Data acquisition method and ion mobility spectrometry-mass spectrometer

Abstract

A data acquisition method, which can be effectively adapted to DDA or DIA, and an ion mobility spectrometry-mass spectrometer (100). The data acquisition method comprises a pre-acquisition step (S1), a parent ion selection step (S2) and a daughter ion acquisition step (S3), wherein the pre-acquisition step (S1) involves obtaining an ion mobility spectrometry-mass spectrometry two-dimensional heat map; and the parent ion selection step (S2) involves configuring an ion mobility analyzer (2) to be in a filter mode, selecting parent ions on the basis of the ion mobility spectrometry-mass spectrum two-dimensional heat map, and correspondingly setting a mobility selection range of the ion mobility analyzer (2) and a mass selection range of a quadrupole mass filter (3). The data acquisition method solves the problems with respect to a limited dynamic range and a low scanning efficiency faced by existing data acquisition methods.

Description

Data acquisition method and ion mobility spectrometry-mass spectrometry Technical Field

[0001] The present invention relates to the field of mass spectrometry, and in particular to a data acquisition method and an ion mobility spectrometer-mass spectrometer. Background Art

[0002] Tandem mass spectrometry has become an important analytical instrument in omics analysis (including metabolomics, proteomics, etc.). For more complex samples in omics analysis, high-throughput, high-sensitivity, and high-coverage data acquisition methods are required.

[0003] The traditional method is data-dependent acquisition (DDA), proposed by Ducret et al. in 1998. This method first performs a scan targeting precursor ions (or "precursor" ions). Then, the more abundant precursor ions are selected and sequentially dissociated in an ion dissociation device to produce daughter ion (or "product" ion) spectra. This method achieves high analyte coverage and remains a widely used acquisition method today.

[0004] Data-independent acquisition (DIA) has seen rapid development in recent years. Rather than targeting specific precursor ions for collisional dissociation and analysis, DIA targets specific mass-to-charge ratio windows or ion clusters across the full mass range, then interprets the resulting complex spectral data. Compared to DDA, DIA can accurately identify relatively low-abundance ions, offering higher sensitivity, dynamic range, analytical throughput, and improved quantification.

[0005] To further improve sensitivity and increase the dimension of orthogonal separation, an increasing number of manufacturers are coupling ion mobility spectrometry (IMS) to tandem mass spectrometry instruments. This allows for further separation of different ions based on their mobility properties, thereby enhancing ion identification capabilities. Several types of IMS have been developed, including drift cell IMS (DT-IMS), traveling wave IMS (TW-IMS), trapped IMS (TIMS), U-shaped IMS (UMA), and differential IMS (DMA). These devices can provide mobility information related to the ion collision cross section (CCS). Another type of device, called differential IMS (DMS) or high-field asymmetric waveform IMS (FAIMS), provides differential ion mobility information rather than mobility information related to the collision cross section. Among them, DT-IMS and TW-IMS work in pulse mode, which slice or accumulate the continuous ion flow through the ion gate or ion trap, and then release it in a pulsed manner for one-time ion mobility analysis; the working mode of TIMS is a special scanning mode, which stores ions in the ion trap, and then scans the ions according to mobility and gradually releases them; the working mode of DMA is a filtering mode, which only allows ions with a specific mobility range in the continuous ion flow to pass through the device within a period of time; the working mode of UMA is more flexible, and can adopt a capture-scan mode similar to TIMS, or a filtering mode similar to DMA; DMS or FAIMS both work in filtering mode, but they do not perform filtering analysis based on mobility, but on differential mobility.

[0006] Regarding the ion mobility spectrometer in filtering mode, in addition to DMA and UMA, Hashimoto et al. also disclosed a device in U.S. Patent 7718960B2. The device has two migration regions, in which there are electric fields and airflows in opposite directions, and the airflow directions in the two migration regions are opposite. The gas in the device is vertically introduced from the connecting region of the two regions. In U.S. Patent 9281170B2, Parks transformed TIMS into an ion mobility analyzer in filtering mode, which also adopts the method of vertically introducing airflow and using a radio frequency quadrupole field to radially confine ions. Both Hashimoto and Parks' high and low pass filter devices have the following problems: first, it is difficult to obtain uniform and reverse airflow, the device is complex, and turbulence is easily introduced; second, this filtering mode cannot achieve mobility analysis with a high duty cycle (i.e., high ion utilization efficiency).

[0007] The tandem mass spectrometry method combined with the ion mobility spectrometry analysis device has been further developed. In U.S. Patent No. 9891194B2, a so-called DDA-PASEF method is disclosed, which uses trapped ion mobility spectrometry (TIMS) as an ion mobility separator, and combined with the parallel accumulation technology (PASEF) disclosed in US9683964B2, dual TIMS can be used (the first TIMS accumulates ions, and the second TIMS serves as an ion mobility separator), and the TIMS scan is synchronized with the quadrupole scan. In theory, 100% ion utilization efficiency can be obtained during DDA acquisition. Furthermore, in the document “Nature Methods, 17, December 2020, 1229–1236”, a DIA-PASEF method is proposed. This method applies the concept of PASEF to the DIA acquisition method, and uses a specially designed mobility spectrometry-quadrupole mass spectrometry scanning process to increase the acquisition efficiency of the traditional DIA method by several times, thereby enabling a deeper analysis of protein samples and better repeatability.

[0008] However, the data collection methods in the prior art still have the following problems:

[0009] First, for omics studies of complex samples, the contents of different components vary greatly, requiring an extremely high dynamic range to achieve good qualitative and quantitative results. In parallel accumulation techniques, to achieve 100% ion utilization efficiency, ions must accumulate in the first TIMS. However, TIMS is developed based on the ion trap principle. Spatial effects can lead to the exclusion of some low-abundance ions, limiting the dynamic range of the coupled instrument by the ion capacity of the first TIMS.

[0010] Secondly, for DIA-PASEF, the quadrupole mass filter has a fixed window width. To synchronize with the migration spectrum, the quadrupole needs to jump between the mass windows. The problem with this method is that it limits the quadrupole scanning speed. At the same time, it lacks the "oversampling" of the mass window in traditional DIA acquisition, resulting in a reduction in ion coverage. Therefore, multiple scans are required to improve ion coverage. The same research group further improved its mobility spectrum-quadrupole scanning strategy in U.S. Patent No. 11474072B2, avoiding the quadrupole mass number jump scan and adopting a sliding method along a fixed mass window while scanning synchronously with the mobility of the TIMS device. This can increase the scanning speed and obtain more correlation information between the parent and daughter ions, making it easier to determine the daughter ion identity. However, this method still requires a very large number of scans, which not only limits the speed of DIA acquisition but also reduces the efficiency of ion utilization.

