the centrifuge tube 14 having the cap 30 having the above configuration, only the sample separation liquid (lower liquid) separated into the lower layer of the liquid separated into the upper liquid and the lower liquid is sucked. The method will be described with reference to FIG.

First, with the cap 30 removed, the sample solution and the organic solvent are dispensed into the centrifuge tube 14, and then the inner tube portion 270 is inserted deep into the centrifuge tube 14 and inserted into the liquid. Then, the cap 30 is attached to the centrifuge tube 14 so that the sealing plug portion 268 is tightly fitted to the upper end of the centrifuge tube 14. In this state, the components in the sample solution are transferred to the organic solvent layer by shaking the centrifuge tube, and then the centrifuge is centrifuged (see Figs. 1 and 2). As shown in 22, the liquid contained between the inner peripheral surface of the centrifuge tube 14 and the outer peripheral surface of the inner tube part 27 of the cap 30 is filled with the upper liquid 27 6 and the lower liquid.

(Sample Separation Solution) At this time, as shown in (a) of FIG. 22, since the lower end of the inner pipe portion 27 of the cap 30 is located near the inner bottom surface of the centrifuge tube 14, the inner pipe portion 27 The lower end of 0 is the upper liquid 2 7

It is located below the boundary surface 280 between 6 and the lower liquid 278. For this reason, the vicinity of the lower end of the inner pipe part 270 is inserted into the lower layer liquid 278. I have.

Next, in order to inhale only the lower layer liquid 278 from the centrifuge tube 14 into the disposable tip 222 of the dispensing nozzle 200, as shown in (a) of FIG. Then, lower the dispensing tip 2 of the dispensing nozzle 200 as shown in (b) of Fig. 22 and insert the through-hole 2 7 of the sealing plug 2 68 of the cap 30 of the centrifuge tube 14. 4 and insert it deeply into the inner pipe section 270. Then, the lower end of the disposable tip 222 presses the closed part 272 at the lower end of the inner pipe part 270 downward, thereby forming a plug as shown in (c) of FIG. The closed part 272 is dropped from the lower end of the inner pipe part 270, and the lower end of the disposable tip 222 is inserted into the lower liquid 278. Thereafter, the syringe 250 (see FIGS. 15 and 17) connected to the dispensing nozzle 200 is driven to drive the disposable tip through the lower end of the disposable tip 222. 2 Inhale liquid into 2 2. At this time, since the lower end of the disposable tip 222 is inserted into the lower liquid 278, only the lower liquid 278 is sucked into the disposable tip 222, and Since the lower end is located below the boundary surface 280 between the upper liquid 276 and the lower liquid 278, a part of the upper liquid 276 mixes with the lower liquid 278 to form a contour. There is no need to worry about producing minerals.

The cap 2 8 2 of the centrifuge tube 14 whose longitudinal section is shown in Fig. 23 is

8 4 force A structure that is placed over the upper end of the centrifuge tube 14 and fits externally in a liquid-tight manner, and the upper end of the inner tube portion 28 6 The part is fixed, and the sealed part 284 and the inner pipe part 286 are integrated. Further, a thin plate-like portion is integrally formed at the lower end of the inner tube portion 286 to form a closed portion 288. The closed part 288 consisting of this thin plate-shaped part is dispensed with the dispensing nozzle 200 By being pressed downward by the lower end of the pochip 222, it can easily burst.

In each of the embodiments described above, the sealing portions 268, 284 constituting the caps 30, 282 and the inner tube portions 270, 286 are separate from each other. The inner tube portions 270 and 286 may be fixed to 688 and 284, and the force for integrating them may be integrally formed, and the hermetic plug portion and the inner tube portion may be integrally formed. Further, the configuration of the closing portion, which closes the lower end of the inner tube portion in a liquid-tight manner and easily falls or ruptures by a downward pressing force, is as follows. The configuration is not limited to the one formed integrally, and for example, a configuration in which the lower end of the inner tube portion is covered with a film and closed in a liquid-tight manner may be used.

