WO2017017787A1 - タンデム型質量分析装置 - Google Patents
タンデム型質量分析装置 Download PDFInfo
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- WO2017017787A1 WO2017017787A1 PCT/JP2015/071382 JP2015071382W WO2017017787A1 WO 2017017787 A1 WO2017017787 A1 WO 2017017787A1 JP 2015071382 W JP2015071382 W JP 2015071382W WO 2017017787 A1 WO2017017787 A1 WO 2017017787A1
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- H—ELECTRICITY
- H01—ELECTRIC ELEMENTS
- H01J—ELECTRIC DISCHARGE TUBES OR DISCHARGE LAMPS
- H01J49/00—Particle spectrometers or separator tubes
- H01J49/004—Combinations of spectrometers, tandem spectrometers, e.g. MS/MS, MSn
- H01J49/0045—Combinations of spectrometers, tandem spectrometers, e.g. MS/MS, MSn characterised by the fragmentation or other specific reaction
- H01J49/0072—Combinations of spectrometers, tandem spectrometers, e.g. MS/MS, MSn characterised by the fragmentation or other specific reaction by ion/ion reaction, e.g. electron transfer dissociation, proton transfer dissociation
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/62—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating the ionisation of gases, e.g. aerosols; by investigating electric discharges, e.g. emission of cathode
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- H—ELECTRICITY
- H01—ELECTRIC ELEMENTS
- H01J—ELECTRIC DISCHARGE TUBES OR DISCHARGE LAMPS
- H01J49/00—Particle spectrometers or separator tubes
- H01J49/0027—Methods for using particle spectrometers
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- H—ELECTRICITY
- H01—ELECTRIC ELEMENTS
- H01J—ELECTRIC DISCHARGE TUBES OR DISCHARGE LAMPS
- H01J49/00—Particle spectrometers or separator tubes
- H01J49/004—Combinations of spectrometers, tandem spectrometers, e.g. MS/MS, MSn
- H01J49/0045—Combinations of spectrometers, tandem spectrometers, e.g. MS/MS, MSn characterised by the fragmentation or other specific reaction
- H01J49/005—Combinations of spectrometers, tandem spectrometers, e.g. MS/MS, MSn characterised by the fragmentation or other specific reaction by collision with gas, e.g. by introducing gas or by accelerating ions with an electric field
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- H—ELECTRICITY
- H01—ELECTRIC ELEMENTS
- H01J—ELECTRIC DISCHARGE TUBES OR DISCHARGE LAMPS
- H01J49/00—Particle spectrometers or separator tubes
- H01J49/02—Details
- H01J49/06—Electron- or ion-optical arrangements
- H01J49/062—Ion guides
- H01J49/063—Multipole ion guides, e.g. quadrupoles, hexapoles
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- H—ELECTRICITY
- H01—ELECTRIC ELEMENTS
- H01J—ELECTRIC DISCHARGE TUBES OR DISCHARGE LAMPS
- H01J49/00—Particle spectrometers or separator tubes
- H01J49/26—Mass spectrometers or separator tubes
- H01J49/34—Dynamic spectrometers
- H01J49/42—Stability-of-path spectrometers, e.g. monopole, quadrupole, multipole, farvitrons
- H01J49/4205—Device types
- H01J49/421—Mass filters, i.e. deviating unwanted ions without trapping
- H01J49/4215—Quadrupole mass filters
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- H—ELECTRICITY
- H01—ELECTRIC ELEMENTS
- H01J—ELECTRIC DISCHARGE TUBES OR DISCHARGE LAMPS
- H01J49/00—Particle spectrometers or separator tubes
- H01J49/26—Mass spectrometers or separator tubes
- H01J49/34—Dynamic spectrometers
- H01J49/42—Stability-of-path spectrometers, e.g. monopole, quadrupole, multipole, farvitrons
- H01J49/4205—Device types
- H01J49/422—Two-dimensional RF ion traps
- H01J49/4225—Multipole linear ion traps, e.g. quadrupoles, hexapoles
Definitions
- CID Collision-Induced Dissociation
- the present invention relates to a mass spectrometer.
- MS / MS analysis which is a method of mass spectrometry, is a useful method for identifying compounds with large molecular weights and analyzing their chemical structures, and has been widely used in various fields in recent years. Yes.
