Sample pretreatment device and method

Abstract

To provide a device capable of shortening time required for pretreatment of a sample.SOLUTION: A sample pretreatment device includes: a container storage part that stores a container for storing supply object liquid; a pipette that suctions and discharges the supply object liquid; a column tube holder 22 for centrifugal separator that is detachably installed on a rotating unit 21 of a centrifugal machine 20, and includes a holding part 2251 for holding a column tube and a waste liquid port 2252; a pipette moving mechanism; and a control unit that controls the pipette moving mechanism and the pipette such that the pipette is moved and the supply object liquid in an amount greater than a predetermined amount is suctioned from the container by the pipette, the pipette is moved and the supply object liquid in the pipette is discharged by the predetermined amount to the column tube, then the pipette is moved and the remaining supply object liquid in the pipette is discharged to the waste liquid port 2252.SELECTED DRAWING: Figure 2

Description

[Technical field]

[0001] The present invention relates to an apparatus and method for pretreating a sample using a reagent before the sample is analyzed by a liquid chromatograph (LC), a liquid chromatograph mass spectrometer (LC-MS), or the like. [Background technology]

[0002] When analyzing biological samples, etc., by LC, LC-MS, etc., a pretreatment is carried out by adding a specific reagent to the sample. Non-Patent Document 1 describes that, as a pretreatment, a treatment is carried out in which a reagent, which is a solution containing an enzyme having the effect of cleaving sugar chains from an antibody, is added to a culture solution containing an antibody, which is a sample, or a reagent having the effect of fluorescently labeling the cleaved sugar chains is added to the culture solution containing the antibody, which is a sample.

[0003] Patent Document 1 describes an apparatus for automatically pretreating samples. This apparatus includes a sample placement section in which multiple sample containers are placed, a reagent placement section in which multiple reagent containers are placed, a dispensing port in which a column tube (described later) is placed, a sampling arm having a pipette (nozzle) at the tip and moving the pipette, and a washing port for disposing of waste liquid in the pipette and washing the pipette. The column tube (described as a "separation device" in Patent Document 1) is formed by a separation membrane having a function of selectively separating specific components and provided at the bottom of a cylindrical container. In this apparatus, the pipette is moved to a sample container (or a reagent container) to aspirate the sample (reagent), then the pipette is moved to the column tube to eject the sample (reagent) in the pipette into the cylindrical container of the column tube, and the pipette is further moved to the washing port to discharge the remaining liquid in the pipette into a waste part in the washing port, and the pipette is washed. These operations are repeated every time one sample or reagent is supplied to the column tube. In the column tube supplied with the sample and reagent, negative pressure is applied from the outside to the bottom, and if the components to be analyzed do not pass through the separation membrane and remain, the liquid that passes through the separation membrane is discarded, and if the components to be analyzed pass through the separation membrane, a collection container is placed outside the separation membrane to collect the liquid.

[0004] When separating a specific component with a column tube, instead of using the negative pressure described in Patent Document 1, centrifugal force (centrifuge) can also be used. For example, Patent Document 2 describes a column tube holder for a centrifuge, which is detachably installed on the rotating part of the centrifuge and includes a plurality of holding parts each holding a column tube, and a storage part receiving and storing liquid discharged from the column tube held in each holding part by centrifugal force. When the component to be analyzed does not pass through the separation membrane and remains, the liquid that has passed through the separation membrane by applying centrifugal force is stored in the storage part and is discarded during maintenance of the centrifuge. When the component to be analyzed passes through the separation membrane, the column tube with a collection container placed outside the separation membrane is held in the holding part and centrifuged, and the liquid is collected in the collection container. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] JP 2017-067484 A [Patent Document 2] Design Registration No. 1736411 [Non-patent literature]

[0006] [Non-Patent Document 1] "Glyco-related products Antibody glyco-analysis kit EZGlyco mAb-N Kit with 2-AB", [online], Sumitomo Bakelite Co., Ltd., [searched September 6, 2023], Internet<URL:https: / / www.sumibe.co.jp / product / s-bio / glycan / ezglyco / > Summary of the Invention [Problem to be solved by the invention]

[0007] When using the column tube holder for a centrifuge described in Patent Document 2, a pipette is moved to each position of a plurality of column tubes in a holding state to supply samples and reagents. In this case, a method called reverse pipetting is used to accurately control the supply amount of the sample / reagent. If the same amount of sample / reagent as the specified supply amount is aspirated into the pipette, even if all the sample / reagent is discharged from the pipette, some of the sample / reagent will adhere to the inner wall surface of the pipette and remain, and the correct amount of sample / reagent cannot be supplied. In contrast, in the reverse pipetting method, a larger amount of sample / reagent than the specified supply amount is aspirated into the pipette, and the supply amount is discharged into one column tube, and the remaining liquid is discarded. This allows the correct amount of sample / reagent to be supplied to the column tube.

