Automated analysis device

Abstract

The present application provides an automatic analysis device capable of selecting whether to perform evaporation concentration for each sample and controlling evaporation concentration of the sample. The automatic analysis device includes an evaporation concentration unit (131) that performs a concentration process of evaporating an extract liquid in which an analysis target component in a sample is extracted to concentrate the analysis target component, an analysis unit (103) that analyzes the analysis target component of the sample, and a control unit (104) that controls operations of the evaporation concentration unit (131) and the analysis unit (103). The control unit (104) determines whether to perform the evaporation concentration process for the analysis target component in the sample, and concentrates the analysis target component in the sample by the evaporation concentration unit (131) for the sample determined to perform the evaporation concentration process. Whether to perform the evaporation concentration process for which sample is stored in a storage unit (107) via an input unit (105), and the control unit (104) selects whether to perform the evaporation concentration for each sample in accordance with the stored content.

Description

TECHNICAL FIELD

[0001] The present application relates to an automatic analysis device that analyzes a sample. BACKGROUND

[0002] As an analysis method of a specific component included in a sample, for example, there is a method using LC-MS (Liquid Chromatography-Mass Spectrometry) in which a liquid chromatograph (LC) and a mass spectrometer (MS) are connected in an online manner.

[0003] The use of this LC-MS is also gradually expanding in the field of clinical examination in which analysis of a biological sample such as blood or urine is performed by an automatic analysis device.

[0004] In a case where a biological sample such as blood or urine (hereinafter, simply referred to as a sample) is analyzed by LC-MS, pretreatment for improving the purification degree of the sample needs to be performed. As the pretreatment of the sample, for example, there are methods such as solid phase extraction (SPE) and liquid-liquid extraction (LLE).

[0005] In particular, SPE is easy to connect with LC-MS in an online manner, and thus, it is possible to integrate and automate the pretreatment based on SPE and the analysis based on LC-MS.

[0006] In the pretreatment of the sample, in order to achieve high-sensitivity detection based on LC-MS, evaporation concentration in which an extract liquid in which an analysis target component included in the sample is extracted is evaporated to increase the concentration of the analysis target component is sometimes performed.

[0007] As a technique of performing such evaporation concentration of the sample, for example, in Patent Literature 1, a device in which evaporation processing of a sample liquid is automatically performed by inserting a container in which the sample is housed into a holder having a heating wire and a coil is disclosed.

[0008] PRIOR ART DOCUMENTS

[0009] PATENT LITERATURE

[0010] Patent Literature 1: Japanese Patent Application Laid-Open No. 2013-527461 SUMMARY

[0011] PROBLEMS TO BE SOLVED BY THE INVENTION

[0012] In the automatic pretreatment device like the above-described prior art, automation based on a batch process is assumed.

[0013] In the batch process, the reaction liquid is processed all at once, and thus, it is not possible to select whether to concentrate or not for each sample.

[0014] However, in an automatic analysis device for clinical examination which continuously processes various samples, it is required to limit detection of an analysis target component in a low concentration region which does not require evaporation concentration, and it is desirable to have a mechanism which can select whether to perform evaporation concentration for each sample and control thereof, and to perform high-sensitivity detection.

[0015] The present application was achieved in view of the above circumstances, and aims to realize an automatic analysis device which can select whether to perform evaporation concentration for each sample and control thereof.

[0016] Means for solving the problem

[0017] In order to achieve the above object, the present application is configured in the following manner.

[0018] In an automatic analysis device, there are an evaporation concentration section which performs concentration processing of evaporating an extracted liquid in which an analysis target component in a sample is extracted to concentrate the analysis target component, an analysis section which analyzes the analysis target component of the sample, and a control section which controls the operation of the evaporation concentration section and the analysis section.

[0019] The control section determines whether to perform evaporation concentration processing on the analysis target component in the sample, and for a sample which is determined to perform evaporation concentration processing, the analysis target component in the sample is concentrated by the evaporation concentration section. BRIEF DESCRIPTION OF DRAWINGS

[0020] Figure 1 is a view which schematically shows the entire structure of an automatic analysis device of Embodiment 1 of the present application.

[0021] Figure 2 is a view which shows an example of a pre-processing step of analysis processing of an automatic analysis device.

[0022] Figure 3 is a view which schematically shows an example of an evaporation concentration mechanism of Embodiment 1 of the present application.

[0023] Figure 4 is a view which shows an ejector pin which pushes up a reaction container from below.

[0024] Figure 5 is a view which schematically shows an evaporation concentration mechanism in Embodiment 2.

[0025] Figure 6 is a view which shows Embodiment 3, and is a view which schematically shows an example in which an evaporation concentration section is provided in one region having an exhaust unit.

[0026] Figure 7 is a view which shows Figure 6A diagram showing a variation of the example shown.

[0027] Figure 8 This is a diagram illustrating an example of the process for calculating the concentration ratio of the refined liquid obtained through the pretreatment process.

[0028] Figure 9 This graph shows a comparison of mass chromatograms derived from the concentration ratio calculation using standard substances, internal standard substances, and standard substances, respectively.

[0029] Figure 10 This graph shows a comparison of mass chromatograms derived from the concentration ratio calculation standard, internal standard, and analyte component, respectively.

[0030] Figure 11 This graph shows a comparison of mass chromatograms derived from the concentration ratio calculation standard, internal standard, and analyte component, respectively. Detailed Implementation

[0031] Hereinafter, embodiments of the present invention will be described with reference to the accompanying drawings. Furthermore, in the embodiments of the present invention, an automated analytical apparatus comprising a sample pretreatment function combination LC-MS (Liquid Chromatography-Mass Spectrometry) as the analytical mechanism will be described as an example. However, the present invention can also be applied, for example, to automated analytical apparatuses that incorporate separation units such as capillary electrophoresis and detectors such as spectrophotometers as analytical mechanisms.