- an ion having a specific mass-to-charge ratio derived from the target compound is selected as a precursor ion among various ions generated from a sample.
- the precursor ions are cleaved by a technique such as contacting with CID gas, and the product ions generated thereby are separated and detected according to the mass-to-charge ratio.
- a mass spectrum (product ion spectrum) indicating the relationship between the mass-to-charge ratio of product ions and the intensity is acquired.
- the product ion peak pattern observed in the mass spectrum and, in some cases, the neutral loss obtained from the mass-to-charge ratio difference between the two peaks are used to estimate the chemical structure of the target compound.
- a well-known mass spectrometer that performs MS / MS analysis is a triple quadrupole mass spectrometer in which a quadrupole mass filter is arranged before and after a collision cell that performs CID.
- the so-called Q-TOF mass spectrometer which replaces the subsequent quadrupole mass filter with a time-of-flight mass analyzer in a triple quadrupole mass spectrometer, is compared to a triple quadrupole mass spectrometer.
- the structure is complicated and expensive, a more accurate mass spectrum can be obtained.
- a mass spectrometer provided with a mass analyzer before and after the collision cell is referred to as a tandem mass spectrometer.
- a mass spectrometer described in Patent Document 1 is used as a device for acquiring a mass spectrum in which various types of product ions generated by cleavage under different collision energies are observed with a small number of analyzes.
- a mass spectrometer described in Patent Document 1 is used.
- an ion trap that temporarily captures ions is provided between the collision cell and the subsequent mass separator (time-of-flight mass analyzer), and is generated by cleavage under different collision energies at different times.
- the accumulated product ions are temporarily accumulated in the ion trap, and then the accumulated ions are discharged all at once from the ion trap and subjected to mass analysis with a time-of-flight mass spectrometer to obtain a mass spectrum.
- the amount of active ions is quite limited.
- the signal intensity per product ion will be sufficiently high, but in general, the more complex the chemical structure of a compound, the more the product Since the number of types of ions increases, the signal intensity per product ion is relatively low, which is disadvantageous for qualitative and structural analysis.
- an object of the present invention is to obtain a mass spectrum in which many types of product ions derived from one compound are observed with high sensitivity. Therefore, an object of the present invention is to provide a tandem mass spectrometer capable of improving the accuracy of compound identification and structural analysis.
- the tandem mass spectrometer which has been made to solve the above problems, has an ion source for ionizing a compound in a sample, and a specific mass-to-charge ratio among the various ions generated.
- a first mass separation unit that sorts ions having precursor ions as precursor ions; an ion cleavage unit that cleaves the precursor ions by collision-induced dissociation; a second mass separation unit that performs mass analysis of various product ions generated by the cleavage and detection
- the first mass separation unit, the ion cleavage unit, or the second mass separation unit is operated as an ion storage unit, or the first mass separation unit and the ion Ions can be temporarily accumulated by an ion accumulating unit provided between the cleaving unit or between the ion cleaving unit and the second mass separation unit.
- tandem mass spectrometer In tandem mass spectrometer, a) a collision energy setting unit for changing the collision energy applied to the precursor ion at the time of collision-induced dissociation in the ion cleavage part in m steps (where m is an integer of 2 or more); b) a mass-to-charge ratio range setting unit for changing the mass-to-charge ratio range of ions accumulated in the ion accumulating unit to n stages (where n is an integer of 2 or more) shifted in the mass-to-charge ratio direction; c) One of the m stages of collision energy is set by the collision energy setting unit, and one of the n stages of mass to charge ratio range is set by the mass to charge ratio range setting unit.
- each part is controlled so as to perform mass analysis of product ions with respect to a predetermined precursor ion under a combination of the mass-to-charge ratio range, and the combination of collision energy and mass-to-charge ratio range
- An analysis control unit that controls the operation of each unit so that mass analysis of product ions is performed at least once for all combinations of m ⁇ n that differ from each other; d)
- One mass spectrum over a predetermined mass-to-charge ratio range is created by integrating the spectrum data obtained by mass analysis of product ions performed at least m ⁇ n times under the control of the analysis control unit.
- a data processing unit It is characterized by having.
- the tandem mass spectrometer which has been made to solve the above problems, includes an ion source that ionizes a compound in a sample, and a specific mass-to-charge ratio among the various ions generated.
