JP2021107830A - Automatic pretreatment apparatus for test specimen - Google Patents

Abstract

To develop a highly versatile automatic pretreatment apparatus for test specimens that can be used for instrument analysis and the like.SOLUTION: An automatic pretreatment apparatus for test specimens adopts mechanisms 3, 7 capable of freely transferring and lifting sample tubes Tu on a predetermined XY plane at the time of performing various treatments such as extracting a predetermined solvent and the like from the test specimen, and further made it an efficient automatic pretreatment apparatus by moving a pipetting device 9 for sorting and supply of the solution as one unit with the movement of the tubes Tu.SELECTED DRAWING: Figure 1

Description

The present invention relates to an automatic pretreatment device for a test sample. In particular, it relates to an automatic pretreatment device for inspection samples, which is preferably used for instrumental analysis.

Extraction operations are often performed to extract the detection target from the test sample, but these operations are often complicated. For example, when analyzing whether or not veterinary drugs, pesticides, etc. remain, the operation of extracting the detection target such as veterinary drugs, pesticides, etc. from food test samples, biopsy samples, etc. is complicated. There are many.
On the other hand, these operations are often performed according to a predetermined procedure, and if these operations can be automated, data with high accuracy and reproducibility of the extraction operation can be obtained. Furthermore, there are merits such as reduction of labor cost, uniformity of extraction operation, and elimination of human error caused by humans.
As described above, it is preferable to automate the pretreatment of the test samples used for various analyzes as much as possible.

For example, the inventors of the present application have studied an automatic pretreatment method in which an extraction operation is automatically performed mechanically for efficient analysis in the analysis of pesticides.

JP 2013-3003

However, although the above literature is suitable for the analysis of pesticides, it has little applicability to other analyzes including veterinary drugs. Therefore, it was decided to develop a more versatile automatic pretreatment device.

As a result of various studies for the above purpose, the sample tube can be freely placed on a predetermined XY plane when various treatments such as extraction of a predetermined solvent or the like are performed as an automatic pretreatment device for a test sample. By adopting a transferable mechanism and moving the pipetting device such as the separation and supply of the solution together with the movement of the tube, it can be made a more efficient device, including veterinary drugs. , Found that it can be used as an automatic pretreatment device for various instrumental analyzes.

That is, as a result of examining the operability of various extraction processes, in the case of automation, the pipetting operation of sucking or discharging the liquid is performed with respect to the pipetting operation such as collecting the supernatant of the solution in the sample tube and adding the solvent. We have found that a method of integrating the device with the movement of the sample tube is effective.

That is, the first invention of the present application is
"1) A transfer axis, a transfer unit having a transfer unit that slides on the transfer axis and can move in a predetermined plane, and a transfer unit.
2) A tube elevating device that is attached to the transfer unit and can raise and lower the sample tube by sandwiching and releasing it.
3) A pipetting device that is attached to the transfer unit, has a predetermined tip that can be attached and detached, and can inhale or discharge liquid, and a pipetting device that can be raised and lowered.
An automatic pretreatment device for test samples. ",.

Further, in the automatic pretreatment apparatus according to claim 1, it has been found that a method of providing a screw cap attachment / detachment device as a sample tube of a screw cap type is effective.
That is, the second invention of the present application is
"In the automatic pretreatment device, the sample tube is a screw cap type sample tube, and further
4) According to claim 1, the screw cap type sample tube mounting portion and a cap attachment / detachment device capable of mounting / detaching the cap by sandwiching / rotating the cap portion of the screw cap and mounting the sample tube in a predetermined place are provided. An automatic pretreatment device for the described test sample. ",.

Next, in the present invention, it is preferable to provide a homogenizer mechanism for homogenizing a sample in a sample tube, a tube shaking mechanism, a dispensing device capable of dispensing a solvent into the tube, and a centrifuge device.

That is, the third invention of the present application is
"The automatic pretreatment device, and 5) a mobile stock section on which the screw cap type sample tube is placed and which can be slidably moved.
6) An elevating homogenizer device that can be inserted into the sample tube housed in the mobile stock unit, and
7) A rack for accommodating the sample tube, a shaking device having a shaking portion capable of shaking the rack, and a shaking device.
8) A solvent dispensing unit having a screw cap type sample tube mounting portion and a dispensing device provided with a pump portion capable of dispensing a solution into the sample tube.
9) A centrifuge for centrifuging the sample tube.
The automatic pretreatment apparatus for a test sample according to claim 2. ",.

Next, in order to carry out the further purification, equipment for column may be provided.
That is, the fourth invention of the present application is
"The automatic pretreatment device further
10) The automatic pretreatment apparatus for an inspection sample according to claim 3, further comprising a column processing unit. ",.

Further, as a concentration step, a mode equipped with a centrifugal evaporator is also possible.
That is, the fifth invention of the present application is
"The automatic pretreatment device further
11) The automatic pretreatment device for a test sample according to claim 4, which is provided with a centrifugal evaporator device. ",.

It is a schematic perspective view of the whole of the automatic pretreatment apparatus of the 1st Embodiment of this invention, and is the enlarged perspective view of the transfer part. It is a top view of the whole of the automatic pretreatment apparatus of the 1st Embodiment of this invention. It is a perspective view of the transfer unit (transfer part, moving shaft, fixed rail). It is a perspective view of the sample tube elevating device ((1) ascending state (2) descending state). It is a front schematic diagram which showed the operation of the sample tube elevating device. It is a perspective view of a pipetting device ((1) ascending state (2) descending state (3) photograph of ascending state (4) photograph of descending state). It is an enlarged view of the chip attachment / detachment part. It is a perspective view of a chip rack ((1) When the chip is not stored (2) Photograph when the chip is stored) It is a front schematic diagram of the process of inserting the tip portion of a tip into a sample tube. It is a perspective view of a cap attachment / detachment device ((1) drawing (2) photograph). It is a front view which showed the operation of a cap attachment / detachment. It is a perspective view which showed the mobile stock part and the homogenizer apparatus group. It is a perspective view of a shaking device ((1) photograph in a substantially horizontal state (2) photograph after rotation (3) perspective view). It is a perspective view of a solvent dispensing unit ((1) drawing (2) photograph). It is a perspective photograph of the stock part (for supplying to a solvent dispensing unit) of a solvent. Pressurizing unit (syringe pump section to the dispensing unit and syringe section for pressurizing the upper part of the column) It is a perspective view of a fixed stock part ((1) drawing (2) photograph). It is a perspective view of the processing part by a column or filtration ((1) drawing (2) photograph (3) whole photograph including solvent stock bottle). It is a perspective view of a centrifugal evaporator.

