Chromatographic mass analysis data processing method and chromatographic mass analysis data processing device

Abstract

The present invention provides a method for determining whether an MS2 spectrum acquired by DDA is a chimeric MS2 spectrum. A first scheme of a chromatogram mass analysis data processing method according to the present invention is a chromatogram mass analysis data processing method for processing an MS2 spectrum in which an MS1 spectrum is acquired by MS1 analysis, one MS1 peak is selected from one or a plurality of MS1 peaks appearing in the acquired MS1 spectrum, and a mass-to-charge ratio range including a mass-to-charge ratio corresponding to the selected one MS1 peak is calculated for a predetermined mass-to-charge ratio range including a mass-to-charge ratio corresponding to the selected one MS1 peak, and a mass-to-charge ratio corresponding to the selected one MS1 peak is calculated for a predetermined mass-to-charge ratio range including a mass-to-charge ratio corresponding to the selected one MS1 peak. The present invention relates to a chromatographic mass analysis data processing method for performing an MS2 spectrum obtained by MS2 analysis using ions belonging to the mass-to-charge ratio range as precursor ions, the method comprising: a process for setting a peak threshold value on the basis of the intensity of the selected one MS1 peak; and a process for comparing the intensity of an MS2 peak appearing in the MS2 spectrum with the peak threshold value, and determining the MS2 spectrum as a chimeric MS2 spectrum when there is an MS2 peak having an intensity greater than the peak threshold value.

Description

Technical Field

[0001] This invention relates to a method and apparatus for processing data obtained through chromatographic quality analysis. Background Technology

[0002] Chromatographic mass analysis (GC-MS) devices are widely used to identify multiple components (compounds) contained in a sample. In a GC-MS device, multiple components in the sample are separated at each retention time in the upstream chromatograph, and then identified by mass analysis at each retention time in the downstream mass analysis unit. Especially when the sample is considered to contain a large number of components, GC-MS devices are sometimes used for more precise component identification. n The mass spectrometer used for analysis (n≥2) serves as the mass analysis unit.

[0003] When analyzing biological samples such as blood, which are believed to contain a large number of components, not only high precision but also comprehensive component identification is required. Data-dependent acquisition (DDA) is known as one of the effective analytical methods for comprehensive component identification.

[0004] In DDA, with each retention time, via MS 1 Analysis and acquisition of MS 1 Spectrum, from this MS 1 MS appearing in the spectrum 1 Peak selection as MS 2 MS of the object being analyzed 1 The peak will have the same MS as the selected peak. 1 The ions with the corresponding m / z values ​​(mass-to-charge ratio) of the peaks were used as precursor ions for MS analysis. 2 Analysis, obtaining MS 2 Spectrum.

[0005] The sample contains components that include isotopic elements. If the components containing isotopic elements are analyzed by MS... 1 Analysis shows that one type of ion can be divided into multiple MS molecules based on its molecular weight. 1 The peak was detected, and MS was obtained. 1 MS peaks arranged at specified mass-to-charge ratio intervals (these sets of peaks are called isotopic distributions) 1 Spectrum. Therefore, in DDA, in order to increase the spectral intensity corresponding to the components of the analyte, sometimes components containing MS are included. 1 Ions with mass-to-charge ratios in the range of spectral isotope distribution numbers in Da (typically 2–4 Da) were used as precursor ions for MS analysis. 2 Analysis (e.g., Patent Document 1).

[0006] Existing technical documents

[0007] Patent documents

[0008] Patent Document 1: Japanese Patent Application Publication No. 2022-079364 Summary of the Invention

[0009] The technical problem that the invention aims to solve

[0010] Previously, in DDA, in 1 MS 2 The precursor ions targeted in the analysis have a narrow mass-to-charge ratio range, therefore the obtained MS 2 If the spectrum does not distinguish between isotopes, it assumes that the ion originates from a single type of ion, i.e., a single component. In DDA, based on this premise, the components are identified according to the mass-to-charge ratio of the precursor ion and the mass-to-charge ratio of the product ion.

[0011] However, when analyzing biological samples such as blood, which are considered to contain a large number of components, multiple types of ions from different components are often mixed within the aforementioned mass-to-charge ratio range of several Da. In this specification, MS analysis using multiple types of ions from different components as precursor ions will be described. 2 The spectrum is called chimeric MS 2 Spectrum. In other words, chimeric MS 2 MS is a technique that uses multiple types of ions from different compositions as precursor ions. 2 MS formed by spectral mixing 2 Spectrum.

[0012] For chimeric MS 2 The above premise does not hold, therefore it is difficult to determine the spectrum based on chimeric MS. 2 The original components were identified by spectroscopic analysis. In addition, only simple observation of the MS obtained via DDA was performed. 2 Spectra are not easily used to determine whether they are chimeric MS. 2 Spectrum. Therefore, in MS 2 The spectrum is a chimeric MS 2 In the case of spectroscopic analysis, there is a risk of incorrect component identification. Based on the inventors' experience, MS data obtained through a series of analyses on biological samples... 2 In the spectrum, approximately 20% to 50% are chimeric MS. 2 Spectrum, chimeric MS 2 The impact of spectra on DDA-based component identification cannot be ignored.

[0013] The technical problem to be solved by the present invention is to provide a method for determining MS obtained through DDA. 2 Is the spectrum a chimeric MS? 2 Spectral methods.

[0014] Solution to the above technical problems

[0015] The first solution of the chromatographic quality analysis data processing method of the present invention, which was completed to solve the above-mentioned technical problems, is to process MS... 2 The method for processing chromatographic quality analysis data of the spectrum, the MS 2 The spectrum was obtained by MS 1 Analysis and acquisition of MS 1 Spectrum, obtained from MS 1 One or more MS appearing in the spectrum 1 Select 1 MS in the peak 1 Peak, for the 1 MS contained in the selection 1 The MS analyzer uses ions within a specified mass-to-charge ratio range, corresponding to the peak, as precursor ions, and performs MS analysis. 2 MS obtained through analysis 2 The chromatographic quality analysis data processing method includes:

[0016] Based on the selected 1 MS 1 Peak intensity, and the setting of peak thresholds;

[0017] The MS 2 MS appearing in the spectrum 2 The intensity of the peak is compared with the peak threshold, and the MS with an intensity greater than the peak threshold is selected. 2 In the presence of a peak, the MS 2 Spectral analysis indicates it is a chimeric MS 2 Spectrum processing.

[0018] The second solution to the chromatographic quality analysis data processing method of the present invention, which was developed to solve the above-mentioned technical problems, is a method for processing MS data. 2 The method for processing chromatographic quality analysis data of the spectrum, the MS 2 The spectrum was obtained by MS 1 Analysis and acquisition of MS 1 Spectrum, obtained from MS 1 One or more MS appearing in the spectrum 1 Select 1 MS in the peak 1 Peak, for the 1 MS contained in the selection 1 The MS analyzer uses ions within a specified mass-to-charge ratio range, corresponding to the peak, as precursor ions, and performs MS analysis. 2 MS obtained through analysis 2 The chromatographic quality analysis data processing method includes:

[0019] Based on the selected 1 MS 1 Peak intensity, and the setting of peak thresholds;

[0020] The MS 2 Multiple MS appearing in the spectrum 2 The sum of the peak intensities is compared to the peak threshold; if the sum is greater than the peak threshold, the MS is... 2 Spectral analysis indicates it is a chimeric MS 2 Spectrum processing.

[0021] The first embodiment of the chromatographic quality analysis data processing apparatus of the present invention, which was completed to solve the above-mentioned technical problems, comprises:

[0022] Input receiving unit, receives data from MS 1 Analysis of the acquired MS 1 One or more MS appearing in the spectrum 1 One MS selected from the peak 1 Peak intensity and MS 2 The input of the spectrum, the MS 2 The spectrum is a 1 MS containing the selected 1 The specified mass-to-charge ratio range, including the peak corresponding to the mass-to-charge ratio, is used by MS analysis using ions belonging to this mass-to-charge ratio range as precursor ions. 2 MS obtained through analysis 2 Spectrum;

[0023] The peak threshold setting unit, based on the selected 1 MS 1 The peak intensity sets the peak threshold;

[0024] The determination unit will determine the MS. 2 MS appearing in the spectrum 2 The intensity of the peak is compared with the peak threshold, and the MS with an intensity greater than the peak threshold is selected. 2 In the presence of a peak, the MS 2 Spectral analysis indicates it is a chimeric MS 2 Spectrum.

