Control method of split liquid chromatograph and split liquid chromatograph

Abstract

The invention provides a method for controlling a split liquid chromatograph and a split liquid chromatograph, which can reduce the number of used recovery containers without performing pre-injection and facilitate the processing of recovered materials in the case of performing automatic splitting accompanying multiple sample injections, and a method for controlling a split liquid chromatograph and a split liquid chromatograph. When a sample component of each sample injection accompanying a plurality of sample injections is divided, during the division of a sample component accompanying a first sample injection, a sample component corresponding to each peak appearing on a chromatogram is taken by the division portion, and the start time of each peak on the chromatogram is stored. When a sample component is separated along with a second or subsequent sample injection among a plurality of sample injections, each time a peak appears on a chromatogram, the peak is used as a determination target peak, and on the basis of the start time of the determination target peak and the start time of each peak, whether the determination target peak is from the same component as one of the respective peaks is determined. On the basis of the determination result, the control unit causes the sorting unit to execute a predetermined sorting operation.

Description

Technical Field

[0001] This invention relates to a control method for a split liquid chromatograph and a split liquid chromatograph. Background Technology

[0002] As an apparatus for separating and individually collecting multiple components contained in a sample, a separating liquid chromatograph is known, which uses a column of a high-performance liquid chromatograph or the like to separate the components over time and then collects the components using a fraction collector.

[0003] The separating liquid chromatograph includes: a liquid chromatograph (LC) section with a pump, column, and detector; a fraction collector located downstream of the LC section; and a control unit for controlling these components. Sample components dissolved and separated over time by the column are sequentially detected by detectors such as a UV-Vis spectrophotometer and then introduced into the fraction collector. In the fraction collector, the internal flow path is switched according to instructions from the control unit, collecting the target components into a recovery container such as a vial.

[0004] Most split-phase liquid chromatographs (HPLC) have an automatic recovery function, known as automatic separation. In automatic separation, the control unit detects peaks appearing on the chromatogram based on the output signal from the detector and controls the fraction collectors to recover the fractions corresponding to the peaks from the dissolution solution from the column into individual recovery containers. Furthermore, the conditions used for peak detection (e.g., threshold values ​​for the signal level of the chromatogram or the slope of the chromatogram curve) are preset by the user.

[0005] [Existing Technical Documents]

[0006] [Patent Literature]

[0007] [Patent Document 1] Japanese Patent Application Publication No. 1-221663 Summary of the Invention

[0008] [The problem the invention aims to solve]

[0009] Furthermore, in the fractional liquid chromatographs described above, multiple injections of the same sample and subsequent automatic separations are sometimes performed. Here, multiple injections of the same sample refer to the sequential injection of multiple samples considered to have the same composition into the LC section, or the injection of the same sample into the LC section multiple times. In such cases, peaks not appearing in the chromatogram obtained with the first injection may sometimes appear in subsequent injections. This is believed to be due to components remaining in the flow path during previous fractional liquid chromatograph use dissolving with subsequent injections, deviations in the preparation of the multiple samples, or contamination with components remaining in the sample container. Therefore, peaks appearing only in subsequent injections originate from inclusions that may not necessarily require separation. However, in conventional fractional liquid chromatographs with automatic separation, the components corresponding to such peaks are also recovered by the fraction collector, leading to problems such as an increased number of recovery containers and more complex handling of the recovered materials.

[0010] Furthermore, in multiple automated fractionation processes involving multiple sample injections of the same content, as described above, it is sometimes desirable to recover the same components contained in each injected sample into a single recovery container. In this case, the operation of the fraction collector is controlled in such a way that the component corresponding to the first appearing peak in the chromatogram obtained with each sample injection is recovered into the first recovery container within the fraction collector, the component corresponding to the second appearing peak is recovered into the second recovery container, and so on, until the component corresponding to the Nth appearing peak is recovered into the Nth recovery container. However, when performing this control, different components are recovered into a single recovery container if the number of peaks appearing in each sample injection is different. Specifically, for example, if the number of peaks appearing with the first sample injection is different... Figure 4 In the case of peaks 1 and 2, peaks 3, 4, and 5 appearing with the second sample injection, and peaks 6, 7, 8, and 9 appearing with the third sample injection, the components corresponding to the initial peaks 1, 3, and 6 in each chromatogram are recovered into the first recovery container in the fraction collector, and the components corresponding to the second peaks 2, 4, and 7 in each chromatogram are recovered into the second recovery container. Therefore, different components (i.e., components with different retention times) are mixed in each recovery container.

[0011] Furthermore, conventional automatic separation liquid chromatographs include those that allow the user to specify the peaks to be separated from the chromatogram obtained by pre-injecting the test sample into the LC unit, and store the signal levels, slopes, and retention times of each peak's start and end points. When the actual sample is separated, separation is performed when the peaks appearing on the chromatogram related to the sample match the stored signal levels, and the slopes and retention times are within a specified range before and after the stored values ​​(see, for example, Patent Document 1). Using such a separation liquid chromatograph, only components corresponding to peaks with retention times close to those appearing during the injection (pre-injection) of the test sample are separated, thus avoiding the problems described above. However, this method, which requires pre-injection, consumes time and sample for pre-injection, making it unsuitable for expensive samples, especially those where it is difficult to ensure the amount injected during pre-injection.

