WO2015189696A1 - クロマトグラフ質量分析装置及びその制御方法 - Google Patents

クロマトグラフ質量分析装置及びその制御方法 Download PDF

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Publication number
WO2015189696A1
WO2015189696A1 PCT/IB2015/001277 IB2015001277W WO2015189696A1 WO 2015189696 A1 WO2015189696 A1 WO 2015189696A1 IB 2015001277 W IB2015001277 W IB 2015001277W WO 2015189696 A1 WO2015189696 A1 WO 2015189696A1
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Prior art keywords
chromatograph
mass
mass spectrometer
ion
time
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Ceased
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English (en)
French (fr)
Japanese (ja)
Inventor
信二 吉岡
博教 山下
彰 前川
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Hitachi High Tech Corp
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Hitachi High Technologies Corp
Hitachi High Tech Corp
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Priority to US15/312,792 priority Critical patent/US9823228B2/en
Priority to DE112015002105.1T priority patent/DE112015002105B4/de
Priority to GB1619450.8A priority patent/GB2549354B8/en
Publication of WO2015189696A1 publication Critical patent/WO2015189696A1/ja
Anticipated expiration legal-status Critical
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    • 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
    • G01N30/86—Signal analysis
    • 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
    • G01N30/86—Signal analysis
    • G01N30/8658—Optimising operation parameters
    • 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
    • G01N30/62—Detectors specially adapted therefor
    • G01N30/72—Mass spectrometers
    • 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
    • G01N30/62—Detectors specially adapted therefor
    • G01N30/72—Mass spectrometers
    • G01N30/7206—Mass spectrometers interfaced to gas chromatograph
    • 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
    • G01N30/86—Signal analysis
    • G01N30/8624—Detection of slopes or peaks; baseline correction
    • G01N30/8631—Peaks
    • 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
    • G01N30/86—Signal analysis
    • G01N30/8624—Detection of slopes or peaks; baseline correction
    • G01N30/8644—Data segmentation, e.g. time windows

Definitions

  • the present invention relates to a chromatographic mass spectrometer and a control method thereof.
  • a mass spectrometer is used as a high-sensitivity detector for a liquid chromatograph.
  • the mass spectrometer for example, there are a quadrupole mass spectrometer, an ion trap mass spectrometer, and a time-of-flight mass spectrometer, which are properly used according to the purpose of measurement.
  • a quadrupole mass spectrometer is often used.
  • the quadrupole mass spectrometer can measure the mass number by two methods of scan measurement and selected ion monitoring (SIM).
  • the scan measurement scanning is performed within a predetermined mass number range, and the spectrum of ions included in the set mass number range is detected.
  • This scanning method is used for qualitative analysis of unknown samples.
  • a chromatogram of ions having a specific mass number designated in advance is selectively detected. This method is used when a component to be analyzed is known and quantitative analysis of the component is performed with high sensitivity.
  • the measurer When determining the mass number for the SIM measurement, it is necessary to scan the target sample in advance, check the mass spectrum of the detected component, and the measurer must determine the mass number that is the target of the SIM measurement. . Thereafter, a mass chromatogram is displayed using the mass number selected by the measurer, and a characteristic mass number is determined for each target peak of each SIM measurement.
  • Patent Document 1 previously determines the mass number corresponding to the target component of quantitative analysis from the scan data of the component to be quantitatively analyzed, and sets it as the mass number at the time of SIM measurement. Thereafter, mass analysis of the mass number corresponding to each target component is performed only for a predetermined time before and after the peak time point, regardless of the difference in target components.
  • the method in which the SIM measurement time is within a predetermined time range before and after the peak time is that the measurer can visually check the chromatogram data when the elution times of the components separated by the liquid chromatograph are different. Since it is necessary to confirm and set the measurement time for each component, it takes time to determine the SIM measurement conditions. Moreover, the technique of Patent Document 1 cannot automatically set an elution time width suitable for each peak even if the elution time width of components separated by a liquid chromatograph varies. Therefore, the present invention provides a chromatograph mass spectrometer apparatus that determines the measurement time used in the SIM measurement for each target component in consideration of baseline information determined from the actual chromatogram peak shape. .
  • the present specification includes a plurality of means for solving the above-mentioned problems. For example, “Generate mass spectrum data and one or more mass chromatogram data based on the detection result of the ion detector”. Then, based on the elution time width appearing in each peak waveform of the generated one or more mass chromatogram data, the measurement time used in the selected ion monitoring is determined for each corresponding ion component ". To do.