- a first mass separation unit that sorts out ions having a precursor ion as a precursor ion; an ion cleavage unit that cleaves the precursor ion by collision-induced dissociation; a second mass separation unit that performs mass analysis of various product ions generated by the cleavage;
- a detector by operating any one of the first mass separation unit, the ion cleavage unit, or the second mass separation unit as an ion accumulation unit, or the first mass separation unit and the Ions can be temporarily stored by the ion storage part provided between the ion cleavage part or between the ion cleavage part and the second mass separation part.
- a collision energy setting unit for changing the collision energy applied to the precursor ion at the time of collision-induced dissociation in the ion cleavage part in m steps (where m is an integer of 2 or more); b) a mass-to-charge ratio range setting unit for changing the mass-to-charge ratio range of ions accumulated in the ion accumulating unit to n stages (where n is an integer of 2 or more) shifted in the mass-to-charge ratio direction; c) One of the m stages of collision energy is set by the collision energy setting unit, and one of the n stages of mass to charge ratio range is set by the mass to charge ratio range setting unit.
- a predetermined mass-to-charge ratio range is obtained by integrating the spectrum data obtained by mass analysis of the product ions that have been performed the number of times greater than the larger value of m or n under the control of the analysis control unit.
- a data processing unit for creating one mass spectrum covering It is characterized by having.
- the mass separation method in the first mass separation unit and the second mass separation unit is not particularly limited, but typically, a quadrupole mass filter, a first mass separation unit, A time-of-flight mass separator may be used as the two mass separator.
- the ion accumulating unit is a three-dimensional quadrupole ion trap or a linear ion trap.
- a predetermined voltage is applied substantially to a rod electrode (or a part of the rod electrode) constituting a quadrupole or a multipole.
- a function equivalent to that of a linear ion trap may be provided.
- the frequency and amplitude of the high-frequency voltage applied to the electrodes constituting the ion trap are changed to form a high-frequency electric field for trapping ions. By doing so, the mass-to-charge ratio range in which ions are stably trapped can be changed.
- the collision energy setting unit is a rod-shaped electrode constituting the quadrupole mass filter.
- the collision energy can be changed by changing the voltage difference between the direct bias voltages applied to the rod-shaped electrodes constituting the ion guides arranged in the collision cell.
- the mass-to-charge ratio range setting unit as described above, the frequency and amplitude of the high-frequency voltage applied to the electrodes constituting the ion trap. Etc. can be changed to change the mass-to-charge ratio range of the accumulated ions. Since both the collision energy and the mass-to-charge ratio range can be changed by changing the voltage, the change can be performed very quickly.
- the analysis control unit performs mass analysis of product ions with respect to a predetermined precursor ion for all combinations of m-stage collision energy and m-stage mass-to-charge ratio range. Each part is controlled to implement.
- the collision energy and the mass-to-charge ratio range setting unit set by the collision energy setting unit are set.
- the mass-to-charge ratio range is fixed, and the first mass separation unit is operated so that ions having a specific mass-to-charge ratio derived from the target compound are selected as precursor ions.
- the precursor ions selected by the first mass separation unit are given predetermined collision energy and reach the ion cleavage unit, and are cleaved by collision-induced dissociation to generate various product ions.
- Product ions enter, for example, an ion accumulating unit disposed between the ion cleavage unit and the second mass separation unit.
- the ion accumulation unit accumulates incident product ions therein for a predetermined time width. Since the collision energy and the mass-to-charge ratio range are fixed during the ion accumulation period, only ions that fall within a predetermined mass-to-charge ratio range among the determined types of product ions are accumulated in the ion accumulation unit.
- this one-time mass analysis provides data showing the product ion spectrum of only a predetermined mass-to-charge ratio range, which is derived from one precursor ion and generated by collision-induced dissociation under a specific collision energy. It is done.
- a predetermined mass-to-charge ratio range which is derived from one precursor ion and generated by collision-induced dissociation under a specific collision energy. It is done.
- the mass-to-charge ratio range is limited, so there are not many types of product ions that can be stored, and the number of ions per type of product ion is sufficient. Can be ensured, and ions can be detected with sufficient intensity.
- the ion intensity of the spectrum data obtained for each mass analysis is sufficiently high.