[0012] The present invention is made in view of the above aspects, and its object is to reduce the number of recovery containers used in the case of automatic separation of multiple sample injections with the same contents by means of a separation liquid chromatograph, without pre-injection, and to facilitate the handling of the recovered material, or to reliably recover the same components into the same container without pre-injection.

[0013] [Technical means to solve the problem]

[0014] The control method for a split liquid chromatograph of the present invention, which addresses the aforementioned problem, is as follows:

[0015] The separating liquid chromatograph includes: a separation column for separating multiple sample components contained in a sample injected into a flow path; a detector disposed downstream of the separation column; a chromatogram generation unit for generating a chromatogram based on the detection result obtained by the detector; and a separation unit for separating sample components corresponding to peaks on the chromatogram from the eluent from the separation column. The control method of the separating liquid chromatograph includes...

[0016] When the sample components are separated during multiple sample injections into the flow path,

[0017] During the sampling of sample components in the first sample injection of the multiple sample injections, the sampling section takes sample components corresponding to each peak appearing on the first chromatogram (chromatogram generated by the chromatogram generation unit) accompanying the first sample injection, and stores the start time of each peak on the first chromatogram.

[0018] During the separation of sample components during the second and subsequent sample injections in the multiple sample injections, whenever a peak appears on the target chromatogram produced by the chromatogram production unit during the second and subsequent sample injections, the peak is designated as the target peak. Based on the start time of the target peak on the target chromatogram and the start time of each peak on the first chromatogram, it is determined whether the target peak originates from the same component as any of the peaks. Based on the determination result, the separation unit performs a predetermined separation operation.

[0019] Furthermore, the separating liquid chromatograph of the present invention, which addresses the aforementioned problem, is as follows:

[0020] It comprises: a separation column for separating multiple sample components contained in a sample injected into a flow path; a detector disposed downstream of the separation column; a chromatogram generation unit for generating a chromatogram based on the detection results obtained by the detector; a separation unit for separating sample components corresponding to peaks on the chromatogram from the eluent from the separation column; and a separation control unit for controlling the separation unit. In the separation liquid chromatograph,

[0021] When the sample components are separated during multiple sample injections into the flow path,

[0022] The dispensing control unit

[0023] The sampling unit is controlled so that, during the sampling of the sample component in the first sample injection of the multiple sample injections, the sample component corresponding to each peak appearing on the first chromatogram (chromatogram generated by the chromatogram generation unit) accompanying the first sample injection is sampled, and the start time of each peak on the first chromatogram is stored.

[0024] Furthermore, the separation unit is controlled so that when separating the sample component during the second and subsequent sample injections in the multiple sample injections, whenever a peak appears on the chromatogram (i.e., the target chromatogram) generated by the chromatogram generation unit during the second and subsequent sample injections, the peak is designated as the target peak. Based on the start time of the target peak on the target chromatogram and the start time of each peak on the first chromatogram, it is determined whether the target peak originates from the same component as any of the peaks, and based on the determination result, a predetermined separation action is performed.

[0025] [The effects of the invention]

[0026] According to the control method of the separation liquid chromatograph of the present invention or the separation liquid chromatograph of the present invention, in the case of automatic separation with multiple sample injections of the same contents, it is possible to reduce the number of recovery containers used without pre-injection and make the processing of the recovered material easier, or to reliably recover the same components into the same container without pre-injection. Attached Figure Description

[0027] Figure 1 This is a diagram showing the schematic structure of the separation liquid chromatograph according to Embodiment 1 of the present invention.

[0028] Figure 2 This is a block diagram showing the main structural components of the control unit in the described embodiment.

[0029] Figure 3 This is a flowchart illustrating the operation of the separation liquid chromatograph in the described embodiment.

[0030] Figure 4 This is an example of a chromatogram produced from multiple sample injections of the same content.

[0031] Figure 5 This is a flowchart illustrating another example of the operation of the separation liquid chromatograph of the present invention.

[0032] Figure 6 This is a diagram showing the schematic structure of the separation liquid chromatograph according to Embodiment 2 of the present invention.

[0033] Explanation of icon numbers

[0034] 10: LC Department

[0035] 20: Automatic sample injection device

[0036] 21: Sample container

[0037] 30: Separation String

[0038] 40: Detector

[0039] 50: Fraction Collector

[0040] 51: Recycling Container

[0041] 53: Dispensing nozzle

[0042] 60: Control / Processing Unit

[0043] 61: LC Control Unit

[0044] 62: Recycling Control Department

[0045] 63: Chromatography Production Department

[0046] 64: Peak Detection Department

[0047] 65: Judgment Department

[0048] 67: Start Time Storage Department Detailed Implementation

[0049] [Implementation Method 1]

[0050] Hereinafter, embodiments for implementing the present invention will be described with reference to the accompanying drawings. Figure 1 This is a schematic structural diagram of a split liquid chromatograph according to an embodiment of the present invention. The split liquid chromatograph includes an LC unit 10, a fraction collector 50, and a control / processing unit 60. Furthermore, in this embodiment, the fraction collector 50 corresponds to the splitting unit of the present invention.

[0051] The LC unit 10 includes a mobile phase container 11, a mobile phase flow path 12, and a mobile phase supply pump 13, an automatic sample injection device 20, a separation column 30, and a detector 40, all respectively disposed on the mobile phase flow path 12. The detector 40 can be any detector used in liquid chromatographs, such as an absorbance detector or a differential refractive index detector.