[0008] Here, when supplying a sample / reagent different from the previous time, it is necessary to discard the remaining liquid and wash the pipette. However, when supplying the same reagent to multiple column tubes each containing a different sample, or when supplying the same sample to multiple column tubes (and then supplying a different reagent to each column tube), it is not necessary to wash the pipette. However, if the same sample / reagent is aspirated again while there is still liquid remaining in the pipette, air may get into the liquid, causing an inaccurate supply amount. For this reason, even if the sample / reagent is the same, it is necessary to discharge the remaining liquid in the pipette to the waste section before aspirating the new liquid. In this case, even though there is no need to wash the pipette, the pipette must be moved to the waste section and the remaining liquid must be discharged, which increases the time required for pretreatment.

[0009] An object of the present invention is to provide an apparatus and method capable of shortening the time required for sample pretreatment. [Means for solving the problem]

[0010] The sample pretreatment device according to the present invention, which has been made to solve the above problems, comprises: a container accommodating section for accommodating a container for storing a supply target liquid, which is a sample and / or a reagent; A pipette that aspirates and discharges the liquid to be supplied; a column tube holder for a centrifuge that is detachably installed on a rotating part of the centrifuge and that has a holding part for holding a column tube and a waste liquid port; a pipette moving mechanism that moves the pipette among the container accommodated in the container accommodation unit, the column tube held in the holding unit, and the waste liquid port; a control unit that controls the pipette moving mechanism and the pipette so as to move the pipette to the container accommodated in the container accommodation unit, to aspirate a larger amount of the liquid to be supplied from the container into the pipette than a predetermined amount, then move the pipette to the column tube held in the holding unit, to discharge the predetermined amount of the liquid to be supplied in the pipette into the column tube, and then move the pipette to the waste liquid port, to discharge the liquid to be supplied remaining in the pipette into the waste liquid port; Equipped with.

[0011] The method for pretreating a sample according to the present invention comprises the steps of: a supply target liquid suction step of moving the pipette to a container storing a supply target liquid, which is a sample and / or a reagent, and suctioning an amount of the supply target liquid from the container into the pipette that is greater than a predetermined amount; a supply target liquid supply step of moving the pipette to a column tube held in a holder of a column tube holder for a centrifuge installed in a rotating part of the centrifuge and discharging the predetermined amount of the supply target liquid in the pipette into the column tube; a residual liquid discharge step of moving the pipette to a waste liquid port of the centrifuge column tube holder and discharging the supply target liquid remaining in the pipette to the waste liquid port; has. Effect of the Invention

[0012] According to the sample pretreatment device and method of the present invention, after the liquid to be supplied is supplied (discharged) from the pipette to the column tube held in the holding part, the liquid to be supplied remaining in the pipette is discharged (discharged) into the waste liquid port. Because the holding part and the waste liquid port are provided in the same centrifuge column tube holder, the distance over which the pipette is moved from the holding part to the waste liquid port can be made shorter than when the waste liquid port is provided outside the centrifuge (such as in a pipette washing part), thereby shortening the time required for pretreatment. The reagent discharged into the waste liquid port can be removed from the rotating part of the centrifuge and collected during maintenance of the device. [Brief description of the drawings]