[0025] The second embodiment of the chromatographic quality analysis data processing device of the present invention, which was completed to solve the above-mentioned technical problems, comprises:

[0026] Input receiving unit, receives data from MS 1 Analysis of the acquired MS 1 One or more MS appearing in the spectrum 1 One MS selected from the peak 1 Peak intensity and MS 2 The input of the spectrum, the MS 2 The spectrum is a 1 MS containing the selected 1 The specified mass-to-charge ratio range, including the peak corresponding to the mass-to-charge ratio, is used by MS analysis using ions belonging to this mass-to-charge ratio range as precursor ions.2 MS obtained through analysis 2 Spectrum;

[0027] The peak threshold setting unit, based on the selected 1 MS 1 The peak intensity sets the peak threshold;

[0028] The determination unit will determine the MS. 2 Multiple MS appearing in the spectrum 2 The sum of the peak intensities is compared to the peak threshold; if the sum is greater than the peak threshold, the MS is... 2 Spectral analysis indicates it is a chimeric MS 2 Spectrum.

[0029] Invention Effects

[0030] Typically, MS n The intensity of a peak appearing in the spectrum (n≥1) represents the detectable amount of an ion with the mass-to-charge ratio corresponding to that peak. Each product ion obtained by dissociating a certain precursor ion constitutes that precursor ion before dissociation; therefore, the detectable amount (MS) of each product ion can be considered as... 2 The peak intensity) is the detection limit (MS) of the precursor ion. 1 Below the peak intensity. Therefore, in MS... 2 In the spectrum, it can be said that it is more compatible with the MS that has been chosen. 1 The intensity of the peak is greater than that of the MS. 2 The product ions corresponding to the peak are not only those selected by the MS. 1 The product ions obtained by the dissociation of the precursor ions corresponding to the peak, the MS 2 The spectrum was obtained by MS 1 Analysis and acquisition of MS 1 Spectrum, obtained from MS 1 One or more MS appearing in the spectrum 1 Select 1 MS in the peak 1 Peak, for the 1 MS contained in the selection 1 MS is performed on ions belonging to a specified mass-to-charge ratio range, including those corresponding to peaks other than the selected peaks, using these ions as precursor ions. 2 MS obtained through analysis 2 Spectrum. Therefore, the selected MS 1 The peak intensity is set as the peak threshold, and the MS is... 2 MS appearing in the spectrum 2 The intensity of the peak is compared with the peak threshold, and the MS with an intensity greater than the peak threshold is... 2 In the presence of the peak, it is possible to [control the MS]. 2 Spectral analysis indicates it is a chimeric MS2 Spectrum. Furthermore, typically, MS n The intensity of peaks appearing in the spectrum (n≥1) may contain a certain degree of error. Therefore, based on the characteristics of the data, by selecting the MS... 1 The peak threshold is set by multiplying the peak intensity by a specified factor, which allows for more accurate determination of the MS. 2 Is the spectrum a chimeric MS? 2 Spectrum.

[0031] When the precursor ion is monovalent, the number of product ions obtained from one precursor ion is one. However, the types of product ions obtained vary depending on which atom constituting the precursor ion is charged. In this case, the detection quantity (MS) of each product ion obtained by dissociating the precursor ion can be considered as... 2 The sum of the peak intensities is the detection limit (MS) of the precursor ion. 1 Below the peak intensity. Therefore, in MS... 2 In the spectrum, in this MS 2 Multiple (or all) MS appearing in the spectrum 2 The sum of the peak intensities is greater than the selected MS. 1 When the peak intensity is greater, it can be said that the MS 2 The spectrum is not only made with the selected MS 1 The spectrum obtained by the dissociation of precursor ions corresponding to the peak, the MS 2 The spectrum was obtained by MS 1 Analysis and acquisition of MS 1 Spectrum, obtained from MS 1 One or more MS appearing in the spectrum 1 Select 1 MS in the peak 1 Peak, for the 1 MS contained in the selection 1 MS is performed within a specified mass-to-charge ratio range, including the peak corresponding to that range, using monovalent ions within that range as precursor ions. 2 MS obtained through analysis 2 Spectrum. Therefore, the selected MS 1 The peak intensity is set as the peak threshold, and the MS is... 2 Multiple MS appearing in the spectrum 2 The sum of the peak intensities is compared to the peak threshold. If the sum is greater than the peak threshold, the MS can be... 2 Spectral analysis indicates it is a chimeric MS 2 Spectrum. Furthermore, typically, MS n The intensity of peaks appearing in the spectrum (n≥1) may contain a certain degree of error. Therefore, based on the characteristics of the data, the peak threshold is set to the selected MS. 1A specified multiple of the peak intensity allows for a more accurate determination of the MS. 2 Is the spectrum a chimeric MS? 2 Spectrum.

[0032] When the precursor ion is multivalent, multiple atoms may become charged and ionized, thus potentially generating multiple product ions from a single precursor ion. For example, when the precursor ion is divalent, the maximum number of product ions obtained from a single precursor ion is two. In this case, if the selected MS... 1 The peak intensity value multiplied by the selected MS 1 Setting the peak threshold based on the valence of the precursor ion corresponding to the peak allows for a more accurate determination of the MS peak. 2 Is the spectrum a chimeric MS? 2 Spectrum. Furthermore, when the precursor ion is multivalent, even if the peak threshold is set to the selected MS value... 1 The peak intensity value, in this MS 2 The spectrum is a chimeric MS 2 Under spectral conditions, the detection amount (MS) of each product ion obtained by dissociating multivalent precursor ions. 2 The sum of the peak intensities (to the detected amount of the precursor ion (selected MS)) is equal to the total peak intensity (to the detected amount of the precursor ion (selected MS). 1 The fact that the peak intensity is greater remains unchanged, therefore it will not be a chimeric MS. 2 MS of the spectrum 2 The spectrum was incorrectly identified as not being a chimeric MS. 2 Spectrum.

[0033] As described above, in the chromatographic quality analysis data processing method and apparatus of the present invention, it is possible to determine the MS obtained by DDA. 2 Is the spectrum a chimeric MS? 2 Spectrum. Attached Figure Description

[0034] Figure 1 This is a schematic diagram of the LC-MS analysis system.

[0035] Figure 2 This is a schematic diagram illustrating the workflow of DDA-based analysis.

[0036] Figure 3 It simulates MS acquired via DDA within a certain retention time. 1 Spectrum and MS 2 A diagram of the spectrum.

[0037] Figure 4 It simulates MS acquired via DDA within a certain retention time. 1 Spectrum and MS 2 A diagram of the spectrum.

[0038] Figure 5 This is a schematic diagram of the input screen that allows users to change the value of the peak threshold.

[0039] Figure 6 It simulates MS acquired via DDA within a certain retention time. 1 Spectrum and MS 2 A diagram of the spectrum.

[0040] Figure 7 This shows that for multiple MS 2 The spectrum will be selected using MS. 1 Peak intensity and MS 2 MS appearing in the spectrum 2 A graph showing the results of plotting the total values ​​of the peak intensities.

[0041] Figure 8 This is a schematic diagram of a modified analytical system for implementing the chromatographic quality analysis data processing method of the present invention. Detailed Implementation

[0042] The LC-MS analysis system capable of implementing the chromatographic quality analysis data processing method of the present invention will be described with reference to the accompanying drawings. Furthermore, the chromatographic quality analysis data processing method of the present invention is specifically implemented by the control / processing unit 4, which will be described later; other configurations are not essential to the present invention and can be appropriately modified.

[0043] Figure 1 This is a schematic diagram of the LC-MS analysis system. (For example...) Figure 1 As shown, the LC-MS analysis system includes: a measurement unit comprising a liquid chromatography unit 1 and a mass analysis unit 2, a control / processing unit 4, an input unit 5, and a display unit 6.