1 Moving shaft 3 Transfer part 5 Fixed rail 6 Transfer unit 7 Tube lifting device 8 Tube holding part 8-1 Tube holding bar 9 Pipetting device 10 Tip tip cover part
11 Chip rack 12 Chip attachment / detachment member 13 Cap attachment / detachment device 15 Cap rotation / movement part 16 Cap elevating part 17 Cap mounting part 18 Cap holding bar 21 Mobile tube mounting part 22 Tube holding part 23 Sliding movable stock part 25 Homogenizer part 26 Slide cover 27 Shaking device 28 Shaking device tube rack 29 Solvent dispensing unit 30 Solvent supply unit 31 Pressurizing unit (syringe unit and syringe pump unit)
31-1 Column pressure syringe 31-2 Syringe pump for solvent dispensing 33 Fixed stock unit 35 Centrifugal separator 37 Centrifugal evaporator device 39 Tube rack 40 Column processing unit 41 Column mounting unit 43 Column solvent supply unit 44 Column pressure supply Part 47 Solvent stock part 49 Centrifugal evaporator 51 Waste liquid bottle 53 Cooling trap 55 Suction pump
Tu screw type sample tube
C cap (screw tube lid)
Ti chip (tip)

The first embodiment of the present invention will be described below. However, it goes without saying that the present invention is not limited to these embodiments.

As shown in FIGS. 1 and 2, the automatic pretreatment apparatus according to the first embodiment of the present application is an automatic pretreatment apparatus for an inspection sample.
1) A movement axis, a transfer unit having a transfer unit that slides on the movement axis and can move in a predetermined plane, and a transfer unit.
2) A tube elevating device that is attached to the transfer unit and can raise and lower the screw cap type sample tube by sandwiching and releasing it.
3) A pipetting device that is attached to the transfer unit, has a predetermined tip that can be attached and detached, and can inhale or discharge liquid, and a pipetting device that can be raised and lowered.
4) A cap attachment / detachment device capable of attaching / detaching the cap by sandwiching / rotating the screw cap type sample tube mounting portion and the screw cap cap portion and placing the cap in a predetermined place.
5) A mobile stock section on which the screw cap type sample tube is placed and can be slidably moved,
6) An elevating homogenizer device that can be inserted into the sample tube housed in the mobile stock unit, and
7) A stock rack for accommodating the sample tube, a shaking device having a shaking portion capable of shaking the rack, and a shaking device.
8) A solvent dispensing unit having a screw cap type sample tube mounting portion and a dispensing device provided with a pump portion capable of dispensing a solution into the sample tube.
9) A centrifuge that centrifuges the sample tube and
10) Column processing unit and
11) Equipped with a centrifugal evaporator device.
An automatic pretreatment device for inspection samples.

Needless to say, the present invention is not limited to the first embodiment of the present invention. FIG. 1 is a schematic perspective view of the entire automatic pretreatment apparatus according to the first embodiment of the present invention. In FIG. 1, some configurations such as a waste liquid bottle and a tube rack are omitted.
Next, FIG. 2 is an overall plan view of the automatic pretreatment apparatus according to the first embodiment of the present invention. Some configurations of the transfer unit and the like are omitted.
Further, various devices and units are connected to a predetermined motor, various power units, a power source, etc. as a power source. These descriptions are omitted.

○ Transfer Unit The transfer unit 6 in the first embodiment of the present invention includes a transfer unit 3, a moving shaft 1, and a fixed rail 5 as shown in FIG. 3, and the transfer unit 3 includes a predetermined moving shaft 1. It is attached to. Further, the moving shaft 1 is mounted so that both ends of the moving shaft 1 can slide and move on a pair of fixed rails 5 orthogonal to the moving shaft 1.
As a result, the moving axis 1 moves in the Y-axis direction as shown in FIG. Further, since the transfer unit 3 can slide on the movement axis 1, it can move in the X-axis direction as shown in FIG.
In this way, the moving axis 1 can move in the Y-axis direction, and the transfer unit 3 can move in the X-axis direction on the moving axis 1, so that it can move freely on the XY plane of FIG. ing.
The transfer unit 3 is equipped with a tube elevating device 7 for sandwiching and releasing the tube Tu, which will be described later, and a pipetting device 9 for sucking or discharging the liquid.

As a result, the tube elevating device 7 and the pipetting device 9 are freely moved in the XY plane shown in FIG. Although not shown in FIG. 3, a drive unit that separately drives the movement of the moving shaft 1 in the Y-axis direction and a drive motor that slides the moving unit 3 on the moving shaft 1 are provided in the transfer unit. It is installed.

○ Sample tube elevating device In the present invention, the test sample is housed in the sample tube Tu having a cap C in a screw type, and various operations such as homogenization and extraction are performed in the tube Tu. Here, various devices are arranged on the XY plane in order to perform various treatments on the sample tube Tu (FIG. 2).

The tube Tu is carried together with the movement of the transfer unit 3 in a state of being sandwiched by the sample tube elevating device 7 mounted on the transfer unit 3 of the transfer unit 6 described above.
Next, after being placed at a predetermined position on a predetermined XY axis, it is necessary to lower the tube Tu by the sample tube elevating device 7 mounted on the above-mentioned transfer unit 3 in order to provide the tube Tu to various devices. There is.

Here, as shown in FIG. 4, the sample tube elevating device 7 is attached to the transfer portion 3, and a columnar holding portion 8 is provided inside a substantially rectangular parallelepiped housing so that the sample tube can be elevated and lowered. It is set freely. For the elevating mechanism, a known method using a pole screw or the like can be used.