- the data processing unit integrates such spectrum data and creates a product ion spectrum over a wide mass-to-charge ratio range. Even if the precursor ions are the same, if the collision energy is different, the type of product ions to be generated and the generation ratio thereof greatly change. In the tandem mass spectrometer of the first aspect, product ions generated under different collision energies can be comprehensively detected over a wide mass-to-charge ratio range. Accordingly, in the product ion spectrum created by the data processing unit, various product ions derived from one target compound are all observed with sufficient sensitivity.
- mass analysis for one combination of collision energy and mass-to-charge ratio range may be executed a plurality of times. That is, for example, all m ⁇ n combinations with different combinations of collision energy and mass-to-charge ratio range were executed L times (where L is an integer of 2 or more), and the m ⁇ n ⁇ L times were executed.
- a product ion spectrum may be obtained by integrating spectral data obtained by mass spectrometry.
- tandem mass spectrometer unlike the first aspect, at least one combination among all combinations of m ⁇ n collision energy and mass-to-charge ratio range. Omit product ion mass spectrometry for. This is because if there is a combination of collision energy and mass-to-charge ratio range where it is assumed that the number of observed product ions is low or very low, performing mass analysis under such conditions is time consuming. Because it is useless.
- the analysis control unit may control each unit so that mass analysis of product ions for such combinations is not performed.
- the result of executing mass analysis the same number of times for all combinations of collision energy and mass-to-charge ratio range is reflected in one mass spectrum.
- the number of executions of mass spectrometry may be different for each combination. That is, for the combination of collision energy and mass-to-charge ratio range that is presumed to yield product ions with high importance for structural analysis of compounds, the number of times of mass analysis is increased compared to other combinations. It is recommended that product ions with high importance are observed with high sensitivity.
- tandem mass spectrometer of the first and second aspects of the present invention a mass spectrum in which product ions in a wide mass-to-charge ratio range derived from one compound are observed with high sensitivity can be efficiently acquired. be able to. Thereby, many pieces of fragment information about the compound, that is, mass information of product ions and neutral loss can be obtained, so that the qualitative and structural analysis accuracy of the compound can be improved.
- tandem mass spectrometer of the second aspect of the present invention it is possible to reduce the time required for the measurement for obtaining the mass spectrum as described above, so that, for example, GC-MS or LC-MS Thus, even when the time devoted to the measurement of ions derived from one compound is short, a mass spectrum having sufficient quality for qualitative and structural analysis of the compound can be obtained. Therefore, it is also suitable for multi-component simultaneous analysis using GC-MS or LC-MS.
- the block diagram of the principal part of the tandem-type mass spectrometer which is one Example of this invention. Explanatory drawing of an example of MS / MS analysis mode in the tandem-type mass spectrometer of a present Example. Explanatory drawing of another example of MS / MS analysis mode in the tandem-type mass spectrometer of a present Example.
- FIG. 1 is a configuration diagram of a main part of a tandem mass spectrometer according to the present embodiment.
- a liquid chromatograph (LC) is connected to the front stage of this mass spectrometer, and an eluate containing a sample whose components are separated by an LC column is introduced into the ion source 1. To do.
- the tandem mass spectrometer of the present embodiment is a collision cell in which an ion source 1, a quadrupole mass filter 2, and an octupole ion guide 31, for example, are disposed inside a vacuum chamber (not shown). 3, an ion trap 4, an orthogonal acceleration reflectron type time-of-flight mass analyzer 5, and an ion detector 55.
- the quadrupole mass filter 2 corresponds to a first mass separation unit
- the collision cell 3 corresponds to an ion cleavage unit
- the ion trap 4 corresponds to an ion accumulation unit
- the time-of-flight mass analyzer 5 corresponds to a second mass separation unit.
- an ion optical element such as an ion guide or an ion lens for efficiently transporting ions to the subsequent stage is provided between the ion source 1 and the quadrupole mass filter 2 or at other appropriate locations.
- an ion optical element such as an ion guide or an ion lens for efficiently transporting ions to the subsequent stage is provided between the ion source 1 and the quadrupole mass filter 2 or at other appropriate locations. In FIG. 1, the description of these elements is omitted.
- the ion source 1 is an ion source by an atmospheric pressure ionization method such as an electrospray ionization (ESI) method or an atmospheric pressure chemical ionization (APCI) method.