[0032] Example

[0033] (Example 1)

[0034] Figure 1 This is a diagram that schematically illustrates the overall structure of the automatic analysis device according to Embodiment 1 of the present invention.

[0035] exist Figure 1 The automatic analysis device 100 includes: a pretreatment unit 101 for pretreating the sample; a separation unit 102 for separating the components in the sample; an analysis unit 103 for analyzing the separated components; a control unit 104 for controlling the overall operation of the device; an input unit 105 for allowing the user to input information into the device; a display unit 106 for displaying information to the user; and a storage unit 107, such as a storage medium, for storing various information related to the control of the automatic analysis device 100.

[0036] The control unit 104, input unit 105, display unit 106, and storage unit 107 constitute a control device for controlling the overall operation of the automatic analysis device 100.

[0037] Further, in the present embodiment 1, the input section 105 and the display section 106 are shown as separate bodies, but for example, the input section 105 and the display section 106 can be integrally configured like a touch panel type monitor.

[0038] The pretreatment section 101 has a transport mechanism 112 that transports a sample container 111 in which a sample that is an analysis target is housed to a sample dispensing position, a reaction container disk 120 that can keep a solution in a reaction container 116 at a constant temperature by mounting the reaction container 116 to a plurality of opening portions 119, a reagent disk 122 that keeps a plurality of reagent containers 121 in which reagents are housed, and a sample dispensing mechanism 113 that dispenses a sample from the sample container 111 transported to the sample dispensing position to the reaction container 116 housed in the opening portion 119 of the reaction container disk 120.

[0039] In addition, the pretreatment section 101 has a reagent dispensing mechanism 123 that dispenses a reagent from the reagent container 121 to the reaction container 116 of the reaction container disk 120, a dispensing tip mounting rack 115 that mounts a disposable dispensing tip 115a of the nozzle mounted to the sample dispensing mechanism 113 that is not used, and a dispensing tip mounting / demounting section 114 that demounts the used dispensing tip 115a from the nozzle of the sample dispensing mechanism 113 and discards it, or mounts the unused dispensing tip 115a to the nozzle.

[0040] In addition, the pretreatment section 101 has a reaction container mounting rack 117 that mounts the reaction container 116 that is not used, and a transport mechanism 118 that performs transport of the unused dispensing tip 115a from the dispensing tip mounting rack 115 to the dispensing tip mounting / demounting section 114, transport of the used reaction container 116 from the opening portion 119 of the reaction container disk 120 to a discarding section (not shown), and transport of the unused reaction container 116 from the reaction container mounting rack 117 to the opening portion 119 of the reaction container disk 120.

[0041] In addition, the pretreatment section 101 has a magnetic separation mechanism 124 that separates a magnetic bead in a solution housed in the reaction container 116 using a magnetic force of a magnet, a transport mechanism 125 that transports the reaction container 116 between the reaction container disk 120 and the magnetic separation mechanism 124, and an evaporation concentration mechanism 131 that performs evaporation concentration of an analysis target component in the solution in the reaction container 116.

[0042] Further, the pretreatment section 101 has a transport mechanism 132 that transports the reaction container 116 between the reaction container disk 120 and the evaporation concentration mechanism 131, a dispensing mechanism 133 for the separation section that dispenses the solution in the reaction container 116 after evaporation concentration to the separation section 102 that separates the components in the sample, and an analysis section 103 that detects and analyzes the components in the solution separated by the separation section 102.

[0043] The magnetic separation mechanism 124 is provided on the rotation track 126 of the reagent dispensing mechanism 123. The reagent dispensing mechanism 123 can discharge a reagent to the reaction container 116 supported by the magnetic separation mechanism 124 or suck the solution in the reaction container 116.

[0044] The reaction container disk 120 functions as an incubator that maintains the temperature of the reaction container 116 provided to the opening portion 119 constant, and incubates the reaction container 116 provided to the opening portion 119 for a certain time.

[0045] The separation section 102 is, for example, an LC (Liquid Chromatography) that has a column or the like as a function of separating the components in the reaction solution dispensed by the dispensing mechanism 133 for the separation section. The separation section 102 separates the components in the reaction solution dispensed from the reaction container 116 by the dispensing mechanism 133 for the separation section, and sequentially introduces the separated components to the analysis section 103.

[0046] The analysis section 103 is, for example, an MS (Mass Spectrometry) that has a dynode or the like as a function of ionizing and mass analyzing the components introduced from the separation section 102. The analysis section 103 ionizes the components introduced from the separation section 102 to detect the ion amount (i.e., the component amount), and outputs the detection result to the control section 104.

[0047] The control section 104 controls the operation of the evaporation concentration mechanism (evaporation concentration section) 131, the operation of the separation section 102, and the operation of the analysis section 103. In addition, the control section 104 calculates the concentration value of the components in the sample using the detection result (ion amount) from the analysis section 103 and the calibration curve obtained in advance, stores the analysis result in the storage section 107 as an analysis result, and causes the analysis result to be displayed on the display section 106.

[0048] As a method of obtaining the calibration curve, for example, first, a standard substance whose concentration is known is analyzed for a plurality of concentrations. Then, the time change (mass chromatogram) of the ion amount, i.e., the ion intensity, is obtained with respect to the m / z (mass / charge ratio) of the ions derived from the standard substance, and the peak area of the mass chromatogram is calculated. The calibration curve is created from the relationship between the area and the concentration of the standard substance.

[0049] By using the calibration curve thus obtained, the component concentration of a sample having the same component as the standard substance and having an unknown concentration can be detected.

[0050] Specifically, the peak area of the mass chromatogram is calculated for a sample of the analysis target, and the component concentration of the analysis target component is determined based on the correspondence between the peak area of the mass chromatogram and the calibration curve.

[0051] Further, when the intensity of the detected ion is normalized based on the intensity of the ion derived from the internal standard substance, comparison between data can be performed with high accuracy. That is, the ion intensity that slightly varies at each analysis can be compared and verified between analyses due to the influence of the pretreatment of the sample, injection of the sample into the LC-MS, ionization in the LC-MS, and the like. This method is called an internal standard method.

