WO2025243767A1 - Système de préparation d'échantillons et procédé de préparation d'échantillons - Google Patents

Système de préparation d'échantillons et procédé de préparation d'échantillons

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Publication number
WO2025243767A1
WO2025243767A1 PCT/JP2025/015725 JP2025015725W WO2025243767A1 WO 2025243767 A1 WO2025243767 A1 WO 2025243767A1 JP 2025015725 W JP2025015725 W JP 2025015725W WO 2025243767 A1 WO2025243767 A1 WO 2025243767A1
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WO
WIPO (PCT)
Prior art keywords
sample
container
sample preparation
stock solution
needle
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
PCT/JP2025/015725
Other languages
English (en)
Japanese (ja)
Inventor
奈津紀 岩田
凌大 金丸
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Shimadzu Corp
Original Assignee
Shimadzu Corp
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Shimadzu Corp filed Critical Shimadzu Corp
Publication of WO2025243767A1 publication Critical patent/WO2025243767A1/fr
Pending legal-status Critical Current
Anticipated expiration legal-status Critical

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Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/04—Preparation or injection of sample to be analysed
    • G01N30/06—Preparation
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/04—Preparation or injection of sample to be analysed
    • G01N30/16—Injection
    • G01N30/18—Injection using a septum or microsyringe
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
    • G01N30/02—Column chromatography
    • G01N30/04—Preparation or injection of sample to be analysed
    • G01N30/24—Automatic injection systems
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/02—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor using a plurality of sample containers moved by a conveyor system past one or more treatment or analysis stations

Definitions

  • the present invention relates to a sample preparation system and a sample preparation method, and more specifically to technology that reduces the time required to prepare samples for liquid chromatographic analysis.
  • the user may need to dilute the original sample solution to prepare the sample for analysis.
  • Non-Patent Document 2 disclose technology in which dilution work is performed by the autosampler of a liquid chromatograph analyzer.
  • an autosampler performs the dilution process.
  • the dilution process involves multiple steps, such as aspirating and dispensing the solution with a needle, moving the needle, and mixing the solution, and the dilution process using an autosampler can take a significant amount of time. Therefore, there is a need to shorten the time required for the dilution process to prepare a sample.
  • the present invention was devised in light of this situation, and its purpose is to provide technology for shortening the time required to prepare samples for liquid chromatographic analysis.
  • a sample preparation system is a sample preparation system that dilutes a stock solution with a diluent to prepare a sample for liquid chromatography analysis.
  • the sample preparation system includes a needle that aspirates and dispenses the stock solution, and a control unit that controls the needle.
  • the control unit causes the needle to perform a first suction to aspirate the required amount of stock solution, a first discharge to dispense the stock solution aspirated by the first suction and the required amount of diluent into a container, a second suction to aspirate the stock solution and diluent from the container at a first speed, and a second discharge to dispense the stock solution and diluent aspirated by the second suction into the container at a second speed that is at least 10 times faster than the first speed.
  • a sample preparation method is a sample preparation method for diluting a stock solution with a diluent to prepare a sample for liquid chromatography analysis.
  • the sample preparation method includes the steps of (a) aspirating a required amount of stock solution, (b) dispensing the aspirated required amount of stock solution and the required amount of diluent into a container, (c) aspirating the stock solution and diluent from the container at a first speed, and (d) dispensing the aspirated stock solution and diluent into the container at a second speed that is at least 10 times the first speed.
  • This disclosure makes it possible to reduce the time required to prepare samples for liquid chromatography analysis.
  • FIG. 1 is a schematic diagram illustrating a configuration of an analysis device according to an embodiment.
  • 2 is a schematic diagram showing a state different from that shown in FIG. 1 in the analysis device according to the embodiment.
  • FIG. FIG. 2 is a schematic diagram illustrating a configuration of a control device according to an embodiment.
  • FIG. 10 is a diagram showing a calibration curve according to Verification Example 1.
  • FIG. 10 is a diagram showing the actual measured values of the dilution ratio in Verification Example 2.