The heater block 2 94 is attached to the fixed frame 29 2, as shown in Fig. 24 and Fig. 25, respectively. The heater block 294 has a plurality of vertical holes 296 into which a plurality of test tubes 24 are fitted from above. The vertical hole 296 is formed so that the inner peripheral surface thereof is in close contact with the outer peripheral surface of the test tube 24. Further, the heater block 294 has a through hole 298 communicating with the bottom of each vertical hole 296, and a push rod 300 is provided in each through hole 298. The push-up plate 302, which is slidably penetrated and fixed to the tip end of the push-up bar 300, reciprocates vertically in the vertical hole 296. . The lower end of each of the plurality of push-up bars 300 is fixed to a common elevating plate 304. Also, above the fixed frame 292, there is a J-Zull head 300 having a plurality of nozzle plugs 300 that abut against the upper end of each test tube 24 to seal the upper end opening hermetically. Has been established. Gas supply for blowing nitrogen gas into the test tube 24 A nozzle 310 and an exhaust hole 312 for discharging waste gas from the inside of the test tube 24 are formed. The nozzle head 306 is connected to the rack 314 and supported by the fixed frame 292. The fixed frame 292 is provided with a motor 3116 for forward and backward rotation, which can be rotated forward and backward. A pinion 318 is fixed to the rotating shaft of the motor 316. 8 and the rack 3 1 4 are screwed together. Further, the rack 314 is connected to the lifting plate 304 via a hook 322 fixed thereto and a connecting member 322 engaged with the hook 322. The reference numeral 3 2 4 is an elevating guide. By driving the lifting drive motor 3 16, the nozzle head 300, the lifting plate 304, the push-up bar 300, and the push-up plate 302 are integrally raised. And descend.

The solvent dispensing stage 68 is provided with a force not shown, which is the same as the shaker 194 installed on the shake stage 58.

As shown in FIG. 26, the injection unit 70 has an injection arm 32 6 that rotates in a horizontal plane with one end as a center. A nozzle holding shaft 330 is attached to the distal end of the nozzle 326 via a slide bearing 328, and the nozzle holding shaft 330 is connected to the injection arm 322 by a compression coil spring 332. It is elastically supported at the tip of 6. A chuck 334 is provided at a lower end portion of the nozzle holding shaft 330, and the injection nozzle 336 is held by the chuck 334. One end of the injection arm 326 is fixed to the arm support shaft 338. The arm support shaft 338 is connected to a ball spline shaft (not shown), and is supported so as to be rotatable about a vertical axis and movable vertically along the vertical axis. Have been. The arm support shaft 338 is moved up and down by the elevation drive mechanism and rotated by the rotation drive mechanism, whereby the injection arm 32 fixed to the arm support shaft 338 is moved. 6 moves up and down and rotates. The configuration of the lifting drive mechanism and the rotary drive mechanism is the same as the lift drive mechanism and the rotary drive mechanism of the solvent dispensing unit 56 shown in FIG. 10, so that illustration and description thereof are omitted. After evaporating the sample separation liquid dispensed in the test tube 24 to dryness, dissolve the residue and inject the organic solvent such as methanol for injection into the analytical instrument into the test tube 24 Although a solvent dispensing unit and an injection unit for aspirating the sample separation liquid from the test tube 24 after that may be provided separately, it is also possible to provide a separate unit (see the operation shown in FIGS. 3 and 4). In this embodiment, the operation of dispensing the organic solvent into the test tube 24 is performed by the injection unit 70 in this embodiment. With this injection unit 70, the operation method from dispensing the solvent into the test tube 24 to inhaling the sample separated solution from the test tube 24 and injecting the sample separated solution into the HPLC column is performed. Explanation will be given based on FIG.

First, the flow path configuration of the injection unit 70 will be described. In addition to the injection nozzle 33, the injection unit 70, the measuring pipe loop 34, and the hexagonal valve 34 2, Syringe for solvent dispensing 3 4 4, 3-way switching valve 3 4 6, Storage container 3 4 8 containing methanol 350 for dissolving the dried residue and injecting it into the HPLC column The flow path is composed of, and pipes. The injection nozzle 3336 is connected to the b-port of the six-way valve 3442 in a flow path, and the metering tube loop 3440 has both ends of the c-port and the e-port of the six-way valve 3442. Each The flow path is connected. The syringe 344 is connected to the port a of the six-way valve 342 and the liquid storage container 348 via the three-way switching valve 346, respectively. The d port of the six-way valve 342 is connected to the HPLC pump, and the f port is connected to the HPLC column.