[0052] The automatic sample injection device 20 includes: a sample container receiving section 22 for receiving a plurality of sample containers 21 containing liquid samples; a suction section 23 for suctioning a predetermined amount of sample from a selected sample container 21; and an injector 24 for injecting the sample suctioned by the suction section 23 into the moving phase flow path 12.

[0053] The fraction collector 50 includes: a container housing 52 that houses a plurality of recovery containers 51 (equivalent to the sorting destination in this invention); a nozzle head 54 with a dispensing nozzle 53 at its lower end; a dispensing valve 55 built into the nozzle head 54 that switches the destination of the liquid delivered from the LC unit 10 to either a drain pipe or a dispensing nozzle 53; and a drive unit (not shown) that moves the nozzle head 54 back and forth, up and down, and left and right.

[0054] The control / processing unit 60 includes a general-purpose computer such as a personal computer or a special-purpose computer or a combination thereof, controls the respective units, and performs prescribed data processing based on the output signal from the detector 40 of the LC unit 10.

[0055] Figure 2The structure of the control / processing unit 60 is shown in the diagram. The control / processing unit 60 includes: an LC control unit 61 that controls the LC unit 10; a recovery control unit 62 (equivalent to the fractionation control unit in this invention) that controls the fraction collector 50; a chromatogram generation unit 63 that generates a chromatogram approximately in real-time based on the output signal from the detector 40; a peak detection unit 64 that detects the start and end points of peaks appearing on the chromatogram over time; and a determination unit 65 that performs a predetermined determination (described later). These are all functional blocks implemented in software by a central processing unit (CPU) installed in the computer constituting the control / processing unit 60, which reads a dedicated program installed in a large-capacity storage device such as a hard disk drive (HDD) installed in the computer into the computer's memory and executes it. The control / processing unit 60 also includes a storage unit 66, in which a start time storage unit 67 is provided (details described later). The function of the storage unit 66 is implemented, for example, by a large-capacity storage device installed in the computer. Furthermore, the computer constituting the control / processing unit 60 is connected to an input unit including a keyboard or mouse, and a display device including a liquid crystal display (both omitted from the illustration).

[0056] Next, refer to Figure 3 The flowchart below explains the operation of the liquid chromatograph in this embodiment.

[0057] When performing automated separation using the separation liquid chromatograph of this embodiment, multiple sample containers 21 (e.g., containers each containing samples of the same composition) are pre-installed in the automated sample injection device 20, and multiple recovery containers 51 for containing various components (sample components) in the sample are installed in the fraction collector 50. Furthermore, by operating the input unit, the user inputs injection conditions indicating the order and quantity of sample collected from the multiple sample containers 21 in the automated sample injection device 20, as well as detection conditions for the peak start and end points in the peak detection unit 64 (e.g., threshold values ​​for the signal level of the chromatogram or threshold values ​​for the slope of the chromatogram curve), and stores them in the storage unit 66.

[0058] When the user instructs the automatic sample collection to begin via the input unit, under the control of the LC control unit 61, a predetermined amount of sample is collected from the pre-designated sample container 21 within the automatic sample injection device 20 and injected into the moving phase flow path 12 via the injector 24 (step 101). Hereinafter, the sample injection in step 101 will be referred to as the "first sample injection".

[0059] The mobile phase, drawn from the mobile phase container 11 by the mobile phase supply pump 13, flows in the mobile phase flow path 12. The sample injected into the mobile phase flow path 12 by the automatic sample injection device 20 is guided to the inlet end of the separation column 30 along with the flow of the mobile phase. Then, during the process of passing through the separation column 30, the sample components are separated and sequentially dissolved from the outlet end of the separation column 30. The liquid flowing out from the outlet end of the separation column 30 (hereinafter referred to as the dissolution liquid) is discharged into the drain pipe via the dispensing valve 55 of the fraction collector 50 after passing through the detector 40. At this time, the output signal of the detector 40 is converted into a digital value by an analog-to-digital (A / D) converter (not shown) and input to the control / processing unit 60.

[0060] In the control / processing unit 60, the chromatogram generation unit 63 starts generating a chromatogram representing the time variation of the detection signal from the detector 40 based on the digital values. Then, the peak detection unit 64 determines whether a peak has appeared on the chromatogram at predetermined time intervals (step 102). Furthermore, the determination of whether a peak has appeared is based on whether the peak start point has been detected. The method for detecting the peak start point is not particularly limited and can be any method known in the past, such as a method based on the signal level of the chromatogram, a method based on the slope of the chromatogram curve, or any method based on both.

[0061] When a peak is determined to have occurred in step 102, the control / processing unit 60 stores the current time (the elapsed time since the first sample injection) as the peak start time in the start time storage unit 67 (step 103). Then, the control / processing unit 60 recovers the portion of the dissolution from the separation column 30 corresponding to the peak into a designated recovery container 51 within the fraction collector 50 (step 104). Specifically, under the control of the recovery control unit 62, the fraction collector 50 moves the nozzle head 54 onto the designated recovery container 51. When the portion of the dissolution corresponding to the start point of the peak reaches the dispensing valve 55, the dispensing valve 55 is switched from the drain pipe side to the dispensing nozzle 53 side, thereby spraying the dissolution from the dispensing nozzle 53 into the designated recovery container 51. Subsequently, when the peak termination point is detected by the peak detection unit 64, at the moment when the portion of the dissolution fluid corresponding to the termination point of the peak reaches the dispensing valve 55, the dispensing valve 55 is switched from the dispensing nozzle 53 side to the drain pipe side, thereby discharging the dissolution fluid from the drain pipe. Furthermore, the method for detecting the termination point of the peak is not particularly limited and can be any method known in the past, such as methods based on the signal level of the chromatogram, methods based on the slope of the chromatogram curve, or any method based on both.

