SAMPLE PRETREATMENT DEVICE, ANALYSIS SYSTEM INCLUDING THE DEVICE, AND AUTOSAMPLER
20220074960 · 2022-03-10
Assignee
Inventors
- Kuniaki KANAMARU (Kyoto-shi, Kyoto, JP)
- Akiko BAMBA (Kyoto-shi, Kyoto, JP)
- Shunsuke ATSUMI (Kyoto-shi, Kyoto, JP)
- Hiroaki MURAKAMI (Kyoto-shi, Kyoto, JP)
- Yosuke IWATA (Kyoto-shi, Kyoto, JP)
Cpc classification
G01N2035/0405
PHYSICS
G01N30/7233
PHYSICS
G01N1/28
PHYSICS
International classification
Abstract
A sample pretreatment device of an aspect of the present invention includes a conveyance mechanism (22) including a Z-axis guide (223) that linearly moves in two axes of an X axis and a Y axis and expands/contracts in a Z-axis direction on a table (21) where a unit that performs each step of pretreatment is arranged. The Z-axis guide (22) is integrally provided with a liquid handler (224) and a grip handler (225) oriented in opposite directions to each other. A side wall of a housing (20) on the autosampler side is provided with a sample conveyance opening (201). When pretreatment of a sample ends, the Z-axis guide (223) is moved to a position close to the opening (201) in a state where a gripper (228) grips a pretreated plate. An arm (227) enters a housing of the autosampler through the opening (201) and an opening formed in the autosampler, and sets a plate at a predetermined position. Thus, the sample pretreatment device can eliminate the labor of the worker setting a pretreated sample in the analysis device.
Claims
1. A sample pretreatment device that performs pretreatment of a sample to be analyzed by an analysis device, the sample pretreatment device comprising: a housing which forms an outer shape of the sample pretreatment device; a sample conveyance opening formed on a wall of the housing on a side where the analysis device is positioned; and a sample conveyance unit which includes a grip unit for gripping a sample container containing a sample having been pretreated in the housing, and which moves the grip unit in the housing and moves the grip unit to an inside of a housing of the analysis device through the sample conveyance opening and an opening formed in a housing of the analysis device.
2. The sample pretreatment device according to claim 1 further comprising: a shutter configured to open or close the sample conveyance opening, wherein the shutter is in an opened state when the sample conveyance unit conveys the sample container from the sample pretreatment device to the analysis device, and the shutter is in a closed state when the sample conveyance unit does not convey the sample container from the sample pretreatment device to the analysis device.
3. The sample pretreatment device according to claim 1, wherein the sample conveyance opening and the opening of the analysis device are set to a size that allows the grip unit and the sample container gripped by the grip unit to pass through in a state where the sample pretreatment device and the analysis device are arranged in a predetermined positional relationship.
4. The sample pretreatment device according to claim 1, wherein the sample conveyance unit has a movable body capable of linearly moving in three axial directions orthogonal to one another in the housing, and the grip unit provided on the movable body.
5. The sample pretreatment device according to claim 4, wherein the grip unit is provided on a surface of the movable body on a side where the analysis device is positioned, and a dispense unit configured to suck and discharge a liquid is provided on a surface of the movable body on a side opposite to a side where the analysis device is positioned.
6. The sample pretreatment device according to claim 5, wherein the sample conveyance unit does not include a rotary drive mechanism that rotates the grip unit and the dispense unit, and the movable body linearly moves in three axial directions orthogonal to one another in a state where the grip unit always exists on a side where the analysis device is positioned and the dispense unit always exists on a side opposite to a side where the analysis device is positioned.
7. The sample pretreatment device according to claim 5, wherein the grip unit includes an arm and a gripper, and a length of the arm and a position of the gripper are set so that when the movable body moves to a predetermined position on a side where the analysis device is positioned, the sample container gripped by the gripper comes to a position corresponding to a location where the sample container is supposed to be arranged in the analysis device.