  • FIG. 10 is a diagram for explaining a sample preparation process.
  • [Configuration of the analyzer] 1 is a diagram showing the configuration of an analytical device 100 according to an embodiment.
  • analytical device 100 will be described using a liquid chromatograph, but is not limited to this, and analytical device 100 may also be, for example, a liquid chromatograph mass spectrometer (LC-MS), a liquid chromatograph tandem mass spectrometer (LC-MS/MS), or a liquid chromatograph inductively coupled plasma mass spectrometer (LC-ICP-MS).
  • LC-MS liquid chromatograph mass spectrometer
  • LC-MS/MS liquid chromatograph tandem mass spectrometer
  • LC-ICP-MS liquid chromatograph inductively coupled plasma mass spectrometer
  • the analytical device 100 includes a mobile phase container 1, a liquid delivery pump 2, an autosampler 3, a column 4, a detector 5, a waste liquid container 6, a control device 7, and a diluent container 8.
  • the analytical device 100 can separate multiple types of components contained in a sample into individual types of components by utilizing differences in interactions with the stationary phase and the mobile phase.
  • the mobile phase container 1 stores solvent M, which is the mobile phase that carries the sample injected into the analytical device 100.
  • Solvent M is, for example, an organic solvent, an aqueous solution (or water), or a mixture of these.
  • the analytical device 100 may be equipped with a single mobile phase container or multiple mobile phase containers.
  • the liquid delivery pump 2 sucks the solvent M stored in the mobile phase container 1 and delivers it at a predetermined flow rate.
  • the flow rate of the solvent M delivered by the liquid delivery pump 2 is constant for each measurement.
  • the analytical device 100 may be equipped with a single liquid delivery pump or multiple liquid delivery pumps.
  • the diluent container 8 stores diluent D, which is a liquid used to dilute the sample stock solution S.
  • the diluent D is, for example, an organic solvent, an aqueous solution (or water), or a mixture of these.
  • the diluent D is also used as a cleaning liquid for the needle 32.
  • the autosampler 3 prepares samples for liquid chromatography analysis. It also introduces the prepared samples into the analytical flow path.
  • the autosampler 3 includes a tray 31, a needle 32, a loop 33, a metering unit 34, an injection port 35, a valve 36, and a low-pressure valve 37.
  • Low-pressure valve 37 is a multi-way switching valve.
  • the common port of low-pressure valve 37 is connected to metering unit 34, and the selected port of low-pressure valve 37 is connected to port 366 of valve 36 and diluent container 8.
  • Tray 31 can accommodate containers containing liquids. Tray 31 includes wells 311 and 312 for arranging the containers. Container C1 containing sample stock solution S is placed in well 311, and empty container C2 is placed in well 312. Sample stock solution S can be, for example, a biological sample, food, beverage, pharmaceutical, pesticide, or liquid derived from the environment.
  • Container C2 contains a sample for liquid chromatography analysis, prepared by diluting sample stock solution S with diluent D. There are no restrictions on the shape or material of container C2, but it is preferable that the shape facilitates stirring of the dispensed liquid. Therefore, container C2 preferably has a relatively small capacity and a rounded bottom.
  • Container C2 is, for example, a polypropylene container with a capacity of 1 mL, or a glass container with a raised bottom and a capacity of 150 ⁇ L.
  • the needle 32 has a tip and a base end, and can aspirate and eject liquid from the tip.
  • the needle 32 is moved vertically and horizontally by a movement mechanism (not shown).
  • Loop 33 is a flow path for retaining the liquid aspirated from the tip of needle 32. Loop 33 is fluidly connected to the base end of needle 32.
  • the metering unit 34 is configured to dispense and suck in liquid or gas.
  • the injection port 35 has an opening for inserting the needle 32.
  • the liquid injected from the injection port 35 is sent to the valve 36.
  • Valve 36 is used to switch the flow path configuration and includes six ports, ports 361 to 366.
  • valve 36 is a six-port valve, but is not limited to this.