[0052] Here, first, the basic procedure of the analysis process will be described.

[0053] Figure 2 is a view showing an example of a pretreatment procedure of the analysis process of the automatic analysis device.

[0054] Before starting the pretreatment, an unused reaction vessel 116 is set to the opening portion 119 on the reaction vessel disk 120 from the reaction vessel mounting rack 117 by the conveyance mechanism 118. In addition, before dispensing of the sample, the sample dispensing mechanism 113 accesses the dispensing tip attachment / detachment portion 114, and the dispensing tip 115a is attached to the tip of the nozzle.

[0055] In the pretreatment, first, the sample containing the analysis target component is sucked from the sample container 111 by the sample dispensing mechanism 113 via the dispensing tip 115a, and is discharged to the reaction vessel 116 of the reaction vessel disk 120 (step S200).

[0056] Further, the sample dispensing mechanism 113, at the end of dispensing of the sample from one sample container 111, discards the used dispensing tip 115a via the dispensing tip attachment / detachment portion 114, and attaches the unused dispensing tip 115a.

[0057] Next, the internal standard substance is sucked from the reagent container 121 of the reagent disk 122 by the reagent dispensing mechanism 123 as a reagent corresponding to the analysis target component, and is discharged to the reaction vessel 116 (step S201).

[0058] Next, a reagent such as a protein removal agent is sucked from the reagent container 121 of the reagent disk 122 by the reagent dispensing mechanism 123, and is discharged to the reaction vessel 116 (step S202).

[0059] Next, the reagent dispensing mechanism 123 draws the suspension of magnetic beads, which is used as a reagent, from the reagent container 121 of the reagent tray 122 and discharges it into the reaction container 116 (step S203).

[0060] Next, the reaction vessel 116, which has been dispensed with the sample, internal standard material, and magnetic beads, is transported by the conveying mechanism 125 to the magnetic separation mechanism 124 for cleaning of the magnetic beads (step S204). In the magnetic separation mechanism 124, the magnetic force of the magnets 201 positioned along the outer surface of the reaction vessel 116 keeps the magnetic beads containing the analyte and internal standard material aggregated on the inner wall surface of the reaction vessel 116. Figure 2 In this state, the solution in the reaction vessel 116 is drawn up and discarded by the reagent dispensing mechanism 123. (This is referred to as magnetic bead group 202).

[0061] At this time, the magnetic beads, the analytical components held by the magnetic beads, and the internal standard substances remain in the reaction vessel 116.

[0062] Next, the cleaning solution used to clean impurities other than the substances (analyte components and internal standard substances) held by the magnetic beads is drawn from the reagent container 121 of the reagent tray 122 by the reagent dispensing mechanism 123 and discharged into the reaction vessel 116.

[0063] At this time, the magnetic constraint of the magnetic bead based on the magnetic force of magnet 201 can also be temporarily released.

[0064] Next, while the magnetic beads are gathered again onto the inner wall of the reaction vessel 116 by the magnet 201, the reagent dispensing mechanism 123 draws up and discards the solution (cleaning solution) from the reaction vessel 116, thereby cleaning the magnetic beads.

[0065] Next, the eluent used as reagents, which is the eluent used to elute the target components and internal standard substances from the magnetic bead group 202, is drawn from the reagent container 121 of the reagent tray 122 by the reagent dispensing mechanism 123 and discharged into the reaction vessel 116 (step S205).

[0066] Next, with the magnetic beads 203, which have been eluted from the analyte and the internal standard substance, gathered on the inner wall of the reaction vessel 116 by the magnetic force of the magnet 201, the solution (purified liquid) of the reaction vessel 116 is drawn up by the reagent dispensing mechanism 123 (step S206) and discharged into an unused reaction vessel 116 in a reaction vessel tray 120 that is different from the reaction vessel 116 disposed in the magnetic separation mechanism 124 (step S207).

[0067] In addition, the purified liquid stored in the reaction vessel 116 of the reaction vessel tray 120 is cultured as needed.

[0068] Next, the reaction vessel 116 containing the purified liquid is conveyed to the evaporation and concentration unit 131 via the conveying mechanism 132, where the components in the purified liquid are evaporated and concentrated (step S208). Furthermore, the detailed structure of the evaporation and concentration unit 131 will be described later.

[0069] Next, by changing the composition of the purified solution to combine the analyte with the separation column (not shown) of the separation section 102, the diluent is drawn from the reagent container 121 of the reagent tray 122 by the reagent dispensing mechanism 123 and discharged into the reaction vessel 116.

[0070] The purified liquid obtained from the above pretreatment process is drawn from the reaction vessel 116 by the dispensing mechanism 133 in the separation section and discharged into the separation section 102. The components separated by the separation section 102 are ionized by the analysis section 103 to detect the ion quantity (i.e., component quantity). The detection result of the analysis section 103 is output to the control section 104, and the concentration value of the component in the sample is calculated using the calibration curve.

[0071] Next, the evaporation concentration process in Embodiment 1 of the present invention will be described. The automatic analysis apparatus of Embodiment 1 includes an evaporation concentration mechanism and a control method capable of selecting whether to perform evaporation concentration based on the analyte component, i.e., the sample. For example, each sample is assigned an identification number, and the input unit 105 stores the information on which sample with which identification number will undergo evaporation concentration treatment in the storage unit 107. Based on the information stored in the control unit 104, the control unit selects whether to perform evaporation concentration treatment based on the sample, and performs sample processing. That is, the control unit 104 determines whether to perform evaporation concentration treatment on the analyte component in the sample, and for samples determined to require evaporation concentration treatment, the evaporation concentration mechanism (evaporation concentration unit) 131 concentrates the analyte component in the sample.

[0072] Figure 3 This is a schematic diagram illustrating an example of the evaporation and concentration mechanism 131 of Embodiment 1 of the present invention.