  • This specification includes the disclosure of Japanese Patent Application No. 2014-121043, which is the basis of the priority of the present application. Effects of the Invention According to the present invention, it is possible to determine the optimum measurement time for each peak waveform without viewing the mass spectrum data. Problems, configurations, and effects other than those described above will become apparent from the following description of embodiments.
  • FIG. 1 is a diagram showing a schematic configuration of a liquid chromatography mass spectrometer.
  • FIG. 2 is a flowchart showing a flow of SIM measurement condition determination processing.
  • FIG. 3 is a diagram showing an example of a condition input screen when searching for molecular ions of the target component.
  • FIG. 4 is a diagram showing an example of a condition input screen when searching for three-component molecular ions.
  • FIG. 5 is a diagram showing a total ion chromatogram of three components.
  • FIG. 6 is a diagram showing a three-component mass spectrum.
  • FIG. 7 is a diagram showing a three-component mass chromatogram.
  • FIG. 8 is a diagram illustrating a baseline determination example of a mass chromatogram.
  • FIG. 9 is a diagram showing a display screen example of SIM condition determination content.
  • FIG. 10 is a diagram for explaining the relationship between a mass chromatogram of samples having different concentrations and an optimum measurement time width.
  • FIG. 1 shows a schematic configuration of a liquid chromatography mass spectrometer. Many of the samples analyzed by the liquid chromatograph mass spectrometer are samples in which a plurality of components are mixed. Therefore, the sample is purified by pretreatment or the like according to each sample and then separated in an LC (liquid chromatograph) unit 101. The separated sample is introduced into the ion source 102 of the mass spectrometer and ionized.
  • LC liquid chromatograph
  • an electrospray ionization method (ESI) or an atmospheric pressure chemical ionization method (APCI) generally used in a liquid chromatograph mass spectrometer is used.
  • the components ionized by the ion source 102 are introduced from the atmospheric pressure into the vacuum, and mass-separated by an MS (mass analysis) unit 103 having a quadrupole mass spectrometer.
  • MS (mass analysis) unit 103 having a quadrupole mass spectrometer.
  • a quadrupole mass spectrometer is used as the MS (mass spectrometry) unit 103.
  • the MS unit 103 may be a triple quadrupole mass spectrometer having a collision cell.
  • the quadrupole mass spectrometer can measure the mass number by two methods of scan measurement and selected ion monitoring (SIM).
  • SIM selected ion monitoring
  • scanning is performed within a predetermined mass number range, and ions included in the set mass number range are detected.
  • This scanning method is used for qualitative analysis of unknown samples.
  • the SIM measurement selectively detects only ions having a specific mass number designated in advance. This method is used when a component to be analyzed is known and quantitative analysis of the component is performed with high sensitivity.
  • the ions mass-separated by the MS (mass spectrometry) unit 103 are sequentially detected by the ion detection unit 104 arranged in the subsequent stage.
  • the operations of the LC (liquid chromatograph) unit 101, the ion source 102, the MS (mass spectrometry) unit 103, and the ion detection unit 104 are controlled by the control unit 105. Control conditions are set in the control unit 105 through the input unit 107. A signal detected by the ion detector 104 is output to the data processor 106. The data processing unit 106 performs data information accumulation, storage, analysis, and the like, and outputs the analyzed data. An analysis operation in the data processing unit 106 is also instructed from the input unit 107.
  • the MS (mass spectrometry) unit 103 obtains the data that is the source of the mass spectrum in accordance with the scanning conditions set by the input unit 107 in synchronization with the start of separation of the sample components by the LC (liquid chromatograph) unit 101. Get continuously. Further, the data processing unit 106 accumulates mass spectrum data indicating the relationship of the ion intensity with respect to the mass-to-charge ratio for each observed retention time.
  • FIG. 2 shows a flow of SIM condition determination / determination processing proposed in this embodiment. Each step shown in FIG. 2 is executed according to a program stored in advance in storage devices of the control unit 105 and the data processing unit 106. The processing shown in FIG.
  • the control unit 105 starts a SIM measurement condition setting process (step 201), and obtains scan data including the target component (step 202). At this time, the control unit 105 controls the MS (mass analysis) unit 103 using a quadrupole mass spectrometer based on the conditions set in the input unit 107 in advance, and executes scan measurement.