[0044] The liquid chromatography section 1 includes: a mobile phase container 10 for storing the mobile phase; a pump 11 for drawing up the mobile phase and delivering it in a substantially constant volume; a syringe 12 for injecting the sample solution into the mobile phase; and a chromatographic column 13 for separating the various components contained in the sample solution at each retention time.

[0045] Mass analysis unit 2 is a four-pole time-of-flight (Q-TOF) mass analyzer, comprising an ionization chamber 201 with an internal atmosphere of approximately atmospheric pressure and a vacuum chamber 20 divided into four compartments. Within the vacuum chamber 20 are a first intermediate vacuum chamber 202, a second intermediate vacuum chamber 203, a first high vacuum chamber 204, and a second high vacuum chamber 205. Each chamber is evacuated by a vacuum pump to increase the vacuum level sequentially. In other words, mass analysis unit 2 employs a multi-stage differential evacuation system.

[0046] An electrospray ionization (ESI) probe 21, which supplies eluent from the outlet of the chromatographic column 13, is disposed in the ionization chamber 201. The ionization chamber 201 is connected to the first intermediate vacuum chamber 202 via a narrow-diameter desolvation tube 22. The first intermediate vacuum chamber 202 and the second intermediate vacuum chamber 203 are connected via a hole formed in the top of the cone-shaped body 24. Ion directors 23 and 25 are disposed in the first intermediate vacuum chamber 202 and the second intermediate vacuum chamber 203, respectively. A quadrupole mass filter 26 and a collision cell 27 containing an ion director 28 are disposed in the first high vacuum chamber 204. Furthermore, multiple electrodes disposed across the first high vacuum chamber 204 and the second high vacuum chamber 205 constitute an ion director 29. Moreover, a time-of-flight mass separator and an ion detector 32, comprising an orthogonal acceleration unit 30 and an ion flight unit 31 with a reflector, are disposed in the second high vacuum chamber 205.

[0047] The control / processing unit 4 includes an analysis and control unit 40, a data storage unit 41, a spectrum generation unit 42, a peak selection / peak threshold setting unit 43, and a hybrid MS. 2 The spectrum determination / processing unit 44, the display processing unit 45, and the input receiving unit 46 are function blocks.

[0048] Typically, the control / processing unit 4 is a personal computer, workstation, or similar entity. The aforementioned functional blocks are implemented by executing one or more dedicated software programs (computer programs) installed on such a computer. These computer programs can be stored on computer-readable, non-transitory recording media such as CD-ROMs, DVD-ROMs, memory cards, or USB dongles and provided to the user. Alternatively, they can be provided to the user via data transmission through communication lines such as the Internet. Alternatively, they can be pre-installed on the computer as part of the system at the time the user purchases the system.

[0049] The analysis and control unit 40 controls the measurement unit, performing analysis on the prepared sample. In this LC-MS analysis system, for analyzing biological samples such as blood, which contain a large number of components and require high-precision and comprehensive component identification, even MS analysis is performed. 2 One particularly effective analytical method for comprehensive component identification is data-dependent acquisition (DDA). The following is a brief overview of the LC / MS analysis process of this LC-MS analysis system.

[0050] In the liquid chromatography section 1, the delivery pump 11 draws mobile phase from the mobile phase container 10 and delivers it to the column 13 at a substantially constant volume. The syringe 12 injects the sample into the mobile phase according to instructions from the analytical control unit 40. The sample, along with the mobile phase, is introduced into the column 13 and separated at each retention time during its passage through the column 13. The eluent eluted from the column 13 is introduced into the ESI probe 21, which sprays the eluent as charged droplets into the ionization chamber 201. As the charged droplets are atomized and the solvent in the droplets vaporizes, the sample components in the droplets become gaseous ions.

[0051] The generated ions are transported to the first intermediate vacuum chamber 202 via the desolvation tube 22, and sequentially pass through the ion guide 23, the cone-shaped body 24, and the ion guide 25 before being introduced into the quadrupole mass filter 26 in the first high vacuum chamber 204. A predetermined voltage is applied to each of the multiple rod electrodes constituting the quadrupole mass filter 26, selecting ions with a specific mass-to-charge ratio corresponding to that voltage, or ions within a specific mass-to-charge ratio range corresponding to that voltage, as precursor ions and allowing them to pass through the quadrupole mass filter 26. A collision gas such as Ar is introduced into the collision cell 27. The precursor ions contact the collision gas and are dissociated through collision-induced dissociation (CID) to generate various product ions. The generated product ions are then transported to the orthogonal acceleration section 30 via the ion guide 29.

[0052] When precursor ions are incident on the collision cell 27, the manner in which they dissociate varies depending on their kinetic energy (collision energy). Therefore, even with the same precursor ions, the type of product ions generated can be varied by appropriately adjusting the collision energy. Furthermore, it is possible to prevent all precursor ions from dissociating, allowing some to remain undissociated. Additionally, it is well known that the collision energy is typically determined by the voltage difference between the DC bias voltage applied to the quadrupole mass filter 26 and the DC voltage applied to the lens electrode disposed at the ion inlet of the collision cell 27.

[0053] In the orthogonal acceleration section 30, ions are accelerated approximately simultaneously in a direction approximately orthogonal to their incident direction (X-axis direction) (Z-axis direction). The accelerated ions travel at a velocity corresponding to their mass-to-charge ratio in the ion flight section 31, as shown in the image. Figure 1 The ions fly back and forth as shown by the double-dotted line and arrive at the ion detector 32. Various ions that start from the orthogonal acceleration unit 30 at approximately the same time arrive at the ion detector 32 in order of increasing mass-to-charge ratio and are detected. The ion detector 32 outputs a detection signal (ion intensity signal) corresponding to the number of ions to the control / processing unit 4.

[0054] In the control / processing unit 4, the data storage unit 41 digitizes the detection signal and then converts the flight time based on the time point when the ion is emitted from the orthogonal acceleration unit 30 into a mass-to-charge ratio, thereby acquiring and storing the MS. 1 Spectral data and MS 2 Mass spectrometry data (in this specification, these are sometimes collectively referred to as mass spectrometry data). In the orthogonal acceleration unit 30, ions are repeatedly ejected into the ion flight unit 31 at a predetermined period. As a result, the data storage unit 41 can repeatedly acquire mass spectrometry data covering a predetermined mass-to-charge ratio range at a predetermined period.

[0055] Next, refer to Figure 2 , Figure 3 The DDA-based analysis actions of this LC-MS analysis system are described. Figure 2 This is a schematic diagram illustrating the workflow of DDA-based analysis. Figure 3 It simulates MS acquired via DDA within a certain retention time. 1 Spectrum and MS 2 A graph of the spectrum. In DDA, it typically has a constant period ( Figure 2 The MS (with time interval Δt) is repeatedly performed across a specified mass-to-charge ratio range. 1 Analysis. In Control / Processing Unit 4, whenever MS is executed... 1 During analysis, the data storage unit 41 stores data via the MS. 1 Analysis of the acquired MS 1 The spectrum data is read from the data storage unit 41 by the spectrum generation unit 42. 1 Spectral data to generate MS 1 Spectrum ( Figure 3 (See above figure), the peak selection / peak threshold setting unit 43 checks the peak selection / peak threshold setting unit in the MS. 1 MS appearing in the spectrum 1 Does the peak meet pre-set specific conditions? These "specific conditions" can be, for example, set to a peak intensity value above a predetermined value. Furthermore, in MS values ​​that meet these specific conditions... 1 When a peak exists, the peak selection / peak threshold setting unit 43 selects the MS that meets the specific conditions. 1 Select 1 MS in the peak 1 Peak (selecting MS in this way) 1 The peak is called the selected MS. 1 (Peak), automatically determining and selecting MS using known methods. 1 The valence of the ions corresponding to the peaks, and based on this selection, MS 1 The peak intensity value is used to set the peak threshold (details about the peak threshold will be described later). Meanwhile, the analysis control unit 40 then connects to the MS... 1 The analysis will be performed within the specified (Da width) mass-to-charge ratio range. Figure 3MS of ions (shaded area in the above figure) as precursor ions 2 Analysis shows that the specified mass-to-charge ratio range includes the selection of MS. 1 The mass-to-charge ratio corresponding to the peak is included. If MS is executed... 2 Analysis will then show the acquired MS data by the data storage unit 41. 2 Spectral data and selection MS 1 The mass-to-charge ratio value corresponding to the peak, and the selection of MS 1 The valence of the ions corresponding to the peaks, the peak threshold, and MS 1 The spectral data is stored accordingly, and the spectrum generation unit 42 reads the MS from the data storage unit 41. 2 Spectral data to generate MS 2 Spectrum. Additionally, the MS generated by the spectrum generation unit 42... 1 Spectrum and MS 2 The spectrum can also be compared with MS. 1 Spectral data and MS 2 The spectral data is correspondingly stored in the data storage unit 41.