[0013] [Figure 1] FIG. 1 is a schematic top view showing an embodiment of a sample pretreatment device according to the present invention. [Diagram 2] FIG. 2 is a top view showing a centrifuge included in the pretreatment device of the present embodiment. [Diagram 3] FIG. 2 is a cross-sectional view taken along line AA showing a centrifugal separator provided in the pretreatment device of the present embodiment. [Figure 4] 3A and 3B are a longitudinal sectional view and a top view showing a carrier (a column tube holder for a centrifuge) of the pretreatment device of the present embodiment. [Diagram 5] 1A is a longitudinal sectional view and FIG. 1B is a perspective view showing a disassembled state of a carrier of a centrifuge. [Figure 6] FIG. 2A is a longitudinal sectional view showing an example of a column tube used in the pretreatment device of the present embodiment, and FIG. 2B is a longitudinal sectional view showing a state in which a collection container is attached to the column tube. [Figure 7] 4 is a diagram showing the state of a carrier during rotation of a rotor in a centrifuge provided in the pretreatment device of the present embodiment. FIG. [Figure 8] FIG. 2 is a vertical cross-sectional view showing a pipette provided in the pretreatment device of the present embodiment. [Figure 9] 3 is a flowchart showing the operation of the pretreatment apparatus of the present embodiment and an embodiment of a sample pretreatment method according to the present invention. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0014] 1 to 9, an embodiment of a sample pretreatment device and method according to the present invention will be described.

[0015] The pretreatment device 1 of this embodiment comprises a sample / reagent container storage section (hereinafter abbreviated as "container storage section") 10, a centrifuge 20, a pipette 30, a pipette moving mechanism 31, a robot arm 40, a waste storage box 50, an unused member storage section 60, a post-pretreatment sample storage section 70, and a control section 80. Note that this pretreatment device 1 does not comprise a washing section for washing the pipette 30, since it uses a pipette 30 with a disposable tip 302 attached to its tip, as described below. Of these components, the robot arm 40 is a well-known one. The other components will be described below.

[0016] The container storage section 10 stores a sample container 11 in which a sample to be pretreated is stored, and a reagent container 12 in which a reagent is stored.

[0017] 2 and 3, the centrifuge 20 is a centrifuge of a swing rotor type, and has a rotor 21 and a carrier 22 held by the rotor 21. The carrier 22 corresponds to the above-mentioned centrifuge column tube holder.

[0018] The rotor 21 has a rotating shaft 211 which is a real axis extending in a substantially vertical direction, and four arms 212 which extend in a substantially horizontal direction from a position near the upper side of the rotating shaft 211 and are provided at 90° intervals around the rotating shaft 211. A motor 24 for rotating the rotating shaft 211 is provided below the rotating shaft 211. The tip of each arm 212 branches into two branches, and each of the branches is provided with a carrier holding pin 23 which extends toward the branch of the adjacent arm 212. The carrier holding pins 23 are disposed opposite each other between the adjacent arms 212.

[0019] The carrier 22 has a main body portion 221 and a main body accommodating portion 222 that accommodates the main body portion 221. A total of two recesses are provided on the side surface of the main body accommodating portion 222. The carrier holding pins 23 provided on each of two adjacent arms engage with these recesses, so that the carrier 22 is held on the rotor 21 so as to be rotatable around a rotation axis 231 shown by a dashed line in Fig. 2. Note that, contrary to the configuration of this embodiment, the carrier holding pins may be provided on the side surface of the main body accommodating portion 222, and a recess may be provided on the branch portion of the arm 212.

[0020] As shown in FIGS. 4 and 5, the main body 221 of the carrier 22 is formed by combining an upper main body member 2211 and a lower main body member 2212.

[0021] The upper main body member 2211 has nine holes that penetrate the front and back (top and bottom) of the plate material 22111, arranged in a line of three vertically and three horizontally in a plan view. Of these nine holes, a central hole 22112, which is a hole arranged in the center both vertically and horizontally, has a larger diameter than the other eight holes and extends with the same diameter throughout the thickness direction of the plate material 22111. In contrast, the other eight holes 225 (described in detail later) extend with the same diameter from the upper surface of the plate material 22111 to the lower surface halfway, and extend so that the diameter continuously decreases toward the lower surface toward the lower surface. An engagement portion 22113 that engages with the upper opening of a box 22121 of the lower main body member 2212 described next is provided on the lower surface of the plate material 22111. On the other hand, the lower main body member 2212 is formed by erecting a cylindrical member 22122 at the center of the bottom of a rectangular parallelepiped box 22121 whose overall planar shape is the same as the plate material of the upper main body member 2211 and whose top surface is open. The main body 221 of the carrier 22 is assembled by inserting the cylindrical member 22122 of the lower main body member 2212 into the central hole 22112 of the upper main body member 2211 and engaging the engagement portion 22113 of the upper main body member 2211 with the top surface opening of the lower main body member 2212.

[0022] The eight holes 225 of the main body portion 221 of the carrier 22 are arranged to have four-fold rotational symmetry around the central hole 22112, so that the main body portion 221 of the carrier 22 may be attached to the main body accommodating portion 222 in any orientation that differs by 90°.