- an ion source using an electron ionization (EI) method or a chemical ionization (CI) method may be used as the ion source 1.
- the ion trap 4 has a three-dimensional quadrupole configuration in which a pair of end cap electrodes 42 and 43 are arranged with a ring electrode 41 interposed therebetween.
- the ion trap 4 only needs to be capable of storing ions therein, and may be a linear ion trap.
- the time-of-flight mass analyzer 5 has an extrusion electrode 51 and a grid electrode 52 as orthogonal ion acceleration units, and has a configuration in which reflectors 54 including a large number of reflection electrodes are arranged in a flight space 53.
- it may not be an orthogonal acceleration type, and may be a linear type or a circular type instead of a reflectron type.
- a predetermined voltage is applied to each rod electrode constituting the quadrupole mass filter 2 from the Q1 driving unit 61.
- a predetermined voltage is applied to each rod electrode constituting the ion guide 31 from the CC drive unit 62.
- a predetermined voltage is applied from the IT driving unit 63 to the ring electrode 41 and the end cap electrodes 42 and 43 constituting the ion trap 4.
- a predetermined voltage is applied from the TOF driving unit 64 to the extrusion electrode 51, the grid electrode 52, the reflector 54, and the like included in the time-of-flight mass analyzer 5.
- These drive units 61 to 64 are controlled by the control unit 7.
- the detection signal obtained by the ion detector 55 is sampled at a predetermined time interval by an A / D converter (not shown), converted into digital data, and input to the data processing unit 8.
- the control unit 7 includes an analysis condition determination unit 71 and an analysis control unit 72 as functional blocks.
- the data processing unit 8 includes a spectrum data integrating unit 81 and a mass spectrum creating unit 82.
- An input unit 91 and a display unit 92 are connected to the control unit 7 as a user interface.
- all or some of the functions included in the control unit 7 and the data processing unit 8 are achieved by executing dedicated software installed on a personal computer (or workstation) on the computer. It can be.
- the eluate from the LC column is introduced into the ion source 1, and the ion source 1 ionizes the compound contained in the introduced eluate.
- Ions derived from the compound generated by the ion source 1 are introduced into the quadrupole mass filter 2 through an ion optical element (not shown).
- the Q1 driving unit 61 applies a voltage to the quadrupole mass filter 2 that allows only ions having a specific mass-to-charge ratio specified in advance to pass. Thereby, only the ions having a specific mass-to-charge ratio among the various ions derived from the sample selectively pass through the quadrupole mass filter 2 as precursor ions.
- the precursor ions are applied with collision energy determined by, for example, a potential difference between the quadrupole mass filter 2 and the ion guide 31 (or an ion incident opening of the collision cell 3), and enter the collision cell 3.
- a CID gas such as He or Ar is introduced into the collision cell 3, and when the precursor ion comes into contact with the CID gas, cleavage occurs and product ions are generated.
- This mode of cleavage depends on CID conditions such as the size of collision energy and CID gas pressure. Therefore, for example, if the magnitude of the collision energy is changed even if the precursor ions are the same, the types of product ions generated by the cleavage and the respective generation amounts change.
- the product ions travel while converging and reach the ion trap 4 by the action of the high-frequency electric field formed by the voltage applied from the CC drive unit 62 to the ion guide 31.
- Product ions are introduced into the internal space of the ion trap 4 through an incident hole formed in the end cap electrode 42, and the product ions are generated by the action of a quadrupole electric field formed by a voltage applied to the ring electrode 41 from the IT driving unit 63. Captured in the internal space.
- Incoming ions are accumulated inside the ion trap 4. After accumulating ions in the internal space of the ion trap 4 for a predetermined time, a predetermined DC voltage is applied from the IT driving unit 63 to the end cap electrodes 42 and 43. As a result, the accumulated ions are ejected almost simultaneously from the ion trap 4 and sent to the ion acceleration unit of the time-of-flight mass analyzer 5.
- the TOF drive unit 64 applies a predetermined voltage to the extrusion electrode 51 and the grid electrode 52 at the timing when the packet-like ions reach the ion acceleration unit, and gives initial energy to each ion, respectively, in a direction substantially orthogonal to the traveling direction. Accelerate to. The accelerated ions pass through the grid electrode 52 and are introduced into the flight space 53, fly back by the action of the reflected electric field formed by the reflector 54, and finally reach the ion detector 55. The ions having substantially the same ion flight start time are separated according to the mass-to-charge ratio during the flight, and reach the ion detector 55 in order from the ions with the lowest mass-to-charge ratio.