[0056] exist Figure 2 In the example shown, MS follows 1 The analysis was performed only once using MS. 2 Analysis is needed, but if there is sufficient time, one more MS can be performed. 1 Analysis was performed using multiple MS analyses targeting different precursor ions. 2 Analysis. In this case, for example, it can be performed in MS. 1 MS appearing in the spectrum 1 A specified number of MSs are selected from the peaks in descending order of intensity. 1 Peak, for this choice MS 1 The peaks were sequentially subjected to MS analysis using ions belonging to a specified mass-to-charge ratio range, including the corresponding mass-to-charge ratio, as precursor ions. 2 Analysis. Additionally, from... Figure 2 It can also be seen that in DDA, there may not necessarily be an MS obtained within a certain retention time. 1 MS corresponding to the spectrum 2 Spectrum.

[0057] Select MS 1 Peaks emerge from multiple MS that constitute the isotopic distribution 1 When a peak is selected, it is usually chosen as the MS. 1 The peak is selected as a single isotope peak within the isotopic distribution (a peak originating from a molecule composed of the isotope with the largest ratio among the isotopes of the elements constituting the sample molecules). In the isotopic distribution, during DDA-based analysis, it is automatically determined which peak is a single isotope peak. In this embodiment, when selecting MS... 1Peaks emerge from multiple MS that constitute the isotopic distribution 1 When a peak is selected, the range of m / z values ​​of a single isotope peak that contains that isotope distribution can be selected.

[0058] When performing LC / MS analysis using DDA as described above on a sample, the MS corresponding to the LC / MS analysis is stored in the data storage unit 41. 1 Spectral data and MS 2 Spectral data. Furthermore, for each selected MS... 1 The peaks are stored in relation to their corresponding mass-to-charge ratio, ion valence, and peak threshold. With this data stored, the control / processing unit 4 performs the following data processing. Several examples of data processing are explained below.

[0059] <First Data Processing Method>

[0060] Reference Figure 4 The first data processing method will be explained. Figure 4 It simulates MS acquired via DDA within a certain retention time. 1 Spectrum and MS 2 A spectrum diagram. (e.g.) Figure 4 As shown, in MS 1 The title section of the spectrum can display and select MS. 1 The mass-to-charge ratio corresponding to the peak, select MS 1 Peak intensity and selection MS 1 The valence of the ions corresponding to the peaks, in MS 2 The title section of the spectrum displays the peak thresholds. In the first data processing method, the MS is embedded... 2 The spectrum determination / processing unit 44 reads the MS data from the data storage unit 41. 2 Spectrum and with the MS 2 The peak thresholds correspondingly stored in the spectrum will be in the MS. 2 MS appearing in the spectrum 2 The peak intensity is compared to the peak threshold. Then, as... Figure 4 As shown in the figure below, MS with an intensity greater than the peak threshold 2 In the presence of a peak, the MS 2 Spectral analysis indicates it is a chimeric MS 2 Spectrum.

[0061] like Figure 4 As shown, it is possible to select MS 1 The peak intensity is set as the peak threshold. Typically, MS nThe intensity of a peak appearing in the spectrum (n≥1) represents the detectable amount of an ion with the mass-to-charge ratio corresponding to that peak. Each product ion obtained by dissociating a certain precursor ion constitutes that precursor ion before dissociation; therefore, the detectable amount (MS) of each product ion can be considered as... 2 The peak intensity) is the detection limit (MS) of the precursor ion. 1 Below the peak intensity. Therefore, in MS... 2 In the spectrum, it can be said that it is more compatible with the MS selection. 1 The intensity of the peak is greater than that of the MS. 2 The product ions corresponding to the peak are not only those selected by MS. 1 The product ions obtained by the dissociation of the precursor ions corresponding to the peak, the MS 2 The spectrum is a combination of inclusion and selection MS 1 The MS spectrum is generated by using ions within a specified mass-to-charge ratio range (including ions corresponding to peaks other than the selected peaks) as precursor ions, within a specified mass-to-charge ratio range. 2 MS obtained through analysis 2 Spectrum. For example, if using Figure 4 The above diagram illustrates the product ions obtained by dissociating the ions corresponding to peak K (however, in actual MS...). 1 Spectrum Figure 4 The spectrum shown above is complex because it is not always clear which peak corresponds to which product ion (the product ion obtained by dissociating the ion from that peak). Therefore, the selection of MS... 1 The peak intensity is set as the peak threshold, and the MS is... 2 MS appearing in the spectrum 2 The intensity of the peak is compared with the peak threshold, and the MS with an intensity greater than the peak threshold is... 2 In the presence of the peak, it is possible to [control the MS]. 2 Spectral analysis indicates it is a chimeric MS 2 Spectrum.

[0062] Alternatively, the peak threshold can be set to select MS. 1 The peak threshold should be 0.8 to 1.2 times the peak intensity. Typically, mass spectrometry intensity values ​​may contain approximately 20% error. Therefore, depending on the characteristics of the data, the peak threshold can be set to the value selected by MS. 1 A more appropriate threshold is to multiply the peak intensity by 0.8 to 1.2. Alternatively, set the peak threshold to the selected MS value. 1 The appropriate level of peak intensity, several times higher than the measured data, was heuristically determined based on the measurement conditions. Therefore, one or more known-homogeneous MS (monitoring parameters) acquired under the same measurement conditions as the data being processed can be used. 2 Based on the spectral data, set an appropriate magnification. Depending on the characteristics of the data, select MS... 1The peak threshold is set by multiplying the peak intensity by a specified factor, which allows for more accurate determination of the MS. 2 Is the spectrum a chimeric MS? 2 Spectrum.

[0063] Chimeric MS 2 The spectrum determination / processing unit 44 can also process the MS of the object being processed. 2 All MS appearing in the spectrum 2 After comparing the peak intensity with the peak threshold, the MS is determined. 2 Is the spectrum a chimeric MS? 2 Spectrum. Alternatively, the MS of the object being processed can be... 2 All MS appearing in the spectrum 2 The intensity of the peaks is compared with a peak threshold in a prescribed order (e.g., the order in which they are stored in the data file), and the peaks with an intensity greater than the threshold are identified. 2 The time point in time when the peak exists, the MS 2 Spectral analysis indicates it is a chimeric MS 2 Spectrum. In this case, the MS can be terminated after the decision. 2 The process of comparing peak intensity with peak threshold can also be continued.

[0064] Chimeric MS 2 The operation of the spectrum determination / processing unit 44 can be performed automatically while storing the data required for determination in the data storage unit 41, or automatically after the operation of the measurement unit, i.e., after the DDA-based analysis is completed, or after the input receiving unit 46 receives the user's input indicating the start of determination and processing from the input unit 5.

[0065] Chimeric MS 2 The spectrum determination / processing unit 44 is capable of determining whether an MS is a chimeric MS. 2 MS of the spectrum 2 Spectrum and / or corresponding MS 2 Spectral data is processed using various methods. For example, it can also be determined to be a chimeric MS. 2 MS of the spectrum 2 Spectral data and MS not identified as chimeric 2 MS of the spectrum 2 The spectral data is stored separately in the data storage unit 41. Alternatively, the data may be classified as chimeric MS. 2 MS of the spectrum 2 Label the spectral data. Alternatively, you can label the MS. 2 MS in spectral data with an intensity greater than the peak threshold 2 The data corresponding to the peak is labeled, or the data is deleted.