[0062] Furthermore, the switching of the dispensing valve 55 as described above is performed at a time that takes into account the time required for the dissolution fluid to reach the dispensing valve 55 from the detector 40.

[0063] Next, it is determined whether a predetermined time has elapsed since the first sample injection (step 105). If it is determined that the predetermined time has not elapsed, the process returns to step 102. Then, if it is determined that a new peak has appeared on the chromatogram (i.e., if it was Yes in step 102), the peak start time of the new peak is recorded (step 103), and the sample component corresponding to the new peak is recovered (step 104). Furthermore, in step 104, the sample components corresponding to each peak on the chromatogram are recovered into different recovery containers 51.

[0064] Then, in step 105, at the time point when it is determined that a predetermined time has elapsed since the first sample injection, the automatic sorting accompanying the first sample injection ends.

[0065] Next, under the control of the LC control unit 61, a predetermined amount of sample is collected from the pre-designated sample container 21 in the automatic sample injection device 20 and injected into the moving phase flow path 12 (step 106). Hereinafter, this will be referred to as "second sample injection".

[0066] When the second sample injection is performed, the chromatogram generation unit 63 starts to generate a chromatogram accompanying the second sample injection, and the peak detection unit 64 determines whether a peak has appeared on the chromatogram at a predetermined time interval (step 107).

[0067] When a peak is detected in step 107 (i.e., when the peak start point is detected by the peak detection unit 64), the determination unit 65 determines, based on the start time of the peak, i.e., the elapsed time from the upcoming sample injection (here, the second sample injection) to the detection of the peak start point, whether the peak is the same as any of the peaks that appeared in the first sample injection (i.e., from the same component) (step 108). Specifically, the peak start time of each peak (hereinafter referred to as the "comparison peak") detected with the first sample injection is read from the start time storage unit 67. If the difference between any of the peak start times and the start time of the peak detected in step 107 (hereinafter referred to as the "determination peak") is within a specified tolerance range, the determination peak is determined to be the same as the peak that appeared in the first sample injection. On the other hand, if the difference between the start time of the determination peak and the start time of each comparison peak is not within the tolerance range, the determination peak is determined to be different from the peak that appeared in the first sample injection. The tolerance value is preset by the user or manufacturer of the split liquid chromatograph of this embodiment and stored in the storage unit 66. Furthermore, in the split liquid chromatograph of this embodiment, as described above, determining whether the target peak is the same as the peak appearing in the first sample injection based on the peak's start time shortens the determination time compared to methods that determine based on waveform processing such as peak shape similarity.

[0068] In step 108, if it is determined that the target peak is the same as the peak that appeared in the first sample injection, the fraction collector 50 recovers the portion of the dissolution from the separation column 30 corresponding to the target peak into a designated recovery container 51 (step 109). At this time, the recovery container 51 is set to be an empty recovery container 51 that was not used during the automatic separation accompanying the first sample injection. Furthermore, the specific recovery method is the same as in step 104, so its description is omitted here.

[0069] On the other hand, in step 108, if it is determined that the target peak is different from the peak that appeared in the first sample injection, the portion of the dissolution corresponding to the target peak is not recovered, but is discharged to the drain pipe via the dispensing valve 55 of the fraction collector 50.

[0070] Then, it is determined whether the prescribed time has elapsed since the second sample injection (step 110). If it is determined that the prescribed time has not elapsed, return to step 107 and repeat steps 107 to 110.

[0071] Then, in step 110, at the point in time when it is determined that a predetermined time has elapsed since the second sample injection, the automatic sorting accompanying the second sample injection ends, and it is determined whether all the pre-specified sample injections have been completed (step 111). Here, if it is determined that all sample injections have not been completed, the process returns to step 106 to perform the third sample injection.

[0072] Then, steps 106 to 111 are repeated until it is determined in step 111 that all sample injections have been completed. The series of processes ends at the time point when it is determined in step 111 that all sample injections have been completed.

[0073] Thus, the separation liquid chromatograph according to this embodiment can prevent the recovery of components detected only in the second and subsequent sample injections during automated separation with multiple sample injections. Therefore, in the case of multiple automated separations with multiple sample injections of the same content, the recovery of impurities can be prevented without pre-injection, the number of recovery containers 51 used can be reduced, and the handling of recovered materials can be kept simple.

[0074] Furthermore, in the described embodiment, sample components are recovered into different recovery containers 51 through automatic separation accompanying the first sample injection and automatic separation accompanying each subsequent sample injection. Thus, for example, when the first sample injection results in... Figure 4 In the case where peaks 1 and 2 appear with the second sample injection, peaks 3 to 5 appear with the second sample injection, and peaks 6 to 9 appear with the third sample injection, peaks 1, 2, 4, 5, 7, and 8 are recovered into different recovery containers 51 (e.g., the first to sixth recovery containers 51 in the fraction collector 50) in the order in which they are detected. Only peaks 3, 6, and 9 appearing in subsequent injections are not recovered and are discarded.