- the time-of-flight spectrum indicating the relationship between the flight time and the signal intensity when the time of ion acceleration in the ion acceleration unit (that is, the time of start of ion flight) is zero is sent from the ion detector 55 to the data processing unit 8. Data is entered. Since the relationship between the mass-to-charge ratio and the flight time can be determined in advance, the mass spectrum can be determined from the time-of-flight spectrum by converting the flight time to the mass-to-charge ratio based on the relationship.
- the mass spectrum thus obtained is a product ion spectrum showing the relationship between the mass-to-charge ratio and the intensity of various product ions generated by cleavage of a specific precursor ion.
- the CID condition such as collision energy
- mass information of various types of product ions derived from the compound it is more convenient to obtain mass information of various types of product ions derived from the compound. Therefore, in the tandem mass spectrometer of the present embodiment, a plurality of collision energies for generating product ions that can be reflected in one product ion spectrum can be set. That is, product ions having different partial structures generated by cleavage under a plurality of different collision energies are reflected in one mass spectrum.
- the ion trap 4 can accumulate ions incident during a certain time width and temporally compress and eject the ions, the amount of ions that can be accumulated due to the space charge effect is limited. There is. In general, since ion traps have a fundamental limitation on the mass-to-charge ratio range that can be captured, it is difficult to comprehensively accumulate ions in a wide mass-to-charge ratio range. Therefore, in the tandem mass spectrometer of the present embodiment, the width of the mass-to-charge ratio of ions accumulated in the ion trap 4 is narrowed (specifically, about 100), and is limited to that width.
- FIG. 2 is an explanatory diagram of the MS / MS analysis mode.
- the analyst sets a plurality of collision energy values (CE values) from the input unit 91 as one of the analysis conditions.
- CE values collision energy values
- the analyst also sets the total mass-to-charge ratio range to be measured from the input unit 91.
- the total mass-to-charge ratio range is set to m / z 10-2000. This may also be determined as a default.
- the analysis condition determination unit 71 divides the total mass-to-charge ratio range set as described above into a plurality of mass-to-charge ratio ranges (m / z ranges) according to a predetermined mass-to-charge ratio width.
- the mass-to-charge ratio width is set to 100
- the m / z range is divided into a total of 21 parts of 10-100, 100-200, 200-300, ..., 1800-1900, 1900-2000.
- the analysis condition determination unit 71 obtains combinations of all set CE values and all m / z ranges. Assuming that the combination of the CE value and the m / z range is (CE value, m / z range), as shown in FIG.
- the analysis condition determining unit 71 sets a period for executing the mass analysis (MS / MS analysis) once under the combination of 63 (CE value, m / z range) described above as one cycle.
- a measurement method is created to do this N cycles for one compound.
- the number of cycles is large, but the measurement time becomes longer as the number of cycles increases. Therefore, the number of cycle repetitions N may be set by an analyst. For example, when a measurement time allowed for one compound is determined in advance, the measurement time and one mass are determined. The number of cycles that can be performed may be automatically calculated from the time for analysis.
- the time width during which one compound is introduced into the ion source 1 differs depending on the separation conditions of the LC or GC. Therefore, it is necessary to set the number of cycles appropriately in consideration of the time width during which the target compound is introduced into the ion source 1.
- the time required for one mass analysis may be constant regardless of the m / z range, but in practice, the smaller the ion mass-to-charge ratio, the shorter the flight time, so the m / z range.
- the lower the mass to charge ratio side the shorter the time allotted for one mass analysis. Therefore, by changing the time allotted to one mass analysis according to the m / z range, it is possible to increase the number of cycles N while keeping the measurement time for one compound the same.
- the time during which the target compound is introduced into the ion source 1 (that is, the retention time) and the mass-to-charge ratio of the precursor ion selected by the quadrupole mass filter 2 are set as the analysis conditions. It can be set as one.
- the measurement method may be determined so that the analysis for a predetermined precursor ion is performed for a predetermined time range when a predetermined time has elapsed since the start of analysis.