  • Port 361 of valve 36 is connected to a drain flow path (not shown)
  • port 362 is connected to injection port 35
  • port 363 is connected to column 4
  • port 364 is connected to liquid delivery pump 2
  • port 365 is connected to loop 33
  • port 366 is connected to low-pressure valve 37.
  • Valve 36 can switch between two states by switching the connections between ports.
  • Figure 2 is a diagram showing a state different from that shown in Figure 1, which is achieved by switching the connections between ports. These two states will be explained using Figures 1 and 2.
  • ports 361 and 362, ports 363 and 364, and ports 365 and 366 are connected.
  • solvent M in mobile phase container 1 is pushed out by liquid delivery pump 2 and delivered to column 4.
  • low-pressure valve 37 and needle 32 are connected. Therefore, liquid can be sucked in and discharged from the tip of needle 32 by metering unit 34.
  • Column 4 is filled with a stationary phase, through which the mobile phase, solvent M, passes. As it passes through column 4, it interacts with the mobile phase and stationary phase, separating the multiple components in the sample in the time direction. Each component elutes from the outlet of column 4 and is introduced into detector 5.
  • the detector 5 sequentially detects the components separated and eluted by the column 4 and transmits the detected data to the control device 7.
  • the solvent flowing out from the detector 5 is discharged into a waste liquid container 6.
  • the solution flowing out from the detector 5 may also be introduced into another analytical device (for example, a mass spectrometer).
  • the control device 7 is connected to the liquid delivery pump 2, autosampler 3, and detector 5, and controls the analytical device 100 while receiving chromatogram data detected by the detector 5. Note that the autosampler 3 is controlled by the control device 7 to prepare samples, and therefore the autosampler 3 and control device 7 correspond to one embodiment of a "sample preparation system.”
  • Figure 3 is a schematic diagram showing the configuration of the control device 7.
  • the control device 7 includes a controller 70, an input unit 75, and an output unit 76.
  • the input unit 75 and the output unit 76 are connected to the controller 70.
  • the control device 7 is, for example, a computer. Note that the control device 7 does not have to be configured by a single computer, but may be configured by multiple computers. The operation of the control device 7 described in this embodiment may be performed in a distributed manner by multiple computers.
  • the controller 70 includes, as its main components, a processor 71, a memory 72, a communication interface (I/F) 73, and an input/output I/F 74. These components are connected to each other via a bus so that they can communicate with each other.
  • the processor 71 is an example of an electrical circuit, and controls the operation of the control device 7 by executing a given program.
  • the program executed by the processor 71 may be stored in the memory 72, or may be stored in a storage device external to the control device 7.
  • the processor 71 is, for example, a CPU (Central Processing Unit).
  • Memory 72 can non-temporarily store programs executed by processor 71 and chromatogram data obtained by detector 5.
  • Memory 72 includes volatile memory (e.g., RAM (Random Access Memory)) and non-volatile memory (e.g., ROM (Read Only Memory), hard disk drive, and solid state drive).
  • volatile memory e.g., RAM (Random Access Memory)
  • non-volatile memory e.g., ROM (Read Only Memory), hard disk drive, and solid state drive.
  • the database and/or program may also be stored in an external storage device accessible by processor 71.
  • the communication I/F 73 is a communication interface for exchanging various data with external devices.
  • the communication I/F 73 is implemented, for example, by a network adapter.
  • the communication method may be wireless communication such as Bluetooth (registered trademark) or wireless LAN, or wired communication using a USB (Universal Serial Bus) or the like.
  • the input/output I/F 74 is an interface for exchanging various types of data between the processor 71 and an external device connected to the input/output I/F 74.
  • the external device includes an input unit 75 and an output unit 76.
  • the input unit 75 accepts information input to the controller 70.
  • This information includes, for example, the dilution ratio, the volume after dilution, the name of the sample, the type of sample, the position of the well in which the container C1 containing the undiluted sample S is placed, the position of the well in which the empty container C2 is placed, the analysis conditions, and analytical conditions.