[0075] However, the present invention is not limited thereto. In the automatic separation process accompanying subsequent sample injections, sample components corresponding to peaks detected during subsequent injections (the target peak) that are determined to be the same as the peak appearing in the first injection (the comparison peak) can be recovered into the same recovery container 51 as the recovery container containing the sample components corresponding to the determined comparison peak. In this case, Figure 4In the example shown, the sample components corresponding to peaks 1, 4, and 7 are collected and recycled into one recycling container 51 (e.g., the first recycling container 51 within the fraction collector 50), while the sample components corresponding to peaks 2, 5, and 8 are collected and recycled into a different recycling container 51 (e.g., the second recycling container 51 within the fraction collector 50). Components corresponding to peaks 3, 6, and 9, which did not appear in the first sample injection, are not recycled and are discarded. Therefore, compared to recycling all sample components corresponding to peaks 1, 4, and 7, and all sample components corresponding to peaks 2, 5, and 8, into different recycling containers 51, the number of recycling containers 51 used can be reduced, and the recycling of different sample components into a single recycling container 51 can be prevented.

[0076] Alternatively, sample components corresponding to peaks that appear with subsequent sample injections but not in the first sample injection may be recovered into recovery container 51. In this case, peaks corresponding to those appearing only in subsequent sample injections (in...) can be recovered. Figure 4 In the example, the sample components (peaks 3, 6, and 9) are all recovered into different recovery containers 51, or peaks equivalent to these peaks that appear together in multiple sample injections after the second injection (in... Figure 4 In the example, the sample components (peaks 3 and 6) are concentrated and recovered into a recovery container 51 (e.g., a third recovery container within the fraction collector 50).

[0077] As mentioned above, refer to Figure 5 The flowchart illustrates an example of a separation operation where components corresponding to peaks that appear only in subsequent sample injections but co-occur in multiple subsequent sample injections are collected and recovered into a recovery container 51. Furthermore, steps 201 to 207 in this flowchart are similar to... Figure 3 Steps 101 to 107 in the flowchart are the same, so the explanation is omitted here.

[0078] exist Figure 5 In the flowchart, when the peak start point on the chromatogram produced with the second or subsequent sample injection is detected (i.e., when step 207 is yes), the time at this moment (the elapsed time since the upcoming sample injection) is stored in the start time storage unit 67 as the peak start time of the peak (step 208).

[0079] Then, the determination unit 65 determines whether the peak detected in step 207 (the target peak) is the same as any of the peaks detected in previous sample injections (the comparison peaks) when this is the Nth sample injection (N is 2 or more). Specifically, if the difference between the start time of any of the comparison peaks stored in the start time storage unit 67 and the start time of the target peak is within a specified tolerance range, the target peak is determined to be the same as the peak that appeared in the previous sample injection. On the other hand, if the difference between the start time of the target peak and the start time of each comparison peak is not within the specified tolerance range, the target peak is determined to be different from the peak that appeared in the previous sample injection.

[0080] In step 209, if it is determined that the target peak is the same as the peak that appeared in the previous sample injection, the fraction collector 50 recovers the sample component corresponding to the target peak into the same recovery container 51 as the recovery container that recovered the comparison peak that was determined to be the same as the target peak (step 210). Furthermore, the method for recovering the sample component is the same as in step 104, so its description is omitted here.

[0081] On the other hand, in step 209, if it is determined that the target peak is different from the peak that appeared in the previous sample injection, the fraction collector 50 recovers the sample component corresponding to the target peak into a new recovery container 51 (when this is the Nth sample injection (N is 2 or more), the recovery container 51 that was not used in the automatic sorting of the sample injections from the first to the N-1th) (step 211).

[0082] Next, it is determined whether a predetermined time has elapsed since the start of the upcoming sample injection (step 212). If it is determined that the predetermined time has not elapsed, the process returns to step 207 and repeats steps 207 to 212. Then, if it is determined in step 212 that the predetermined time has elapsed since the start of the upcoming sample injection, the process proceeds to step 213, where it is determined whether all pre-specified sample injections have been completed. If it is determined that all sample injections have not been completed, the process returns to step 206 and repeats steps 206 to 213. A series of processes are completed at the point in step 213 when it is determined that all sample injections have been completed.

[0083] By performing the actions shown above, for example, in Figure 4In the scenario shown, peaks 1, 4, and 7 are recycled into one recycling container 51 (e.g., the first recycling container 51 within the fraction collector 50), peaks 2, 5, and 8 are recycled into another recycling container 51 (e.g., the second recycling container 51 within the fraction collector 50), peaks 3 and 6 are recycled into yet another recycling container 51 (e.g., the third recycling container 51 within the fraction collector 50), and peak 9 is recycled into yet another recycling container 51 (e.g., the fourth recycling container within the fraction collector 50).

[0084] Furthermore, in a split liquid chromatograph, it is ideal for the user to pre-select whether components corresponding to peaks identified as identical are collected together into a single recovery container 51, or collected separately into different recovery containers 51. Additionally, when collecting components corresponding to peaks identified as identical into a single recovery container 51, it is ideal for the user to pre-select whether to recover peaks appearing only in subsequent sample injections. Moreover, when recovering peaks appearing only in subsequent sample injections, it is ideal for the user to pre-select whether components corresponding to peaks appearing only in subsequent sample injections that co-occur in multiple subsequent sample injections are collected together into a single recovery container 51, or collected separately into different recovery containers 51.

[0085] Furthermore, the present invention is shown as an example of a separating liquid chromatograph with a structure that directly uses the fraction collector 50 to separate the dissolution solution from the separation column 30. However, the invention is not limited to this and can also be applied to a separating liquid chromatograph with a structure that temporarily captures various sample components contained in the dissolution solution in a trapping column for later recovery. This structure will be described below.