- the retention time of the compound is unknown, so normal mass spectrometry without selection of a precursor ion or cleavage operation is required.
- the measurement method may be determined so that the MS / MS analysis is subsequently performed with the peak as a target.
- a function is known as automatic MS n, for example.
- the analysis control unit 72 performs the analysis according to the measurement method determined by the analysis condition determining unit 71.
- the target compound is known
- Various product ions generated thereby are accumulated in the ion trap 4.
- Product ions in a specific mass-to-charge ratio range having a narrow mass-to-charge ratio range accumulated in this way are separated for each mass-to-charge ratio by the time-of-flight mass analyzer 5 and detected by the ion detector 55.
- the time-of-flight spectrum data input to the data processing unit 8 for one mass analysis is only for product ions corresponding to one collision energy value and one mass-to-charge ratio range. Therefore, if a mass spectrum is created based on this time-of-flight spectrum data, it is a mass spectrum in a narrow mass-to-charge ratio range as shown in FIG. As described above, when mass analysis is performed on each of 63 (CE value, m / z range) combinations for one target compound, 63 mass spectra with different observed peaks are obtained. (However, these mass spectra are not actually created, and if a mass spectrum is created, such a spectrum waveform is an example).
- the spectrum data integration unit 81 integrates time-of-flight spectrum data sequentially input for one target compound as described above. As a result, time-of-flight spectral data over a wide range of time-of-flight corresponding to a wide mass-to-charge ratio range of m / z 10-2000 can be obtained, and the spectrum data includes -10V, -20V, and -30V. Information on product ions generated by cleavage under two different CE values is included. If the time-of-flight spectrum data obtained by N-cycle mass spectrometry is accumulated, the mass spectrum creation unit 82 creates one mass spectrum based on the accumulated time-of-flight spectrum data (see FIG. 2C). ), For example, on the screen of the display unit 92. In this mass spectrum, product ions over a wide mass-to-charge ratio range of m / z / 10-2000 were observed, and the product ions were generated under three different CE values.
- mass information of various fragments of one target compound can be obtained from the mass spectrum, for example, the structure of the target compound can be estimated by obtaining a partial structure from the mass information of the fragments.
- Comprehensive mass information about various fragments can be obtained, which facilitates structure estimation and improves accuracy.
- the MS / MS analysis mode of the above embodiment is performed under a combination of brute force (CE value, m / z range) for the set CE value and m / z range. For this reason, it is excellent in that product ions over a wide mass-to-charge ratio range obtained under a plurality of collision energies can be detected comprehensively, that is, without being missed. On the other hand, if no product ions are observed in a certain m / z range, it can be said that the measurement is a waste of time.
- FIG. 3 is an explanatory diagram of such MS / MS analysis mode.
- the total number of combinations is 21, which is 1/3 of the total number of combinations in the MS / MS analysis mode shown in FIG. Therefore, in this case, if the number of cycles N is the same as the MS / MS analysis mode shown in FIG. 2, the measurement time is shortened to about 1/3. Further, if the measurement time is the same, the number of cycles can be increased by about three times, and the intensity of the same product ion is added accordingly, so that the sensitivity is increased.
- CE value with the smallest collision energy -10V
- the mass-to-charge ratio range is limited to m / z 1000-2000, respectively, and combinations of (CE value, m / z range) are defined. Also good.
- the number of executions of mass spectrometry for one compound differs depending on the combination of (CE value, m / z range), but there is no inconvenience in creating a mass spectrum for that compound.
- product ions having a specific mass-to-charge ratio are detected is important in the identification and structural analysis of compounds. For example, for two different compounds A and B having substantially the same molecular weight, if product ions a and b having specific mass-to-charge ratios are detected, it is determined that the compound A is a compound A. In such a case, under the condition of combining the m / z ranges (CE value, m / z range) including those product ions so that the detection sensitivity of such specific one or more product ions is high. It is preferable that the number of executions of the mass analysis is greater than the number of executions of the mass analysis under other combinations.
- the mass-to-charge ratio of product ions important for identifying a compound is 231 and 562, ( ⁇ 10, m / z 200-300), ( ⁇ 20, m / z 200) -300), (-30, m / z 200-300), (-10, m / z 500-600), (-20, m / z 500-600), (-30, m / z 500-600 )
- Mass analysis was performed 5 times under the conditions of 6 combinations of (CE value, m / z range), and under the conditions of other combinations of (CE value, m / z range) What is necessary is just to perform mass spectrometry twice each.