[0023] These eight holes 225 can be used as either a holder 2251 for holding the column tube 90 described later or a waste liquid outlet 2252 for disposing of residual liquid (waste liquid) in the pipette 30. Each of these eight holes 225 can be used as either a holder 2251 or a waste liquid outlet 2252, but the role of the holes 225 arranged at each position when the main body 221 of the carrier 22 is attached to the main body accommodating section 222 is determined in advance. The reason for specifying "the holes 225 arranged at each position when the main body 221 is attached to the main body accommodating section 222" here is that the main body 221 may be attached to the main body accommodating section 222 in any orientation that differs by 90° as described above, and therefore the positions of the holes 225 are not specified before attachment. Having defined the role of each hole 225 in this manner, during operation of the pretreatment device 1, the control unit 80 controls each part of the pretreatment device 1 so that the hole 225 designated as the holding portion 2251 holds the column tube 90, and the hole 225 designated as the waste liquid outlet 2252 discharges the remaining liquid in the pipette 30.

[0024] As an example in this embodiment, as shown in FIG. 2, of the eight holes 225, when the main body portion 221 is attached to the main body accommodating portion 222, a total of six holes 225 arranged in two rows of three holes each (not sandwiching the central hole 22112) in a direction perpendicular to the carrier's rotation axis 231 are defined as the holding portion 2251, and the two holes 225 arranged in the same direction, sandwiching the central hole 22112, are defined as the waste liquid port 2252.

[0025] The space of the box 22121 of the main body 221 of the carrier 22 is referred to as the "waste liquid storage section 226" below, since the box 22121 and the hole 225 have the above-mentioned structure and are therefore in communication with the hole 225. In other words, the waste liquid storage section 226 is in communication with both the holding section 2251 and the waste liquid port 2252. Therefore, the waste liquid storage section 226 functions as a common storage section for the liquid (described later) that has passed through the separation membrane 93 of the column tube 90 during centrifugation and the liquid discarded to the waste liquid port 2252.

[0026] As shown in FIG. 6(a), the column tube 90 has a cylindrical sample storage section 91, a lid section 92 attached to the upper end of the sample storage section 91, and a separation membrane 93 provided at the lower end of the sample storage section 91. A rubber membrane is provided on a part of the lid section 92, and a sample or a reagent can be supplied into the sample storage section 91 by piercing the membrane with a pipette 30. The separation membrane 93 has a very small pore size so that the liquid to be separated can pass through only when the contents in the sample storage section 91 are subjected to centrifugal force toward the separation membrane 93 by the centrifuge. When the component to be analyzed does not pass through the separation membrane 93 during centrifugation, the column tube 90 holds the component in the holding section 2251 as it is, and when the component to be analyzed passes through the separation membrane 93, a collection container 95 is attached to the lower side of the column tube 90 (see FIG. 6(b)) and the component to be analyzed is held in the holding section 2251. In the former case, the liquid that has passed through the separation membrane 93 flows into the waste liquid storage section 226 that communicates with the holding section 2251, and is stored in the waste liquid storage section 226. An unused column tube 90 is stored in the unused member storage section 60, and is transported to the holding section 2251 by the robot arm 40.

[0027] During operation of the centrifuge 20, the rotor 21 is rotated at high speed by the motor 24, and centrifugal force acts on the carrier 22, causing it to rotate around the rotation axis 231, and the carrier 22 is tilted in a direction in which the bottom of the carrier 22 and the separation membrane 93 side of the column tube 90 held by the holding part 2251 are on the outside of the rotation of the rotor 21. At this time, the cylindrical sample storage part 91 of the column tube 90 becomes approximately horizontal (FIG. 7), and centrifugal force toward the separation membrane 93 acts on the contents of the column tube 90. As a result, the liquid to be separated passes through the separation membrane 93. If the collection container 95 is not attached to the lower side of the column tube 90, the liquid that passes through the separation membrane 93 flows into the waste liquid storage part 226 and is stored in the waste liquid storage part 226. In addition, the remaining liquid in the pipette 30 discharged (discharged) to the waste liquid port 2252 also flows into the waste liquid storage part 226 and is stored in the waste liquid storage part 226. A user of the apparatus removes the main body portion 221 of the carrier 22 from the main body accommodation portion 222 at an appropriate timing, such as during maintenance of the pretreatment device 1, and recovers the liquids stored in the waste liquid storage portions 226.