[0066] The display processing unit 45 reads MS data from the data storage unit 41.1 Spectrum and / or MS 2 The spectrum is displayed on display unit 6. At this time, if the MS is embedded... 2 The spectrum determination / processing unit 44 will determine it as a chimeric MS. 2 MS of the spectrum 2 Spectral data and MS not identified as chimeric 2 MS of the spectrum 2 Spectral data are stored separately in data storage unit 41, or for MSs determined to be chimeric 2 MS of the spectrum 2 If the spectral data is tagged, the display processing unit 45 can determine that it is a spliced ​​MS without reading it from the data storage unit 41. 2 MS of the spectrum 2 Spectral data, or even if read from data storage unit 41 and determined to be a chimeric MS 2 MS of the spectrum 2 Spectral data is also not displayed on display unit 6. This eliminates the need for the user to rely on the embedded MS. 2 Attempts at component identification using spectra are futile. Furthermore, if chimeric MS... 2 The spectral determination / processing unit 44 pairs MS with intensities greater than the peak threshold. 2 If the data corresponding to the peak is labeled or deleted, the display processing unit 45 can prevent the display of MS with an intensity greater than the peak threshold. 2 Peaks (i.e., MS peaks corresponding to ions originating from the following components) 2 Peak, this component and selected MS 1 The peaks correspond to ions from different components, or the MS signal is displayed after the intensity is set to 0. 2 Spectrum. Therefore, in the chimeric MS 2 In the spectrum, it is possible to reduce and select MS 1 The contribution of different components from which the ions corresponding to the peaks originate can improve MS performance. 2 The purity of the spectrum. The result is that the chimeric MS... 2 Identifying the components in the spectrum becomes easier.

[0067] The display processing unit 45 can perform the above-mentioned processing automatically, or it can be performed after the input receiving unit 46 receives the user's input indicating that the input unit 5 intends to start the display processing.

[0068] The display processing unit 45 is capable of displaying, for example, Figure 5 The input screen shown in (a) allows the user to change the value of the peak threshold. The input receiving unit 46 accepts various inputs from the user through the input unit 5 and outputs the content of the input to each unit. For example, in... Figure 5 In the input screen shown in (a), which allows the user to change the peak threshold value, if the user sets the peak threshold to MS... 1The input receiving unit 46 updates the value of the peak multiplied by a specified multiplier (the × mark in the figure indicates mode selection) and changes the value of either the "Multiplier" or "Peak Threshold" column, then displays the changed parameters. Next, when the user presses the "Reprocess" button, the input receiving unit 46 can send data to the embedded MS. 2 The spectrum determination / processing unit 44 outputs the modified peak threshold and sends it to the embedded MS. 2 The spectrum determination / processing unit 44 outputs a signal indicating that determination and processing should be performed again. (Mixed MS) 2 If the spectrum determination / processing unit 44 receives the signal, it can perform the determination and processing as described above again. By performing such an operation, it is possible to perform determination and processing that effectively utilizes the user's cognition, and to more accurately determine whether it is a fused MS. 2 Spectrum. Furthermore, the peak threshold can be easily and repeatedly fine-tuned, making it very convenient for users ultimately performing component identification.

[0069] In addition, the initial value of the peak threshold can be automatically set to the selected MS. 1 The intensity of the peak can also be preset by the user using the methods described above.

[0070] <Second Data Processing Method>

[0071] Reference Figure 6 The second data processing method will be explained. Figure 6 It simulates MS acquired via DDA within a certain retention time. 1 Spectrum and MS 2 A spectrum diagram. (e.g.) Figure 6 As shown, in MS 1 The title section of the spectrum can display and select MS. 1 The mass-to-charge ratio corresponding to the peak, select MS 1 Peak intensity and selection MS 1 The valence of the ions corresponding to the peaks, in MS 2 The title section of the spectrum displays peak thresholds. In the second data processing method, the MS is embedded... 2 The spectrum determination / processing unit 44 reads the MS data from the data storage unit 41. 2 Spectrum and with the MS 2 The peak thresholds correspondingly stored in the spectrum are calculated in this MS. 2 Multiple occurrences in the spectrum (in) Figure 6 (Middle is all) MS 2 The sum of the peak intensities is compared to the peak threshold. Then, as... Figure 6 As shown in the figure below, when the total value is larger than the peak threshold, the MS... 2 Spectral analysis indicates it is a chimeric MS 2 Spectrum.

[0072] like Figure 6 As shown, it is possible to select MS 1 The peak intensity is set as the peak threshold. Typically, the precursor ion is monovalent; in this case, one product ion is obtained from one precursor ion. However, the type of product ion obtained varies depending on which atom constituting the precursor ion is charged. In this case, the detectable amount (MS / s) of each product ion obtained by dissociating the precursor ion can be considered as... 2 The sum of the peak intensities is the detection limit (MS) of the precursor ion. 1 Below the peak intensity. Therefore, in MS... 2 In the spectrum, in this MS 2 Multiple (or all) MS appearing in the spectrum 2 The sum of the peak intensities is greater than the selected MS. 1 When the peak intensity is greater, it can be said that the MS 2 The spectrum is not only made with the selected MS 1 The spectrum obtained by the dissociation of precursor ions corresponding to the peak, the MS 2 The spectrum is a combination of inclusion and selection MS 1 The specified mass-to-charge ratio range, including the peak corresponding to the mass-to-charge ratio, is used by MS with monovalent ions belonging to this mass-to-charge ratio range as precursor ions. 2 MS obtained through analysis 2 Spectrum. For example, if using Figure 6 The above diagram illustrates the product ions obtained by dissociating the ions corresponding to peak K (however, in actual MS...). 1 Spectrum Figure 6 The spectrum shown above is complex because the product ion obtained by dissociating the ion corresponding to which peak is not necessarily clear. Therefore, MS will be selected. 1 The peak intensity is set as the peak threshold, and the MS is calculated. 2 Multiple MS appearing in the spectrum 2 The sum of peak intensities is compared to a peak threshold. If the sum is greater than the peak threshold, the MS can be... 2 Spectral analysis indicates it is a chimeric MS 2 Spectrum.

[0073] Alternatively, as described in the first processing method, the peak threshold can also be set to select MS. 1 The value is obtained by multiplying the peak intensity by a specified factor (preferably 0.8 to 1.2).

[0074] When the precursor ion is multivalent, multiple atoms may become charged and ionized, thus potentially generating multiple product ions from a single precursor ion. For example, when the precursor ion is divalent, the maximum number of product ions obtained from a single precursor ion is two. In this case, if the peak threshold is set to MS... 1 The peak intensity value multiplied by the selected MS 1 The value obtained by determining the valence of the precursor ion corresponding to the peak can more accurately determine the MS. 2 Is the spectrum a chimeric MS? 2 Spectroscopy. Furthermore, the valence of precursor ions depends to some extent on the type of sample and the type of ionization method used, and therefore can be predicted to a certain extent based on experience. For example, when measuring peptides by ionization via ESI, precursor ions with valences of 2 to 4 are frequently generated, and occasionally precursor ions with valences of 5 or higher are also generated. Moreover, in the case of matrix-assisted laser desorption / ionization (MALDI), monovalent ions are generated with a high probability, regardless of the sample being measured. Therefore, when setting the peak threshold to select MS... 1 The peak intensity value multiplied by the selected MS 1 When the value is obtained from the valence of the precursor ion corresponding to the peak, the valence of the precursor ion is predicted and determined based on the type of sample and the type of ionization method of the sample. Thus, even when the valence of the precursor ion is unclear, the peak threshold can be set with higher precision.