Further, regarding the columnar holding portion 8, three sample tube holding bars 8-1 are provided around the circumference, and as shown in FIG. 5, the upper part of the sample cap C can be held and released. It is configured.

The tube Tu in the embodiment of the present invention discloses a screw-type cap tube Tu, but the present invention is not necessarily limited to this, and a cap-type tube that can be opened and closed by pressing or a cap C that can be opened and closed without a cap C is released. It is also possible to perform a pretreatment operation with the state tube. In this case, the device to be provided differs depending on the form of each tube.

○ Pipetting device As shown in FIG. 5, a pipetting device 9 which is attached to the transfer unit 3 to allow a predetermined tip to be attached and detached and to suck or discharge a liquid is provided. The pipetting device 9 is mounted on the transfer unit 3 described above, and is mounted adjacent to the sample tube lifting device 7 described above.

In the present invention, there is a description that the above-mentioned sample tube elevating device 7 and the pipetting device 9 are attached to the transfer unit 3, but the description of this attachment does not necessarily mean that the transfer unit 3 and the sample tube elevating device 3 are attached. It is not intended that the device 7 and the pipetting device 9 are clearly separated as members.
For example, even when these are integrally configured, each of the transfer unit 3, the sample tube elevating device 7, and the pipetting device 9 referred to in the present invention is provided.

As shown in FIG. 6, the pipetting device 9 is provided with a columnar holding portion inside a substantially rectangular parallelepiped housing, and is set so as to be able to move up and down. For the elevating mechanism, a known method using a pole screw or the like can be used.
As shown in FIG. 7, the pipetting device 9 has a predetermined fitting portion with respect to the tip Ti, and the tip Ti is attached / detached at the chip attachment / detachment portion provided above the fitting portion. ..

In addition, a conduit for pulling the inside is provided in the center of the fitting portion of the tip, and the conduit is configured to communicate with the suction device via a tube or the like, and the tip Ti. It is configured so that the solution can be sucked inside.

Then, a tip Ti of a predetermined size is attached to the fitting portion at the tip of the pipetting device 9 by pressing, so that the tip of the pipetting portion can suck the solution in the same manner as a normal Pipetman (registered trademark). It is configured.
That is, it is configured so that the inside can be sucked as a pulling pressure so that the solution can be sucked with the tip Ti attached. Further, by releasing the pressure, the sucked solution can be discharged.

Further, in the first embodiment of the present invention, the type to which the tip Ti is attached is disclosed, but the present invention is not limited to the configuration, and for example, a pipette made of glass or plastic is attached. Of course, the configuration may be such that
Further, when the chip Ti is used, it is necessary to replace the chip when handling different solutions, and the predetermined chip rack 11 shown in FIG. 8 is installed for the replacement. As a result, the tip Ti at the tip can be replaced.

The operation of the pipetting device 9 is that when the solution is supplied or sucked to the predetermined tube Tu, the cap C is removed while the device 9 is located above the target tube Tu. The pipetting device 9 descends into the opening of the tube Tu in the state, and the tip portion is inserted through the opening of the tube Tu. Then, it is configured to discharge or suck the solution.

Further, in the pipetting operation, it is preferable to prevent sagging due to liquid leakage from the tip of the tip Ti at the tip in a state where the solution to be dispensed is sucked.
Therefore, as shown in FIG. 6 (3), when the pipetting device is raised upward in the sucked state and moved via the movement of the transfer unit 3, the tip Ti is sagging due to liquid leakage from the tip end portion. It is preferable to move the transfer unit 3 with the chip tip cover portion 10 covering the lower part of the chip Ti tip portion so that the liquid can be received even if the liquid leaks.

Next, in the present invention, one of the two layers of the solution may be collected for purification of the sample or the like. In the case of the two layers, the inside of the tube Tu becomes a two-layer solution, and an interface between the layers may occur.

When the solution is sucked and discharged using the pipetting device 9, the tip Ti with the tube Tu slanted is lowered vertically downward so as not to suck the interface layer between the layers of the solution. The method is preferably adopted. That is, specifically, as shown in FIG. 9 below, when the tip Ti is inserted into the tube Tu, the tube Tu is tilted in advance and the tip Ti is lowered in this state. The tip Ti is made to penetrate into the lower layer while avoiding the interface as much as possible by lowering the tip Ti while avoiding the interface as much as possible so that the tip of the tip Ti follows the wall surface of the tube Tu. be able to.
As shown on the right side of FIG. 9, by sucking the solution with the tip of the tip Ti sufficiently inserted into the solution of the lower layer, the solution of the lower layer is sucked reasonably while avoiding the suction of the interface. It can be recovered.

When sucking and discharging the solution by the pipetting device 9, the tube Tu can be carried out by the mobile tube mounting portion 21 of the cap attachment / detachment device 13 described later. The tube mounting portion 21 is configured so that the tube Tu can be tilted at the time of the above-mentioned pipetting.
In this way, the sample can be set to be slanted so that the interface is not sucked by the pipetting operation. Further, a waste liquid bottle 51 is installed in a substantially central portion of FIG. 2 in order to discard the solution that becomes a residue by the pipetting operation.

In this way, when the solution is sucked from the chip Ti by the pipetting device 9 and collected, a new tube Tu for discharging the collected solution is placed in a predetermined rack and the used chip Ti is removed. It is preferable to prepare it in a chip disposal bottle or the like.
Next, the first embodiment of the present invention further has the following configuration.

-Cap attachment / detachment device The cap attachment / detachment device 13 in the first embodiment is a device that rotates the screw cap C to remove the cap C from the tube Tu or attach the cap C as necessary. As shown in FIG. 10, in the first embodiment, the cap attachment / detachment device 13 includes a movable tube mounting portion 21, a substantially rectangular parallelepiped cap rotating / moving portion 15, and a disk-shaped cap mounting portion. It has a part 17.

In the tube mounting portion 21, a tube Tu with a cap to be attached or detached from the cap C is mounted to support the lower portion of the tube Tu so that it cannot rotate.
Next, the cap rotating / moving portion 15 approaches the solution tube Tu with a cap carried to the tube mounting portion 21 for attaching / detaching the cap C, and the solution with a cap under the cap C is attached / detached. The tube Tu is positioned so that it is located.