[0028] The pipette 30 can be freely moved horizontally and vertically within the pretreatment device 1 by a pipette moving mechanism 31, which is a known moving mechanism. The pipette 30 is composed of a cylindrical main body 301 and a disposable tip 302 attached to the tip of the main body (FIG. 8). Unused disposable tips 302 are stored in an unused member storage section 60, and the pipette moving mechanism 31 moves the main body 301 of the pipette 30 to the unused member storage section 60 and attaches it there. The pipette 30 is also connected to an aspirating / discharging mechanism (not shown) that aspirates gas within the pipette 30 and discharges gas into the pipette 30. With the tip of the pipette 30 inserted into the liquid, the aspirating / discharging mechanism aspirates the gas within the pipette, thereby aspirating the liquid into the pipette 30. The aspirating / discharging mechanism discharges gas into the pipette, thereby discharging the liquid from the pipette 30.

[0029] The waste collection box 50 is a box in which waste such as used disposable tips 302 is disposed. The waste collection box 50 can be pulled out from the pretreatment device 1. The user of the device collects waste from the waste collection box 50 as needed.

[0030] As described above, the unused member storage section 60 stores unused disposable tips 302, column tubes 90, etc. The pre-processed sample storage section 70 stores column tubes 90 that store samples for which pre-processing has been completed, and collection containers 95. The column tubes 90 and collection containers 95 are transferred from the holder 2251 to the pre-processed sample storage section 70 by the robot arm 40.

[0031] Hereinafter, the sample pretreatment method of this embodiment will be described while explaining the operation of the pretreatment device 1 of this embodiment using the flowchart of Fig. 9. A user of the pretreatment device 1 previously accommodates the sample container 11 storing the sample to be pretreated and the reagent container 12 storing the reagent in the container storage section 10, and also accommodates unused disposable tips 302 and column tubes 90 in the unused member storage section 60. In addition, the main body section 221 is stored in the main body storage section 222 of the carrier 22 of the centrifuge 20.

[0032] Pre-processing is started when a user executes a predetermined operation on the pre-processing device 1. The control unit 80 controls the operation of each unit of the pre-processing device 1 to execute the following operations.

[0033] First, the robot arm 40 transfers the column tubes 90 (or column tubes 90 with collection containers 95 attached) stored in the unused component storage section 60 one by one to each holding section 2251 of the carrier 22 of the centrifuge 20, and holds them in the holding sections 2251 (step 1).

[0034] Next, the pipette moving mechanism 31 moves the pipette 30 to the unused member storage section 60, and attaches an unused disposable tip 302 to the tip of the body 301 of the pipette 30 (step 2).

[0035] Next, the pipette moving mechanism 31 moves the pipette 30 to one of the sample containers 11 housed in the container housing 10. Then, the pipette 30 aspirates the sample in the sample container 11 in an amount larger than the specified amount to be supplied to the column tube 90 (step 3, which corresponds to the "supply target liquid aspirating step" described later). Furthermore, the pipette moving mechanism 31 moves the pipette 30 to one of the column tubes 90 held in each holder 2251, lowers the pipette 30 to pierce the membrane of the cover 92 of the column tube 90, and discharges the specified amount of sample. As a result, the specified amount of sample is supplied into the sample housing 91 of the column tube 90 (step 4, which corresponds to the "supply target liquid supplying step" described later).

[0036] Next, the pipette moving mechanism 31 raises the pipette 30 to remove it from the lid 92 of the column tube 90, and moves it to the waste liquid outlet 2252 in the same carrier 22 as the holder 2251 holding the column tube 90. Then, the remaining liquid of the sample in the pipette 30 is discharged to the waste liquid outlet 2252 and discarded (step 5, which corresponds to the "residual liquid discharge step" described below). Through the operations up to this point, the operation of supplying the sample stored in one sample container 11 to one column tube 90 is completed.