8. The sample pretreatment device according to claim 5, wherein when an inside of the housing is viewed from top, a dispense-operable region, which is a range in which the dispense unit is movable, and a conveyance-operable region in which the grip unit can perform a conveyance operation are set so as to partially overlap with each other, and a unit requiring both a dispense operation and a sample conveyance operation in a pretreatment step is arranged in the overlap region, a unit requiring only a dispense operation in a pretreatment step is arranged in the dispense-operable region other than the overlap region, and a unit requiring only a sample conveyance operation in a pretreatment step is arranged in the conveyance-operable region other than the overlap region.
9. The sample pretreatment device according to claim 8, wherein a unit requiring both a dispense operation and a sample conveyance operation in a pretreatment step includes at least one of a vacuum suction filtration unit and a constant-temperature shake unit, a unit requiring only a dispense operation in a pretreatment step includes a reagent reservoir unit, and a unit requiring only a sample conveyance operation in a pretreatment step includes at least one of a sealer unit and a storage area.
10. An analysis system including the sample pretreatment device according to any one of claims 1 to 9 and the analysis device, wherein the analysis device includes an analysis device body unit configured to perform analysis, and an autosampler configured to store a plurality of samples to be analyzed in the analysis device body unit, to sequentially select the plurality of samples, to collect selected samples, and to feed them to the analysis device body unit, and the sample conveyance unit of the sample pretreatment device conveys samples to the autosampler.
11. The analysis system according to claim 10, wherein the analysis device body unit is a liquid chromatograph or a liquid chromatograph mass spectrometer.
12. The analysis system including the sample pretreatment device according to any one of claims 1 to 9 and the analysis device, wherein a housing of the sample pretreatment device and a housing of the analysis device are configured by one common housing.
13. An autosampler used in the analysis system according to claim 10, the autosampler comprising: a housing; and a sample reception opening formed on a wall of the housing on a side where the sample pretreatment device is positioned, wherein the grip unit of the sample conveyance unit configured to grip a sample container containing a sample having been pretreated in the housing of the sample pretreatment device is moved to an inside of the housing of the autosampler through the sample conveyance opening and the sample reception opening.
14. The autosampler according to claim 13 further comprising: a shutter configured to open or close the sample reception opening, wherein the shutter is in an opened state when the sample conveyance unit conveys the sample container from the sample pretreatment device to the autosampler, and the shutter is in a closed state when the sample conveyance unit does not convey the sample container from the sample pretreatment device to the autosampler.
Description
BRIEF DESCRIPTION OF DRAWINGS
[0064]
[0065]
[0066]
[0067]
[0068]
DESCRIPTION OF EMBODIMENTS
[0069] An LC-MS analysis system that is an embodiment of the sample pretreatment device according to the present invention and the analysis system including the sample pretreatment device will be described below in detail with reference to the accompanying drawings.
[0070]
[0071] The LC-MS analysis system of the present embodiment includes an analysis device 1 and a sample pretreatment device 2. The analysis device 1 includes an autosampler 11, a liquid chromatograph (LC) 12, and a mass spectrometry (MS) 13. These units are independent units, and are connected by appropriate piping and wiring that are not illustrated.
[0072] In the present system, the analysis device 1 and the sample pretreatment device 2 are arranged in a predetermined positional relationship. The predetermined positional relationship means, as described in detail later, a position at which the sample container containing the sample pretreated in the sample pretreatment device 2 can be conveyed to the analysis device 1. As shown in
[0073] The autosampler 11 is a device capable of containing a plurality of microplates (microwell plates or deep-well plates), and repeats, in accordance with a predetermined program, an operation of sucking a predetermined amount of a liquid sample stored in one well of one of the microplates and feeding the liquid sample to the LC 12. The microplate has plural wells. Here, the number of wells is, for example, 8×12=96, as an example.