First, in order to dispense methanol into the test tube 24, as shown in (a) of FIG. 27, a syringe 34 4 and an injection nozzle 33 36 are passed through a six-way valve 34 2. When the three-way switching valve 3446 is switched and operated, the methanol is sucked into the syringe 344 from the liquid storage container 348, and the methanol is removed from the syringe 344. It is sent to the injection nozzle 336, and then, for example, 0.1 ml of methanol is discharged from the injection nozzle 336 into the test tube 24. Next, switch the 6-way valve 3 4 2

27 As shown in (b), 7-way valve 3 4 2 and metering tube loop 3 4

After the injection nozzle 3336 and the syringe 34 are in communication with each other through 0, the liquid in the test tube 24 is agitated with a shaker (not shown). Test tube into injection nozzle 3 3 6

Aspirate the sample lysis solution in 24 and introduce a fixed amount of sample lysis solution into the metering tube loop 340. Next, switch the 6-way valve 3 4 2

2 As shown in (c) of 7, the hexagonal valve 3 4 2 and the metering tube loop 3 4

The state is such that the HPLC pump and the HPLC column are in communication with each other via 0, and the syringe 344 and the injection nozzle 336 are in communication with each other via the six-way valve 342. Then, a certain amount of the sample solution retained in the measuring tube loop is injected into the column by the HPLC pump.At the same time, the injection nozzle 336 is moved to the washing tank 352, and the three-way switching is performed. Switch valve 3 4 6 and syringe Drive 3 4 to send methanol from the liquid storage container 3 4 8 to the injection nozzle 3 3 6, discharge the methanol from the tip end of the injection nozzle 3 3 6, and perform the injection. Rinse nozzle 3 3 6 and pipes with methanol.

Next, an example of an operation of automatically measuring the concentration of a specific component substance (for example, a drug) contained in a sample, for example, a frozen serum, by an automatic concentration measuring device having the above-described configuration will be described. explain.

First, a plurality of sample tubes 10 with lids containing frozen serum are set in the sample tube holder 74 of the circular turntable 48 of the automatic solvent extraction unit 36. After the frozen serum is thawed and homogenized, the sample tube 10 is moved from the top of the circular table 48 to the sample tube suction stage 54 by the cap attaching / detaching mechanism 13, and the sample is sucked. The sample tube 10 is fixed on the stage 54. Then, the cap 12 of the sample tube 10 is removed by the cap attaching / detaching mechanism 1388. Next, the arm 84 of the sample dispensing unit 52 is moved in the Y-axis direction, the dispensing head 86 is moved in the X-axis direction, and the dispensing nozzle 88 is moved in the Z-axis direction. 4 Attach the disposable tip 1 16 held by the disposable tip holding portion 76 at the tip of the dispensing nozzle 88. Next, the arm 84, the dispensing head 86, and the dispensing nozzle 88 of the sample dispensing unit 52 are moved in the Y-axis direction, the X-axis direction, and the Z-axis direction, respectively. The tip (lower end) of the 8 disposable chip 1 16 is immersed in the sample solution (melted serum) in the sample tube 10 fixed on the sample suction stage 54 (see the two-dot chain line in Fig. 7). ) Inhale the sample solution into the disposable tip 1 16. And arm of sample dispensing unit 52, dispensing head 86 and dispensing nozzle The nozzle 8 8 is moved in the Y-axis direction, X-axis direction, and Ζ-axis direction, and the tip of the dispensing tip 1 16 of the dispensing nozzle 8 8 Insert into the centrifuge tube 14 held in 8 (see the two-dot chain line in FIG. 8), and discharge the sample liquid sucked into the disposable tip 116 into the centrifuge tube 14. After that, the used disposable tip 1 16 is discarded to the dumping port 73, the cap 12 is attached to the re-sample tube 10 by the cap attaching / detaching mechanism 1 38, and then the sample tube 10 is turned circularly. Return to sample tube holder 7 4 of table 4 8.