[0011] Third, TIMS uses a balance between axial gas flow and the electric field to control ion ejection. If ions are to be ejected sequentially, they can only be ejected in descending order of collision cross section (CCS) (from low to high ion mobility). This limits scanning efficiency and can also cause some unwanted ions to be ejected, affecting analysis. While this issue can be addressed through dynamic adjustment of the scan rate or further ion clearing mechanisms, it still increases the complexity of the device.

[0012] In addition, patent CN113495112A proposes an ion mobility spectrometer operating in filtering mode. Typically, the device uses a UMA as a mobility analyzer and can be combined with a mass spectrometer for DDA or DIA analysis. However, the patent does not specify how to improve the utilization efficiency and scanning efficiency of ions in the DDA and DIA processes, thereby improving the coverage of omics.

[0013] Summary of the Invention

[0014] To address the above issues, the present invention provides a data acquisition method that can be effectively adapted to DDA or DIA, solving the problems of limited dynamic range, slow scanning speed, and low omics coverage faced by existing data acquisition methods.

[0015] A first aspect of the present invention provides a data acquisition method for an ion mobility spectrometer-mass spectrometer, wherein the ion mobility spectrometer-mass spectrometer comprises an ion source, an ion mobility analyzer, a quadrupole mass filter, an ion dissociation device, and a mass spectrometer connected in sequence. The data acquisition method comprises the following steps:

[0016] In the survey scan step, the parent ions obtained by ionization in the ion source are sequentially passed through an ion mobility analyzer and a mass spectrometer to obtain a two-dimensional thermal map of the ion mobility spectrum and mass spectrum of the parent ions;

[0017] a precursor ion selection step, configuring the ion mobility analyzer in filtering mode, selecting precursor ions based on the ion mobility spectrum-mass spectrum two-dimensional heat map, and setting the mobility selection range of the ion mobility analyzer and the mass selection range of the quadrupole mass filter accordingly;

[0018] In the daughter ion collection step, the selected parent ions enter the ion dissociation device for dissociation to produce daughter ions, which enter the mass spectrometry device for analysis to obtain the ion mobility spectrum-mass spectrometry data of the daughter ions.

[0019] The parent ion selection step and the daughter ion acquisition step are repeated to obtain ion mobility spectrum-mass spectrum data of daughter ions corresponding to multiple groups of parent ions.

[0020] According to the data acquisition method provided by the present invention, by first carrying out a pre-collection to obtain the ion mobility spectrum-mass spectrum two-dimensional heat map of parent ion, it is possible to determine trend line (trend line), i.e. the relationship between ion mobility and mass number, the determination of trend line helps to select more suitable mobility selection range and mass selection range in the different stages of the same cycle in the parent ion selection step. In addition, pre-collection step can also obtain the intensity information and mobility information of parent ion, and the intensity information of parent ion is needed in implementing DDA analysis, and the mobility information of parent ion can be used for calculating collision cross-sectional area CCS, to help qualitative analysis. After pre-collection step, carry out the selection and cascade analysis of parent ion, in the parent ion acquisition step, by ion mobility analyzer being configured to filter mode, ion mobility analyzer does not have dynamic range limitation in theory, can effectively improve dynamic range, avoids dynamic range being limited to ion trap capacity. In addition, filter mode can select a wider mobility window, and when carrying out DIA or DDA analysis, wide window can better cover target ion. Furthermore, during repeated precursor ion selection, the mobility window and mass window can be simultaneously slid and scanned to ensure both ion coverage and scan speed. Furthermore, the filter mode allows for arbitrary selection of filtering channels, enabling both step-by-step and skipped scanning, which not only increases scan speed but also avoids interference from unwanted ions.

[0021] Through the above methods, this data acquisition method can be effectively adapted to DDA or DIA analysis, solving the problems faced by existing data acquisition methods, such as limited dynamic range, slow scanning speed, and low omics coverage.

[0022] In an optional technical solution of the present invention, the ion mobility analyzer is a U-shape ion mobility analyzer (UMA).

[0023] The U-shaped ion mobility analyzer (IMA) operates in filter-scan mode by controlling the balance between the electric field and airflow, allowing only ions within a specific mobility range to pass through at a given time. Ion mobility analysis is then performed by scanning. Filtered ions are not lost during the scanning process but are instead stored in the device's storage area, effectively preserving them and releasing them from the IMA at the appropriate time, thereby effectively improving ion utilization efficiency.

[0024] In an optional technical solution of the present invention, the period of the ion mobility analyzer scanning the mobility selection range is synchronized with the period of the quadrupole mass filter scanning the mass selection range; the ion mobility analyzer scans multiple mobility selection ranges in a manner in which the mobility at least partially changes from high to low within one analysis cycle.

[0025] Furthermore, in an optional technical solution of the present invention, the ion mobility analyzer scans multiple mobility selection ranges in a manner of high to low mobility, and the quadrupole mass filter scans multiple mass selection ranges in a manner of small to large mass-to-charge ratio.

[0026] During the daughter ion collection step, since smaller parent ions usually correspond to smaller daughter ions, and the retention time (dwell time) of small daughter ions in the ion dissociation device is also shorter, their transmission in the ion dissociation device is not easily interfered with by subsequent larger daughter ions, and therefore will not affect the quality of their migration spectrum (such as avoiding migration spectrum peak mismatch, etc.), and can also speed up the channel scanning speed of DDA or DIA.

[0027] Furthermore, in an optional technical solution of the present invention, during the precursor ion selection step, the mass selection range of the quadrupole mass filter is a mass window of fixed width, and the quadrupole mass filter sequentially scans different mass windows from small to large within a single analysis cycle. This data acquisition method can efficiently acquire data suitable for DIA analysis.

[0028] Furthermore, in an optional technical solution of the present invention, in the precursor ion selection step, the mass selection range of the quadrupole mass filter overlaps with the mass selection range of the next repeated precursor ion selection step. This data acquisition method can improve the analytical coverage of DDA and DIA methods.

[0029] Furthermore, in an optional technical solution of the present invention, in the precursor ion selection step, the mobility selection range of the ion mobility analyzer overlaps with the mobility selection range of the next repeated precursor ion selection step. This data acquisition method can further improve the analytical coverage, ion utilization efficiency, and acquisition speed of DDA and DIA methods.