[0037] If there is another column tube 90 to which a sample should be supplied (YES in step 6), proceed to step 7. If the sample to be supplied to the next column tube 90 is different from the sample supplied to the other column tube 90 immediately before (NO in step 7), the pipette moving mechanism 31 moves the pipette 30 to the waste storage box 50, discards the used disposable tip 302 in the waste storage box 50, moves the pipette 30 to the unused member storage section 60, and replaces the disposable tip 302 by attaching a new unused disposable tip 302 to the tip of the body 301 of the pipette 30 (step 8). Then, the operations of steps 3 to 5 are performed to supply a new sample to the other column tube 90. The used disposable tip 302 discarded in the waste storage box 50 in step 8 is collected by the user when the pretreatment device 1 is stopped, but since the remaining liquid of the sample in the pipette 30 is discharged to the waste liquid port 2252 in step 5, it is possible to prevent the user from accidentally touching the remaining liquid when collecting the sample.

[0038] On the other hand, if the sample to be supplied next to the column tube 90 is the same as the sample previously supplied to another column tube 90 (YES in step 7), step 7 is not performed, i.e., the disposable tip 302 is not replaced, and the operations of steps 3 to 5 are performed to supply the same sample to the other column tube 90.

[0039] After the sample has been supplied to all of the target column tubes 90 through the above operations (NO in step 6), the process proceeds to step 9.

[0040] In step 9, the pipette moving mechanism 31 moves the pipette 30 to one of the reagent containers 12 housed in the container housing 10. The pipette 30 then aspirates the reagent from the reagent container 12 in an amount larger than the specified amount to be supplied to the column tube 90 (step 9 above corresponds to the "supply target liquid aspirating step" described below, as with step 3). Furthermore, the pipette moving mechanism 31 moves the pipette 30 to one of the column tubes 90 held in each holder 2251, lowers the pipette 30 to pierce the membrane of the cover 92 of the column tube 90, and dispenses the specified amount of reagent. Through the above operations, the specified amount of reagent is supplied into the sample housing 91 of the column tube 90 (step 10, which corresponds to the "supply target liquid supplying step" described below, as with step 4).

[0041] Next, the pipette moving mechanism 31 raises the pipette 30 to remove it from the lid 92 of the column tube 90, and moves it to the waste liquid outlet 2252 in the same carrier 22 as the holder 2251 holding the column tube 90. Then, the remaining liquid of the reagent in the pipette 30 is discharged to the waste liquid outlet 2252 and discarded (Step 11, which corresponds to the "residual liquid discharge step" described below, as in Step 5). This completes the operation of supplying the reagent stored in one reagent container 12 to one column tube 90.

[0042] If there is another column tube 90 to which the reagent should be supplied (YES in step 12), proceed to step 12. If the reagent to be supplied next to the column tube 90 is different from the reagent supplied to the other column tube 90 immediately before (NO in step 13), replace the disposable tip 302 by the same operation as in step 8 (step 14. Note that, for the same reason as in the case of the sample, this prevents the user from accidentally touching the remaining liquid of the reagent when collecting the used disposable tip 302). Then, execute the operations of steps 9 to 11 to supply a new reagent to the other column tube 90. On the other hand, if the reagent to be supplied next to the column tube 90 is the same as the reagent supplied to the other column tube 90 immediately before (YES in step 13), skip step 14 (replacement of the disposable tip 302) and execute the operations of steps 9 to 11 to supply the same reagent to the other column tube 90.

[0043] After the reagent has been supplied to all of the target column tubes 90 through the above operations (NO in step 12), the rotor 21 of the centrifuge 20 is rotated to perform centrifugation (step 15).

[0044] With the operations up to this point, one pretreatment operation of mixing with reagent and centrifuging is completed for each sample. Depending on the contents of the pretreatment, further treatment using other reagents may be performed, and in such a case (YES in step 16), the procedure returns to step 9 and the operations from step 9 to step 15 are performed. On the other hand, if treatment using other reagents is not performed (NO in step 16), the robot arm 40 transfers the column tubes 90 held in each holder 2251 to the post-pretreatment sample storage section 70 (step 17), completing the series of operations.

[0045] According to the sample pretreatment device and method of this embodiment, since the holding portion 2251 and the waste liquid outlet 2252 are provided in the same carrier 22 (centrifuge column tube holder) of the centrifuge 20, when the same sample or the same reagent is continuously supplied to another column tube 90 without replacing the disposable tip 302, the distance over which the pipette 30 is moved to discard the liquid remaining in the pipette 30 can be shortened, thereby shortening the time required for pretreatment.

[0046] The present invention is not limited to the above-described embodiment, and various modifications are possible.