  • the analysis conditions include, for example, the flow rate of the mobile phase and the temperature of the column oven.
  • the analytical conditions are conditions used when analyzing the obtained chromatogram, and include, for example, conditions for waveform processing and component identification.
  • the input unit 75 is configured, for example, by a touch panel, a mouse, and/or a keyboard.
  • the output unit 76 displays information in accordance with instructions from the controller 70. This information may be, for example, chromatogram data detected by the detector 5, a created calibration curve, or the calculated concentration of components contained in the sample.
  • the output unit 76 may be, for example, a liquid crystal display capable of displaying images.
  • Non-Patent Documents 1 and 2 disclose technology in which the dilution process is performed by an autosampler in a liquid chromatograph analytical device.
  • Non-Patent Document 1 and Non-Patent Document 2 the user places the sample stock solution and the dilution solution used for dilution into the device, and the autosampler then performs the dilution process. This reduces the burden on the user required for dilution.
  • dilution involves multiple steps, such as aspirating and dispensing the solution with a needle, moving the needle, and mixing the solutions, and can be very time-consuming.
  • the needle performs three suction and discharge operations each to mix the undiluted sample solution and diluted solution dispensed into the container. If only one suction and discharge operation each is performed to mix the sample, the time required for the dilution process is shortened, but the undiluted sample solution and diluted solution may not mix uniformly within the sample. In such cases, the accuracy of measuring the components contained in the sample may decrease. Therefore, it is desirable to shorten the time required to prepare a sample in which the undiluted sample solution and diluted solution are mixed uniformly.
  • sample dilution system Therefore, in the sample preparation system including the autosampler 3 and the control device 7 according to the embodiment, the sample stock solution S and the diluent D are aspirated at a first speed, and the aspirated sample stock solution S and the diluent D are discharged into the container at a second speed that is at least 10 times faster than the first speed. This reduces the number of times the aspirating and dispensing steps are performed to mix the sample. Furthermore, by discharging the sample into the container at the second speed, the sample stock solution S and the diluent D are uniformly mixed in the discharged sample.
  • the sample preparation system according to the embodiment can shorten the time required to prepare the sample without reducing the degree of mixing of the sample stock solution S and the diluent D.
  • the sample preparation system can reduce the number of times the sample is dispensed to mix it, thereby improving the reproducibility of the dilution process.
  • some of the sample may remain on the inner wall of the needle.
  • the prepared sample will differ from the dilution ratio and/or volume of the diluted sample desired by the user. Therefore, by reducing the number of times the sample is aspirated and dispensed, the variation in the dilution ratio among multiple samples diluted in the dilution process can be reduced, improving reproducibility.
  • Sample preparation is performed by a sample preparation system consisting of an autosampler 3 and a control device 7.
  • the control device 7 receives information necessary for the dilution process from the user via the input unit 75.
  • Information necessary for the dilution process includes, for example, the dilution ratio, the volume after dilution, the position of the well in which the container C1 containing the sample stock solution S is placed, and the position of the well in which the empty container C2 is placed.
  • the control device 7 calculates the required amount of sample stock solution S and the required amount of diluent D based on the information required for the dilution process received from the user.
  • the user places a container containing sample stock solution S and an empty container in the wells of tray 31 according to the information entered into input unit 75.
  • container C1 containing sample stock solution S is placed in well 311
  • empty container C2 is placed in well 312.
  • control device 7 sends diluent D from the diluent container 8 and retains the diluent D in the loop 33 and the piping from the loop 33 to the tip of the needle 32.
  • the control device 7 moves the needle 32 to the container C1 using a moving mechanism (not shown).
  • the control device 7 switches the selected port so that the common port of the low-pressure valve 37 is connected to the port 366 of the valve 36.
  • the control device 7 controls the metering unit 34 to aspirate the calculated required amount of the sample stock solution S from the container C1 into the needle 32.
  • control device 7 After the control device 7 has aspirated the sample stock solution S into the needle 32, it controls the metering unit 34 to aspirate air into the needle 32.