Specifically, the tube mounting portion 21 is formed with a holding portion 22 of the tube Tu as shown in FIG. 11, and the tube Tu is inserted at intervals of the holding portion 22.

The lower portion and the bottom portion of the holding portion 22 are configured so that the lower portion of the tube Tu can be supported, fixed, and released, and when fixed, the rotation of the tube Tu is prevented. In this state, the cap rotating / moving portion 15 approaches from the upper part of the tube Tu, the cap elevating portion 16 is lowered, the three cap holding bars 18 are lowered, the cap C is held, and then the cap C is rotated. You can remove the cap C with.

Further, with respect to the removed screw type cap C, the removal of the cap C is completed by placing the cap C on the disk-shaped mounting portion 17.

On the other hand, when attaching the cap C, the tube Tu from which the cap C has been removed is placed on the upper part of the tube with the disc-shaped cap C sandwiched, and then the cap C is rotated and opened by the cap C. Close the part and complete the closure with cap C. The cap rotating / moving portion 15 approaches the upper portion while sandwiching the lower portion of the tube Tu, the cap elevating portion 16 descends, the cap C of the tube Tu is sandwiched, and the cap C is rotated. remove. Next, the cap C portion is placed on the upper part of the mounting portion 17 of the disk stand, and the process proceeds to the next step.

In the first embodiment of the present invention, the following configuration may be further provided.

-Mobile Stock Unit In the first embodiment of the present invention, a slide-movable stock unit 23 on which a plurality of sample tubes Tu of the screw cap can be placed is provided (FIGS. 1 and 2). In the present invention, as shown in FIG. 12, it is provided in the region in front of the homogenizer device 25.

○ Homogenizer device In the first embodiment of the present invention, an elevating homogenizer device 25 as shown in FIG. 13 that can be inserted into the sample tube Tu is provided.
The homogenizer device 25 is configured to be able to move up and down in the vertical and vertical directions, and in front of the homogenizer device 25, the tube Tu housed in the tube rack, which is a mobile stock portion 23 that can be positioned directly under the homogenizer device, moves. Then, it is carried so as to be located in the lower part of the homogenizer device 25.
In this state, the homogenizer device 25 descends vertically downward, passes through the hole of the slide cover 26 for preventing liquid dripping, the tip of the homogenizer is inserted into the tube Tu, and the homogenizer is operated. By doing so, solid matter and the like can be crushed.
After homogenization, the homogenizer device 25 is raised and allowed to stand still on the tube Tu for a short time so that the liquid on the homogenizer blade falls into the tube Tu. In addition, the slide cover can be moved to prevent liquid dripping from the homogenizer device. Further, the mobile stock portion (tube rack) 25 is moved and returned to its original position.

As shown in FIG. 12, the homogenization according to the embodiment of the present invention is arranged in a row in the Y-axis direction with respect to the first, third, fifth, seventh, ninth, and eleventh tubes Tu in order from the upper direction (back side of the paper surface) in the Y-axis direction. And a row of homogenizers in the Y-axis direction with respect to the second, fourth, sixth, eighth, tenth, and twelfth tube Tus are arranged in parallel, and the homogenizer rows for these tube Tus are arranged so as to be sequentially performed. Has been done.

○ Shaking device In the first embodiment of the present invention, the shaking device 27 is provided. In the shaking device 27, the sample tube Tu can be accommodated and shaken.

In the first embodiment of the present invention, the shaker 27 as shown in FIG. 13 is provided, and in the shaker 27, the tube Tu housed in the tube rack 28 of the shaker 27 is provided. On the other hand, before shaking, the slide cover having a plurality of L-shaped portions slides in the longitudinal direction of the tube rack so that the cover portion is located on the upper portion of the tube Tu. As a result, the tube Tu does not fall even if the rack 28 rotates, and the rack 28 rotates left and right as shown in FIG. 14 while the tube Tu is held by the rack 28, and the inside of the sample tube Tu. It is possible to shake and mix the solution of.

○ Solvent Dispensing Unit In the first embodiment of the present invention, as shown in FIG. 14, a solvent (solution) dispensing device 29 is provided. The dispensing device 29 is provided with a solvent supply unit 30 capable of dispensing a solvent (solution) to the sample tube Tu by moving the screw cap type sample tube mounting unit 21 in the cap attachment / detachment device 15. There is.
Regarding the solvent dispensing unit 29, a plurality of solvent supply units 30 are connected to a solvent stock bottle arranged in the solvent stock unit 47 by a flexible thin tube (FIG. 15), and the solvent in the stock unit 47 is connected. The solvent supplied from the stock bottle is dispensed.
The sample tube mounting portion 21 can be moved from the cap attachment / detachment device 15 to the solvent supply portion 30 because the solvent is often dispensed with the screw type cap C removed. be.

That is, in the sample tube Tu, the tube mounting portion 21 moves in a state where the cap C is removed from the tube Tu by the above-mentioned cap attachment / detachment device 13, and the sample tube Tu in which the cap C is removed and the upper part is open is released. It is moved and configured to be located below the tip of the solvent supply 30 of the solvent dispensing unit 29.
The solvent dispensing unit 29 according to the first embodiment of the present invention shown in FIG. 14 is configured to have six supply units 30 so that any of the six types of solvents can be dispensed. In order to dispense the solvent, the tube mounting portion 21 described above moves to the lower portion of the tip portion of any of the supply portions 30, and any one of the supply portions 30 slides in the direction of the arrow. , It will be located at the top of the tube Tu and is configured to be able to supply the solvent.

In addition, in order to make the amount of liquid in the tube Tu constant, it is possible to control the volume using an optical sensor so that the insufficient solution can be additionally supplied.
For example, when it is necessary to adjust the volume to 30 ml, if the solution in the tube Tu is 28 ml, an additional 2 ml can be supplied so as to be 30 ml by using an optical sensor.

Further, regarding the solvent (solution) to be dispensed by the dispensing device 29, there is a stock section 47 provided with a plurality of solvent stock bins as shown in FIG. 15, and the syringes shown in FIGS. 17 to 17 are present. The solvent is sucked and discharged by the operation of the pump unit 31-2.