Next, the processing turntable 50 is rotated to move the centrifuge tube 14 held by the centrifuge tube holding unit 78 and containing the sample solution to the solvent dispensing position. Then, the dispensing arm 140 of the solvent dispensing unit 56 is rotated in the Θ direction (rotated in a horizontal plane), then lowered, and the liquid sending tube fixed to the nozzle portion 142 is rotated. — Insert the tips of the pieces 144, 144, and i48 into the centrifuge tube 14 held by the centrifuge tube holder 78 of the processing turntable 50 (the two-dot chain line in FIG. 10). Dispense ethyl acetate (organic solvent), methanol and pH buffer into centrifuge tube 14). Next, the arm 84, the dispensing head 86, and the chuck unit 90 of the sample dispensing unit 52 are moved in the Y-axis direction, the X-axis direction, and the Z-axis direction, respectively. Activate 90 to grip the centrifuge tube cap 16 held in the cap holding section 80 of the processing turntable 50 with a pair of chuck claws 1 18, 1 18, and from the cap holding section 80 Remove cap 16. Then, the arm 84, the dispensing head 86, and the chuck unit 90 of the sample dispensing unit 52 are moved in the Y-axis direction, the X-axis direction, and the Z-axis direction, respectively, and then the chuck unit 90 is moved. To operate the centrifuge tube holder of the processing turntable 50 Attach cap 16 to centrifuge tube 14 held in.

When the cap 16 is attached to the centrifuge tube 14, the sample dispensing unit 5 2 is held while holding the cap 16 with the pair of chuck jaws 1 18 and 1 18 of the chuck unit 90. After moving the arm 84, dispensing head 86 and chuck unit 90 in the Y-axis direction, X-axis direction and Z-axis direction, the chuck unit 90 is operated and the processing turn is performed. The centrifuge tube 14 held in the centrifuge tube holder 78 of the table 50 is moved to the shaking stage 58, and the centrifuge tube 14 is set in the shaker 1994. Then, the shaker 1994 is driven to shake the centrifuge tube 14, and the target component substance in the sample solution is transferred into the organic solvent in the centrifuge tube 14. After shaking, the arm 84, dispensing head 86 and chuck unit 90 of the sample dispensing unit 52 are moved in the Y-axis direction, X-axis direction and Z-axis direction, and the chuck is moved. Activate the unit 90, move the centrifuge tube 14 from the shaking stage 58 to the centrifuge 60, and set the centrifuge tube 14 in the centrifuge 60. Then, the liquid is centrifuged by driving the centrifuge 60. After the centrifugation is completed, the arm 196 of the separated solution dispensing unit 62, the dispensing head 198 and the chuck unit (not shown) of the centrifuge tube transfer ffi are moved in the Y-axis direction and the X-axis direction. And the Z-axis direction, and operate the chuck unit to remove the centrifuge tube 14 from the centrifuge 60 and move the centrifuge tube 14 to the separation liquid suction stage 64 to fix the centrifuge tube.

Fix to 2 2 8 Subsequently, the cap 16 is removed from the centrifuge tube 14 by the cap removal unit 22. In this operation example, the sample separation liquid is separated into the upper layer side in the centrifuge tube 14 by centrifugation, and the sample separation liquid is separated into the lower layer side in the centrifuge tube 14 by centrifugation. Do In this case, there is no need to remove the cap 30 from the centrifuge tube 14 as shown in FIGS. 5 and 22.

When the cap 16 is removed from the centrifuge tube 14, the arm 1 96 of the separation liquid dispensing unit 62 is moved in the Y-axis direction, the dispensing head 198 is moved in the X-axis direction, and the dispensing nozzle 2 The disposable tip 22 is held in the disposable tip holder 22 of the disposable tip rack 72 by displacing the disposable tip 22 in the Z-axis direction. . Next, the arm 196, the dispensing head 198, and the dispensing nozzle 200 of the separation liquid dispensing unit 62 are moved in the Y-axis direction, the X-axis direction, and the Z-axis direction, respectively. The tip (lower end) of the disposable tip 222 of the dispensing nozzle 200 is connected to the centrifuge tube 14 fixed to the centrifuge tube fixing unit 222 of the separated liquid suction stage 64. Immerse in the pull separation solution and aspirate the sample separation solution into the disposable tip 222 (see Figs. 14 to 16 and Figs. 18 and 19). Subsequently, the arm 196, the dispensing head 198, and the dispensing nozzle 200 of the separation liquid dispensing unit 62 are moved in the Y-axis direction, the X-axis direction, and the Z-axis direction, respectively. The tip of the disposable tip 222 of the dispensing nozzle 200 is placed in the test tube 2 held at the dispensing position A (see FIG. 2) of the test tube holder 82 of the processing turntable 50. Into the test tube 24 (see Fig. 15), and discharge the sample separation liquid sucked into the disposable tip 222 into the test tube 24. After that, the used disposable tip 222 is discarded to the dumping pot 73. Further, the injection unit 70 is operated, and the liquid remaining in the centrifuge tube 14 is sucked into the injection nozzle 336, and then discarded into the washing tank 352. Subsequently, the separation liquid dispensing unit 62 was operated, and the cap 16 was attached to the centrifuge tube 14 using the chuck unit for transferring the centrifuge tube. Discard centrifuge tube 14 to dumping port 73.