[0030] Furthermore, in an optional technical solution of the present invention, the parent ion selection step, the daughter ion collection step, and the repeated parent ion selection step and daughter ion collection step are recorded as the first scan; thereafter, the parent ion selection step, the daughter ion collection step, and the repeated parent ion selection step and daughter ion collection step are performed (N-1) times and recorded as the second, third, and Nth scans; wherein in at least two consecutive scans, the mobility selection ranges of the parent ions corresponding to the same mass selection range overlap. By having the mass window and mobility window in the next parent ion selection step overlap with the mass window and mobility window of the previous step, "oversampling" of the ions can be achieved, thereby achieving better coverage.

[0031] Furthermore, in an optional technical solution of the present invention, the mobility selection range of the ion mobility analyzer in the parent ion selection step is different from the mass selection range in the next repeated parent ion selection step; or the mass selection range of the quadrupole mass filter in the parent ion selection step is different from the mass selection range in the next repeated parent ion selection step. In other words, both the mobility window and the mass window in the parent ion selection step can be changed in real time to achieve more efficient scanning.

[0032] In an optional technical solution of the present invention, during the parent ion selection step, parent ions outside the current mobility selection range are at least partially stored in the ion storage area of ​​the ion mobility analyzer. When the parent ion selection step is repeated, parent ions in at least a portion of the ion storage area are selected and released to the ion dissociation device. Using the ion storage area of ​​the ion mobility analyzer to temporarily store ions and then releasing them to the ion dissociation device at the appropriate time can effectively improve ion utilization efficiency and increase the duty cycle of the combined instrument.

[0033] In an optional technical solution of the present invention, in the precursor ion selection step, multiple groups of precursor ions whose signal intensities in the ion mobility spectrometry-mass spectrometry two-dimensional heat map are selected, or multiple groups of precursor ions whose signal intensities in the ion mobility spectrometry-mass spectrometry two-dimensional heat map are selected. This data acquisition method can accurately screen target precursor ions and efficiently acquire data suitable for analysis using the DDA method.

[0034] In an alternative embodiment of the present invention, the mass selection range in the precursor ion selection step is the full mass range, and this range remains constant in each product ion collection step. This data collection method can efficiently collect data suitable for full mass range DIA analysis methods.

[0035] A second aspect of the present invention provides an ion mobility spectrometer-mass spectrometer using the data acquisition method of the present invention.

[0036] In an optional technical solution of the present invention, the mass analysis device is one or more of a quadrupole mass analysis device, a time-of-flight mass spectrometer, a Fourier transform mass spectrometer, an ion trap mass spectrometer, and a magnetic mass spectrometer.

[0037] In an optional technical solution of the present invention, the ion dissociation device is one or more of a collision-induced dissociation device, an electron dissociation device, an infrared photon dissociation device, an ultraviolet photon dissociation device, and a free radical dissociation device. BRIEF DESCRIPTION OF THE DRAWINGS

[0038] FIG1 is a schematic structural diagram of an ion mobility spectrometer-mass spectrometer provided in an embodiment of the present invention;

[0039] FIG2 is a schematic structural diagram of a U-shaped ion mobility analyzer in an ion mobility spectrometer-mass spectrometer according to an embodiment of the present invention;

[0040] FIG3 is a flow chart of a data acquisition method provided by an embodiment of the present invention;

[0041] FIG4 is a schematic diagram of a trend line determined based on an ion mobility spectrometry-mass spectrometry two-dimensional heat map according to an embodiment of the present invention;

[0042] 5 is a schematic diagram of a scan adapted to a DDA method according to an embodiment of the present invention, wherein a mobility selection range and a mass selection range are selected and scanned simultaneously;

[0043] FIG6 is a schematic diagram of a scanning method adapted to the DDA method according to a preferred embodiment of the present invention, wherein a mobility selection range and a mass selection range are selected and scanned simultaneously;

[0044] FIG7( a ) is a schematic diagram of a first scan adapted to a DIA method according to an embodiment of the present invention, in which a mobility scan and a mass selection window are scanned synchronously;

[0045] FIG7( b ) is a schematic diagram of a first scan and a second scan adapted to a DIA method according to an embodiment of the present invention, in which a mobility scan and a mass selection window are scanned synchronously;

[0046] FIG8( a ) is a schematic diagram of a first scan of a preferred data acquisition method adapted to the DIA method according to a preferred embodiment of the present invention, wherein the mobility selection range and the mass selection range are selected and scanned simultaneously;

[0047] FIG8( b ) is a schematic diagram of the first scan, the second scan, and the third scan of a preferred data acquisition method adapted to the DIA method according to a preferred embodiment of the present invention, wherein the mobility selection range and the mass selection range are selected and scanned simultaneously;

[0048] FIG9 is a schematic diagram of a first scan and a second scan of another preferred data acquisition method adapted for the DIA method according to a preferred embodiment of the present invention, wherein the mobility selection range and the mass selection range are selected and scanned simultaneously;

[0049] 10( a ) and ( b ) are schematic diagrams of the first scan and the first three scans, respectively, of another preferred data acquisition method adapted for the DIA method according to a preferred embodiment of the present invention, wherein the mobility selection range and the mass selection range are selected and scanned simultaneously;

[0050] FIG11 is a schematic diagram of the ion control process of the U-shaped ion mobility analyzer in a preferred embodiment of the present invention within one cycle.

[0051] Figure markings: 100-ion mobility spectrometer-mass spectrometer; 1-ion source; 2-ion mobility analyzer; 201-ion inlet; 202-ion transfer port; 203-ion outlet; 204-first ion storage area; 205-second ion storage area; 3-quadrupole mass filter; 4-ion dissociation device; 5-mass spectrometry device; 6-ion optical device; CH1-first channel; CH2-second channel; T-path. DETAILED DESCRIPTION

[0052] The following will be combined with the accompanying drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the embodiments described are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0053] <Terms and their explanations>

[0054] The term "ion mobility spectrometer-mass spectrometer" is not limited to the combination of ion mobility spectrometer and mass spectrometer, and can also be combined with equipment such as chromatography to form a chromatography-ion mobility spectrometer-mass spectrometer.

[0055] The phrase "sequentially connected" does not necessarily prohibit the addition of other components between components; the connections between components can be indirect. For example, "an ion source, an ion mobility analyzer, a quadrupole mass filter, an ion dissociation device, and a mass spectrometer connected sequentially" means that the ion mobility analyzer is positioned after the ion source, and the quadrupole mass filter is positioned after the ion mobility analyzer. However, other components can be positioned between the ion mobility analyzer and the ion source, and similarly, between the ion mobility analyzer and the quadrupole mass filter.