  • the amount of air aspirated into the needle 32 depends on the inner diameter of the needle 32, and the needle 32 is caused to aspirate air so that the distance from the tip of the needle 32 to the liquid surface of the sample stock solution S inside the needle 32 is a predetermined distance.
  • the predetermined distance is, for example, 4 to 5 mm.
  • the volume of air aspirated is, for example, 0.1 to 1 ⁇ L.
  • the control device 7 moves the needle 32 to the container C2.
  • the control device 7 then controls the measuring unit 34 to dispense the aspirated sample stock solution S and the calculated required amount of diluent D from the needle 32 into the container C2.
  • control device 7 causes the needle 32 to aspirate and dispense the sample stock solution S and diluent D contained in the container C2 once each, thereby mixing the sample stock solution S and diluent D.
  • the control device 7 causes the needle 32 to aspirate the sample stock solution S and diluent D contained in container C2.
  • air is aspirated to prevent the aspirated sample stock solution S and diluent D from mixing with the diluent D held in the loop 33.
  • the amount of air is, for example, 5 ⁇ L.
  • the speed at which the needle 32 aspirates the sample stock solution S and diluent D contained in container C2 is defined as the first speed.
  • the first speed is, for example, preferably 3 ⁇ L/sec or less, and more preferably 1 to 2 ⁇ L/sec.
  • control device 7 causes the needle 32 to eject the sample stock solution S and diluent D aspirated at the first speed into the container C2 at a second speed that is at least 10 times faster than the first speed.
  • the second speed is, for example, 30 to 150 ⁇ L/sec, and preferably 30 to 40 ⁇ L/sec.
  • the sample preparation system can prepare a sample to be subjected to liquid chromatography analysis.
  • the prepared sample is aspirated through needle 32 and held in loop 33.
  • needle 32 is connected to injection port 35, and with ports 364 and 365 connected (as shown in Figure 2), solvent M is delivered from delivery pump 2, and the sample held in loop 33 is introduced into column 4 together with solvent M.
  • the sample preparation method disclosed herein can reduce the number of times that the suction and discharge steps are performed to mix the undiluted sample solution and the diluted solution when preparing a sample for liquid chromatography analysis. This reduces the time required to prepare the sample.
  • the diluent D for diluting the sample stock solution S is supplied from the diluent container 8, but this is not limited to this.
  • a container containing diluent D may be placed in a well provided on the tray 31, and the diluent D in that container may be used to dilute the sample stock solution S.
  • the control device 7 dispenses the required amount of diluent D into the container C2 using the needle 32, then causes the needle 32 to suck the sample stock solution S and ejects the sample stock solution S into the container C2.
  • the analytical device 100 includes a mobile phase container 1 that stores a solvent M used as the mobile phase, and a diluent container 8 that stores a diluent D used to dilute the stock sample solution S, but these may be the same container.
  • the solvent M used as the mobile phase and the diluent D used to dilute the stock sample solution S are the same.
  • sample preparation is performed by a sample preparation system consisting of an autosampler 3 and a control device 7, but this is not limited to this. Sample preparation may also be performed, for example, using a dispenser that allows the user to dispense liquid.
  • the sample preparation system dilutes a standard sample to prepare a plurality of samples for liquid chromatographic analysis, and checks the linearity of the measured values of those samples in the liquid chromatographic analysis.
  • Verification Example 1 a 250 mg/L aqueous caffeine solution is used as the sample stock solution. Ultrapure water is used as the dilution solvent.
  • the sample stock solution is diluted at dilution rates of 500x, 200x, 100x, 50x, 20x, 10x, and 5x to prepare samples using a sample preparation system consisting of an autosampler 3 and a control device 7.
  • the prepared samples are introduced into a column 4, and chromatogram data for each sample is created by the control device 7 based on the data obtained by the detector 5.
  • the area values in the created chromatogram data are used as the measured values for each sample.
  • the control device 7 receives the dilution rate and the volume of the diluted sample from the user via the input unit 75.
  • the volume of the diluted sample is, for example, 100 ⁇ L.