Next, the processing turntable 50 is rotated, and the test tube 24 is moved to the evaporating and drying stage 66, and the test tube 24 is held in the state shown in FIGS. 24 and 25. The test tube 2 is heated from the surroundings by the hot block 294, and the gas supply nozzle 308 of the nozzle head 306 is supplied to the inside of the test tube 224 from the nozzle 310. Nitrogen gas is blown through the opening, and waste gas is exhausted through the exhaust holes 312, thereby evaporating the sample separation liquid in the test tube 24 and drying the sample separation liquid. Next, the arm 196 of the separation liquid dispensing unit 62, the dispensing head 198 and the centrifuge tube transfer tube unit are moved in the Y-axis direction, the X-axis direction, and the Z-axis direction, respectively. At the same time, the chuck unit is operated to remove the test tube 24 from the removal position B of the test tube holding portion 82 of the processing turntable 50 (see FIG. 2), and transfer the test tube 24 to the solvent dispensing stage 68. And fix it on a machine (not shown). Then, the injection unit 70 is operated, and an organic solvent, for example, methanol is dispensed into the test tube 24 by the operation shown in FIG. Subsequently, the shaker is driven to shake the test tube 24, and the dried residue is dissolved in methanol. C When the shaking is completed, the injection unit 70 is operated to activate the injection unit 70. By the operation shown in (b) of (a), the sample solution in which the component substances are dissolved in the organic solvent is sucked into the injection nozzle 336 from the test tube 24, and the sucked sample solution is sucked. Inject a predetermined amount of へ into an HPLC column and measure the concentration of the component substances. Then, the separation liquid dispensing unit 62 is operated, and the used test tubes are transferred using the chuck unit for transferring the centrifuge tubes.

Discard 2 4 to dumping port 73. In addition, the injection unit 7 0 is operated, and the injection nozzle 3336 and the inside of the pipe are washed with methanol by the operation shown in (c) of Fig. 27. Figures 28 and 29 show flowcharts of this series of operations.

In the above-described embodiment, the sample separation liquid dispensed into the container (test tube 24) is configured by the apparatus configuration including the evaporating and drying stage 66 and the solvent dispensing stage 68 provided with a shaker. After evaporating to dryness, the organic solvent is dispensed into the container, the container is shaken, and dissolved in an organic solvent to prepare a sample solution to be injected into analytical instruments such as HPLC. With the equipment configuration equipped with a concentration stage without the solvent dispensing stage 68, a part of the organic solvent of the sample separated solution dispensed into the container is evaporated, and the concentrated sample separated solution is evaporated. It may be prepared. When the sample separation liquid obtained by the solvent extraction has a necessary concentration, the evaporating and drying stage and the concentration stage may not be particularly provided. Furthermore, according to an apparatus configuration provided with an evaporating and drying stage 66 without the solvent dispensing stage 68, all of the organic solvent of the sample separation liquid dispensed into the container is evaporated. An automatic extraction device that finally obtains the dried residue is used to prepare a sample liquid to be injected into analytical equipment such as an HPLC or gas chromatograph from the dried residue obtained by the automatic extraction device. You may. In addition, an automatic concentration measuring device is configured without the separation liquid dispensing unit 62 in addition to the evaporating and drying stage 66, the concentration stage and the solvent dispensing stage 68, and the separation liquid dispensing unit 62 By using an injection unit having a function as described above, the sample separated solution separated in a centrifuge tube may be directly sucked from the centrifuge tube and injected into an analytical instrument such as HPLC. Industrial applicability

INDUSTRIAL APPLICABILITY The automatic extraction device and the automatic concentration measurement device according to the present invention are used in a clinical test center or a laboratory of a pharmaceutical company which needs to perform a large amount of sample analysis processing at one time. For labs of pharmaceutical companies, for example, this will result in significant improvements in work efficiency and work space.