[0056] "Filtering mode" is an operating mode that can be applied to an ion mobility analyzer, similar to the filtering mode of a general quadrupole mass filter. In the ion mobility analyzer, the characteristic of this operating mode is that, within a certain time period, only ions within a specific mobility range are allowed to pass through the ion mobility analyzer, while ions outside the mobility range are not allowed to pass through the ion mobility analyzer. In the present invention, the physical quantity analyzed or measured by the ion mobility analyzer is limited to the ion mobility under low field, and the ion mobility is directly related to the collision cross section (CCS) of the ion, that is, the ion mobility analyzer can be used to obtain CCS information. The ion mobility analyzer operating in "filtering mode" continuously screens out ions within a specific mobility range while maintaining continuous ion injection at the sample inlet.

[0057] There are at least two types of "filter modes": one is static, allowing only ions within a certain mobility range to pass through for an extended period of time, commonly referred to as filter-selective ion monitoring (SIM) mode; the other is dynamic scanning, allowing ions within different mobility ranges to pass through sequentially at different times, known as filter-scan mode. The filter-scan mode is preferred. In filter-scan mode, the ion mobility analyzer can directly receive a continuous ion stream and perform mobility analysis without storing or packaging the ions. Therefore, there is no ion storage capacity issue, resulting in a higher dynamic range and better quantification.

[0058] Unlike differential ion mobility spectrometry (DMS) or high-field asymmetric waveform ion mobility spectrometry (FAIMS), the "ion mobility spectrometry" in the present invention is only used to measure ion mobility (ion mobility) under low field, and this ion mobility is directly related to the collision cross section (CCS) of the ion; although DMS and FAIMS can also work in filtering mode, they can only measure the difference in ion mobility caused by the difference between high and low fields, that is, differential ion mobility (differential mobility), and cannot obtain information on ion mobility and collision cross section, and the resolution of DMS and FAIMS is usually low and the ion utilization efficiency is also very low.

[0059] This embodiment provides a data acquisition method and an ion mobility spectrometer-mass spectrometer 100 using the data acquisition method.

[0060] <Ion Mobility Spectrometry-Mass Spectrometry 100>

[0061] 1 , the ion mobility spectrometer-mass spectrometer 100 provided in this embodiment includes an ion source 1 , an ion mobility analyzer 2 , a quadrupole mass filter 3 , an ion dissociation device 4 and a mass spectrometer 5 connected in sequence.

[0062] In this embodiment, the ion source 1 includes an ion source 1 selected from the group consisting of: (i) an electrospray ionization ("ESI") ion source; (ii) an atmospheric pressure photoionization ("APPI") ion source; (iii) an atmospheric pressure chemical ionization ("APCI") ion source; (iv) a matrix-assisted laser desorption ionization ("MALDI") ion source; (v) a laser desorption ionization ("LDI") ion source; (vi) an atmospheric pressure ionization ("API") ion source; (vii) an on-silicon desorption ionization ("DIO") ion source. S”) ion source; (viii) electron impact (“EI”) ion source; (ix) chemical ionization (“CI”) ion source; (x) field ionization (“FI”) ion source; (xi) field desorption (“FD”) ion source; (xii) inductively coupled plasma (“ICP”) ion source; (xiii) fast atom bombardment (“FAB”) ion source; (xiv) liquid secondary ion mass spectrometry (“LSIMS”) ion source; (xv) electrospray desorption ionization (“DESI”) ion source; (xvi) nickel-63 radioactive ion source; (xvii) atmospheric pressure matrix-assisted laser desorption ionization ion source; (xviii) thermal spray ion source; (xix) atmospheric sampling glow discharge ionization ("ASGDI") ion source; (xx) glow discharge ("GD") ion source; (xxi) impactor ion source; (xxii) direct analysis in real time ("DART") ion source; (xxiii) laser spray ionization ("LSI") ion source; (xxiv) sonic spray ionization ("SSI") ion source; (xxv) matrix assisted inlet ionization ("MAII") ion source; (xxvi) solvent assisted inlet ionization ("SAII") ion source; (xxvii) Penning ionization ion source; (xxviii) laser ablation electrospray ionization ("LAESI") ion source; (xxix) He plasma (HePl) ion source. More preferably, a chamber pressure or real-time ion source such as desorption electrospray ionization ("DESI") ion source, matrix-assisted laser desorption ionization ion source ("MALDI"), direct analysis in real time ion source ("DART"), laser ablation electrospray ionization ("LAESI") ion source is used.

[0063] The ion mobility analyzer 2 is a U-type ion mobility analyzer 2 .

[0064] Referring to Figure 2 , the U-shaped ion mobility analyzer 2 includes a first channel CH1 and a second channel CH2 arranged in parallel and side by side. Each of the first channel CH1 and the second channel CH2 includes a set of electrode arrays arranged opposite each other. An ion inlet 201 is provided on the electrode array near the front end of the first channel CH1, through which a continuous ion flow can enter the first channel CH1. Ion transfer ports 202 are correspondingly provided on the electrode array near the rear end of the first channel CH1 and the electrode array near the front end of the second channel CH2. Within the first channel CH1, when the continuous ion flow moves along a path T in the longitudinal direction (left-right direction in Figure 2 ) to the ion transfer port 202, it can be transferred from the first channel CH1 to the second channel CH2 via the ion transfer port 202. After the direction of movement reverses, the continuous ion flow continues along the longitudinal path T until it reaches the vicinity of the ion outlet 203, exiting the second channel CH2 through the ion outlet 203 and being transported to the lower stage. The ion outlet 203 is provided on the electrode array near the rear end of the second channel CH2.

[0065] Airflow enters the first channel CH1 and the second channel CH2 from the same end along the length, flows along the length, and exits from the other end of the first channel CH1 and the second channel CH2 along the length. A DC electric field is applied to the electrode array along the length, exerting an electric force along the length on the ions within it. The direction of this electric force is opposite to the direction of the force exerted by the airflow on the ions. The balance between the airflow force (the magnitude of which depends on the CCS, airflow velocity, etc.) and the electric field force is used to screen out ions that meet the target mobility range and move along path T. Ions that do not meet the target mobility range leave path T and are removed from both ends of the first channel CH1 or the second channel CH2, where they are lost, or transported to the end of the first channel CH1 or the second channel CH2 for storage (to be described in detail later).