[0074] The sample pretreatment device 2 has a function of carrying out a pretreatment of a liquid sample, and automatically setting, at a predetermined position (shown by a reference sign 113 in
[0075]
[0076] As shown in
[0077] A space in the housing 20 above the table 21 is provided with a conveyance mechanism 22. The conveyance mechanism 22 includes an X-axis guide rail 221 extending in the X-axis direction and attached near the top surface in the housing 20, and a Y-axis guide rail 222 extending in the Y-axis direction and movable in the X-axis direction along the X-axis guide rail 221. The Y-axis guide rail 222 is attached with a Z-axis guide 223 movable in the Y-axis direction along the guide rail 222. The Z-axis guide 223 is attached with a grip handler 225 provided on the surface on the side (right in
[0078] Here, a rotary drive mechanism that rotates the liquid handler 224 and the grip handler 225, which is often provided in the conventional device, in order to downsize the conveyance mechanism. Therefore, the liquid handler 224 and the grip handler 225 do not rotationally move. The grip handler 225 always exists on the side where the autosampler 11 is positioned, and the liquid handler 224 always exists on the side opposite to the side where the autosampler 11 is positioned.
[0079] The grip handler 225 has a pair of arms 227 separated by a predetermined distance in the Y-axis direction and movable so that the distance varies in a predetermined range. The tip of each arm 227 is attached with a gripper 228 that holds and grips a microplate or a chip rack described later. That is, the grip handler 225 conveys a microplate or a chip rack. The length of the arm 227 in the X-axis direction (i.e., direction in which the autosampler 11 is positioned) and the position at which the gripper 228 is formed are set so that when the guide rail 222 reaches the end portion on the autosampler 11 side in the X-axis direction, the microplate or the like gripped by the gripper 228 is at a position facing the upper portion of the microplate installation region, which is a predetermined position of the autosampler 11.
[0080] On the other hand, the liquid handler 224 detachably has a chipset 226 to which a plurality (eight in this example) of pipetting chips (hereinafter referred to simply as a “chip”) are coupled, and performs dispense by sucking a liquid sample or a reagent into each chip of the chipset 226 and discharging the sucked liquid. The chipset 226 has the eight chips coupled at the same pitch as adjacent eight wells of the microplate so that a liquid can be simultaneously dispensed into the eight wells. A liquid sample or a reagent adheres to the chip, and hence, in order to avoid contamination, the chip unit is disposable and normally replaced automatically after each use.
[0081] The temperature-control plate 23 temporarily stores, at a predetermined temperature, a large number of liquid samples to be pretreated. On the temperature-control plate 23, for example, a vial rack 32 (or other labware) that holds plural small vials each containing a liquid sample is placed. When the liquid sample is a biological sample such as blood, the temperature-control plate 23 is set to a low temperature in the range of, for example, about 0 to 10° C. in order to prevent denaturalization of the components in the sample before pretreatment.
[0082] The reagent reservoir unit 24 holds a plurality of reagent reservoirs containing various kinds of reagent used for pretreatment. The reagents to be used vary depending on the purpose and type of pretreatment.
[0083] The waste box 25 has a waste port 251 opening upward and is for discarding used chips. As shown in
[0084] The constant-temperature shake unit 26 shakes a microplate (filter-type microplate described later) 30a (or other labware) placed on the upper surface of the constant-temperature shake unit 26 while holding the microplate 30a at a predetermined temperature, thereby stirring the liquid in each well of the microplate 30a and accelerating the mixing and chemical reaction of the liquid sample with the reagent.
[0085] The vacuum suction filtration unit 29 includes a vacuum manifold, and performs filtration by performing vacuum suction to the microplate 30a corresponding to vacuum suction filtration, for example, to remove unnecessary components and the like from the liquid sample.
[0086] The sealer unit 27 substantially seals each well of a microplate 30b by attaching a predetermined sealing material to the upper surface of the microplate 30b containing the pretreated liquid sample. For convenience of explanation, a general microplate before the sealing material is attached by the sealer unit 27 is denoted by a reference sign 30b, and a general microplate after the sealing material has been attached is denoted by a reference sign 30c.
[0087] The storage area 28 includes a shelf-like first region 281 that stores a plurality of unused microplates (filter-type microplates and normal microplates) 30d and a shelf-like second region 282 that stores a plurality of chip racks 31b containing an unused chip.