Claims

Scope of the request. A sample holding unit for holding a plurality of sample containers containing a liquid sample, an extraction container holding unit for holding a plurality of extraction containers, and taken out of the sample holding unit or holding the sample. A predetermined amount of a liquid sample is sucked from the sample container held in the extracting portion, and the sucked liquid sample is taken out of the extracting container holding portion or the extracting container held in the extracting container holding portion. Sample dispensing means for discharging into the extraction container, extraction solvent dispensing means for discharging a predetermined amount of the organic solvent for extraction into the extraction container, and a target component in the liquid sample contained in the extraction container A component substance transfer means for transferring the substance into the organic solvent for extraction; a storage container holding section for holding a plurality of storage containers; A predetermined amount of the sample separated liquid dissolved in the container is sucked, and the sucked sample separated liquid is taken out of the holding container holding portion or discharged into the holding container held by the holding container holding portion. An automatic extraction device for a component substance in a liquid sample, comprising: a dispensing means; 2. The automatic extraction device for component substances in a liquid sample according to claim 1, further comprising evaporating and drying means for evaporating the sample separated liquid contained in the storage container to dry the sample separated liquid. A dissolving solvent dispensing means for discharging a predetermined amount of the organic solvent for dissolution into the storage container, and dissolving means for dissolving the dried residue in the organic solvent for dissolution in the storage container are provided. 3. The automatic extraction device for a component substance in a liquid sample according to claim 2. The dissolution means shakes the container! The automatic extraction device for component substances in a liquid sample according to claim 3, which is a machine. 3. The liquid sample according to claim 2, wherein the evaporating and drying means comprises a heater for heating the container from the periphery thereof, and nitrogen gas supply means for blowing nitrogen gas into the container through an upper opening of the container. Automatic extraction device for constituent substances inside. The automatic extraction of the component substances in the liquid sample according to claim 1, wherein a concentration means for evaporating a part of the organic solvent of the sample separated liquid contained in the container and concentrating the sample separated liquid is provided. apparatus. 7. The liquid sample according to claim 6, wherein the concentrating means comprises a heater for heating the container from the periphery thereof, and nitrogen gas supply means for blowing nitrogen gas into the container through an upper opening thereof. Automatic extraction equipment for component substances. The separation liquid dispensing means sucks a predetermined amount of the sample separation liquid contained in the extraction container from the lower end port, and holds the dispensing nozzle for discharging the sample separation liquid from the lower end port, and the dispensing nozzle. A nozzle holding means, and a lower position where the lower end of the dispensing nozzle is immersed in the sample separation liquid in the extraction container and an upper position where the lower end of the dispensing nozzle is separated upward from the extraction container. A nozzle elevating means for elevating and lowering between the positions, a nozzle moving means for moving the nozzle holding means between a position immediately above the extraction container and a dispensing position, and a lower end opening into the dispensing nozzle. A syringe for allowing a predetermined amount of the sample separation liquid in the extraction container to be sucked from the container, discharging the sample separation liquid in the dispensing nozzle from the lower end port at the dispensing position, and driving the syringe. A syringe drive means, the silicate and Syringe control means for controlling the Nji driving means, the configured claims preceding claim provided with an automatic extractor component material in a liquid sample. When the lower end of the dispensing nozzle rises above the sample separation liquid when the lower end of the dispensing nozzle is pulled up from the sample separation liquid in the extraction container, the dispensing nozzle Continue to inhale a small amount of air from the lower end of the navel to generate air bubbles inside the sample separation liquid in the dispensing nozzle. 9. The automatic extraction device for component substances in a liquid sample according to claim 8, further comprising a means for generating bubbles. 0. When the predetermined amount of the sample separation liquid is sucked into the dispensing nozzle with the lower end immersed in the sample separation liquid in the extraction container, the upper end of the sample separation liquid A liquid level sensor, which is disposed at a height position where the upper end of the sample separation liquid has reached the height position, and photoelectrically detects whether the upper end of the sample separation liquid has reached the height position. 9. The apparatus for automatically extracting a component substance in a liquid sample according to claim 8, wherein the syringe control means controls the syringe driving means to stop driving the syringe.