[0047] For example, in the above embodiment, a common hole 225 is used for the holding part 2251 and the waste liquid port 2252, but the holding part 2251 and the waste liquid port 2252 may each be a hole of a different shape. Also, in the above embodiment, the holding part 2251 and the waste liquid port 2252 are connected to a common waste liquid storage part 226, but separate waste liquid storage parts may be provided so that the holding part 2251 and the waste liquid port 2252 are connected to different waste liquid storage parts.

[0048] In the above embodiment, the pipette 30 is equipped with a disposable tip 302. Instead of using such a disposable tip 302, a washing unit for washing the pipette 30 may be provided in the pretreatment device. In that case, when the same sample or the same reagent is continuously supplied to another column tube 90, the sample or reagent is supplied to one column tube 90, the remaining liquid in the pipette 30 is discarded to the waste liquid port 2252, and then the sample or reagent is supplied to the other column tube 90 without washing the pipette 30. On the other hand, when a sample or reagent is supplied to a certain column tube 90 and then another sample or reagent is supplied to another column tube 90, the remaining liquid in the pipette 30 after the previous supply is discarded to the waste liquid port 2252, and then the pipette 30 is washed in the washing unit.

[0049] In the above embodiment, a sample is first supplied to the column tube 90, a reagent is then supplied to the column tube 90, and centrifugation is then performed, but instead, a reagent may first be supplied to the column tube 90 and centrifuged to cause the reagent to permeate the separation membrane 93, and then a sample may be supplied to the column tube 90 and centrifugation may be performed. In that case, a different reagent may be further supplied to the sample to be centrifuged and then centrifugation may be performed.

[0050] [Aspects] It will be apparent to those skilled in the art that the above-described exemplary embodiments are illustrative of the following aspects.

[0051] (Item 1) A sample pretreatment device according to one aspect of the present invention comprises: a container accommodating section for accommodating a container for storing a supply target liquid, which is a sample and / or a reagent; A pipette that aspirates and discharges the liquid to be supplied; a column tube holder for a centrifuge that is detachably installed on a rotating part of the centrifuge and that has a holding part for holding a column tube and a waste liquid port; a pipette moving mechanism that moves the pipette among the container accommodated in the container accommodation unit, the column tube held in the holding unit, and the waste liquid port; a control unit that controls the pipette moving mechanism and the pipette so as to move the pipette to the container accommodated in the container accommodation unit, to aspirate a larger amount of the liquid to be supplied from the container into the pipette than a predetermined amount, then move the pipette to the column tube held in the holding unit, to discharge the predetermined amount of the liquid to be supplied in the pipette into the column tube, and then move the pipette to the waste liquid port, to discharge the liquid to be supplied remaining in the pipette into the waste liquid port; Equipped with.

[0052] (Item 6) A method for pretreating a sample according to one aspect of the present invention comprises: a supply target liquid suction step of moving the pipette to a container storing a supply target liquid, which is a sample and / or a reagent, and suctioning an amount of the supply target liquid from the container into the pipette that is greater than a predetermined amount; a supply target liquid supply step of moving the pipette to a column tube held in a holder of a column tube holder for a centrifuge installed in a rotating part of the centrifuge and discharging the predetermined amount of the supply target liquid in the pipette into the column tube; a residual liquid discharge step of moving the pipette to a waste liquid port of the centrifuge column tube holder and discharging the supply target liquid remaining in the pipette to the waste liquid port; has.

[0053] According to the sample pretreatment device according to paragraph 1 and the sample pretreatment method according to paragraph 6, the liquid to be supplied is supplied (discharged) from the pipette to the column tube held in the holding part, and then the liquid to be supplied remaining in the pipette is discharged (discharged) to the waste liquid port. Since the holding part and the waste liquid port are provided in the same centrifuge column tube holder, the distance over which the pipette is moved from the holding part to the waste liquid port can be made shorter than when the waste liquid port is provided outside the centrifuge (such as in a pipette washing part), thereby shortening the time required for pretreatment. The reagent discharged to the waste liquid port can be removed from the rotating part of the centrifuge and collected during maintenance of the device.

[0054] (Item 2) The sample pretreatment device according to item 2 is the sample pretreatment device according to item 1, further comprising a waste liquid storage section communicating with both the holding section and the waste liquid port.

[0055] According to the sample pretreatment device of paragraph 2, two types of liquid to be disposed of, i.e., the liquid remaining in the pipette when supplied to the column tube and the liquid that has passed through the separation membrane of the column tube during centrifugation, can be stored in a common waste liquid storage section, thereby simplifying the configuration of the device.