[0088] At a predetermined position on the right side wall of the housing 20 facing the autosampler 11, a sample conveyance opening 201 is formed, and the sample conveyance opening 201 is openable/closable by a shutter 202. The sample conveyance opening 201 is typically formed in a rectangular shape, and the longitudinal and lateral lengths of the sample conveyance opening 201 are set so as to allow the arm 227, the gripper 228, the microplate held by the gripper 228, and the like to pass through.
[0089] As described above, the storage area 28 shown in
[0090] The units on the table 21 described above can be roughly divided into the following three, those requiring handling of both the liquid handler 224 and the grip handler 225 (group A), those requiring only handling by the grip handler 225 (group B), and those requiring only handling by the liquid handler 224 (group C).
[0091] Specifically, the sealer unit 27 and the storage area 28 fall into the group B, the reagent reservoir unit 24 falls into the group C, and the other blocks, except them and the temperature-control plate 23, fall into the group A. In the sample pretreatment device 2 of the present embodiment, the blocks requiring handling by the liquid handler 224, i.e., the group A and the group C, are arranged in the range of La in
[0092] In the sample pretreatment device 2 of the present embodiment, the liquid handler 224 and the grip handler 225 are attached on surfaces of one Z-axis guide 223 opposite to each other, and a rotary drive mechanism that rotates the liquid handler 224 and the grip handler 225 is not provided. Therefore, there are restrictions on the movable range of the liquid handler 224 and the movable range of the grip handler 225 on the X-axis. On the other hand, by appropriately arranging the blocks included in the groups A to C on the table 21 as described above, the liquid handler 224 and the grip handler 225, having the restrictions on the movable range as described above, are each made possible to perform necessary handling. This simplifies the structure of the conveyance mechanism 22, which is advantageous in reducing the installation floor space, compared with the case where the liquid handler 224 and the grip handler 225 are operated separately or one of them is not a linear drive system but a rotary drive system.
[0093] Here, the configuration of the autosampler 11 that receives the microplate 30c containing the pretreated sample from the sample pretreatment device 2 will be described.
[0094] As shown in
[0095] Similar to the sample conveyance opening 201, the sample reception opening 111 is also provided with an openable/closable shutter 112 so that, except when the microplate 30c is conveyed, the shutter 112 can close the sample reception opening 111. When the shutters 202 and 112 are both in an opening state, the space in the housing 20 of the sample pretreatment device 2 and the space in the housing 110 of the autosampler 11 substantially communicate with each other through the sample conveyance opening 201 and the sample reception opening 111.
[0096] Next, an example of the operation from when the sample pretreatment device 2 carries out pretreatment to when the microplate containing the pretreated liquid sample is conveyed from the sample pretreatment device 2 to the autosampler 11 will be described with reference to the flowchart shown in
[0097] The worker opens the door body 203 of the sample pretreatment device, and places, on the temperature-control plate 23, the vial rack 32 in which plural vials each containing a liquid sample have been set. Then, the worker closes the door body 203, and instructs the start of treatment by an operation unit that is not illustrated. The temperature-control plate 23 cools the sample contained in each vial to an appropriate temperature within a range of about 0 to 10° C. This suppresses denaturalization of the components in the sample.
[0098] In the sample pretreatment device 2, the liquid handler 224 mounted with the unused chipset 226 is moved to a predetermined position on the vial rack 32, and the liquid handler 224 sucks the liquid sample from each of the plurality (eight) of vials in the vial rack 32 and dispenses the liquid sample into each well of the filter-type microplate 30a placed on the constant-temperature shake unit 26 (step S1). The filter-type microplate used herein is, for example, a microplate having a structure in which, even if an organic solvent is injected inside, the solution does not drop unless vacuum-sucked, e.g., Impact (registered trademark) commercially available from Phenomenex Inc.
[0099] At the time of work of step S1, the constant-temperature shake unit 26 is stopped, and its temperature is kept low so as to suppress denaturalization of the sample. The liquid handler 224 dispenses the liquid sample into all the wells of the filter-type microplate 30a by repeating the operation of discarding the used chipset 226 into the waste box 25 and mounting the unused chipset 226 to perform the dispense.