1. The nozzle raising / lowering means is composed of a stepping motor whose driving amount is controlled by the number of pulses, and lowers the dispensing nozzle to immerse the lower end of the dispensing nozzle into the sample separation liquid in the extraction container. A nozzle detecting means is provided for detecting whether the lower end of the dispensing nozzle has reached the reference height position when the dispensing-nozzle has reached the reference height position. A signal having a certain number of pulses is input to the stepping motor from the time point when is detected.

10. The automatic extraction device for component substances in a liquid sample according to item 10.

2. The automatic extraction device for component substances in a liquid sample according to claim 8, wherein the dispensing nozzle is configured using a disposable suction pipe.

3. The apparatus for automatically extracting a component substance in a liquid sample according to claim 1, further comprising a water or aqueous solution dispensing means for discharging a predetermined amount of water or an aqueous solution into the extraction container.

4. The automatic extraction device for component substances in a liquid sample according to claim 1, wherein a cap attaching / detaching means for attaching / detaching a cap to / from the extraction container is provided.

5. The apparatus for automatically extracting a component substance in a liquid sample according to claim 1, wherein the component substance transfer means is a shaker that shakes the extraction container.

6. The liquid according to claim 1, wherein a centrifugal separator is provided for centrifuging the liquid in which the target component substance is transferred from the liquid sample into the organic solvent for extraction by the component substance transfer means. Automatic extraction equipment for component substances in samples.

7. An automatic extraction device for a component substance in a liquid sample according to any one of claims 1 or 3 to 16;

Concentration measuring means for measuring the concentration of the component substances in the liquid sample, A liquid injecting means for inhaling a component solution obtained by dissolving a target component substance in an organic solvent from the container, and injecting the inhaled component solution by a predetermined amount into the concentration measuring means;

An automatic concentration measuring device for a component substance in a liquid sample, comprising:

8. The automatic concentration measuring apparatus for a component substance in a liquid sample according to claim 17, wherein the liquid injection means has a measuring tube for holding a predetermined amount of the component solution to be injected into the concentration measuring means. .

9. A sample holder for holding a plurality of sample containers containing a liquid sample,

A container holding unit for holding a plurality of containers,

A predetermined amount of a liquid sample is sucked from a sample container taken out of the sample holding unit or held in the sample holding unit, and the sucked liquid sample is taken out of the container holding unit or transferred to the container holding unit. Sample dispensing means for discharging into the held container,

Extraction solvent dispensing means for discharging a predetermined amount of the organic solvent for extraction into the container,

Component substance transfer means for transferring the target component substance in the liquid sample contained in the container into the organic solvent for extraction,

A concentration measuring means for measuring the concentration of the component substance in the liquid sample; and a sample separation liquid in which the target component substance is dissolved in an organic solvent separated in the container, and the sucked sample separation liquid is collected. Separation liquid injection means for injecting only a fixed amount into the concentration measurement means;

0. Dispense water or aqueous solution to discharge a predetermined amount of water or aqueous solution into the container The apparatus for automatically measuring the concentration of a component substance in a liquid sample according to claim 19, wherein means is provided.

10. The automatic concentration measuring device for component substances in a liquid sample according to claim 19, wherein a cap attaching / detaching means for attaching / detaching the cap to / from the container is provided.

2. The method according to claim 19, wherein a centrifuge is provided for centrifuging a liquid in which a target component substance is transferred from the liquid sample into the organic solvent for extraction by the component substance transfer means. Automatic concentration measuring device for component substances in liquid samples.