[0056] (Item 3) The sample pretreatment device according to item 3 is the sample pretreatment device according to item 1 or 2, wherein the holding part and the waste liquid outlet are holes of the same shape.

[0057] According to the sample pretreatment device of the third aspect, since the holding part and the waste liquid outlet are formed of holes of the same shape, each hole can be used as either the holding part or the waste liquid outlet.

[0058] (Item 4) The sample pretreatment device according to item 4 is the sample pretreatment device according to item 3, wherein the holes are arranged rotationally symmetrically in the centrifuge column tube holder.

[0059] According to the sample pretreatment device of item 4, since the holes are arranged in rotational symmetry, the centrifuge column tube holder can be attached to the rotating part of the centrifuge in any of a number of rotationally symmetric orientations.

[0060] (Item 5) The sample pretreatment device according to item 5 is the sample pretreatment device according to any one of items 1 to 4, wherein the tip of the pipette is a disposable tip.

[0061] In the sample pretreatment device according to paragraph 5, the tip of the pipette is a disposable tip, so by replacing the disposable tip every time the reagent or sample to be aspirated by the pipette is changed, there is no need to wash the pipette. In addition, before replacing the disposable tip, the reagent remaining in the disposable tip is discharged to the waste liquid outlet, so that no reagent remains in the used disposable tip. This prevents the user of the device from touching the reagent when collecting the used disposable tip. [Explanation of symbols]

[0062] 1...Pretreatment device 10... Sample / reagent container storage section (container storage section) 11...Sample container 12...Reagent container 20…Centrifuge 21...Rotor 211...Rotation axis 212…Arm 22…Career 221…Main body 2211…Main body upper part 22111…Plate material 22112…Central hole 22113…Engagement part 2212…Bottom part 22121...box 22122…Cylindrical material 222...Main body storage section 225...hole 2251...Holding part 2252… Wastewater outlet 226...Waste liquid storage section 23…Carrier retaining pin 231... Rotating shaft 24…Motor 30…Pipette 301...Main body 302…Disposable tip 31...Pipette movement mechanism 40…Robot arm 50…Waste storage box 60: Unused material storage section 70: Pre-processed sample storage section 80...Control unit 90…Column tube 91...Sample storage section 92…Lid part 93...Separation membrane 95…Collection container

Claims

1. a container accommodating section for accommodating a container for storing a supply target liquid, which is a sample and / or a reagent; A pipette that aspirates and discharges the liquid to be supplied; a column tube holder for a centrifuge that is detachably installed on a rotating part of the centrifuge and that has a holding part for holding a column tube and a waste liquid port; a pipette moving mechanism that moves the pipette among the container accommodated in the container accommodation unit, the column tube held in the holding unit, and the waste liquid port; a control unit that controls the pipette moving mechanism and the pipette so as to move the pipette to the container accommodated in the container accommodation unit, to aspirate a larger amount of the liquid to be supplied from the container into the pipette than a predetermined amount, then move the pipette to the column tube held in the holding unit, to discharge the predetermined amount of the liquid to be supplied in the pipette into the column tube, and then move the pipette to the waste liquid port, to discharge the liquid to be supplied remaining in the pipette into the waste liquid port; A sample pretreatment device comprising:

2. The sample pretreatment device according to claim 1 , further comprising a waste liquid storage section communicating with both said holding section and said waste liquid outlet.

3. 3. The sample pretreatment device according to claim 1, wherein the holding portion and the waste liquid outlet are holes of the same shape.

4. The sample pretreatment device according to claim 3 , wherein the holes are arranged rotationally symmetrically in the centrifuge column tube holder.

5. The sample pretreatment device according to claim 1 or 2, wherein the tip of the pipette is a disposable tip.

6. a supply target liquid suction step of moving the pipette to a container storing a supply target liquid, which is a sample and / or a reagent, and suctioning an amount of the supply target liquid from the container into the pipette that is greater than a predetermined amount; a supply target liquid supply step of moving the pipette to a column tube held in a holder of a column tube holder for a centrifuge installed in a rotating part of the centrifuge and discharging the predetermined amount of the supply target liquid in the pipette into the column tube; a residual liquid discharge step of moving the pipette to a waste liquid port of the centrifuge column tube holder and discharging the supply target liquid remaining in the pipette to the waste liquid port; The method for pretreatment of a sample comprises the steps of:

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