[0073] exist Figure 3 In this process, the evaporation and concentration mechanism 131 has multiple container receiving sections 301 that receive and hold the refined liquid obtained by the above-mentioned pretreatment process. The reaction container 116 is delivered to the container receiving section 301 and distributed to one of the container receiving sections 301 of the evaporation and concentration section 302 where evaporation and concentration are performed, or to the container receiving section 301 of the standby section 303 where evaporation and concentration are not performed.

[0074] Here, the evaporation and concentration unit 131 has a heating unit 304 for heating the reaction vessel 116 and an exhaust unit 305 for extracting vapor from the reaction vessel 116. As examples of the heating unit 304, Peltier elements and heaters that can control the temperature of the analytical apparatus are listed.

[0075] Furthermore, as an example of the exhaust unit 305, a vacuum pump, etc., whose operation can be controlled by the analysis device, is also listed. Additionally, a valve 306 may be provided between the opening of the exhaust unit 305 and the drain pipe.

[0076] Two examples are given below as methods for distributing the reaction vessel 116 to either the container receiving section 301 of the standby section 303 or the container receiving section 301 of the evaporation and concentration section 302.

[0077] The first example is a method for making the evaporation and concentration mechanism 131 movable. First, the control unit 104 moves the evaporation and concentration mechanism 131 so that the point where the conveying mechanism 132 accesses the evaporation and concentration mechanism 131 is aligned with the container receiving unit 301.

[0078] For samples that the control unit 104 determines will not undergo evaporation and concentration, the reaction vessel 116 is transported to the container receiving unit 301 on the standby unit 303 by the conveying mechanism 132. After a certain period of standby time, the purified liquid is introduced into the separation unit 102 by the dispensing mechanism 133 of the separation unit.

[0079] In the case of evaporation and concentration, the reaction vessel 116 is conveyed to the container receiving section 301 on the evaporation and concentration section 302 using the conveying mechanism 132. At this time, the exhaust unit 305 is positioned near the heating section 304 of the evaporation and concentration section 302 and at a certain distance away from the heating section 304 (first position). Next, the exhaust unit (exhaust section) 305 is moved so that it is positioned above the reaction vessel 116 positioned on the heating section 304 and in close contact with the reaction vessel 116 (position in close contact with the heating section 304: second position). In this case, the evaporation and concentration mechanism 131 can also be moved. Furthermore, the reaction vessel 116 is heated by the heating unit (heating section) 304 for a certain period of time and the exhaust is performed with the valve 306 open, thereby performing evaporation and concentration.

[0080] To improve evaporation efficiency, the evaporation and concentration section 302 can be raised relative to the exhaust unit 305, or the exhaust unit 305 can be lowered relative to the reaction vessel 116, so that the opening of the reaction vessel 116 is in close contact with the exhaust unit 305.

[0081] However, in either case, the reaction vessel 116 is in close contact with the container receiving portion 301, therefore, it is considered that the reaction vessel 116 may not be able to detach when being removed from the container receiving portion 301. As a countermeasure, such as... Figure 4 As shown, a top ejector pin 401 can also be provided for pushing the reaction vessel 116 upward from below. By setting the top ejector pin 401 as a mechanism that moves up and down simultaneously with the exhaust unit 305, the reduction of drive units such as motors can be achieved.

[0082] The second example is a method for fixing the position of the evaporation and concentration mechanism 131. In this case, the control unit 104, depending on whether evaporation and concentration of a sample is to be performed, uses the conveying mechanism 132 to transport the reaction vessel 116 to the container receiving unit 301 of the evaporation and concentration unit 302 or the standby unit 303. Then, relative to the reaction vessel 116 on the evaporation and concentration unit 302, the exhaust unit 305 is moved such that it is positioned above the reaction vessel 116.

[0083] Furthermore, the reaction vessel 116 is heated and vented for a certain period of time to achieve evaporation and concentration. The purified liquid in the reaction vessel 116 is then introduced into the separation section 102 via the separation section dispensing mechanism 133, which is accessible through the container receiving section 301 of the evaporation and concentration mechanism 131.

[0084] also, Figure 3 The number of container receiving sections 301 is one example, and can be varied considering factors such as the number of samples that the automatic analyzer can process per hour (throughput) and the time required for evaporation and concentration. Assuming the throughput is set to 100 samples / hour (36 seconds / sample) and the time required for evaporation and concentration is set to 108 seconds (36 seconds × 3), if three container receiving sections 301 are provided in both the evaporation and concentration section 302 and the standby section 303, then even when the process is performed continuously, the evaporation and concentration mechanism 131 can process the reaction containers 116 sequentially without idle time, achieving a throughput of 100 samples / hour. In this case, an exhaust unit 305 is required for each of the three container receiving sections 301 in the evaporation and concentration section 302.

[0085] When venting is performed by keeping the reaction vessel 116 in close contact with the venting unit 305, the valve 306 is opened during the steam absorption process and closed after the absorption is completed. This prevents the reaction vessel 116 from leaving the venting unit 116.

[0086] Additionally, consider the condensed steam dripping from the exhaust unit 305 after the steam absorption process is completed.

[0087] As a countermeasure, heating the exhaust unit 305 itself is listed. Furthermore, as a countermeasure, after the exhaust is completed and the exhaust unit 305 leaves the reaction vessel 116, empty suction based on the exhaust unit 305 is implemented.

[0088] As described above, according to Embodiment 1 of the present invention, the sample requiring evaporation and concentration is moved to the evaporation and concentration section 302, and the sample not requiring evaporation and concentration is moved to the standby section 303 where no evaporation and concentration is performed. For the sample requiring evaporation and concentration, evaporation and concentration are performed in the evaporation and concentration section 302, and the sample is moved to the separation section 102 together with the sample not requiring evaporation and concentration. Then, in the separation section 102, the separated components are moved to the analysis section 103 for analysis.

[0089] Therefore, an automated analytical device is available that can select whether to perform evaporation and concentration based on the sample, enabling control over the evaporation and concentration of the sample.