[0066] In the filter-SIM mode, the DC electric field E1 of the first channel CH1 and the DC electric field E2 of the second channel CH2 can be kept unchanged, and the difference ΔE between E1 and E2 is kept fixed, thereby screening out ions with a fixed mobility range or fixed mobility (ΔE=0).

[0067] For the filter-scan mode, the difference ΔE between E1 and E2 can be kept constant while E1 and E2 are changed synchronously, for example, by scanning from low to high, thereby scanning different mobility ranges or mobility values ​​(ΔE=0) with a fixed mobility window, for example, allowing ions to pass through in order of mobility from high to low. Continuing with reference to FIG1 , the ion dissociation device 4 is a collision-induced dissociation device, an electron dissociation device, an infrared photon dissociation device, an ultraviolet photon dissociation device, or a free radical dissociation device. Preferably, the ion dissociation device 4 is a collision-induced dissociation device.

[0068] The mass spectrometer 5 can be one or more of a quadrupole mass spectrometer, a time-of-flight mass spectrometer, a Fourier transform mass spectrometer, an ion trap mass spectrometer, and a magnetic mass spectrometer. Optionally, the mass spectrometer 5 is a quadrupole mass spectrometer or a time-of-flight mass spectrometer, and together with the quadrupole mass filter 3, forms a tandem mass spectrometer in the form of a Q-TOF or triple quadrupole.

[0069] In this embodiment, the ion mobility spectrometer-mass spectrometer 100 is further provided with an ion optical device 6 between the ion source 1 and the U-shaped ion mobility analyzer 2, and between the U-shaped ion mobility analyzer 2 and the quadrupole mass filter 3, for performing functions such as ion convergence, guidance, or storage. It should be noted that the multi-stage vacuum structure and the corresponding configuration of the ion optical device 6 in FIG1 are merely exemplary. In other embodiments of the present invention, the configuration of the multi-stage vacuum structure and the ion optical device 6 can be adjusted according to actual needs, and the present invention is not limited thereto.

[0070] <Data Collection Method>

[0071] Referring to FIG3 , the data acquisition method provided by the present invention comprises the following steps:

[0072] In the pre-collection step S1 , the parent ions obtained by ionization in the ion source 1 are sequentially passed through the ion mobility analyzer 2 and the mass spectrometer 5 to obtain a two-dimensional thermal map of the ion mobility spectrum and mass spectrum of the parent ions.

[0073] Figure 4 shows an example of a two-dimensional ion mobility spectrum-mass spectrum heat map obtained in pre-acquisition step S1. After pre-acquisition step S1, this ion mobility spectrum-mass spectrum heat map is analyzed to identify trend lines. This trend line allows for more precise differentiation of substances of different charge numbers and types, enabling more targeted selection of target precursor ions for DDA analysis and more accurate determination of mobility and mass selection ranges for DIA analysis.

[0074] In addition, in the pre-collection step S1, the intensity information and mobility information of the parent ion can also be obtained. The intensity information of the parent ion is required in the implementation of DDA analysis, and the mobility information of the parent ion can be used to calculate the collision cross section area CCS to assist qualitative analysis.

[0075] After completing the pre-acquisition step S1 and determining the trend line, the parent ion selection and cascade analysis are performed. Specifically, first, based on the ion mobility spectrum-mass spectrum two-dimensional heat map and its trend line, the mobility selection range of the ion mobility analyzer 2 and the mass selection range of the quadrupole mass filter 3 in different time periods within a cycle are determined.

[0076] Then, the parent ion selection step S2 and the daughter ion collection step S3 are repeatedly performed to obtain ion mobility spectrum-mass spectrum data of multiple groups of daughter ions corresponding to the parent ions.

[0077] Among them, the parent ion collection step S2 is to configure the ion mobility analyzer 2 to the filtering mode, select the parent ion according to the ion mobility spectrum-mass spectrum two-dimensional thermal map, and set the mobility selection range of the ion mobility analyzer 2 and the mass selection range of the quadrupole mass filter 3 accordingly.

[0078] After the parent ion collection step S2, or simultaneously with the parent ion collection step S2, the daughter ion collection step S3 is performed. In the daughter ion collection step S3, the selected parent ions enter the ion dissociation device 4 for dissociation to produce daughter ions, which then enter the mass spectrometry device 5 for analysis to obtain ion mobility spectrum-mass spectrum data of the daughter ions.

[0079] After all parent ions have been selected and the corresponding daughter ion collection step S3 has been performed, all of the above parent ion collection steps S2 and daughter ion collection steps S3 may be performed again one or more times. In particular, in the repeated collection steps, certain parameter settings may be changed compared to the previous selection and collection, such as changing the mobility selection range or window size, or the mass selection range or window size, for parent ion selection, or changing the order of parent ion selection, or changing the collision energy or retention time (dwell time) of the ion dissociation device 4, etc., so that more information can be obtained by changing the parameters to improve the coverage and depth of the analysis.

[0080] <Mobility selection range and quality selection range>

[0081] In the present embodiment, mobility selection range and quality selection range can be determined according to the analytical method used by coupling instrument, when for example coupling instrument adopts DDA data analysis method, with reference to Figure 5, it is possible to select many groups of parent ions in ion mobility spectrum-mass spectrum two-dimensional heat map where signal intensity is higher than threshold value, or, select many groups of parent ions in ion mobility spectrum-mass spectrum two-dimensional heat map where signal intensity is relatively high. That is, select several groups of parent ions with the strongest or relatively strong signal intensity to analyze. In the process of selecting parent ion, trend line can provide reference information for the charge situation of parent ion, so that parent ion (such as single-charged parent ion is removed) can be selected more accurately. Of course, in some other embodiments, the relatively low parent ion in abundance can also be selected to be analyzed as target.

[0082] Preferably, when using DDA data analysis method, refer to Figure 6. The number 1-17 in Figure 6 is the number of parent ions selected (17 kinds of parent ions in this example, i.e. top17), and for the curve as shown in Figure 5 of trend line, there is a situation of fluctuation. In an embodiment of the present invention, because the UMA operating in filter mode can freely select mobility window, it is possible to achieve the curve setting mobility selection range and quality selection range following fluctuation, and do not affect scanning speed. Specifically, mobility selection range and / or quality selection range may also have fluctuation or callback, such as when the overall system has a trend, in some areas, the trend is opposite, which is also one of the advantages of filter mode, i.e., window scanning does not need to be unidirectional and the size of window may also be dynamically changed and adjusted. In the present embodiment, since migration spectrum and subsequent quadrupole all operate in filter mode, the size of mobility window and quality window may be dynamically changed according to heat map, and scanning line may be along a straight line, or along a broken line or curve. This scanning mode may improve the coverage for point of interest / region in omics analysis. Similarly, the ion mobility spectrometer-mass spectrometer 100 coupled with the UMA operating in the filtering mode can also realize a data acquisition method in which the selection of the mobility selection range in a partial region is opposite to the overall change trend.