  • the measurement is performed under the same conditions as the separation conditions when the quantitative analysis is actually performed using the SIM scan. If scan data has already been acquired before this setting is made, it is also possible to select the acquired scan data and use it for subsequent flowchart analysis.
  • a condition for extracting ions of each component from the data of the scan-measured target sample is input to the control unit 105 through the input unit 107 (condition setting screen) (step 203).
  • FIG. 3 shows a configuration example of the condition setting screen.
  • the setting item 301 corresponds to a channel for performing SIM measurement, and is the number of components for actual quantitative analysis. When three-component SIM measurement is performed, three settings are required as “Channel No.”.
  • the name of a component that is actually subjected to quantitative analysis is input.
  • the component name may be input manually by a measurer or selected from component names prepared in advance.
  • the molecular weight corresponding to the component of the setting item 302 is set. The molecular weight specifies the non-ion pattern of molecular ions when ions are actually generated by electrospray ionization (ESI) or atmospheric pressure chemical ionization (APCI).
  • Item 304 is a designation input field for proton addition ions
  • Item 305 is a designation input field for sodium addition ions
  • Item 306 is a designation input field for ammonium addition ions.
  • An item 307 is a designation input field for designating an ion species that does not correspond to the items 304 to 306, and is used for setting an additional mass number that designates the ion species.
  • the measurer can arbitrarily set the additional mass number.
  • the control unit 105 adds “+1.0” to the mass number input to the corresponding channel when the measurer specifies the proton addition ion in the item 304 (when the check box is checked).
  • the measurer designates sodium addition ions in item 305 “+23.0” is added to the mass number input to the corresponding channel.
  • the control unit 105 sets the mass number of the molecular ion calculated according to the setting on the condition setting screen (FIG. 3) as the mass number condition for mass chromatogram extraction.
  • FIG. 3 shows an example in which one additional ion is selected for each channel, a plurality of additional ions can be selected.
  • the control unit 105 selects the chromatogram data having the maximum peak intensity from the plurality of mass chromatogram data in the subsequent step 205, and thereafter Process.
  • FIG. 4 shows an example of a condition input screen used when searching for three-component molecular ions.
  • a SIM condition setting screen when performing quantitative analysis for three components of testosterone, methyl testosterone, and progesterone is shown.
  • Testosterone, methyl testosterone, and progesterone are input as the component names of the item 401.
  • a molecular weight of 288.4 is input for testosterone, a molecular weight of 302.4 for methyltestosterone, and a molecular weight of 314.5 for progesterone.
  • FIG. 4 shows an example of a condition input screen used when searching for three-component molecular ions.
  • a SIM condition setting screen when performing quantitative analysis for three components of testosterone, methyl testosterone, and progesterone.
  • Testosterone, methyl testosterone, and progesterone are input as the component names of the item 401.
  • a molecular weight of 288.4 is input for testosterone, a molecular weight of 302.4 for methyltestosterone,
  • the control unit 105 extracts a mass chromatogram from the actual scan data according to the setting conditions received in step 203 (mass number information of molecular ions corresponding to each channel). In the case of FIG.
  • the mass number calculated in step 203 can be given a width in the front-rear direction.
  • the selection range specified in the program is used as the selection range when extracting the mass chromatogram. The selection range is determined based on the mass accuracy of the mass spectrometer to be used. In this embodiment, a selection width of ⁇ 0.2 is used.
  • a mass chromatogram is extracted in a range of ⁇ 0.2 with respect to the mass number of molecular ions of each component calculated in step 203. Therefore, for “Channel No. 1”, a mass chromatogram having a mass width of 289.2 to 289.6 is extracted. For “Channel No. 2”, a mass chromatogram having a mass width of 303.2 to 303.6 is extracted. For “Channel No. 3”, a mass chromatogram having a mass width of 315.3 to 315.7 is extracted.
  • FIG. 5 is an example of total ion chromatogram data obtained by scanning and measuring the three components of testosterone, methyl testosterone, and progesterone using the liquid chromatography mass spectrometer shown in FIG. As shown in FIG.
  • a three-component peak waveform is detected in the mass chromatogram.
  • the peak waveform 501 corresponds to the testosterone peak
  • the peak waveform 502 corresponds to the methyl testosterone peak
  • the peak waveform 503 corresponds to the progesterone peak in order from the peak with the earlier elution time.
  • the control unit 105 extracts a mass chromatogram from the scan data shown in FIG. 5 using the mass number information of the molecular ions in Step 203.
  • FIG. 6 is a diagram showing three-component mass spectrum data from the scan data of FIG.