[0100] The liquid handler 224 then sucks a prescribed amount of a predetermined reagent from the reagent reservoir placed in the reagent reservoir unit 24 and dispenses it into a predetermined well in the filter-type microplate 30a, thereby adding the reagent to the liquid sample (step S2). This reagent is, for example, a reagent for binding to a specific component in a liquid sample by a chemical reaction to precipitate the specific component, or for denaturing and precipitating protein when the removal target is the protein.
[0101] After the reagent is added to the liquid samples in all the wells of the filter-type microplate 30a, the constant-temperature shake unit 26 shakes the filter-type microplate 30a while appropriately raising the temperature, thereby stirring, for a predetermined length of time, the liquid sample to which the reagent has been added (step S3). This accelerates the chemical reaction between the liquid sample and the reagent. In some cases, the reaction is further accelerated by standing under a constant temperature for a predetermined time after stirring (step S4).
[0102] After that, the grip handler 225 is moved to a position where the filter-type microplate 30a on the constant-temperature shake unit 26 can be gripped, and the filter-type microplate 30a is gripped and conveyed to the vacuum suction filtration unit 29. The vacuum suction filtration unit 29 filters the liquid sample contained in each well of the filter-type microplate 30a by vacuum suction filtration, and drips the filtrate into the general microplate 30b having been installed in advance in the lower stage of the vacuum suction filtration unit 29, thereby removing the precipitated solid matter (step S5). This removes an unnecessary component and the like in the liquid sample from the liquid sample. Then, the grip handler 225 conveys, to a predetermined position of the sealer unit 27, the microplate 30b in which the liquid sample after the unnecessary component has been removed is stored. The sealer unit 27 attaches a sealing material so as to substantially seal each well of the microplate 30b (step S6). Thus, the series of pretreatment ends.
[0103] The grip handler 225 grips the microplate 30c on which the seal is applied, and conveys the microplate, to the inside of the housing 110 of the autosampler 11, through the sample conveyance opening 201 in which the shutter 202 is opened and the sample reception opening 111 in which the shutter 112 is opened (step S7). As shown in
[0104] It is preferable that each of the shutters 202 and 112 is opened only when the microplate 30c is conveyed from the sample pretreatment device 2 to the autosampler 11, and the sample conveyance opening 201 and the sample reception opening 111 are closed in other periods. Thus, it is possible to prevent temperature change due to intrusion of outside air or the like when the temperature of the internal space of the autosampler 11 or the internal space of the sample pretreatment device 2 is controlled. It is also possible to suppress the electric power consumption for temperature control.
[0105] As described above, in the LC-MS analysis system of the present embodiment, the microplate containing the liquid sample for which a predetermined pretreatment has been performed in the sample pretreatment device 2 can be automatically conveyed from the sample pretreatment device 2 to the autosampler 11. Thus, it is possible to reduce the labor of the worker due to this conveyance. Even in a case of use of a reagent or the like harmful to the human body, it is possible to reduce the risk of the worker coming into contact with such a reagent at the time of conveyance work of the microplate. Furthermore, since the configuration of the conveyance mechanism 22 is simple as described above, and the work and treatment by the liquid handler 224 and the grip handler 225 are performed by one X, Y, and Z three-axis drive system, it is possible to reduce the floor space of the sample pretreatment device 2 on the table 21. Thus, it is possible to suppress the installation floor space of the sample pretreatment device 2 and install the sample pretreatment device 2 and the analysis device 1 on a dedicated table with a top plate having a relatively small floor space.
[0106] Of course, the pretreatment performed in the sample pretreatment device 2 is not limited to those described above, and appropriate treatments can be combined in accordance with the purpose or the like. For example, not for the purpose of removing an unnecessary component or the like in the sample but for the purpose of labeling or derivatizing a specific component (or a specific site of a specific component) in the sample, the pretreatment may be performed so that a predetermined reagent is added to the sample and shaking is performed in order to accelerate the reaction of the labeling or derivatizing of the sample.