[0083] For another example, when using DIA data analysis, referring to Figure 7(a), fixed-width mass windows and ion mobility windows can be selected stepwise, and the period of scanning the mobility selection range by the ion mobility analyzer 2 is synchronized with the period of scanning the mass selection range by the quadrupole mass filter 3. The advantages of using a filtering mode for ion mobility spectrum scanning during DIA acquisition are twofold: a higher dynamic range and the ability to select a wider mobility window, increasing scanning speed. Furthermore, due to the mass window filtering properties of the quadrupole mass filter, the mass window can be relatively wide (e.g., 25 Da) and slid during the scan, meaning that the mass window in the next parent ion selection step overlaps with the current one. This allows for "oversampling" of ions, thereby improving the coverage of the DIA analysis. A more important benefit of "oversampling" is that it can obtain more information. For example, based on the intensity distribution of ions within several adjacent windows, the mass range of the parent ion can be further confirmed. This narrows the actual mass window of the parent ion, reducing the difficulty of subsequent DIA processing. Specifically, the ion mobility analyzer 2 scans and releases ions of different mobilities in a sequence from high to low mobility (i.e., collision cross-section area (CCS) from large to small) within a single analysis cycle. The quadrupole mass filter 3 scans sequentially from small to large mass windows within a single analysis cycle. The complexity of substances in omics analysis is typically high, and a single scan cannot provide adequate coverage of the ion mobility spectrum-mass spectrometry heat map. Therefore, multiple scans are typically required, such as the second and third scans shown in Figure 7(b), with each scan covering a different region of the heat map to ultimately achieve good coverage.

[0084] Similarly, during DIA analysis, the size of the mobility window and mass window in the precursor ion selection step, as well as the shape of the scan line, can be dynamically changed in real time to further improve the efficiency and speed of the scan.

[0085] In addition, when performing the daughter ion collection step S3, since smaller parent ions usually correspond to smaller daughter ions, and the retention time of small daughter ions in the ion dissociation device 4 is also shorter, their transmission in the ion dissociation device 4 is not easily interfered with by subsequent larger daughter ions, and therefore will not affect the quality of their migration spectrum (such as avoiding migration spectrum peak mismatch, etc.), and can also speed up the channel scanning speed of DDA or DIA.

[0086] Preferably, when adopting DIA data analysis method, refer to Figure 8 (a). Because ion mobility analyzer 2 operates in filtering mode in the embodiment of the present invention, its mobility window size can be arbitrarily selected, preferably wider mobility window and slide this window in scanning, that is, the mobility window (and / or mobility selection range) in the next parent ion selection step overlaps with the mobility window (and / or mobility selection range) of this time. Synchronously coordinating scanning with the mass window " oversampling " in above-mentioned Fig. 6 can further improve the efficiency of " oversampling ", and therefore can further improve the coverage of DIA analysis. In addition, adopting wider mobility window, the scanning speed of mobility can be improved, although this often means lower migration spectrum resolution, but as mentioned above, more information about mobility range can be obtained during mobility " oversampling", that is, by the intensity distribution of parent ions in adjacent windows, determining and narrowing the mobility range of parent ions, so that the migration spectrum resolution of loss can be compensated. This mode can be realized by collocation U-type ion mobility analyzer, and can not be realized in TIMS and other prior art. Furthermore, Figure 8(b) shows that during the second and third scans, the mobility window still overlaps with the first scan, which allows for multiple “oversampling” to maximize analysis coverage.

[0087] Preferably, when using the DIA data analysis method, refer to Figure 9. In this example, the first scan still uses the oversampling mode of Figure 8(a), but during the second scan, the mobility window does not substantially overlap with the first scan. This approach slightly reduces the degree of "oversampling" (similar to that in Figure 6(b) and, therefore, similar to that in Figure 6(b)), but significantly reduces the number of scans, thereby increasing the overall DIA analysis speed and the sampling frequency of the mass spectrometer on the chromatogram.

[0088] Preferably, when using the DIA data analysis method, refer to Figures 10(a) and (b). In this example, during the first scan (Figure 10(a)), within the adjacent mobility-mass selection regions, the mobility windows overlap, but the mass windows do not. That is, the mobility window slides, but the mass window jumps; similarly, during the second and third scans, the mobility window may or may not overlap with the previous scan (indicated as overlapping in Figure 10(b)). This scanning method can still maintain fairly good coverage; at the same time, since the mass windows do not overlap in this method, the complexity of the product ion spectrum is reduced, thereby partially reducing the difficulty of data post-processing.

[0089] In some embodiments, in the parent ion selection step S2, the mass selection range is the full mass range, and remains unchanged in each daughter ion collection step S3. This data collection method passes all parent ions into the ion dissociation device 4 for fragmentation in one cycle, and performs data analysis on all daughter ions, which can efficiently collect data applicable to full mass range DIA analysis methods (such as MS E Method) data.

[0090] <Advantages of the Data Acquisition Method with a U-Type Ion Mobility Analyzer>

[0091] Particularly, in this embodiment, the ion mobility analyzer 2 is a U-shaped ion mobility analyzer 2 .

[0092] First, in the U-shaped ion mobility analyzer 2 working in the filtering mode, ions in different ion mobility ranges can be released to the lower level in a continuous manner, one after another, end to end, to obtain a continuous ion flow. In theory, there is no dynamic range limitation, which can effectively improve the dynamic range and avoid the dynamic range being limited by the ion trap capacity.

[0093] It should be noted that differential ion mobility spectrometry (DMS) or high-field asymmetric waveform ion mobility spectrometry (FAIMS), which measure differential ion mobility, can also be used in conjunction with mass spectrometry. Their acquisition modes for DDA or DIA are well known in the industry, and DMS or FAIMS also operate in "filter mode." However, in the present invention, ion mobility spectrometry is limited to measuring ion mobility at low fields, not differential ion mobility. Problems with DMS or FAIMS when used in conjunction with mass spectrometry and in such acquisition modes include low resolution, low ion utilization efficiency, and an inability to provide collision cross-section information, thus providing very limited assistance for proteomics analysis.