  • FIG. 6 shows mass spectrum data of testosterone, methyltestosterone, and progesterone in the lower direction from the uppermost stage.
  • the peak 601 has a mass number of 289.5, which is consistent with the mass number 289.4 ⁇ 0.2 range of the testosterone protonated ions calculated in step 203.
  • the peak 602 has a mass number of 303.5, which is consistent with the range of the mass number of the proton addition ion of methyltestosterone calculated in step 203 of 303.4 ⁇ 0.2.
  • the peak 603 has a mass number of 315.6, which coincides with the range of the mass number of 315.5 ⁇ 0.2 of the proton-added ion of progesterone calculated in Step 203.
  • FIG. 7 is a diagram showing three-component mass chromatogram data when extraction is performed in step 204.
  • FIG. 7 shows mass chromatogram data of testosterone 701, methyl testosterone 702, and progesterone 703 from the top to the bottom.
  • FIG. 7 shows the result of extraction using the mass number information of molecular ions calculated in step 203 and the mass width of ⁇ 0.2.
  • both the start time and the end time are extended with the same time width, but different time widths may be extended between the start time and the end time.
  • the control unit 105 performs peak determination of the mass chromatogram data of each component extracted in step 204. In peak determination, the control unit 105 performs peak determination using a threshold value determined for signal strength, for example.
  • the control unit 105 determines that a peak is detected when a peak where a signal intensity higher than a preset threshold is detected is detected. When a peak having a signal intensity lower than the threshold value is detected, the control unit 105 returns to step 203 and executes the processing from step 204 again on other additional ion species. Regarding the chromatogram peak detected in step 205, the control unit 105 performs baseline determination in step 206.
  • FIG. 8 shows an example of baseline determination data performed on the testosterone mass chromatogram peak. Since various methods have already been proposed for determining the baseline 801, a detailed description thereof will be omitted.
  • the base line 801 is determined as a straight line connecting the start point 802 and the end point 803 of the peak waveform detected in the determination process. For example, in the baseline determination process, first, the peak waveform peak is detected, and then the peak start point 802 and end peak 803 are sequentially detected. In the next step 207, the control unit 105 acquires each time information of the peak start point 802 and the end point 803 from the information of the baseline 801. This time information becomes basic information for determining a measurement time width when performing SIM measurement of the corresponding component. Thus, in this embodiment, the baseline 801 is determined for each peak waveform, and the measurement time width (start time and end time) is determined individually and automatically based on the information.
  • the start time and end time of the baseline 801 obtained in step 207 are indicated by the corresponding channel number.
  • the start time and end time for performing the SIM measurement may be set as they are, but may be combined with the automatic extension function of the measurement time.
  • the control unit 105 sets a time point that is ⁇ 0.1 minute to the start time 802 of the baseline 801 as a new start time
  • the measurement time may be automatically extended with a time point of +0.1 minute as compared to the end time 803 as a new end time. Due to the presence of this automatic extension function, even if the elution time width of the chromatographic peak changes depending on the concentration (when the peak shape changes due to so-called tailing, etc.), the time width originally required for the measurement time width does not become insufficient. You can The relationship between the measurement peak waveform and the optimum measurement time when measuring the same sample having a concentration difference will be described later.
  • step 208 the control unit 105 registers the time information (start time 802 and end time 803) determined in step 207 from the mass ion mass number information calculated in step 203 as SIM measurement condition information of each component. .
  • the control unit 105 ends the SIM measurement condition setting process (step 209).
  • FIG. 9 shows an example of a screen displayed as an interface screen after the SIM measurement condition setting process is completed.
  • the information on the display screen is generated based on the SIM condition information of each component set and registered in step 208.
  • item 901 is a channel number.
  • Item 902 is the SIM measurement start time
  • item 903 is the SIM measurement end time
  • item 904 is the arrangement of the measurement time of each channel in the entire analysis time.
  • the measurement time of testosterone (SIM measured mass number 289.5) is 1.6 minutes from 3.9 minutes to 5.5 minutes
  • the measurement time of methyl testosterone (SIM measured mass number 303.5) Is 1.7 minutes from 4.8 minutes to 6.5 minutes
  • the measurement time of progesterone (SIM measured mass number 315.6) is 1.9 minutes from 7.0 minutes to 8.9 minutes.
  • the measurement time of each component is represented by a horizontal bar on the time axis. [Relationship between sample concentration difference and optimum measurement time]
  • both peak waveforms 1001 and 1002 are chromatograms measured for samples of the same component.