  • the control device 7 Based on the received dilution rate and the volume of the diluted sample, the control device 7 calculates the required volume of stock solution and the required volume of diluent. For example, if the dilution rate is 100 times and the volume of the diluted sample is 100 ⁇ L, the required volume of stock solution is calculated to be 1 ⁇ L and the required volume of diluent is calculated to be 99 ⁇ L.
  • Figure 4 shows the caffeine concentration of each sample and the area value of the chromatogram obtained by liquid chromatography analysis.
  • the caffeine concentrations of samples obtained by diluting a 250 mg/L caffeine aqueous solution at dilution rates of 500, 200, 100, 50, 20, 10, and 5 correspond to 0.5 mg/L, 1.25 mg/L, 2.5 mg/L, 5 mg/L, 12.5 mg/L, 25 mg/L, and 50 mg/L, respectively.
  • the horizontal axis represents the caffeine concentration
  • the vertical axis represents the area value, which is the measurement value of the corresponding sample.
  • Figure 4 also shows a linear equation that expresses the relationship between caffeine concentration and the measurement value.
  • R is the correlation coefficient of this linear equation, and the closer the contribution rate or coefficient of determination, expressed as the square of the correlation coefficient, is to 1, the stronger the positive correlation.
  • a sample preparation system including an autosampler 3 and a control device 7 prepares a sample by diluting a stock sample solution containing component X at a 50-fold dilution rate.
  • the prepared sample is introduced into a column 4, and chromatogram data for each sample is created by the control device 7 based on the data obtained by the detector 5.
  • the area value of the peak derived from component X in the created chromatogram data is taken as the measured value of component X in each sample.
  • the six samples prepared with the first speed set to 1 ⁇ L/sec and the second speed set to 35 ⁇ L/sec were grouped together as Group 1. Furthermore, the six samples prepared with the first speed set to 1 ⁇ L/sec and the second speed set to 1 ⁇ L/sec were grouped together as Group 2.
  • Figure 5 shows the dilution ratios calculated from the chromatogram data obtained by analyzing samples from each group.
  • the vertical axis represents the ratio of the actual dilution ratio of the prepared sample to the set dilution ratio (50x). Therefore, the closer the value on the vertical axis is to 100%, the better the dilution accuracy relative to the set dilution ratio.
  • the average measured dilution rate in Group 1 was 42.9x, which is 85.8% of the set 50x dilution rate.
  • the average measured dilution rate in Group 2 was 42.2x, which is 84.5% of the set 50x dilution rate. Therefore, Group 1, which has a faster second speed, is diluted at a dilution rate closer to the set dilution rate than Group 2, which has a faster second speed.
  • the error bars on each graph indicate the variability in the measured dilution ratio for each group.
  • the variability in the measured dilution ratio for Group 1 is smaller than the variability in the measured dilution ratio for Group 2.
  • Group 1, which has a faster second speed has smaller variability in the dilution ratio of the prepared samples than Group 2, which has a faster second speed, and the reproducibility of the dilution process is higher.
  • the sample preparation system disclosed herein can dilute the sample at a dilution ratio closer to the set dilution ratio than a sample preparation system with a slower second speed, and furthermore, the reproducibility of the sample dilution process is high.
  • [Processing flow] 6 shows a process for preparing a sample for liquid chromatographic analysis by diluting a stock solution of the sample.
  • steps S10 and S12 relate to the operation of the control device 7 itself.
  • steps S14 to S22 relate to the operation performed by the autosampler 3 controlled by the control device 7.
  • step S10 the control device 7 receives the dilution rate and the volume of the diluted sample from the user via the input unit 75.
  • step S12 the control device 7 calculates the amount of sample stock solution S and the amount of diluent D required to prepare the sample based on the dilution ratio and volume of the diluted sample received in step S10.
  • step S14 the control device 7 controls the autosampler 3 to move the needle 32 to the container C1 containing the sample stock solution S, and causes the needle 32 to aspirate the required amount of stock solution S calculated in step S12. In one embodiment, this operation is the first aspirate.