This syringe pump unit 31-2 is a device for supplying a solvent such as a buffer stored in a predetermined solvent stock bottle into a tube Tu as needed.
A solvent for supply exists in the solvent stock bottle, and is set to be sucked and discharged by the syringe pump 31-2.
Further, the tube mounting portion 21 in the solvent dispensing unit 29 described above is also used in the mobile tube mounting portion 21 when performing pipetting operations such as suction and discharge of the solution by the pipetting device 9 described above. As shown in FIG. 9, the tube Tu is also configured to be tilted at a predetermined angle.

○ Centrifugal device In the present invention, the centrifugal device 35 for centrifuging the sample tube Tu is provided. Centrifugation This allows the residue and supernatant to be separated in a homogenized solution or the like. In the present invention, it is preferable to use a swing type rotor.

Further, in the centrifuge device 35, it is preferable that the tube elevating device 7 described above vertically lowers the tube Tu when the sample tube Tu is inserted into the rotor. Therefore, it is preferable that the tube accommodating portion of the rotor faces in the vertical direction while the rotor has stopped rotating.
Further, the first embodiment of the present invention has the following configuration.

-Fixed Stock Unit In the first embodiment of the present invention, a rack-type fixed stock unit 33 on which the sample tube Tu can be placed is provided. Further, the stock portion 33 is also a cooling unit so as to be held at a low temperature by a cooling trap 53 (FIG. 17).
A sample tube Tu can be placed on the stock section as appropriate. Further, it can be usefully used when holding the sample tube Tu or the organic solvent having a relatively high volatility after the centrifugation operation in a state where the cap C is removed.

○ Treatment unit by column In the first embodiment of the present invention, further purification can be performed by adding a purification step by column or filtration, which is preferable. In the first embodiment of the present invention, as shown in FIG. 18, a processing unit 40 for processing by a column is provided.
In the column processing unit, the column mounting unit 41 and the column solvent supply unit 43 provided with a syringe or the like for supplying a buffer or the like from the upper part of the column slide from the horizontal direction and are located on the upper part of the column. ..
Further, in order to promote the passage of liquid in the column, a column pressure supply unit 44 is also provided, which sends air or the like from the upper part of the mounted column to supply pressure to the column. FIG. 16 shows a pressure supply syringe 31-1 that supplies air pressure to the pressure supply unit.

○ Centrifugal evaporator device In the present invention, the centrifugal evaporator 49 can be used, and the final preparation of the sample can be made by adding a drying / concentrating step. In the first embodiment of the present invention, the configuration including the centrifugal evaporator device 49 is shown (FIG. 19).
The centrifugal evaporator device 49 is connected to a suction pump 55 installed on the right side surface of the automatic pretreatment device, stores a tube Tu with an open upper part, and volatilizes the solvent by centrifuging in a degassed state. Can be allowed to dry.
A sample can be finally completed by supplying a predetermined solvent to the tube Tu after drying and dissolving the contents.

In the centrifugal evaporator device 49, it is preferable that the above-mentioned tube elevating device 7 vertically lowers the tube Tu when the sample tube Tu is inserted into the rotor. Therefore, in the horizontal state, the tube receiving portion of the rotor is in the vertical direction. When the rotor or the centrifugal evaporator device 49 itself is tilted, it is preferable that the tube Tu is lowered vertically so as to be inserted into the tube receiver and accommodated in the rotor.

〇 Others Other than those shown in Fig. 2, waste liquid bottles 51, etc. for discarding the solution to be removed in the tube rack part for mounting a new unused screw type tube Tu and the pipetting operation are appropriately used as necessary. It can be provided. The screw type tube Tu can also be placed on the fixed stock portion 33 or the mobile stock portion 23 described above.

Next, a processing example of the inspection sample in the automatic pretreatment apparatus according to the first embodiment of the present invention will be shown. Since the present disclosure is merely a processing example, the present disclosure is not limited to this processing example.

In particular, the case of extracting, purifying, and drying veterinary drugs from foods will be described with reference to the operating procedures shown in Tables 1 and 2. However, it goes without saying that each setting condition is not limited to the ones shown below.
In the next pretreatment method for analysis of veterinary drugs, pretreatment is performed by three steps of extraction → purification → concentration.
This flow is shown in Tables 1 and 2 for hydrophilicity and lipophilicity, respectively. These are steps of eluting veterinary drugs from samples such as agricultural products and foods into an extraction solvent.

本発明の第一の実施態様においては、以下のように自動前処理装置が稼働して、機器分析（LC-MS/MS）に供するサンプルを自動で調製することができる。
スクリュー式のチューブTuに対して、試料（食品、農作物等の粉砕しているものが好ましい）を添加する。当該試料の重量については記録しておく。

In the first embodiment of the present invention, the automatic pretreatment apparatus is operated as follows, and a sample to be subjected to instrumental analysis (LC-MS / MS) can be automatically prepared.
Add a sample (preferably crushed food, crops, etc.) to the screw-type tube Tu. Record the weight of the sample.

Further, in the case of the first embodiment of the present invention, the screw type cap C can be left attached to the tube Tu.

In this first embodiment, 12 samples can be processed. The sample can be attached by arranging a tube Tu with a cap C in which a solid or liquid sample is contained in the mobile stock portion 23 shown in FIGS. 1 and 2. In this state, the automatic pretreatment by the automatic pretreatment apparatus of the first embodiment can be started.

1) When extracting hydrophilic veterinary drugs (1) Extraction process Movement and transfer of the moving axis 1 on the fixed rail 5 in the Y-axis direction with respect to the tube Tu with the cap C containing the sample. Through the movement in the Z-axis direction on the moving shaft 1 of the part 3, the tube elevating device 7 approaches the sample tube Ti with the cap C, and the sample cap holding bar 18 descends downward with respect to the upper part of the sample tube Ti. Then, the upper part of the sample tube Tu is sandwiched.

Then, in the pinched state, the pinching bar 18 rises upward, and subsequently moves via the moving shaft 1 and the fixed rail 5, and the sample tube Tu containing the sample is placed on the tube of the cap attachment / detachment device 13. Move to the place.