[0086] [Implementation Method 2]

[0087] Figure 6 The figure shows the main structural components of a split liquid chromatograph according to a second embodiment of the present invention. As shown in the figure, the split liquid chromatograph of this embodiment, in addition to including the same LC section 110, fraction collector 150, and control / processing section 160, also includes a component capture section 200 having multiple capture columns 270, and a solvent supply section 300 supplying dissolution solvent to each capture column 270. In this embodiment, the component capture section 200 corresponds to the splitting section in the present invention, and the capture columns 270 correspond to the splitting destination in the present invention. Furthermore, in Figure 6 In China, for the sake of Figure 1 For identical or corresponding components shown, the last two digits of the symbol are marked, and the description is omitted where appropriate.

[0088] The component capture unit 200 includes: multiple flow paths, each equipped with one of the multiple capture columns 270; a drainage flow path 280, without a capture column 270; and a flow path switching unit 201, which switches which of the multiple capture columns 270 and the drainage flow path 280 the dissolution solution from the separation column 130 is directed to. Furthermore, each capture column 270 is filled with a capturing agent to capture sample components. Figure 6 The diagram shows a structure with 6 collection tubes 270, but the number of collection tubes 270 provided in the component collection section 200 is not limited to this, and may be 2 or more and 5 or less or 7 or more.

[0089] The solvent supply unit 300 includes: a solvent supply pump 302 for drawing dissolution solvent contained in a solvent container 301; and a switching valve 303 disposed in the flow path between the LC unit 110 and the component capture unit 200. The switching valve 303 selectively switches which of the dissolution solution from the LC unit 110 or the dissolution solvent supplied by the solvent supply pump 302 is sent to the component capture unit 200.

[0090] In this type of separating liquid chromatograph, firstly, with the eluent discharged into the drain pipe via switching valve 303, flow path switching unit 201, drain flow path 280, and dispensing valve 155, the sample is injected by the automatic sample injection device 120. Then, when the portion of the eluent from the separation column 130 corresponding to the start point of the peak on the chromatogram reaches the flow path switching unit 201, the flow path switching unit 201 is switched to select any one of the trapping columns 270. Subsequently, when the portion corresponding to the end point of the peak reaches the flow path switching unit 201, the flow path switching unit 201 is switched to select the drain flow path 280. Thus, the sample component corresponding to the peak is captured in the trapping column 270 (hereinafter referred to as the capture process). After the capture process is completed, the designated capture column 270 is selected by the flow path switching unit 201, and then the solvent supply pump 302 is driven and the switching valve 303 is switched, thereby allowing the dissolution solvent to flow into the designated capture column 270 and causing the sample components to dissolve from the capture column 270 (hereinafter referred to as the dissolution process). In the dissolution process, the liquid dissolved from the capture column 270 (containing the dissolution solvent of the sample components) is introduced into the fraction collector 150 and recovered into the recovery container 151 corresponding to each capture column 270.

[0091] In the fractionating liquid chromatograph of this embodiment, the capture step is performed as follows: Figure 3 The process shown in the flowchart or as... Figure 5 The process is shown in the flowchart. In this case, in... Figure 3Steps 104 and 109 of the flowchart, and Figure 5 In steps 204 and 210 or 211 of the flowchart, the portion of the dissolution from the separation column 130 that corresponds to the peak on the chromatogram is introduced into any collection column 270 instead of being recycled to the recovery container 151.

[0092] For example, in the capture process, such as Figure 3 In the case of the process shown in the flowchart, in step 104 of this flowchart, portions of the dissolution from the separation column 130 based on the first sample injection, corresponding to each peak, are introduced into different trapping columns 270. Thus, the sample components corresponding to each peak are captured in different trapping columns 270. Then, in the subsequent step 109, only portions of the dissolution from the separation column 130 based on subsequent sample injections, corresponding to peaks determined to be the same as those appearing in the first sample injection (comparison peak), are introduced into trapping columns 270 (either the same trapping column 270 as the trapping column capturing the sample components corresponding to the same comparison peak, or a different trapping column 270). Portions of peaks determined to be different from those appearing in the first sample injection are discharged to the drain pipe via the drain path 280 of the component capture section 200 and the dispensing valve 155 of the fraction collector 150.

[0093] On the other hand, in the capture process, such as Figure 5 In the case of the process shown in the flowchart, in step 204 of this flowchart, the portions of the eluent from the separation column 130 that are dissolved during the first sample injection, corresponding to each peak, are introduced into different trapping columns 270. Then, in the subsequent step 209, the portions of the eluent from the separation column 130 based on the second and subsequent sample injections that are corresponding to peaks determined to be the same as those appearing in the previous sample injections (comparison peaks) are introduced into the same trapping column 270 as the trapping column that captures the sample components that are corresponding to those determined to be the same comparison peaks. The portions of peaks that are corresponding to peaks determined to be different from those appearing in the previous sample injections are introduced into a new trapping column 270 (when this is the Nth sample injection (N is 2 or more), the trapping column 270 that was not used in the trapping process accompanying the first to the N-1th sample injections).

[0094] After multiple sample injections and accompanying capture processes, the dissolution process is performed to dissolve the sample components captured by each capture column 270, which are then collected using the fraction collector 150. Consequently, each recovery container 151 within the fraction collector 150 contains a liquid containing the sample components dissolved from different capture columns 270.