[0090] In addition, Figure 3 In the example shown, the standby section 303 is configured to be adjacent to the evaporation and concentration section 302. However, the standby section 303 can also be configured to be formed of a heat-insulating material, which can limit the heat effect from the evaporation and concentration section 302. In addition, the standby section 303 and the evaporation and concentration section 302 can also be configured separately from each other in a way that eliminates the heat effect.

[0091] (Example 2)

[0092] Next, Embodiment 2 of the present invention will be described.

[0093] The difference between Example 2 and Example 1 lies in the structure of the evaporation and concentration mechanism; all other structures are the same. Therefore, illustrations and detailed descriptions of parts other than the evaporation and concentration mechanism are omitted.

[0094] Figure 5 This is a schematic diagram illustrating the evaporation and concentration mechanism 131 in Example 2.

[0095] exist Figure 5 In the process, the evaporation and concentration mechanism 131 has a container support 501 that receives and supports the reaction vessel 116 containing the purified liquid obtained through the above-mentioned pretreatment process, and controls whether to perform evaporation and concentration according to the reaction vessel 116 being transported to the container support 501.

[0096] Here, the evaporation and concentration mechanism 131 in Embodiment 2 includes: a heating unit 502 for heating the reaction vessel 116; a movable unit (movable part) 503 for controlling the contact / non-contact of the heating unit 502 with respect to the reaction vessel 116; an exhaust unit 305 for drawing vapor from the reaction vessel 116; and a valve 306 located between the opening of the exhaust unit 305 and the drain pipe.

[0097] First, the reaction vessel 116 is conveyed to the container support 501 of the evaporation and concentration mechanism 131 using the conveying mechanism 132. When evaporation and concentration are not performed, the heating unit (heating section) 502 is located in a non-heating position, not in contact with the reaction vessel 116 and not heating the extract within the reaction vessel 116. When evaporation and concentration are performed, the heating unit 502 moves towards the reaction vessel 116 via the movable unit (movable section) 503, moving to a position in contact with the reaction vessel 116 (heating position), and heats the extract within the reaction vessel 116.

[0098] Furthermore, the exhaust unit 305 is moved to the upper part of the reaction vessel 116 to perform heating for a certain period of time and exhaust with the valve 306 open.

[0099] Without evaporating and concentrating the purified liquid in the reaction vessel 116, the movable unit 503 maintains the heating unit 502 at a distance of more than a certain interval from the reaction vessel 116, and maintains the reaction vessel 116 and the heating unit 502 in a state where they do not come into contact with each other for a certain period of time.

[0100] Furthermore, after a certain period of time, the purified liquid in the reaction vessel 116 is introduced into the separation unit 102 through the dispensing mechanism 133 in the separation unit.

[0101] like Figure 5 As shown, unlike Example 1, the standby section is set up separately from the evaporation section. By moving the heating unit 502, contact with or non-contact with the reaction vessel 116 can be performed, and evaporation concentration can be performed according to the reaction vessel 116.

[0102] According to Embodiment 2 of the present invention, in addition to achieving the same effects as in Embodiment 1, the contact between the heating unit 502 and the reaction vessel 16 can be released via the movable unit 503. Therefore, there is no need to consider the configuration. Figure 4 The ejector pin shown.

[0103] Furthermore, in Example 2, a standby section for the reaction vessel is not required, thus enabling the evaporation and concentration mechanism 131 to be miniaturized.

[0104] (Example 3)

[0105] Next, Embodiment 3 of the present invention will be described.

[0106] The difference between Example 3 and Example 1 lies in the structure of the evaporation and concentration mechanism; all other structures are the same. Therefore, illustrations and detailed descriptions of parts other than the evaporation and concentration mechanism are omitted.

[0107] In Example 3, by setting the evaporation and concentration section 302 in a region with an exhaust unit, evaporation and concentration can be performed on multiple reaction vessels 116 at one time.

[0108] Figure 6 This diagram illustrates Example 3, schematically showing an example where the evaporation and concentration section 302 is disposed in a region having an exhaust unit. Additionally, Figure 7 It means Figure 6 A diagram showing a variation of the example shown.

[0109] In Example 3, except Figure 1 In addition to the structure shown, the device also includes: an evaporation and concentration zone 601, opening and closing units (opening and closing sections) 602 and 603 separating the pretreatment section 101, and an exhaust unit (exhaust section) 604. The exhaust unit (exhaust section) 604 exhausts air from the evaporation and concentration zone 601.

[0110] Examples of opening / closing units 602 and 603 include opening / closing devices whose opening and closing can be controlled by an automatic analysis device. Examples of exhaust units 604 include vacuum pumps, etc., whose operation can be controlled by an automatic analysis device.

[0111] exist Figure 6 The method for making the evaporation and concentration section 302 movable is shown in the figure.

[0112] First, during the period when the opening and closing unit 602 is open and the evaporation and concentration area 601 is open, in order to receive the reaction vessel 116 for which evaporation and concentration are performed, the evaporation and concentration section 302 is moved to a point where the conveying mechanism 132 can access the vessel receiving section 301.

[0113] Furthermore, the reaction container 116 is moved into the container receiving section 301 by the conveying mechanism 132. If the reaction container 116 is continuously moved into multiple container receiving sections 301, the above operation is repeated.

[0114] Next, after moving the evaporation and concentration section 302 into the evaporation and concentration area 601, the opening and closing unit 602 is closed, so that the evaporation and concentration area 601 is isolated from the outside air, and the exhaust unit 604 is activated.

[0115] After evaporation and concentration for a certain period of time, the opening and closing unit 603 is opened, and the conveying mechanism 132 moves the evaporation and concentration section 302 to a point where the separation section dispensing mechanism 133 can access the reaction vessel 116. Then, the purified liquid in the reaction vessel 116 is introduced into the separation section 102 through the separation section dispensing mechanism 133.