The holding cap rotating portion 15 of the cap attachment / detachment device slides horizontally with respect to the sample tube Tu placed on the tube mounting portion 17, and the cap portion of the tube Tu is rotated to remove the cap C. The cap C is placed on the disc-shaped cap mounting plate 17. The tube Tu containing the sample placed on the tube mounting portion of the attachment / detachment device slides in the Y-axis direction and is carried to a position for dispensing.

With respect to the tube Tu that has moved to the position for dispensing, one of the dispensing devices 29, the supply unit 30, is displaced in the X-axis direction so that it is located at the upper part of the tube Tu. Then, 15 ml of McKillpein buffer is poured from the upper supply unit 30. Next, the tube Tu containing the sample mounted on the tube mounting portion 21 of the attachment / detachment device slides in the reverse direction in the Y-axis direction and is returned to the original position.
The tube Tu after the addition of the buffer is sandwiched by the tube elevating device 7 from the tube mounting portion 21 which has returned to the original position in the same manner as described above, and is placed on the transferable stock portion 23 in front of the homogenizer device 25.
The operation is repeated, and the McKillbane buffer is sequentially poured into the tube Tu containing the samples placed in the mobile stock unit 23 in order, and the McKillbane buffer is added to the tube Tu containing all the samples. It is added and arranged in alignment with the mobile stock unit 23.

The mobile stock portion 23 slides to move each tube Tu to which the buffer has been added to the lower portion of the homogenizer device 25.

The homogenizer portion 25 is lowered, the tip portion of the homogenizer 25 enters the liquid surface of the tube Tu, and homogenization is started. Homogenization is performed for 2 minutes, and after homogenization, the homogenize portion 25 rises and the tip of the homogenizer separates from the sample tube Tu. The homogenization is performed in two rows depending on the number of samples.

The homogenized sample tube Tu is transferred by the tube elevating device 7 to the cap attachment / detachment portion 13, and the cap C is attached in the reverse manner of the previous step. Then, the sample tube Tu after the cap is attached is transferred to the centrifuge device 35 by the tube elevating device 7.

This operation is repeated an even number of times, and the homogenized sample tube Tu is placed on the centrifuge 35 so as to be a target. When the sample to be processed is an odd number, it is preferable to insert an empty tube Tu to make an even number.

The tube Tu after centrifugation is once sequentially transported to and arranged in the cooling storage rack 33. Meanwhile, the empty tube Tu is aligned with the rack 27 of the shaker with the cap C open. After the tube Tu after centrifugation placed on the cooling rack is sequentially removed from the cap C by the cap attachment / detachment device 13, the tube mounting portion 21 sucks / discharges (pipetting portion), and the supernatant liquid is collected. Then, it is added to an empty tube Tu prepared in advance in the rack 27 of the shaking device.
That is, the collected liquid is transferred to the tube Tu placed on the rack 27 of the shaking device. In the present embodiment, the sucked supernatant is carried by the suction / discharge mechanism by the pipetting device 9 mounted on the transfer unit and discharged into the new sample tube Tu.

The McKillpain buffer was poured into the sample tube Tu of the residue from which the buffer had been removed by the pipetting device 9 in the same manner as described above, and transferred to the homogenizing device 25 for homogenization (1 minute). After that, centrifugation is performed.

After homogenization + centrifugation, the supernatant liquid generated by the second homogenization + centrifugation is sucked into the tube Tu containing the supernatant liquid first, and is discharged by the pipetting operation.

By the two extraction operations, the tube Tu containing the recovered buffer is placed in a position for dispensing, and the McKillbane buffer is added and volumetrically prepared so that the volumetric flask is 30 ml. An optical sensor is used to accommodate a 30 ml line.

(2) Purification step If these operations are repeated and a veterinary drug is present, the fraction containing the component is recovered. Next, 20 ml of water-saturated hexane for purification is added to the recovered solution in 30 ml. The cap C is closed, and the tube Tu after the cap is closed is housed in the rack 28 of the shaking device 27. After repeating the storage a predetermined number of times, the rack 28 of the shaking device 27 repeatedly rotates and reciprocates back and forth to perform shaking.

The sample tube Tu after shaking is centrifuged in the same manner as described above, and then the upper hexane layer is removed by the pipetting device 9. The underlying McKillbane buffer layer is then recovered by a pipetting operation.

At this time, as shown in FIG. 9, in order to avoid suction at the interface between the hexane layer and the buffer layer in the pipetting operation, the sample tube Tu is tilted slightly, and the pipetting device is tilted from above in the tilted state. The chip Ti is operated so that the mounting portion is lowered downward and the chip Ti is inserted into the buffer layer so that suction at the interface with the hexane layer does not occur.

While the above purification operation with hexane is being carried out, in the column processing unit 40, the mounted column is placed, and a buffer or the like is supplied to the column from the upper part of the column. The column solvent supply unit 43 slides from the horizontal direction and moves to the upper part of the column to supply 5 ml of methanol from the attached supply syringe. In this way, the column layer is washed by passing the liquid at atmospheric pressure. Next, 5 ml of distilled water is passed through at atmospheric pressure.

Next, the pipetting device 9 approaches, and after mounting a new chip Ti, 20 ml of the lower layer of the solution in the tube after removing the hexane layer is taken and donated to the column (OASIS HLB). As the column, a column in which 5 ml of methanol and 5 ml of distilled water are passed under atmospheric pressure is used as conditioning in advance.

Next, 20 ml of the lower layer of the solution in the tube Tu is taken and fed to the column (OASIS HLB), and after passing the liquid under atmospheric pressure, air pressure is applied from the upper part to completely pass the column. Next, 5 ml of distilled water is provided to wash the column. After passing the liquid by atmospheric pressure, air pressure is applied again from the upper part to further pass the liquid. The tube Tu containing the waste liquid that has passed through the column is removed by the tube elevating device 7.
After that, a recovery tube Tu is placed in the lower part of the column, 5 ml of ethanol is added as an eluate, pressure is applied, and then the sample solution containing a hydrophilic veterinary drug is passed through the tube. Collect in Tu. Then, the liquid is further passed under air pressure to elute the ethanol recovery liquid in the column.