[0095] The above description provides specific examples for implementing the present invention, but the present invention is not limited to the described embodiments, and appropriate modifications are permissible within the scope of the present invention. For example, in the described embodiments, all samples installed in the automatic sample injection device 20 and the automatic sample injection device 120 are subjected to... Figure 3 or Figure 5 The automatic sampling shown in the flowchart is not limited to this; it can also be performed on two or more samples pre-specified by the user from among multiple samples installed in the automatic sample injection device 20 and automatic sample injection device 120. Figure 3 or Figure 5 The flowchart shows the automatic sorting process, which performs the same automatic sorting on other samples as before.

[0096] In addition, in the above embodiment, in each of the multiple sample injections, samples are collected from different sample containers 21 and injected into the injector 24, but alternatively, in each of the multiple sample injections, samples are collected from the same sample container 21 and injected into the injector 24.

[0097] [Way]

[0098] It will be clear to those skilled in the art that the exemplary embodiments described are specific examples of the following methods.

[0099] (Item 1) One aspect of the present invention provides a control method for a split liquid chromatograph, which is as follows:

[0100] The separating liquid chromatograph includes: a separation column for separating multiple sample components contained in a sample injected into a flow path; a detector disposed downstream of the separation column; a chromatogram generation unit for generating a chromatogram based on the detection result obtained by the detector; and a separation unit for separating sample components corresponding to peaks on the chromatogram from the eluent from the separation column. The control method of the separating liquid chromatograph includes...

[0101] When the sample components are separated during multiple sample injections into the flow path,

[0102] During the sampling of sample components in the first sample injection of the multiple sample injections, the sampling section takes sample components corresponding to each peak appearing on the first chromatogram (chromatogram generated by the chromatogram generation unit) accompanying the first sample injection, and stores the start time of each peak on the first chromatogram.

[0103] During the separation of sample components during the second and subsequent sample injections in the multiple sample injections, whenever a peak appears on the target chromatogram produced by the chromatogram production unit during the second and subsequent sample injections, the peak is designated as the target peak. Based on the start time of the target peak on the target chromatogram and the start time of each peak on the first chromatogram, it is determined whether the target peak originates from the same component as any of the peaks. Based on the determination result, the separation unit performs a predetermined separation operation.

[0104] (Item 2) The control method for the fractionating liquid chromatograph in Item 2 is based on the control method for the fractionating liquid chromatograph in Item 1, wherein,

[0105] For the predetermined separation action, if the determination determines that the target peak and any of the peaks on the first chromatogram come from the same component, the sample component corresponding to the target peak is separated; if the determination determines that the target peak and the peaks on the first chromatogram do not come from the same component, the sample component corresponding to the target peak is not separated.

[0106] (Item 3) The control method for the fractionating liquid chromatograph in Item 3 is based on the control method for the fractionating liquid chromatograph in Item 1, wherein,

[0107] For the predetermined separation action, if the determination determines that the target peak and any of the peaks on the first chromatogram originate from the same component, the sample component corresponding to the target peak is separated into a separation destination that is the same as the separation destination for the sample component corresponding to the peak determined to originate from the same component as the target peak among the multiple separation destinations contained in the separation unit. If the determination determines that the target peak and the peaks on the first chromatogram do not originate from the same component, the sample component corresponding to the target peak is not separated, or is separated into a separation destination that is different from the separation destination for the sample component corresponding to the peak.

[0108] (Item 4) The fractional liquid chromatograph for item 4 is as follows:

[0109] It comprises: a separation column for separating multiple sample components contained in a sample injected into a flow path; a detector disposed downstream of the separation column; a chromatogram generation unit for generating a chromatogram based on the detection results obtained by the detector; a separation unit for separating sample components corresponding to peaks on the chromatogram from the eluent from the separation column; and a separation control unit for controlling the separation unit. In the separation liquid chromatograph,

[0110] When the sample components are separated during multiple sample injections into the flow path,

[0111] The dispensing control unit

[0112] The sampling unit is controlled so that, during the sampling of the sample component in the first sample injection of the multiple sample injections, the sample component corresponding to each peak appearing on the first chromatogram (chromatogram generated by the chromatogram generation unit) accompanying the first sample injection is sampled, and the start time of each peak on the first chromatogram is stored.

[0113] Furthermore, the separation unit is controlled so that when separating the sample component during the second and subsequent sample injections in the multiple sample injections, whenever a peak appears on the chromatogram (i.e., the target chromatogram) generated by the chromatogram generation unit during the second and subsequent sample injections, the peak is designated as the target peak. Based on the start time of the target peak on the target chromatogram and the start time of each peak on the first chromatogram, it is determined whether the target peak originates from the same component as any of the peaks, and based on the determination result, a predetermined separation action is performed.

[0114] (Item 5) The fractionating liquid chromatograph of item 5 is based on the fractionating liquid chromatograph of item 4, wherein,

[0115] For the predetermined separation action, if the determination determines that the target peak and any of the peaks on the first chromatogram come from the same component, the sample component corresponding to the target peak is separated; if the determination determines that the target peak and the peaks on the first chromatogram do not come from the same component, the sample component corresponding to the target peak is not separated.