  • the sample corresponding to the peak waveform 1001 corresponds to the case where the concentration is higher than the sample corresponding to the peak waveform 1002.
  • the end point 1004 of the peak waveform 1002 of the low concentration sample is about 61 seconds
  • the end point 1005 of the peak waveform of the high concentration sample is about 69 seconds, which is about 8 seconds with respect to the end point 1004.
  • the end point of the measured peak waveform may be different. This means that the measurement time width determined when the low concentration sample is subjected to the SIM measurement cannot be used as it is for the SIM measurement of the high concentration sample. If the measurement time for SIM measurement determined for a low-concentration sample is applied as it is to the measurement time for SIM measurement of a high-concentration sample, the measurement time ends during the elution of the high-concentration sample, as shown in FIG.
  • the measurement time (or measurement time width) for the low concentration sample is not an optimum value as the measurement time (or measurement time width) for the high concentration sample.
  • the baseline 801 is determined for each measurement sample to determine the measurement time (or measurement time width), or the measurement time (or measurement time width) for SIM measurement determined for a certain concentration. ) For the start time and end time respectively.
  • the fixed time is extended in both directions before and after the measurement time, but it is also possible to provide a setting function that can extend the measurement time only for the start time or only for the end time.
  • the control unit 105 determines the end time of the determined SIM measurement. Set an extension time of +10 seconds.
  • the controller 105 automatically sets the SIM measurement time to 71 seconds obtained by adding 10 seconds to the end time 61 seconds determined for the low concentration sample, the end point 1005 of the baseline of the high concentration sample ( 69 seconds) is included in the automatically set measurement time, so that the entire elution time width of the high concentration sample can be measured by SIM.
  • the extended time can be provided in common for each component, or can be provided for each component as described above. In the present embodiment, it has been explained as a technical effect that the SIM measurement can be performed accurately regardless of the change in the measurement time (or measurement time width) between the same samples having different concentrations.
  • the separation column used in the liquid chromatograph The expansion function of the measurement time (or measurement time width) is also effective for changes in peak shape due to deterioration of the peak.
  • the mass number and the measurement time (or measurement time width) when each component designated as the measurement target is measured by SIM, and the corresponding scan data are obtained. Can be determined automatically. This eliminates the need for the measurer to visually check the mass spectrum data, and also significantly reduces the work load for setting the measurement time (measurement time width) according to the molecular ion species to be measured. can do.
  • the measurement time (or measurement time width) of each component is calculated based on the baseline information individually determined for the peak waveform. Therefore, even when the elution time width is different for each component, the optimum SIM measurement time (or measurement time width) for each component can be easily determined.
  • the SIM measurement time (or measurement time width) determined for the measured sample is extended by the above-described measurement time extension function. Depending on the application, the time until the SIM measurement result is obtained may be shortened.
  • the present invention may be applied to a gas chromatography mass spectrometer.
  • the measurement time of each component is determined by performing mass chromatogram detection, baseline determination, and the like.
  • the measurement time may be determined for the peak waveform corresponding to each component concurrently (in real time) with the detection operation of the scan data of the target component. If this function is used, the time required for setting the SIM measurement conditions can be further reduced.
  • control unit 105 in the above-described embodiment may be realized as, for example, an integrated circuit or other hardware.
  • Information such as programs, tables, and files used to realize the functions of the control unit 105 is stored in a storage device such as a memory, a hard disk, an SSD (Solid State Drive), or a storage medium such as an IC card, an SD card, or a DVD. can do.
  • Control lines and information lines indicate what is considered necessary for the description, and do not represent all control lines and information lines necessary for the product. In practice, it can be considered that almost all components are connected to each other.

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PCT/IB2015/001277 2014-06-12 2015-07-29 クロマトグラフ質量分析装置及びその制御方法 Ceased WO2015189696A1 (ja)

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US15/312,792 US9823228B2 (en) 2014-06-12 2015-07-29 Chromatograph mass spectrometer and control method therefor
DE112015002105.1T DE112015002105B4 (de) 2014-06-12 2015-07-29 Chromatograph-Massenspektrometer und Verfahren zu dessen Steuerung
GB1619450.8A GB2549354B8 (en) 2014-06-12 2015-07-29 Chromatograph mass spectrometer and control method therefor

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JP2014121043A JP6226823B2 (ja) 2014-06-12 2014-06-12 クロマトグラフ質量分析装置及びその制御方法

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