[0075] As an example, Figure 7 The diagram shows the results for multiple MS. 2 The spectrum will be measured by selecting MS under the condition that the peptide is ionized by ESI. 1 Peak intensity and MS 2 MS appearing in the spectrum 2 The result is plotted by grouping the total values ​​of peak intensities. In this embodiment, it is compared with the selected MS. 1 The peaks correspond to ions with valences ranging from 2 to 4. In the plot enclosed by solid lines, in MS... 2 MS appearing in the spectrum 2 The total peak intensity is much greater than the corresponding MS selection. 1 A peak intensity four times greater (shown in the figure as a straight line with a slope of 4 passing through the origin for reference) can be considered to correspond to the MS values ​​of these plots. 2 The spectrum is a chimeric MS 2 Spectrum. If confirmed by other known methods, most of these are chimeric MS. 2 Spectrum. In the plot surrounded by dashed lines, in MS 2 MS appearing in the spectrum 2The sum of the peak intensities is the corresponding MS selection. 1 The peak intensity is approximately four times or less, which can be considered to correspond to the MS values ​​of these plots. 2 The spectrum is not a chimeric MS 2 Spectrum. If confirmed by other known methods, most of these are not chimeric MS. 2 Spectrum.

[0076] Chimeric MS 2 The spectrum determination / processing unit 44 can determine the MS of the object being processed. 2 All MS appearing in the spectrum 2 The sum of the peak intensities is compared to the peak threshold. Alternatively, the MS of the processed object can be... 2 All MS appearing in the spectrum 2 A specified number of MS peaks 2 The sum of the peak intensities is compared to the peak threshold. In this case, a specified number of MS values ​​are used. 2 Peaks can be selected in descending order of intensity, or in the order they are stored in the data file, or multiple combinations can be prepared (in which case, multiple MS tests can be performed). 2 (Comparison of the sum of peak intensities with the peak threshold).

[0077] Furthermore, when the precursor ion is multivalent, even if the peak threshold is set to the selected MS... 1 The peak intensity value, in this MS 2 The spectrum is a chimeric MS 2 Under spectral conditions, the detection amount (MS) of each product ion obtained by dissociating multivalent precursor ions. 2 The sum of the peak intensities (to the detected amount of the precursor ion (selected MS)) is equal to the total peak intensity (to the detected amount of the precursor ion (selected MS). 1 The fact that the peak intensity is greater remains unchanged, therefore the MS can be... 2 Spectral analysis indicates it is a chimeric MS 2 Spectrum.

[0078] As described in the first processing method, the embedded MS 2 The operation of the spectrum determination / processing unit 44 can be performed automatically while storing the data required for determination in the data storage unit 41, or automatically after the operation of the measurement unit, i.e., after the DDA-based analysis is completed, or after the input receiving unit 46 receives the user's input indicating that the determination / processing should begin at the input unit 5.

[0079] Chimeric MS 2 The spectrum determination / processing unit 44 is capable of determining whether an MS is a chimeric MS. 2 MS of the spectrum 2 Spectrum and / or corresponding MS 2The spectral data was processed using various methods. The details are the same as those for the first processing method, therefore, they are omitted here.

[0080] The display processing unit 45 reads MS data from the data storage unit 41. 1 Spectrum and / or MS 2 The spectrum is displayed in display section 6. The details are the same as in the first processing method, therefore, the explanation is omitted.

[0081] As described in the first processing method, the display processing unit 45 can perform the above-mentioned processing automatically, or it can be performed after the input receiving unit 46 receives the user's input indicating that the input unit 5 intends to start the display processing.

[0082] The display processing unit 45 is capable of displaying, for example, Figure 5 (b) shows the input screen for the user to change the peak threshold value. The input receiving unit 46, similar to the first analysis method, accepts various inputs from the user via the input unit 5 and outputs the content of these inputs to each unit. For example, in... Figure 5 In the input screen shown in (b) where the user changes the peak threshold value, when the user sets the peak threshold to MS... 1 The peak intensity multiplied by the specified magnification and summed with the selected MS 1 The input receiving unit 46 updates the other value and displays the changed peak threshold when the value of either the "Magnification" or "Peak Threshold" column is changed, based on the valence of the ion corresponding to the peak (the × mark in the figure indicates mode selection). The processing after the user presses the "Reprocess" button is the same as the first processing method, therefore, the explanation is omitted.

[0083] In addition, similar to the first processing method, the initial value of the peak threshold can be automatically set to the selected MS. 1 The intensity of the peak can also be preset by the user using the methods described above.

[0084] [Variation Example]

[0085] As described above, the chromatographic quality analysis data processing method of the present invention is specifically implemented by the control / processing unit 4. Other components are not essential to the present invention and can be appropriately modified. That is, in the chromatographic quality analysis data processing method of the present invention, the measuring unit is not necessary and can be provided instead, for example... Figure 8 The data management computer 7 shown contains past DDA-based analysis results, especially MS data. 1 Spectral data and MS 2 Spectral data, and then, will be compared with each selected MS 1 The mass-to-charge ratio corresponding to the peak, and the selection of MS 1The valence of the ions corresponding to the peaks and the peak thresholds are stored accordingly. The data storage unit 41 reads and saves this data from the data management computer 7, thereby enabling, for example, the first processing method and the second processing method described above to be implemented.

[0086] For the same MS 2 Spectral data can be processed using both the first and second data processing methods. This allows for more reliable splicing of MS data. 2 Spectral determination.

[0087] [plan]

[0088] The exemplary embodiments described above are specific examples of the following solutions, which will be obvious to those skilled in the art.

[0089] (Item 1) A method for processing chromatographic quality analysis data according to one aspect of the present invention involves processing MS... 2 The method for processing chromatographic quality analysis data of the spectrum, the MS 2 The spectrum was obtained by MS 1 Analysis and acquisition of MS 1 Spectrum, obtained from MS 1 One or more MS appearing in the spectrum 1 Select 1 MS in the peak 1 Peak, for the 1 MS contained in the selection 1 The MS analyzer uses ions within a specified mass-to-charge ratio range, corresponding to the peak, as precursor ions, and performs MS analysis. 2 MS obtained through analysis 2 The chromatographic quality analysis data processing method includes:

[0090] Based on the selected 1 MS 1 Peak intensity, and the setting of peak thresholds;

[0091] The MS 2 MS appearing in the spectrum 2 The intensity of the peak is compared with the peak threshold, and the MS with an intensity greater than the peak threshold is selected. 2 In the presence of a peak, the MS 2 Spectral analysis indicates it is a chimeric MS 2 Spectrum processing.

[0092] (Item 10) A chromatographic quality analysis data processing apparatus according to one aspect of the present invention comprises:

[0093] Input receiving unit, receives data from MS 1 Analysis of the acquired MS 1 One or more MS appearing in the spectrum 1 One MS selected from the peak1 Peak intensity and MS 2 The input of the spectrum, the MS 2 The spectrum is a 1 MS containing the selected 1 The specified mass-to-charge ratio range, including the peak corresponding to the mass-to-charge ratio, is used by MS analysis using ions belonging to this mass-to-charge ratio range as precursor ions. 2 MS obtained through analysis 2 Spectrum;

[0094] The peak threshold setting unit, based on the selected 1 MS 1 The peak intensity sets the peak threshold;

[0095] The determination unit will determine the MS. 2 MS appearing in the spectrum 2 The intensity of the peak is compared with the peak threshold, and the MS with an intensity greater than the peak threshold is selected. 2 In the presence of a peak, the MS 2 Spectral analysis indicates it is a chimeric MS 2 Spectrum.

[0096] Typically, MS n The intensity of a peak appearing in the spectrum (n≥1) represents the detectable amount of an ion with the mass-to-charge ratio corresponding to that peak. Each product ion obtained by dissociating a certain precursor ion constitutes that precursor ion before dissociation; therefore, the detectable amount (MS) of each product ion can be considered as... 2 The peak intensity) is the detection limit (MS) of the precursor ion. 1 Below the peak intensity. Therefore, in MS... 2 In the spectrum, it can be said that it is more compatible with the MS that has been chosen. 1 The intensity of the peak is greater than that of the MS. 2 The product ions corresponding to the peak are not only those selected by the MS. 1 The product ions obtained by the dissociation of the precursor ions corresponding to the peak, the MS 2 The spectrum was obtained by MS 1 Analysis and acquisition of MS 1 Spectrum, obtained from MS 1 One or more MS appearing in the spectrum 1 Select 1 MS in the peak 1 Peak, for the 1 MS contained in the selection 1 MS is performed on ions belonging to a specified mass-to-charge ratio range, including those corresponding to peaks other than the selected peaks, using these ions as precursor ions. 2 MS obtained through analysis 2Spectrum. Therefore, according to the chromatographic quality analysis data processing method involved in item 1 and the chromatographic quality analysis data processing device involved in item 10, the selected MS... 1 The peak intensity is set as the peak threshold, and the MS is... 2 MS appearing in the spectrum 2 The intensity of the peak is compared with the peak threshold, and the MS with an intensity greater than the peak threshold is... 2 In the presence of the peak, it is possible to [control the MS]. 2 Spectral analysis indicates it is a chimeric MS 2 Spectrum.