  • step S16 the control device 7 causes the needle 32 to suck in air so that the distance from the tip of the needle 32 to the liquid surface of the sample stock solution S is a predetermined distance.
  • the predetermined distance is, for example, 4 to 5 mm. This prevents the stock solution S sucked in in step S14 from leaking out of the needle 32.
  • step S18 the control device 7 moves the needle 32 to the empty container C2 and dispenses the required amount of concentrate S aspirated in step S14 and the required amount of diluent D calculated in step S12.
  • the diluent D is supplied to the needle 32 from the diluent container 8 via the measuring unit 34. This operation is the first dispense in one embodiment.
  • step S20 the control device 7 causes the needle 32 to aspirate the sample stock solution S and diluent D from container C2 at a first speed.
  • the first speed is preferably 3 ⁇ L/sec or less, and more preferably 1 to 2 ⁇ L/sec. In one embodiment, this operation is the second aspirate.
  • step S22 the control device 7 ejects the sample stock solution S and diluent D aspirated in step S20 through the needle 32 at a second speed that is at least 10 times the first speed.
  • the second speed is, for example, 30 to 150 ⁇ L/sec, and preferably 30 to 40 ⁇ L/sec. In one embodiment, this operation is the second ejection.
  • the sample preparation system dilutes a stock solution with a diluent to prepare a sample for liquid chromatography analysis.
  • the sample preparation system includes a needle that aspirates and dispenses the stock solution, and a control unit that controls the needle.
  • the control unit may cause the needle to perform a first suction to aspirate a required amount of the stock solution, a first discharge to dispense the stock solution aspirated by the first suction and a required amount of the diluent into a container, a second suction to aspirate the stock solution and the diluent from the container at a first speed, and a second discharge to dispense the stock solution and the diluent aspirated by the second suction into the container at a second speed that is at least 10 times faster than the first speed.
  • the preparation system described in paragraph 1 can reduce the time required to prepare samples for liquid chromatography analysis.
  • the needle may be fluidly connected to a metering unit that supplies the diluent from a storage container that stores the diluent.
  • the diluent for diluting the sample is supplied from a metering unit that is fluidly connected to the needle.
  • control unit may cause the needle to aspirate and dispense the required amount of diluent into the container before performing the first suction.
  • a diluent for diluting the sample stock solution is prepared in an empty container before the needle aspirates the sample stock solution.
  • control unit may cause the needle to aspirate air after performing the first suction.
  • control unit when the control unit causes the needle to aspirate air, the control unit may cause the needle to aspirate air so that the distance from the tip of the needle to the liquid surface of the stock solution aspirated by the first suction is a predetermined distance.
  • a predetermined distance is provided between the liquid surface of the undiluted sample solution in the needle and the tip of the needle, thereby preventing the undiluted sample solution from leaking out of the needle.
  • control unit may receive the dilution rate and the volume after dilution, and calculate the required volume of the original solution and the required volume of the diluted solution.
  • the required volume of concentrate and the required volume of diluted solution are calculated based on the dilution ratio and volume after dilution received by the control device.
  • the second rate may be 30 to 40 ⁇ L/sec.
  • the sample can be mixed by being ejected into the container at a rate of 30 to 40 ⁇ L/second.
  • the sample preparation method is a method for diluting a stock solution with a diluent to prepare a sample for liquid chromatography analysis.
  • the sample preparation method may include the steps of aspirating a required amount of the stock solution, dispensing the aspirated required amount of the stock solution and the required amount of the diluent into a container, aspirating the stock solution and the diluent from the container at a first speed, and dispensing the aspirated stock solution and the diluent into the container at a second speed that is at least 10 times faster than the first speed.
  • the sample preparation method described in paragraph 8 can shorten the time required to prepare samples for liquid chromatography analysis.
  • the sample preparation method described in Clause 8 may further include a step of aspirating and discharging the required volume of dilution liquid before the step of aspirating the required volume of stock solution is performed.