3. A dispensing nozzle that sucks a predetermined amount of liquid contained in the liquid container from the lower end port and discharges the liquid from the lower end port;

Nozzle holding means for holding the dispensing nozzle,

A nozzle elevating means for raising and lowering the nozzle holding means between a lower position where the lower end of the dispensing nozzle is immersed in the liquid in the liquid container and an upper position where the lower end of the dispensing nozzle is separated upward from the liquid container. Means,

A nozzle moving means for moving the nozzle holding means between a position immediately above the liquid container and a dispensing position;

A syringe for sucking a predetermined amount of the liquid in the liquid container into the dispensing nozzle from a lower end thereof, and discharging the liquid in the dispensing nozzle from the lower end of the dispensing nozzle at the dispensing position;

Syringe driving means for driving the syringe;

Syringe control means for controlling the syringe drive means;

In a liquid dispensing device equipped with

When the lower end of the dispensing nozzle is pulled out of the liquid in the liquid container, the lower end of the dispensing nozzle is moved into the dispensing nozzle when the lower end of the dispensing nozzle comes out of the liquid. Equipped with an air bubble generating means that keeps inhaling a small amount of air from the lower end port to generate air bubbles inside the liquid inside the dispensing nozzle and keep this state until just before the liquid inside the dispensing nozzle is discharged. Liquid dispensing device characterized by the above-mentioned.

4. The liquid dispensing device according to claim 23, wherein the bubble generating means is a control circuit provided in the syringe control means and controlling the syringe driving means so as to drive the syringe at a low speed.

5. The bubble generation means

A low-speed syringe, low-speed syringe driving means for driving the low-speed syringe at low speed, and low-speed syringe control means for controlling the low-speed syringe driving means;

Flow path switching means for selectively connecting a syringe for sucking a liquid into the dispensing nozzle and discharging the liquid from the lower end of the dispensing nozzle and the low-speed syringe to the dispensing nozzle;

The liquid dispensing device according to claim 23, comprising:

6. A dispensing nozzle that sucks a predetermined amount of liquid stored in the liquid container from the lower end port and discharges the liquid from the lower end port.

Nozzle holding means for holding the dispensing nozzle,

A predetermined amount of liquid in the liquid container is introduced into the dispensing nozzle from a lower end port thereof. A syringe for discharging the liquid in the dispensing nozzle from the lower end port at the dispensing position,

Syringe driving means for driving the syringe;

Syringe control means for controlling the syringe drive means;

In a liquid dispensing device equipped with

The upper end of the liquid is at the height where the upper end of the liquid is located when a predetermined amount of liquid is sucked into the dispensing nozzle in a state where the lower end is immersed in the liquid in the liquid container. A liquid level sensor that photoelectrically detects whether or not the liquid has reached the position, and controls the syringe driving means by the syringe control means based on a detection signal of the liquid level sensor to drive the syringe. A liquid dispensing device characterized by stopping the operation. 7. The nozzle raising / lowering means is composed of a stepping motor whose driving amount is controlled by the number of pulses, and is used to lower the dispensing nozzle to lower the lower end of the dispensing nozzle into the liquid in the liquid container. Nozzle detecting means is provided to detect whether the lower end of the dispensing nozzle has reached the reference height 懨, and the nozzle detecting means detects that the lower end of the dispensing nozzle has reached the reference height position. 27. The liquid dispensing device according to claim 26, wherein a signal having a fixed number of pulses is input to said steving motor from the time when said liquid is dispensed.

8. The liquid dispensing apparatus according to claim 27, wherein the nozzle detecting means is configured by a photoelectric sensor that photoelectrically detects a lower end of the dispensing nozzle. 9. In a sedimentation tube for centrifugal separation of a liquid, comprising: a container body having a tubular shape with an open upper surface; and a cap covering the upper surface opening of the container body, A hermetic plug having a through hole at the center thereof and closely fitting to the upper end of the container main body; a tubular having an outer diameter smaller than the inner diameter of the container main body; an upper end having a through hole of the hermetic plug; An inner tube portion having a length such that the lower end is located near the inner bottom surface of the container body when the sealing portion is closely fitted to the upper end portion of the container body;

A sedimentation tube for centrifugal separation of liquid, characterized in that the inner tube has a lower end liquid-tightly closed and a closing part which easily falls off or ruptures by a downward pressing force.

0. The liquid according to claim 29, wherein the closing portion is a plug inserted into the lower end of the inner tube portion of the cap or a thin plate portion integrally formed at the lower end of the inner tube portion of the cap. Centrifuge sedimentation tube.