[0097] (Item 2) The chromatographic quality analysis data processing method involved in Item 2 is, in addition to the chromatographic quality analysis data processing method involved in Item 1, further comprising: processing MS data with an intensity greater than the peak threshold. 2 The peak intensity is set to 0.

[0098] According to the chromatographic quality analysis data processing method involved in item 2, in the chimeric MS 2 In the spectrum, it is possible to reduce and select MS 1 The contribution of different components from which the ions corresponding to the peaks originate can improve MS performance. 2 The purity of the spectrum. The result is that the chimeric MS... 2 Identifying the components in the spectrum becomes easier.

[0099] (Item 4) A method for processing chromatographic quality analysis data according to one aspect of the present invention involves processing MS... 2 The method for processing chromatographic quality analysis data of the spectrum, the MS 2 The spectrum was obtained by MS 1 Analysis and acquisition of MS 1 Spectrum, obtained from MS 1 One or more MS appearing in the spectrum 1 Select 1 MS in the peak 1 Peak, for the 1 MS contained in the selection 1 The MS analyzer uses ions within a specified mass-to-charge ratio range, corresponding to the peak, as precursor ions, and performs MS analysis. 2 MS obtained through analysis 2 The chromatographic quality analysis data processing method includes:

[0100] Based on the selected 1 MS 1 Peak intensity, and the setting of peak thresholds;

[0101] The MS 2 Multiple MS appearing in the spectrum 2The sum of the peak intensities is compared to the peak threshold; if the sum is greater than the peak threshold, the MS is... 2 Spectral analysis indicates it is a chimeric MS 2 Spectrum processing.

[0102] (Item 11) A chromatographic quality analysis data processing apparatus according to one aspect of the present invention comprises:

[0103] Input receiving unit, receives data from MS 1 Analysis of the acquired MS 1 One or more MS appearing in the spectrum 1 One MS selected from the peak 1 Peak intensity and MS 2 The input of the spectrum, the MS 2 The spectrum is a 1 MS containing the selected 1 The specified mass-to-charge ratio range, including the peak corresponding to the mass-to-charge ratio, is used by MS analysis using ions belonging to this mass-to-charge ratio range as precursor ions. 2 MS obtained through analysis 2 Spectrum;

[0104] The peak threshold setting unit, based on the selected 1 MS 1 The peak intensity sets the peak threshold;

[0105] The determination unit will determine the MS. 2 Multiple MS appearing in the spectrum 2 The sum of the peak intensities is compared to the peak threshold; if the sum is greater than the peak threshold, the MS is... 2 Spectral analysis indicates it is a chimeric MS 2 Spectrum.

[0106] When the precursor ion is monovalent, the number of product ions obtained from one precursor ion is one. However, the types of product ions obtained vary depending on which atom constituting the precursor ion is charged. In this case, the detection quantity (MS) of each product ion obtained by dissociating the precursor ion can be considered as... 2 The sum of the peak intensities is the detection limit (MS) of the precursor ion. 1 Below the peak intensity. Therefore, in MS... 2 In the spectrum, in this MS 2 Multiple (or all) MS appearing in the spectrum 2 The sum of the peak intensities is greater than the selected MS. 1 When the peak intensity is greater, it can be said that the MS 2 The spectrum is not only made with the selected MS 1 The spectrum obtained by the dissociation of precursor ions corresponding to the peak, the MS 2 The spectrum was obtained by MS 1Analysis and acquisition of MS 1 Spectrum, obtained from MS 1 One or more MS appearing in the spectrum 1 Select 1 MS in the peak 1 Peak, for the 1 MS contained in the selection 1 MS is performed within a specified mass-to-charge ratio range, including the peak corresponding to that range, using monovalent ions within that range as precursor ions. 2 MS obtained through analysis 2 Spectrum. Therefore, according to the chromatographic quality analysis data processing method involved in item 4 and the chromatographic quality analysis data processing device involved in item 11, the selected MS... 1 The peak intensity is set as the peak threshold, and the MS is... 2 Multiple MS appearing in the spectrum 2 The sum of the peak intensities is compared to the peak threshold. If the sum is greater than the peak threshold, the MS can be... 2 Spectral analysis indicates it is a chimeric MS 2 Spectrum.

[0107] (Item 3) The chromatographic quality analysis data processing method involved in Item 3 is, in the chromatographic quality analysis data processing method described in Item 1 or Item 2, further comprising: processing the MS... 2 Multiple MS appearing in the spectrum 2 The sum of the peak intensities is compared to the peak threshold; if the sum is greater than the peak threshold, the MS is... 2 Spectral analysis indicates it is a chimeric MS 2 Spectrum processing.

[0108] According to the chromatographic quality analysis data processing method described in item 3, by performing both the determinations in the method described in item 1 and the determinations in the method described in item 4, the chimeric MS can be performed more reliably. 2 Spectral determination.

[0109] (Item 5) The chromatographic quality analysis data processing method involved in Item 5 is, in any one of the chromatographic quality analysis data processing methods described in Items 1 to 4, wherein the peak threshold is the selected MS... 1 Peak intensity.

[0110] (Item 6) The chromatographic quality analysis data processing method involved in Item 6 is, in any one of the chromatographic quality analysis data processing methods described in Items 1 to 4, wherein the peak threshold is the selected MS... 1 The value is obtained by multiplying the peak intensity by a specified multiplier.

[0111] (Item 7) The chromatographic quality analysis data processing method involved in Item 7 is, in the chromatographic quality analysis data processing method described in Item 6, the specified magnification is in the range of 0.8 to 1.2 times.

[0112] Typically, in MS n The intensity of peaks appearing in the spectrum (n≥1) may contain a certain degree of error (typically around 20%). Based on the chromatographic quality analysis data processing methods described in items 5–7, and according to the characteristics of the data, the selected MS... 1 Multiplying the peak intensity by a specified factor, particularly by 0.8 to 1.2, and setting this value as the peak threshold allows for more accurate determination of the MS. 2 Is the spectrum a chimeric MS? 2 Spectrum.

[0113] (Item 8) The chromatographic quality analysis data processing method involved in Item 8 is, in the chromatographic quality analysis data processing method described in Item 3 or Item 4, wherein the peak threshold is the selected MS... 1 The peak intensity multiplied by the selected MS 1 The value is obtained by considering the valence of the ion corresponding to the peak.

[0114] When the precursor ion is multivalent, multiple atoms may become charged and ionized, thus potentially generating multiple product ions from a single precursor ion. For example, when the precursor ion is divalent, the maximum number of product ions obtained from a single precursor ion is two. In this case, if the selected MS... 1 The peak intensity value multiplied by the selected MS 1 Setting the peak threshold based on the valence of the precursor ion corresponding to the peak allows for a more accurate determination of the data processing performed in the chromatographic quality analysis data processing methods described in item 3 or 4, based on MS. 2 The determination is based on the comparison between the sum of peak intensities and the peak threshold. That is, according to the chromatographic quality analysis data processing method described in item 8, it is possible to more accurately determine the chromatographic quality analysis data processing performed in item 3 or 4 based on MS. 2 The determination is based on the comparison between the sum of peak intensities and the peak threshold. Additionally, when the precursor ion is multivalent, even if the peak threshold is set to the selected MS value... 1 The peak intensity value, in this MS 2 The spectrum is a chimeric MS 2 Under spectral conditions, the detection amount (MS) of each product ion obtained by dissociating multivalent precursor ions. 2 The sum of the peak intensities (to the detected amount of the precursor ion (selected MS)) is equal to the total peak intensity (to the detected amount of the precursor ion (selected MS). 1 The fact that the peak intensity is greater remains unchanged, therefore it will not be a chimeric MS. 2MS of the spectrum 2 The spectrum was incorrectly identified as not being a chimeric MS. 2 Spectrum.