[0116] (Item 6) The fractionating liquid chromatograph of item 6 is based on the fractionating liquid chromatograph of item 4, wherein,

[0117] For the predetermined separation action, if the determination determines that the target peak and any of the peaks on the first chromatogram originate from the same component, the sample component corresponding to the target peak is separated into a separation destination that is the same as the separation destination for the sample component corresponding to the peak determined to originate from the same component as the target peak among the multiple separation destinations contained in the separation unit. If the determination determines that the target peak and the peaks on the first chromatogram do not originate from the same component, the sample component corresponding to the target peak is not separated, or is separated into a separation destination that is different from the separation destination for the sample component corresponding to the peak.

Claims

1. A control method of a fraction collection liquid chromatograph, wherein the fraction collection liquid chromatograph has: a separation column that separates a plurality of sample components contained in a sample injected into a flow path; a detector that is provided downstream of the separation column; a chromatogram production section that produces a chromatogram based on a detection result obtained by the detector; and a fraction collection section that fraction collects sample components corresponding to peaks on the chromatogram from an eluate from the separation column, in the control method of the fraction collection liquid chromatograph, when fraction collection of sample components for each sample injection accompanying a plurality of sample injections to the flow path is performed, at the time of fraction collection of sample components accompanying a first sample injection among the plurality of sample injections, fraction collecting, by the fraction collection section, sample components corresponding to each peak that appears on a chromatogram produced by the chromatogram production section, i.e., a first chromatogram, accompanying the first sample injection, and storing a start time of the each peak on the first chromatogram, at the time of fraction collection of sample components accompanying a second or later sample injection among the plurality of sample injections, whenever a peak appears on a chromatogram produced by the chromatogram production section, i.e., a target chromatogram, accompanying the second or later sample injection, the peak is taken as a determination target peak, it is determined, based on a start time of the determination target peak on the target chromatogram and the start time of the each peak on the first chromatogram, whether the determination target peak comes from the same component as any of the each peak, and based on a result of the determination, the fraction collection section is caused to perform a predetermined fraction collection operation.

2. The control method of a fraction collection liquid chromatograph according to claim 1, wherein, for the predetermined fraction collection operation, in a case where it is determined in the determination that the determination target peak comes from the same component as any of the each peak on the first chromatogram, fraction collecting sample components corresponding to the determination target peak, and in a case where it is determined that the determination target peak does not come from the same component as the each peak on the first chromatogram, not fraction collecting sample components corresponding to the determination target peak.

3. The control method of a fraction liquid chromatograph according to claim 1, wherein, for the predetermined fraction collection operation, in a case where it is determined in the determination that the determination target peak comes from the same component as any of the each peak on the first chromatogram, fraction collecting sample components corresponding to the determination target peak into a fraction collection destination among a plurality of fraction collection destinations contained in the fraction collection section that is the same as a fraction collection destination into which sample components corresponding to a peak among the each peak that is determined to come from the same component as the determination target peak are fraction collected, and in a case where it is determined that the determination target peak does not come from the same component as the each peak on the first chromatogram, not fraction collecting sample components corresponding to the determination target peak, or fraction collecting into a fraction collection destination among the plurality of fraction collection destinations that is different from a fraction collection destination into which sample components corresponding to the each peak are fraction collected.

4. A fraction collection liquid chromatograph having: a separation column that separates a plurality of sample components contained in a sample injected into a flow path; a detector provided downstream of the separation column; a chromatogram production section that produces a chromatogram based on a detection result obtained by the detector; a fraction collection section that fraction collects sample components corresponding to peaks on the chromatogram from an eluate from the separation column; and a fraction collection control section that controls the fraction collection section, in the fraction collection liquid chromatograph, when fraction collection of sample components for each of a plurality of sample injections to the flow path is performed, the fraction collection control section controls the fraction collection section so that, at the time of fraction collection of sample components for a first sample injection among the plurality of sample injections, sample components corresponding to each peak appearing on a chromatogram produced by the chromatogram production section in association with the first sample injection, that is, a first chromatogram, are fraction collected, and start times of the each peak on the first chromatogram are stored, and controls the fraction collection section so that, at the time of fraction collection of sample components for a second or later sample injection among the plurality of sample injections, whenever a peak appears on a chromatogram produced by the chromatogram production section in association with the second or later sample injection, that is, a target chromatogram, the peak is taken as a determination target peak, it is determined, based on a start time of the determination target peak on the target chromatogram and the start times of the each peak on the first chromatogram, whether the determination target peak comes from the same component as any of the each peak, and based on a determination result, a predetermined fraction collection operation is performed.

5. The fraction collection liquid chromatograph of claim 4 wherein, for the predetermined fraction collection operation, in a case where it is determined in the determination that the determination target peak comes from the same component as any of the each peak on the first chromatogram, sample components corresponding to the determination target peak are fraction collected, and in a case where it is determined that the determination target peak does not come from the same component as the each peak on the first chromatogram, sample components corresponding to the determination target peak are not fraction collected.

6. The fraction collection liquid chromatograph of claim 4 wherein, for the predetermined fraction collection operation, in a case where it is determined in the determination that the determination target peak comes from the same component as any of the each peak on the first chromatogram, sample components corresponding to the determination target peak are fraction collected into a fraction collection destination among a plurality of fraction collection destinations contained in the fraction collection section that is the same as a fraction collection destination into which sample components corresponding to a peak among the each peak that is determined to come from the same component as the determination target peak are fraction collected, and in a case where it is determined that the determination target peak does not come from the same component as the each peak on the first chromatogram, sample components corresponding to the determination target peak are not fraction collected, or are fraction collected into a fraction collection destination among the plurality of fraction collection destinations that is different from a fraction collection destination into which sample components corresponding to the each peak are fraction collected.

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