  • a diluent for diluting the undiluted sample solution is prepared in an empty container before aspirating the undiluted sample solution.
  • the sample preparation method described in clause 8 or clause 9 may further include a step of aspirating air after the step of aspirating the required amount of stock solution has been performed.
  • the air may be aspirated so that the distance from the tip of the needle aspirating the stock solution to the liquid surface of the stock solution aspirated by the first suction is a predetermined distance.
  • the container may have a round bottom.
  • the container in which the sample is prepared has a round bottom, which improves the efficiency of stirring the sample.
  • the container may be a polypropylene container with a capacity of 1 mL, or a glass container with a raised bottom with a capacity of 150 ⁇ L.
  • the container in which the sample is prepared is suitable for mixing the sample, thereby improving the efficiency of stirring the sample.
  • the second rate may be 30 to 40 ⁇ L/sec.
  • the sample can be mixed by discharging the sample into the container at a rate of 30 to 40 ⁇ L/second.
  • each step may be performed by an autosampler that prepares the sample for liquid chromatographic analysis.
  • the sample preparation method described in paragraph 15 can be performed using an autosampler that prepares samples for liquid chromatography analysis. This eliminates the need for the user to perform the dilution work manually, thereby reducing the burden on the user associated with the dilution work.

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  • Physics & Mathematics (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Immunology (AREA)
  • Pathology (AREA)
  • Automatic Analysis And Handling Materials Therefor (AREA)

Abstract

Dans un système de préparation d'échantillon selon la présente invention, une solution mère est diluée avec un diluant pour préparer un échantillon pour une analyse de chromatographe en phase liquide. Le système de préparation d'échantillon est pourvu d'une aiguille et d'une unité de commande qui commande l'aiguille. L'unité de commande amène l'aiguille à exécuter une première aspiration pour aspirer la solution mère, une première décharge pour décharger la solution mère aspirée et le diluant dans un récipient, une seconde aspiration pour aspirer la solution mère et le diluant du récipient à une première vitesse, et une seconde décharge pour décharger la solution mère aspirée et le diluant dans le récipient à une seconde vitesse qui est au moins 10 fois la première vitesse.
PCT/JP2025/015725 2024-05-22 2025-04-23 Système de préparation d'échantillons et procédé de préparation d'échantillons Pending WO2025243767A1 (fr)

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JP2024083546 2024-05-22
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JP2024-138487 2024-08-20

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Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH08211073A (ja) * 1995-02-01 1996-08-20 Jasco Corp システムコントローラーを用いたオートサンプラー
JP2016166876A (ja) * 2012-12-19 2016-09-15 株式会社日立ハイテクノロジーズ 自動分析装置および分析方法
WO2017006477A1 (fr) * 2015-07-09 2017-01-12 株式会社島津製作所 Dispositif de prétraitement, et système d'analyse le comportant
WO2021260994A1 (fr) * 2020-06-23 2021-12-30 株式会社日立ハイテク Dispositif d'analyse automatique et procédé d'analyse

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH08211073A (ja) * 1995-02-01 1996-08-20 Jasco Corp システムコントローラーを用いたオートサンプラー
JP2016166876A (ja) * 2012-12-19 2016-09-15 株式会社日立ハイテクノロジーズ 自動分析装置および分析方法
WO2017006477A1 (fr) * 2015-07-09 2017-01-12 株式会社島津製作所 Dispositif de prétraitement, et système d'analyse le comportant
WO2021260994A1 (fr) * 2020-06-23 2021-12-30 株式会社日立ハイテク Dispositif d'analyse automatique et procédé d'analyse

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
ANONYMOUS: "Automation of Pipetting Using an Autosampler", AGILENT TRUSTED ANSWERS, 22 January 2020 (2020-01-22), pages 1 - 8, XP093377897, Retrieved from the Internet <URL:https://www.agilent.com/cs/library/technicaloverviews/Public/technicaloverview-injector-program-1260-infinity-ii-vialsampler-multisampler-5994-1704ja-jp-agilent.pdf> *

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