[0116] Next, regarding Figure 7 The following is an explanation of the variant examples shown. Figure 7The example shown is an example of a method for fixing the installation position of the evaporation and concentration section 302.

[0117] First, while the opening / closing unit 602 is open, the reaction vessel 116 for evaporation and concentration is moved into the container receiving section 301 of the evaporation and concentration section 302 using the conveying mechanism 132. If the reaction vessel 116 is being moved into multiple container receiving sections 301 consecutively, the above operation is repeated.

[0118] Next, after closing the opening and closing unit 602, the exhaust unit 604 is activated. After evaporation and concentration for a certain period of time, the opening and closing unit 603 is opened, and the purified liquid in the reaction vessel 116 is introduced into the separation unit 102 through the dispensing mechanism 133 in the separation unit.

[0119] According to Embodiment 3 of the present invention, in addition to achieving the same effect as Embodiment 1, it is possible to provide only one exhaust unit 604.

[0120] (Example 4)

[0121] Next, Embodiment 4 of the present invention will be described.

[0122] First, the basic principle of the concentration ratio calculation process in Example 4 will be explained.

[0123] The concentration ratio calculation process in Example 4 is used to calculate the concentration ratio of the sample that underwent evaporation and concentration treatment, or to determine the presence or absence of abnormalities in the evaporation and concentration treatment. The determination process in Example 4 can also be applied to any of Examples 1 to 3.

[0124] In this invention, a sample intended for evaporation and concentration treatment is mixed with a sample not subjected to evaporation and concentration treatment, and the mixture is continuously processed at a certain period (a certain interval). Therefore, Example 4 is an example used to confirm whether the concentration treatment has been reliably performed, to determine cases of abnormal concentration, and to ensure reliability.

[0125] Figure 8 This is a diagram illustrating an example of the process for calculating the concentration ratio of the refined liquid obtained through the pretreatment process. Additionally, Figures 9-11 This is a schematic diagram illustrating an example of mass chromatograms obtained from the analytical section for the standard substance used in concentration ratio calculations, the internal standard substance, the standard substance, and the analyte. The vertical axis represents ionic strength, and the horizontal axis represents the LC retention time. Furthermore, Figure 9 (A) is a mass chromatogram derived from the standard substance used in the concentration ratio calculation. Figure 9 (B) is a mass chromatogram derived from internal standard substances. Figure 9 (C) indicates a mass chromatogram derived from a standard substance.

[0126] in addition, Figure 10 and Figure 11 Each of them has (A) a mass chromatogram derived from the standard substance used for concentration ratio calculation, (B) a mass chromatogram derived from the internal standard substance, and (C) a mass chromatogram derived from the analyte component. These are explanatory diagrams for comparison with the judgment criteria.

[0127] exist Figure 8 First, the reaction vessel 116 containing the purified liquid obtained through the pretreatment process is transported to the evaporation and concentration unit 131 to evaporate and concentrate the analyte component 801 in the purified liquid (step S208).

[0128] Next, in order to change the composition of the purified solution so that the analytical component 801 is combined with the separation column of the separation section 102, the diluent is drawn from the reagent container 121 of the reagent tray 122 and discharged into the reaction vessel 116 by the reagent dispensing mechanism 123 (step S801).

[0129] Furthermore, a concentration ratio calculation standard substance 802 of a predetermined known concentration is added to the reagent container 121 containing the diluent of Example 4. As the concentration ratio calculation standard substance, a substance selected that is simultaneously detected with the analyte component is chosen within the range of the LC holding time for detecting the analyte component of the sample being analyzed.

[0130] In addition, standard substances with known concentrations are analyzed according to the analytical processing procedures, and the mass chromatograms of the standard substances are obtained in advance (refer to...). Figure 9 (etc.), which are recorded in the storage unit 107 as reference data for judgment.

[0131] Here, when a mass chromatogram is obtained by analyzing a sample of the analyte according to the analytical processing procedure, data containing peaks of the analyte component originating from the sample or from an internal standard substance are obtained (acquired data), and data containing peaks from a standard substance used for concentration ratio calculation are obtained within the same LC retention time range (refer to data). Figure 10 and Figure 11 ).

[0132] When comparing mass chromatograms of various components and substances obtained from other sources, the comparisons are performed after data standardization. For example, data standardization is achieved by comparing peaks with the same retention time with each other, and calculating the percentage of the peak area of ​​the obtained data when the peak area of ​​the benchmark data is set to 100%.

[0133] For example, if the peak area of ​​the acquired data is 97% of the peak area of ​​the benchmark data for judgment, it is determined that there is a 3% difference. Here, as an indicator of the degree of inconsistency between the benchmark data for judgment and the acquired data, the difference ratio is defined by the following mathematical formula (1).

[0134] Difference ratio (%) = |1 - (obtained data) / (benchmark data for judgment)| × 100…(1)

[0135] In the evaporation concentration anomaly determination process of this embodiment 4, the difference ratio given by the above mathematical formula (1) is compared with a preset difference ratio threshold, and the presence or absence of an evaporation concentration anomaly is determined based on the comparison result. That is, the difference ratio is calculated simultaneously with the concentration of the component to be analyzed, and compared with the difference ratio threshold, thereby temporarily determining whether an evaporation concentration anomaly exists. The determination process is performed by the control unit 104.

[0136] The differential ratio threshold, which serves as the judgment criterion, is preset before the analysis and processing and the evaporation-concentration anomaly judgment processing, and is stored in the storage unit 107, just like the judgment reference data. Alternatively, it can be set to be input appropriately by the operator. It is known that the standard measurement error of LC-MS is approximately 5 to 10%. Therefore, in this embodiment 4, the case where the differential ratio threshold is set to 15% will be illustrated.

[0137] That is, when the difference ratio exceeds 15%, an anomaly of evaporation and concentration is determined. Furthermore, since the measurement error is considered to be dependent on the device, it is also possible to further improve the accuracy of the determination of anomalies of evaporation and concentration by setting a difference ratio threshold according to the device.