[0115] (Item 9) The chromatographic quality analysis data processing method involved in Item 9 is, in the chromatographic quality analysis data processing method described in Item 8, as the selected MS 1 The valence of the ion corresponding to the peak is a value predicted based on the type of sample and the type of ionization method used for that sample.

[0116] The valence of precursor ions depends to some extent on the type of sample and the type of ionization method used for that sample, and therefore can be predicted to some extent based on experience. Therefore, according to the chromatographic quality analysis data processing method described in Item 9, by predicting and determining the valence of precursor ions based on the type of sample and the type of ionization method used for that sample, peak thresholds can be set with higher precision even when the valence of the precursor ions is unclear.

[0117] Explanation of reference numerals in the attached figures

[0118] 1. Liquid Chromatography Section

[0119] 10. Mobile Phase Container

[0120] 11 Liquid delivery pump

[0121] 12 syringes

[0122] 13 chromatographic column

[0123] 2 Quality Analysis Department

[0124] 20 vacuum chambers

[0125] 201 Ionization Chamber

[0126] 202 First Intermediate Vacuum Chamber

[0127] 203 Second Intermediate Vacuum Chamber

[0128] 204 High Vacuum Chamber No. 1

[0129] 205 Second High Vacuum Chamber

[0130] 21ES I probe

[0131] 22 desolventizing tube

[0132] 23, 25, 28, 29 Ion Guides

[0133] 24 cone body

[0134] 26 Quadruple Filter

[0135] 27 Collision Pool

[0136] 30 Orthogonal Accelerators

[0137] 31 Ion Flight Division

[0138] 32 Ion Detector

[0139] 4 Control / Processing Unit

[0140] 40 Analysis and Control Department

[0141] 41 Data Storage Department

[0142] 42 Spectrum Generation Section

[0143] 43 Peak Selection / Peak Threshold Setting Section

[0144] 44-chip MS 2 Spectrum Determination / Processing Department

[0145] 45 Display Processing Unit

[0146] 46 Input Receiving Unit

[0147] 5 Input Section

[0148] 6 Display Section

[0149] 7. Computers for data management.

Claims

1. A method for processing chromatographic quality analysis data, which is used to process MS data. 2 The method for processing chromatographic quality analysis data of the spectrum, the MS 2 The spectrum was obtained by MS 1 Analysis and acquisition of MS 1 Spectrum, obtained from MS 1 One or more MS appearing in the spectrum 1 Select 1 MS in the peak 1 Peak, for the 1 MS contained in the selection 1 The MS analyzer uses ions within a specified mass-to-charge ratio range, corresponding to the peak, as precursor ions, and performs MS analysis. 2 MS obtained through analysis 2 The chromatographic quality analysis data processing method includes: based on the selected 1 MS 1 intensity of the peak, processing of setting a peak threshold said MS 2 MS present in the spectrum 2 the intensity of the peak is compared to the peak threshold, and the MS having an intensity greater than the peak threshold 2 the MS is determined to be a chimeric MS 2 the spectrum is determined to be a chimeric MS 2 processing of the spectrum.

2. The method of processing chromatographic mass analysis data according to claim 1, wherein, Further comprising: MS having an intensity greater than the peak threshold 2 processing in which the intensity of the peak is changed to 0.

3. The method of processing chromatographic mass analysis data according to claim 1, wherein, Further comprising: said MS 2 a plurality of MSs present in the spectrum 2 the total value of the intensities of the peaks is compared with the peak threshold value, and in the case where the total value is greater than the peak threshold value, the MS 2 the spectrum is determined to be a chimeric MS 2 the processing of the spectrum.

4. A chromatographic mass spectrometric data processing method of processing MS 2 spectra, the chromatographic mass spectrometric data processing method of processing MS 2 spectra, the chromatographic mass spectrometric data processing method of processing MS 1 spectra, the chromatographic mass spectrometric data processing method of processing MS 1 spectra, the chromatographic mass spectrometric data processing method of processing MS 1 spectra, the chromatographic mass spectrometric data processing method of processing MS 1 spectra, the chromatographic mass spectrometric data processing method of processing MS 1 spectra, the chromatographic mass spectrometric data processing method of processing MS 1 spectra, the chromatographic mass spectrometric data processing method of processing MS 2 spectra, the chromatographic mass spectrometric data processing method of processing MS 2 spectra, the chromatographic mass spectrometric data processing method of processing MS based on the selected 1 MS 1 intensity of the peak, a process of setting a peak threshold said MS 2 a plurality of MSs present in the spectrum 2 the total value of the intensities of the peaks is compared with the peak threshold value, and in the case where the total value is greater than the peak threshold value, the MS 2 the spectrum is determined to be a chimeric MS 2 the processing of the spectrum.

5. The method of processing chromatographic mass analysis data according to any one of claims 1 to 4, characterized in that, The peak threshold is the selected MS 1 Intensity of the peak.

6. The method of processing data of a chromatographic mass analysis according to any one of claims 1 to 4, characterized in that, The peak threshold is the selected MS 1 A value obtained by multiplying the intensity of a peak by a prescribed multiple.

7. The method of processing chromatographic mass analysis data according to claim 6, wherein, The prescribed magnification is in the range of 0.8 to 1.2 times.

8. The method of processing data of a chromatographic mass analysis according to claim 3 or 4, characterized in that, The peak threshold is the selected MS 1 The value obtained by multiplying the intensity of the peak by the valence of the ion corresponding to the selected MS 1 The value obtained by multiplying the intensity of the peak by the valence of the ion corresponding to the selected MS 9. The method of processing chromatographic mass analysis data according to claim 8, wherein, As with the selected MS 1 The valence of the ion to which the peak corresponds, using a value predicted based on the kind of the sample and the kind of the ionization method of the sample.

10. A chromatographic mass analysis data processing apparatus, characterized by, Possessing: The input accepting section accepts an input of a spectrum from a mass spectrometer (MS) 1 1 The input accepting section accepts an input of a spectrum from a mass spectrometer (MS) 1 1 The input accepting section accepts an input of a spectrum from a mass spectrometer (MS) 1 1 The input accepting section accepts an input of a spectrum from a mass spectrometer (MS) 1 1 The input accepting section accepts an input of a spectrum from a mass spectrometer (MS) 1 2 The input accepting section accepts an input of a spectrum from a mass spectrometer (MS) 1 2 The input accepting section accepts an input of a spectrum from a mass spectrometer (MS) 1 1 The input accepting section accepts an input of a spectrum from a mass spectrometer (MS) 1 2 The input accepting section accepts an input of a spectrum from a mass spectrometer (MS) 1 2 The input accepting section accepts an input of a spectrum from a mass spectrometer (MS) 1 a peak threshold setting section sets a peak threshold based on the selected one MS 1 the intensity of the peak is set as the peak threshold The determination unit determines whether the MS 2 The MS appearing in the spectrum 2 The intensity of the peak is compared with the peak threshold value, and in the case where the MS 2 The MS having a peak with an intensity greater than the peak threshold value is determined as a chimeric MS 2 The spectrum is determined as a chimeric MS 2 The spectrum.

11. A chromatographic mass analysis data processing apparatus, characterized by, Possessing: input accepting section, accepting an input of a spectrum from a mass spectrometer (MS) 1 MS acquired by analyzing the input 1 one or more MS appearing in the spectrum 1 one MS selected from the peaks 1 intensity of the peak and the MS 2 input of the spectrum, the MS 2 the spectrum is a spectrum acquired by analyzing a sample containing the selected one MS 1 a predetermined mass-to-charge ratio range including the mass-to-charge ratio corresponding to the selected one MS 2 MS acquired by analyzing the input 2 spectrum; a peak threshold setting section sets a peak threshold based on the selected one MS 1 the intensity of the peak is set as the peak threshold The determination unit determines whether the MS 2 The MS 2 The determination unit compares the total value of the intensities of the peaks appearing in the spectrum with the peak threshold value, and in the case where the total value is larger than the peak threshold value, determines that the MS 2 The determination unit determines that the MS 2 The determination unit determines that the MS

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