[0094] Specifically, the ion mobility spectrometer-mass spectrometer 100 provided in embodiments of the present invention has a higher resolution than both DMS and FAIMS. For example, during DDA or DIA analysis, even with a relatively fast full-spectrum scan of 100 ms, the resolution is above 60. This mobility spectrometry separation capability not only enables the generation of highly accurate CCS values ​​but also greatly facilitates the analysis of complex protein samples. Conventional DMS and FAIMS, on the other hand, have very low resolution and are unable to provide collision cross section information.

[0095] In addition, because the ion mobility spectrometry-mass spectrometry two-dimensional heat map is first collected during the analysis process, it is also beneficial for the valence analysis of protein or peptide ions. Since the mobility is related to the charge number of the ion, ions of different valence states will be distributed in different areas of the two-dimensional heat map. For DDA or DIA, protein or peptide ions with a valence of +2 or higher are usually selected for cascade analysis; especially for DDA, it is more necessary to select the parent ions with a valence of +2 and higher in real time. Therefore, the collection of the ion mobility spectrometry-mass spectrometry two-dimensional heat map makes the selection of ions of different valence states more convenient and accurate. As for DMS and FAIMS, although there are reports that DMS or FAIMS can also filter out +1 ions under specific working conditions, its selectivity and accuracy are much lower than the method of using ion mobility under low field for selection.

[0096] Ion mobility analyzers working in "filter mode", such as DMA (differential mobility analyzer, note that although the name contains differential, it does not actually measure differential ion mobility, and the name is derived for historical reasons), use orthogonal airflow and electric field to separate ions, but the resolution is usually low and high ion utilization efficiency cannot be obtained. In the present invention, a U-shaped ion mobility analyzer UMA (U-shaped mobility analyzer) is preferred, which can measure ion mobility under low field and has high mobility spectrum resolution and ion utilization efficiency. Secondly, the U-shaped ion mobility analyzer 2 working in filter mode can form a mobility window that allows ions to pass through by setting the field strength difference between the two channels. Due to the flexibility of the electric field setting, different ion mobility windows can be scanned step by step as described above, or different ion mobility windows can be skipped (for example, in the DDA method, a specific ion mobility or ion mobility window is selected based on the ion mobility of the parent ion), which can improve the scanning efficiency and the spectral quality of the ion mobility spectrum.

[0097] Furthermore, as disclosed in patent CN115223844A, the U-shaped ion mobility analyzer 2 can store ions in different areas by controlling the balance between the electric field and the airflow. Specifically, in the parent ion selection step S2, the parent ions that are not within the current mobility selection range are at least partially stored in the ion storage area of ​​the ion mobility analyzer 2. When the parent ion selection step S2 is repeated, the parent ions in at least part of the ion storage area are selected and released to the ion dissociation device 4. By temporarily storing ions in the ion storage area of ​​the ion mobility analyzer 2 and then releasing them to the ion dissociation device 4 at the appropriate time, the ion utilization efficiency can be effectively improved (theoretically, the ion utilization efficiency can reach 100%), thereby improving the duty cycle of the coupled instrument.

[0098] Specifically, referring to Figure 11 , the U-shaped ion mobility analyzer 2 can scan the electric field strength E(t) within the ion mobility analyzer 2 during an operating cycle from t0 to t1, thereby allowing ions of different mobilities to pass through the ion mobility analyzer 2 in sequence. In filter-scan mode, the ion mobility analyzer 2 operates periodically. This description uses the DC electric field strength E0-E1 corresponding to the full mobility range as an example.

[0099] At the beginning of each cycle, at time t=t0, the DC electric field E of the first channel CH1 is S1 The configuration shown in Figure 11(a) is generally low, equal to E0, with no electric field gradient. This is because the electric field force within the entire first channel CH1 is relatively low. Ions of nearly the entire mobility range can migrate longitudinally within the first channel CH1 toward the ion transfer port 202. The ion transfer port 202 remains open, continuously transporting ions of nearly the entire mobility range to the second channel CH2. The DC electric field in the second channel CH2 is configured to be relatively low on the left side, equal to E0 + ΔE, and then suddenly increase to an electric field distribution of E1 on the right side of the ion transfer port 202 (the first ion storage region 204). This electric field configuration ensures that only ions with high mobility (i.e., ions with a small collision cross section CCS) entering the second channel CH2 are transported longitudinally through the second channel CH2 and to the next stage via the ion outlet 203. All other ions with larger collision cross sections CCS are temporarily stored in the first ion storage region 204 to the right of the ion transfer port 202.

[0100] Next, referring to FIG11(b), when synchronously raising E S1 and E S2 During the process, the electric field strength of the main part of the first channel CH1 and the second channel CH2 is always kept at a difference of ΔE, so as to scan the full mobility range with a specific mobility window. S1 and E S2 The increase can be a synchronous gradual increase or a synchronous step-by-step increase. While the electric field strength of the main body increases, the electric field strength on the right side of the first ion storage region 204 is stably maintained at E1, and the electric field strength on the left side of the second ion storage region 205 is stably maintained at E0. Therefore, the electric field strength gradient within the first ion storage region 204 decreases, and the corresponding covered ion mobility range decreases. At t=t0, ions with a larger CCS that are not within the target mobility range will enter the target mobility range due to mobility scanning, and can move to the left along the length direction and be transported to the subsequent stage through the ion outlet 203. The electric field strength gradient within the second ion storage region 205 increases, and the corresponding covered ion mobility range increases, gradually allowing ions with a larger CCS range to be stored in the second ion storage region 205.

[0101] In some embodiments, ΔE can also be 0. The size of ΔE determines the size of the mobility window of the migration spectrum, and its value is related to the resolution of the migration spectrum and the ion pass rate. The smaller ΔE is, the higher the resolution is, but the pass rate is often lower. In an actual scanning process, due to the continuous enhancement of the field strength, the field strength experienced by the ions in the first channel CH1 and the second channel CH2 is not the same, so ΔE is not necessarily set to a positive value. When ΔE = 0, or even a negative value close to 0, the ions still have a certain pass rate. Experiments have found that ΔE = 0 (that is, the mobility window is set to 0) can achieve a higher resolution, but still have a higher ion pass efficiency, and can obtain more accurate mobility and CCS values, which is beneficial for the identification of compounds such as proteins and peptides.

[0102] Referring to FIG11(c), at t=t1, only the ions with the largest CCS can pass through the ion mobility analyzer 2, while the remaining ions with smaller CCS are stored in the second ion storage region 205 of the first channel CH1. At t=t0 in the next cycle, due to the re-application of the DC electric field shown in FIG11(a), almost all ions in the second ion storage region 205 are rapidly transferred to the second channel CH2. Among them, ions that meet the target mobility range corresponding to the electric field shown in FIG11(a) can be transported to the subsequent stage through the second channel CH2 and the ion outlet 203, while most of the other ions are transferred to the first ion storage region 204.