[0138] For example, in Figure 10 In the case illustrated in (A), when comparing the judgment reference data (dashed line) with the peak data obtained from the standard reference for concentration ratio calculation, the peak area obtained from the standard reference for concentration ratio calculation is reduced in the data obtained during sample analysis compared to the judgment reference data.

[0139] Therefore, if the difference ratio exceeds the difference ratio threshold (15%), it is determined to be an anomaly of reduced evaporation of the purified liquid during the evaporation and concentration process. This is because, when the evaporation is reduced, the total volume of the purified liquid after adding a diluent containing a standard substance for calculating the concentration ratio is greater than normal, and the concentration of the standard substance for calculating the concentration ratio is lower. Meanwhile, as... Figure 10 As shown in (B), the concentrations of the analyte and internal standard substances in the purified solution decrease, resulting in a reduced peak area. Therefore, as... Figure 10 As shown in (C), the concentration of the analyte component is calculated less.

[0140] In this case, the data derived from the analyte can also be standardized using the internal standard method, either by using data from internal standard materials or by using data from standard materials calculated based on the concentration ratio, thereby performing correction.

[0141] In addition, Figure 11 In the case illustrated in (A), when comparing the judgment reference data (dashed line) with the peak data obtained from the standard reference for concentration ratio calculation, the peak area obtained from the standard reference for concentration ratio calculation increases in the data obtained during sample analysis compared to the judgment reference data.

[0142] Therefore, if the difference ratio exceeds the difference ratio threshold (15%), it is considered an anomaly indicating an increase in the evaporation of refined liquid during the evaporation and concentration process. This is because, when the evaporation rate increases, such as... Figure 11 As shown in (B), the total volume of the purified solution after adding the diluent containing the standard substance for concentration ratio calculation is less than normal, while the concentration of the standard substance for concentration ratio calculation is higher. Simultaneously, the concentrations of the analyte and the internal standard substance in the purified solution increase, and the peak area increases. Therefore, as... Figure 11 As shown in (C), the concentration of the analyte component is calculated more.

[0143] In this case, the data derived from the analyte can also be standardized using the internal standard method, either by using data from internal standard materials or by using data from standard materials calculated based on the concentration ratio, thereby performing correction.

[0144] The control unit 104 determines whether the evaporation and concentration process is sufficient. If it is determined to be insufficient, the control unit 104 can enable the display unit 106 to display an alarm indicating that the evaporation and concentration process is insufficient.

[0145] Here, a general explanation is given of the method for selecting the standard substance used in the concentration ratio calculation.

[0146] In MS, stable isotopic compounds of the analyte that have been labeled with isotopes or compounds with similar chemical and physical properties (analogous compounds) are generally used as internal standard substances.

[0147] Therefore, this point was taken into consideration when selecting the standard material for calculating the concentration ratio. The standard material was selected as the one that could be captured by magnetic beads during sample pretreatment, and that could be fully separated from the peaks of the analyte and the internal standard material in the mass chromatogram, and that could be detected within the retention time range of the LC measured for the analyte.

[0148] First, the chemical properties of the substances selected as standard substances for concentration ratio calculation were studied.

[0149] In this Example 4, the analyte is captured through hydrophobic interaction with the magnetic beads. Furthermore, in LC, a reversed-phase column is typically used in the separation column. The reversed-phase column is essentially maintained based on hydrophobic interactions.

[0150] Therefore, for example, it is preferable that the standard material used for concentration ratio calculation has the same degree of hydrophobicity as the analyte component, which is captured by the magnetic beads and the reversed-phase column. For example, ionic compounds in their non-dissociated state are generally highly hydrophobic, thus strengthening the retention of the reversed-phase column.

[0151] The difference between the dissociated and non-dissociated states arises from the relationship between the pH of the mobile phase in LC and the pKa of the compound. The pKa of the analyte and internal standards are generally set to a value ±2 or greater than the pH of the mobile phase, thereby ensuring stable performance. That is, considering the above, it is important to select substances that are sufficiently separated from the peaks originating from the analyte and internal standards in the chromatogram, and that are detected within the retention time range of the LC used for measuring the analyte, based on the equilibrium between the pKa of the analyte, internal standards, and the mobile phase, as standards for calculating the concentration ratio.

[0152] Next, the molecular weights of the substances selected as standard substances for concentration ratio calculation were studied.

[0153] When the m / z of ions derived from the concentration ratio calculation standard overlaps to a degree that is unrecognizable by the mass resolution of MS, there is a possibility of falsely identifying an increase in peak intensity of ions derived from the concentration ratio calculation standard. In this case, it is difficult to determine whether an anomaly in the fractionation rate exists.

[0154] The typical mass resolution of an MS is around 1 (m / z). Therefore, the peak m / z differences between the mass chromatograms derived from the analyte and internal standards, as well as the standards used for concentration ratio calculation, should be at least 1 [Da], and preferably more than 3 [Da].

[0155] In particular, for accurate analysis of samples containing a large number of impurities, such as biological samples, the MS detector preferably uses an MS / MS apparatus capable of detecting product ions. When using MS / MS, even if the m / z of the precursor ions of each substance are the same, it is acceptable as long as the m / z of the product ions are different.

[0156] According to Embodiment 4 of the present invention, a diluent containing a standard substance is added to the extract that has been concentrated by the evaporation and concentration unit 131. The signal amount of the standard substance is detected by the analysis unit 103, and the concentration ratio of the extract is calculated based on the detected signal amount. Therefore, by mixing the heated sample and the unheated sample and concentrating the heated sample, and by continuously performing the treatment at a certain period, it is confirmed whether the concentration treatment has been reliably performed, and a determination is made in case of concentration abnormality. Therefore, reliability can be ensured.

[0157] The present invention, constructed as described above, achieves the following effects.