[0107] In the sample pretreatment device in the embodiment described above, the liquid handler 224 and the grip handler 225 are provided so as to be integrally movable in the Z-axis direction with respect to the Z-axis guide 223, but the liquid handler 224 and the grip handler 225 may be independently movable in the Z-axis direction with respect to the Z-axis guide 223. In that case, although the number of drive mechanisms increases, the restriction of the movable range of each of the liquid handler 224 and the grip handler 225 decreases, and it becomes possible to utilize the space in the housing 20 more effectively.
[0108] It is preferable to provide the shutter 202 that opens/closes the sample conveyance opening 201 of the sample pretreatment device 2 and the shutter 112 that opens/closes the sample reception opening 111 of the autosampler 11. However, these are not essential elements. In particular, since the entire interior of the autosampler 11 is often temperature-controlled, it is desirable that the sample reception opening 111 of the autosampler 11 is provided with the shutter 112. On the other hand, in the sample pretreatment device 2, partial temperature control is performed by the temperature-control plate 23 or the like, but the entire interior of the sample pretreatment device 2 is not necessarily temperature-controlled (of course, it may be temperature-controlled). Therefore, the sample conveyance opening 201 of the sample pretreatment device 2 may not have the shutter 202, but it is preferable to have the shutter 202 in order to prevent dust and the like from entering from the outside as much as possible.
[0109] While the positional relationship between the sample pretreatment device 2 and the analysis device 1 is not limited to that shown in
[0110] The system may be configured such that a rack changer is arranged close to the sample pretreatment device, and a microplate and other labware having been pretreated in the sample pretreatment device are conveyed to the rack changer as described above and further transferred from the rack changer to the autosampler.
[0111] The housing of the sample pretreatment device and the housing of the analysis device (autosampler or integrated liquid chromatograph) may be configured by one common housing. Even in a case of where the housing is thus commonalized, it is necessary to temperature-control the inside of the autosampler, and hence a wall that separates the sample pretreatment device from the autosampler is formed between them, and the wall is provided with an opening serving as both a sample conveyance opening and a sample reception opening. The opening is provided with a shutter that opens when the sample is conveyed and closes at other times. In such a configuration, it can be said that the wall between the sample pretreatment device and the autosampler is a part of the housing of the sample pretreatment device and the analysis device (or autosampler). In this case, it is not necessary to arrange the sample pretreatment device and the analysis device in the predetermined positional relationship as described above.
[0112] The above-described embodiment is merely an example of the present invention, and it is natural that any changes, modifications, and additions made as appropriate within the scope of the present invention are included in the scope of the claims.
REFERENCE SIGNS LIST
[0113] 1 . . . Analysis Device [0114] 11 . . . Autosampler [0115] 20, 110 . . . Housing [0116] 111 . . . Sample Reception Opening [0117] 112, 202 . . . Shutter [0118] 12 . . . Liquid chromatograph (LC) [0119] 13 . . . Mass Spectrometry (MS) [0120] 2 . . . Sample Pretreatment Device [0121] 201 . . . Sample Conveyance Opening [0122] 203 . . . Door Body [0123] 21 . . . Table [0124] 22 . . . Conveyance Mechanism [0125] 221 . . . X-Axis Guide Rail [0126] 222 . . . Y-Axis Guide Rail [0127] 223 . . . Z-Axis Guide [0128] 224 . . . Liquid Handler [0129] 225 . . . Grip Handler [0130] 226 . . . Chipset [0131] 227 . . . Arm [0132] 228 . . . Gripper [0133] 23 . . . Temperature-Control Plate [0134] 24 . . . Reagent Reservoir Unit [0135] 25 . . . Waste Box [0136] 251 . . . Waste Port [0137] 26 . . . Constant-Temperature Shake Unit [0138] 27 . . . Sealer Unit [0139] 28 . . . Storage Area [0140] 29 . . . Vacuum Suction Unit [0141] 30a . . . Filter-Type Microplate [0142] 30b, 30c . . . Microplate [0143] 30d . . . Unused Microplate [0144] 31a, 31b . . . Chip Rack [0145] 32 . . . Vial Rack