(3) Concentration step The sample tube elevating device 7 approaches the open tube Tu containing the ethanol recovery liquid containing hydrophilic veterinary drugs, carries it to the centrifugal evaporator 37, and stores it in the rotor. .. Place a blank of the same weight in the target area to balance the centrifuge.
While starting the suction (vacuum) pump for depressurization and degassing, centrifuge is performed to volatilize the solvent and dry under reduced pressure.

Next, 1 ml of 50% methanol is added (0.5 ml in the case of a dried product), the cap C is closed by the cap removing device 13, shaken by the shaking device 27 for 3 minutes, and stored at 4 ° C.

2) When extracting lipophilic veterinary drugs (1) Extraction step A sample tube elevating device 7 carries a 50 ml tube Tu containing 1.5 g of a powder or paste-like sample to a cap attachment / detachment device 13, and the cap C Open. The tube Tu after opening is transferred to the solution dispensing device 29, and acetonitrile is dispensed and added. The sample tube elevating device 7 transfers the dispensed tube Tu to the mobile stock section (slide rack) 23 under the homogenizing device 25, and the mobile stock section (slide rack) 23 is slid to the bottom of the homogenizing device 25. Then, the homogenizing device 25 is lowered and inserted into the liquid surface to crush and homogenize the solid matter.

After homogenization, the homogenizer device 25 rises and the slide rack 23 slides back to its original position. The tube Tu on the returned slide rack 23 is transferred to the centrifuge device 35 after the cap C is attached by the cap attachment / detachment device 13, and centrifugation is performed for a predetermined time.

After the centrifugation is completed, the tube elevating device 7 transfers the cap to the tube mounting portion 21 of the cap attachment / detachment device 13, and after the cap C is removed, the supernatant is sucked by the pipetting device 9 to which the tip Ti is attached. Qing is collected in a new tube Tu.
Next, the original tube Tu was moved to the solvent supply unit 30 of the solution dispensing device 29, and after acetonitrile was dispensed again, the tube Tu was homogenized by the tube elevating device 7 of the homogenizing device 25. It is reprinted below and homogenized as described above.

After the homogenization is followed by centrifugation, it is sucked by pipetting by the pipetting device 9 as before, and the supernatant is added to the new tube Tu.
By the two extraction operations, the tube Tu containing the recovered buffer is moved to the position for dispensing under the solvent dispensing unit, and the McKillbane buffer is added so that the volumetric flask is 30 ml. Will be done. An optical sensor is used to accommodate a 30 ml line.
Then, after closing the cap C and centrifuging, the cap C is opened and the centrifugal supernatant is collected in a new tube Tu. Repeat the above operation.

(2) Purification step If these operations are repeated and a veterinary drug is present, the fraction containing the component is recovered. Next, 20 ml of acetonitrim saturated hexane for purification is added to the recovered acetonitrile solution contained in 30 ml by using the solvent dispensing unit 29 and the pipetting device 9. The cap C is closed, and the tube Tu after closing the cap is housed in the tube rack 28 of the shaking device 27. After repeating the storage a predetermined number of times, the tube rack 28 repeatedly rotates and reciprocates left and right to shake.

The sample tube Tu after shaking is centrifuged in the same manner as described above, and then the upper hexane layer is removed by the pipetting device 9.
Then, 15 ml of acetonitrile-saturated hexane is added, the volume is adjusted to 50 ml, the cap C is rotated and attached by the cap attachment / detachment device 13, and then shaken in the mixer section for 10 minutes. After that, centrifugation is performed at 3000 rpm for 3 minutes by a centrifuge, the cap C is removed by the cap attachment / detachment device 13, and the upper hexane layer is removed.

Then, 10 ml of the lower layer is taken into a new tube Ti. At this time, in order to avoid suction at the interface between the hexane layer and the buffer layer in the pipetting operation, the sample tube Tu is tilted slightly, and the tip Ti of the pipetting device is mounted from above in the tilted state. Is manipulated so that the tip Ti is inserted into the buffer layer so that suction at the interface with the hexane layer does not occur. 10 ml of the lower acetonitrile layer is dispensed and dispensed into a new tube Tu.

(3) Concentration step The sample tube Tu elevating device approaches the open tube Tu containing the recovery liquid containing the new oil-based veterinary drug, carries it to the centrifugal evaporator 37, and stores it in the rotor. Place a blank of the same weight in the target area to balance the centrifuge.
While degassing by activating the suction (vacuum) pump 55 for depressurization, centrifugation is performed to volatilize the solvent and dry under reduced pressure.
Add 0.5 ml of 50% methanol, close cap C, shake with a mixer for 3 minutes, and store at 4 ° C.

Specific processing examples of veterinary drugs will be described below. However, it goes without saying that the present invention is not limited to these examples.
The following is an example of analysis of veterinary drugs.
In the case of the above-mentioned refining steps (hydrophilic and lipophilic)
[Example 1] (Automatic pretreatment device sample: Pig hydrophilic)
As a sample, pig muscle was used, and the hydrophilic veterinary drug shown in Table 3 was added to the sample, and a recovery test was carried out.

具体的には、豚の筋肉（生）（フードプロセッサーにより均一なペースト状にしたもの：500g）に表３に記載の４２種の親水性の動物用医薬品を添加し、添加・回収試験用の食品サンプルとした。

Specifically, 42 kinds of hydrophilic veterinary drugs shown in Table 3 are added to pig muscle (raw) (uniform paste made by a food processor: 500 g) for addition / recovery test. It was used as a food sample.

Using the food sample, the extraction operation was performed according to the above-mentioned procedure for the hydrophilic sample using the automatic pretreatment apparatus of the present invention.

The sample after extraction was analyzed by LC / MS / MS under the following conditions.
<LC-MS / MS measurement conditions>
Column: Octadecylsilylated silica gel Column temperature: 40 ° C
Mobile phase: Liquid A (0.1% formic acid aqueous solution) and liquid B (0.1% formic acid-containing methanol) are sent with a concentration gradient.
Mobile phase flow rate: 0.3 mL / min Ionization mode: ESI
Injection volume: 5 μL
As a result of the analysis, the number of veterinary drugs having a recovery rate of 70% or more and less than 120% was evaluated. The results are shown in Table 5.