[0158] In an automated analytical apparatus that integrates pretreatment and LC-MS-based analysis, automation of the process based on batch processing is envisioned. In batch processing, the reaction solution is processed simultaneously, thus making it impossible to select whether or not to concentrate based on the sample. However, in automated analytical apparatuses for clinical examinations that continuously process various samples, the requirement to detect analytes in low-concentration regions is limited; therefore, it is preferable to have a mechanism and control that allows for selective evaporation and concentration based on the sample.

[0159] In contrast, this invention includes an evaporation concentration mechanism 131, which performs evaporation concentration by evaporating the extract containing the analyte to prepare a sample solution before introducing the sample solution into the separation unit 102 (liquid chromatography). The evaporation concentration mechanism 131 is configured to have a container receiving section 301 that receives a reaction vessel 116 containing the extract, and has a mechanism and control method for determining whether evaporation concentration is performed based on the reaction vessel 116 arriving at the container receiving section 301. Therefore, it is possible to select whether to perform evaporation concentration based on the sample, and an automated analytical device capable of controlling the evaporation concentration of the sample can be realized.

[0160] Furthermore, the present invention is not limited to the embodiments and modifications described above, but includes various other modifications. For example, the embodiments described above are detailed for ease of understanding and illustration of the invention, and are not limited to having all the structures described.

[0161] In the examples above, examples of concentration treatment involving both heating and venting were shown; however, examples of concentration treatment involving either heating or venting can also be embodiments of the present invention. This is because sample concentration treatment can be performed solely by heating or solely by venting.

[0162] Furthermore, the aforementioned structures and functions can also be implemented, for example, by designing part or all of them in an integrated circuit. Alternatively, the aforementioned structures and functions can be implemented in software by a processor interpreting and executing programs that implement each function.

[0163] Alternatively, the analysis unit 103 can be an optical analysis device, other than a quality analysis device.

[0164] Symbol Explanation

[0165] 100…Automatic analysis device, 101…Pretreatment unit, 102…Separation unit, 103…Analysis unit, 104…Control unit, 105…Input unit, 106…Display unit, 107…Storage unit, 111…Sample container, 112…Transfer mechanism, 113…Sample dispensing mechanism, 114…Dispensing pipette tip assembly / disassembly unit, 115…Dispensing pipette tip holder, 115a…Dispensing pipette tip, 116…Reaction vessel, 117…Reaction vessel holder, 118…Transfer mechanism, 119…Opening, 120…Reaction vessel tray, 121…Reagent container, 122…Reagent tray, 123…Reagent dispensing mechanism, 124…Magnetic separation mechanism, 125…Transfer… 126… Rotary track, 131… Evaporation and concentration mechanism, 132… Conveying mechanism, 133… Dispensing mechanism for separation section, 201… Magnet, 202… Magnetic bead assembly, 203… Magnetic bead assembly, 301… Container receiving section, 302… Evaporation and concentration section, 303… Standby section, 304… Heating unit, 305… Exhaust unit, 306… Valve, 401… Top pin, 501… Container support section, 502… Heating unit, 503… Movable unit, 601… Evaporation and concentration zone, 602… Opening and closing unit, 603… Opening and closing unit, 604… Exhaust unit, 801… Analytical component, 802… Standard substance for concentration ratio calculation.

Claims

1. An automatic analysis device, comprising: The evaporation and concentration unit performs a concentration process that concentrates the analyte by evaporating the extract from the sample. The analytical section analyzes the analyte components of the sample; and The control unit controls the operation of the evaporation and concentration unit and the analysis unit. Its features are, The control unit determines whether to perform evaporation and concentration treatment on the analyte in the sample. For samples determined to require evaporation and concentration treatment, the evaporation and concentration unit concentrates the analyte in the sample. The extract contains an internal standard substance. A diluent containing a standard substance for calculating the concentration ratio is added to the extract that has undergone the concentration treatment in the evaporation and concentration section. The analytical unit detects the signal quantities of the analyte component, the internal standard substance, and the standard substance used to calculate the concentration ratio. The control unit performs correction by standardizing data derived from the analyte component using data from the internal standard material, or by standardizing data derived from the concentration ratio calculation standard material.

2. The automatic analysis device according to claim 1, characterized in that, The evaporation and concentration section has: A heating unit that heats the extract to evaporate it; and The exhaust section draws in the steam generated by the heating section.

3. The automatic analysis device according to claim 2, characterized in that, The evaporation and concentration section also has a standby section, which enables the control section to determine that the sample is in standby mode and will not undergo evaporation and concentration.

4. The automatic analysis device according to claim 2, characterized in that, The exhaust section moves between a first position, which is a certain distance away from the heating section, and a second position, which is close to the heating section. When the evaporation and concentration are not being performed, the control unit is positioned at the first location with the exhaust section in place. When the evaporation and concentration are performed, the control unit moves the exhaust section to the second position.

5. The automatic analysis device according to claim 2, characterized in that, The extract is stored in a container. The heating part includes: a container receiving part that receives the container; and an ejector pin that pushes the container into the container receiving part from below.

6. The automatic analysis device according to claim 2, characterized in that, The extract is stored in a container. The automatic analysis device includes a movable part that allows the heating part to move between a non-heated position where it does not contact the container and does not heat the extract in the container, and a heated position where it contacts the container.

7. The automatic analysis device according to claim 2, characterized in that, The heating section includes an opening and closing section disposed within the evaporation and concentration area to open and close the evaporation and concentration area, and an exhaust section to exhaust gas from the evaporation and concentration area.

8. The automatic analysis device according to claim 1, characterized in that, The automatic analysis device has a display unit, and the control unit determines whether the evaporation and concentration treatment is sufficient based on the concentration ratio of the extract. If it is determined that the evaporation and concentration treatment is insufficient, the display unit displays an alarm indicating that the evaporation and concentration treatment is insufficient.

9. The automatic analysis device according to claim 2, characterized in that, The sample to be evaporated and concentrated was mixed with the sample not to be evaporated and concentrated and then processed continuously at certain intervals.

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