[0103] Because ions that are not within the target mobility range can still be stored in the first ion storage area 204 or the second ion storage area 205, and the stored ions can be released for utilization at the appropriate time, ions within the entire mobility range can basically be efficiently utilized, almost reaching 100% ion utilization efficiency, and achieving a higher dynamic range.

[0104] The stored ion operation mode described in Figure 11 can be applied to both DDA and DIA data acquisition. For example, in the DDA acquisition mode shown in Figure 6, when a specific precursor ion is selected (i.e., the UMA selects a specific mobility window, and the quadrupole selects a corresponding mass window), other unselected precursor ions can be temporarily stored in the UMA's ion storage area. When the precursor ion selection step is repeated, the previously stored precursor ions are reselected and released. This allows for a 100% duty cycle within a single DDA scan.

[0105] Similarly, the DIA acquisition methods shown in Figures 7 through 10 can also be operated in a similar manner to achieve a 100% duty cycle within a single UMA scan. For the entire DIA acquisition process, the duty cycle is determined by the ratio of the area covered by a single UMA scan to the area covered by the entire acquisition process, so the duty cycle is generally less than 100%. However, due to the use of a filtering mode with a wider mobility window (as shown in Figure 9), the duty cycle is still higher than that of the TIMS PASEF mode.

[0106] The above are only preferred embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A data acquisition method, applied to an ion mobility spectrometer-mass spectrometer, characterized in that: The ion mobility spectrometer-mass spectrometer comprises an ion source, an ion mobility analyzer, a quadrupole mass filter, an ion dissociation device and a mass spectrometer connected in sequence, and the data acquisition method comprises the following steps: A pre-collection step, wherein the parent ions obtained by ionization of the ion source are sequentially passed through the ion mobility analyzer and the mass spectrometer to obtain a two-dimensional heat map of the ion mobility spectrum-mass spectrum of the parent ions; A parent ion selection step, configuring the ion mobility analyzer to a filtering mode, selecting parent ions according to the ion mobility spectrum-mass spectrum two-dimensional heat map, and correspondingly setting the mobility selection range of the ion mobility analyzer and the mass selection range of the quadrupole mass filter; In the daughter ion collection step, the selected parent ions enter the ion dissociation device for dissociation to generate daughter ions, and the daughter ions enter the mass spectrometry device for analysis to obtain ion mobility spectrum-mass spectrum data of the daughter ions. Repeat the parent ion selection step and the daughter ion collection step, and set a different mobility selection range or a different mass selection range from the previous step in each repeated step to obtain ion mobility spectrum-mass spectrum data of daughter ions corresponding to multiple groups of parent ions.

2. The data collection method according to claim 1, characterized in that: The mobility selection range of the ion mobility analyzer in the parent ion selection step overlaps with the mobility selection range in the next repeated parent ion selection step.

3. The data collection method according to claim 1, characterized in that: The mass selection range of the quadrupole mass filter in the parent ion selection step overlaps with the mass selection range in the next repeated parent ion selection step.

4. The data collection method according to claim 1, characterized in that: The parent ion selection step, the daughter ion collection step, and the repeated parent ion selection step and daughter ion collection step are recorded as the first scan; then the parent ion selection step, the daughter ion collection step, and the repeated parent ion selection step and daughter ion collection step are performed (N-1) times, and recorded as the second, third and Nth scans; wherein in at least two scans that occur successively, the mobility selection ranges of the parent ion selections corresponding to the same mass selection range overlap.

5. The data collection method according to claim 1, characterized in that: The ion mobility analyzer is a U-type ion mobility analyzer.

6. The data collection method according to claim 1, characterized in that: The period of scanning the mobility selection range by the ion mobility analyzer is synchronized with the period of scanning the mass selection range by the quadrupole mass filter; The ion mobility analyzer scans a plurality of selected mobility ranges in a manner that the mobility at least partially changes from high to low within one analysis cycle.

7. The data collection method according to claim 6, characterized in that: The ion mobility analyzer scans a plurality of mobility selection ranges in a manner of mobility from high to low, and the quadrupole mass filter scans a plurality of mass selection ranges in a manner of mass-to-charge ratio from small to large.

8. The data collection method according to claim 6, characterized in that: In the precursor ion selection step, the mass selection range of the quadrupole mass filter is a mass window with a fixed width.

9. The data collection method according to claim 1, characterized in that: In the parent ion selection step, parent ions that are not within the current mobility selection range are at least partially stored in the ion storage area of ​​the ion mobility analyzer. While repeating the precursor ion selection step, at least a portion of the precursor ions in the ion storage region are selected and released to the ion dissociation device.

10. The data collection method according to claim 1, characterized in that: The mobility selection range of the ion mobility analyzer in the parent ion selection step is different from the mobility selection range in the next repeated parent ion selection step; or, the mass selection range of the quadrupole mass filter in the parent ion selection step is different from the mass selection range in the next repeated parent ion selection step.

11. The data collection method according to claim 1, characterized in that: In the parent ion selection step, multiple groups of parent ions whose signal intensities are higher than a threshold value in the ion mobility spectrum-mass spectrometry two-dimensional heat map are selected, or multiple groups of parent ions whose signal intensities are relatively high in the ion mobility spectrum-mass spectrometry two-dimensional heat map are selected.

12. The data collection method according to claim 1, characterized in that: In the precursor ion selection step, the mass selection range is the full mass range and remains unchanged in each product ion collection step.

13. The data collection method according to claim 1, characterized in that: In the pre-collection step, the ion mobility analyzer operates in a filtering mode or a trapping mode.

14. An ion mobility spectrometer-mass spectrometer, using the data acquisition method according to any one of claims 1 to 13.

15. The ion mobility spectrometer-mass spectrometer according to claim 14, characterized in that: The mass analysis device is one or more of a quadrupole mass analysis device, a time-of-flight mass spectrometer, a Fourier transform mass spectrometer, an ion trap mass spectrometer, and a magnetic mass spectrometer.

16. The ion mobility spectrometer-mass spectrometer according to claim 14, characterized in that: The ion dissociation device is one or more of a collision induced dissociation device, an electron dissociation device, an infrared photon dissociation device, an ultraviolet photon dissociation device, and a free radical dissociation device.

Images