[Comparative Example 1] (Manual sample: Pig hydrophilic)
The same as that carried out by the automatic pretreatment apparatus in Example 1 by adding the 42 kinds of lipophilic veterinary drugs shown in Table 3 prepared in Example 1 and using the food sample for the addition / recovery test. A technician manually extracted veterinary drugs according to the procedure described in. Other conditions are the same as those shown in Example 1.

[Example 2] (Automatic pretreatment device sample: shrimp hydrophilic)
Using shrimp as a sample, the hydrophilic veterinary drugs shown in Table 3 were added to the sample, and a recovery test was carried out.

Specifically, 42 kinds of lipophilic veterinary drugs shown in Table 3 were added to shrimp (a uniform paste made by a food processor: 500 g) to prepare a food sample for addition / recovery test.

Using the food sample, the extraction operation was performed according to the procedure of the lipophilic sample described above using the automatic pretreatment apparatus of the present invention. Other conditions are the same as those shown in Example 1.

[Comparative Example 2] (Manual sample: Shrimp hydrophilic)
Using the shrimp prepared in Example 2, 42 kinds of lipophilic veterinary drugs shown in Table 3 were added, and the automatic pretreatment apparatus was carried out in Example 2 using the food sample for the addition / recovery test. A technician manually extracted veterinary drugs in the same procedure as. Other conditions are the same as those shown in Example 2.

[Example 3] (Automatic pretreatment device sample: pig lipophilic veterinary drug)
As a sample, the pig muscle (raw) prepared in Example 1 was used, and the lipophilic veterinary drug shown in Table 4 was added to the sample, and a recovery test was carried out.

具体的には、豚の筋肉（生）（フードプロセッサーにより均一なペースト状にしたもの：500g）に表４に記載の154種の親油性の動物医薬品を添加し、添加・回収試験用の食品サンプルとした。

Specifically, 154 lipophilic veterinary drugs shown in Table 4 are added to pig muscle (raw) (uniform paste made by a food processor: 500 g), and foods for addition / recovery tests are added. It was used as a sample.

Using the food sample, the extraction operation was performed according to the procedure of the lipophilic sample described above using the automatic pretreatment apparatus of the present invention.

The sample after extraction was analyzed by LC / MS / MS under the following conditions.
<LC-MS / MS measurement conditions>
Column: Octadecylsilylated silica gel Column temperature: 40 ° C
Mobile phase: Liquid A (10% methanol containing 10 mmol / L ammonium acetate) and liquid B (methanol containing 10 mmol / L ammonium acetate) are fed with a concentration gradient.
Mobile phase flow rate: 0.3 mL / min Ionization mode: ESI
Injection volume: 5 μL
As a result of the analysis, the number of veterinary drugs having a recovery rate of 70% or more and less than 120% was evaluated. The results are shown in Table 5.

[Comparative Example 3] (Manual sample: Pork lipophilic)
The pig muscle (raw) prepared in Example 3, the 154 lipophilic veterinary drugs shown in Table 4 were added, and the food sample for the addition / recovery test was used, and the automatic pretreatment apparatus in Example 3 was used. The technician manually extracted the veterinary drug by following the same procedure as that performed by the technician.
Other conditions are the same as those shown in Example 3.

[Example 4] (Sample: Shrimp lipophilic)
As a sample, shrimp was used, and the lipophilic veterinary drug shown in Table 4 was added to the sample, and a recovery test was carried out.

Specifically, 154 kinds of lipophilic veterinary drugs shown in Table 4 were added to shrimp (a uniform paste made by a food processor: 500 g) to prepare a food sample for addition / recovery test.

Using the food sample, the extraction operation was performed according to the procedure of the lipophilic sample described above using the automatic pretreatment apparatus of the present invention. Other conditions are the same as those shown in Example 3.

[Comparative Example 4] (Manual sample: Shrimp lipophilic)
Using the shrimp prepared in Example 4, 154 kinds of lipophilic veterinary drugs shown in Table 4 were added, and the automatic pretreatment apparatus was carried out in Example 4 using the food sample for the addition / recovery test. A technician manually extracted veterinary drugs in the same procedure as. Other conditions are the same as those shown in Example 4.

結果として、手作業に比較して本自動前処理装置を利用すると、70％以上120％未満の回収率を得ることができる動物用医薬品の数が向上した。

As a result, the number of veterinary drugs that can obtain a recovery rate of 70% or more and less than 120% has been improved by using this automatic pretreatment device as compared with manual work.

Claims (4)

1) A movement axis, a transfer unit having a transfer unit that slides on the movement axis and can move in a predetermined plane, and a transfer unit.
2) A tube elevating device that is attached to the transfer unit and can raise and lower the screw cap type sample tube by sandwiching and releasing it.
3) A pipetting device that is attached to the transfer unit, has a predetermined tip that can be attached and detached, and can inhale or discharge liquid, and a pipetting device that can be raised and lowered.
4) A cap attachment / detachment device capable of attaching / detaching the cap by sandwiching / rotating the screw cap type sample tube mounting portion and the screw cap cap portion and placing the cap in a predetermined place.
5) A mobile stock section on which the screw cap type sample tube is placed and can be slidably moved,
6) An elevating homogenizer device that can be inserted into the sample tube housed in the mobile stock unit, and
7) A rack for accommodating the sample tube, a shaking device having a shaking portion capable of shaking the rack, 8) a mounting portion for the screw cap type sample tube, and dispensing a solution into the sample tube. A solvent dispensing unit with a dispensing device, which is equipped with a capable pump section, and
9) A centrifuge for centrifuging the sample tube.
Automatic pretreatment device for inspection samples. The automatic pretreatment device further
10) The automatic pretreatment apparatus for an inspection sample according to claim 1, further comprising a column processing unit. The automatic pretreatment device further
11) The automatic pretreatment device for a test sample according to claim 2, further comprising a centrifugal evaporator device. The automatic pretreatment device according to any one of claims 1 to 3, wherein the automatic pretreatment device for a test sample is an automatic pretreatment device for a test sample used for analysis of a veterinary drug.

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