- the tandem mass spectrometer of the above embodiment has a configuration in which a three-dimensional quadrupole ion trap is combined with a so-called Q-TOF mass spectrometer, but the present invention is a triple quadrupole mass spectrometer. It is also applicable to a configuration in which an ion trap is combined.
- the ion guide 31 disposed in the collision cell 3 has a function of accumulating ions, or the orthogonal ion acceleration of the time-of-flight mass analyzer 5 is performed.
- the part may be provided with a function of accumulating ions, and the characteristic analysis as described above may be executed using the ion accumulation function.
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Abstract
Description
a)前記イオン開裂部における衝突誘起解離の際にプリカーサイオンに付与される衝突エネルギをm段階(ただしmは2以上の整数)に変更する衝突エネルギ設定部と、
b)前記イオン蓄積部に蓄積するイオンの質量電荷比範囲を質量電荷比方向にずらしたn段階(ただしnは2以上の整数)に変更する質量電荷比範囲設定部と、
c)前記衝突エネルギ設定部によりm段階の衝突エネルギのうちの一つを設定するとともに、前記質量電荷比範囲設定部によりn段階の質量電荷比範囲のうちの一つを設定し、その衝突エネルギと質量電荷比範囲とを組み合わせた条件の下での所定のプリカーサイオンに対するプロダクトイオンの質量分析を実施するように各部の動作を制御するものであって、衝突エネルギと質量電荷比範囲との組み合わせが相違するm×n個の全ての組み合わせについて、それぞれ少なくとも1回ずつプロダクトイオンの質量分析を実施するように各部の動作を制御する分析制御部と、
d)前記分析制御部による制御の下で少なくともm×n回実施されたプロダクトイオンの質量分析により得られたスペクトルデータを積算することで、所定質量電荷比範囲に亘る一つのマススペクトルを作成するデータ処理部と、
を備えることを特徴としている。
a)前記イオン開裂部における衝突誘起解離の際にプリカーサイオンに付与される衝突エネルギをm段階(ただしmは2以上の整数)に変更する衝突エネルギ設定部と、
b)前記イオン蓄積部に蓄積するイオンの質量電荷比範囲を質量電荷比方向にずらしたn段階(ただしnは2以上の整数)に変更する質量電荷比範囲設定部と、
c)前記衝突エネルギ設定部によりm段階の衝突エネルギのうちの一つを設定するとともに、前記質量電荷比範囲設定部によりn段階の質量電荷比範囲のうちの一つを設定し、その衝突エネルギと質量電荷比範囲とを組み合わせた条件の下での所定のプリカーサイオンに対するプロダクトイオンの質量分析を実施するように各部の動作を制御するものであって、m段階の衝突エネルギのそれぞれに対しn段階の質量電荷比範囲のうちのnよりも小さい任意の整数の質量電荷比範囲を、少なくともn段階の質量電荷比範囲が少なくとも一回ずつ選択されるようにそれぞれ組み合わせ、そのm又はnのいずれか大きいほうの値以上である全ての組み合わせについて、それぞれ少なくとも1回ずつプロダクトイオンの質量分析を実施するように各部の動作を制御する分析制御部と、
d)前記分析制御部による制御の下でm又はnのいずれか大きいほうの値以上の回数だけ実施されたプロダクトイオンの質量分析により得られたスペクトルデータを積算することで、所定質量電荷比範囲に亘る一つのマススペクトルを作成するデータ処理部と、
を備えることを特徴としている。
なお、3次元四重極型、リニア型のいずれのイオントラップにおいても、イオンを捕捉するための高周波電場を形成するべくイオントラップを構成する電極に印加される高周波電圧の周波数や振幅などを変更することによって、イオンが安定的に捕捉される質量電荷比範囲を変えることができる。
このように実質的に有用なスペクトルデータが得られない衝突エネルギと質量電荷比範囲との組み合わせに対する質量分析を実行しないことで、同等の品質のプロダクトイオンスペクトルを得るのに必要な測定時間を短縮することができるし、また、同じ測定時間であればより高感度のプロダクトイオンスペクトルを得ることができる。
LCのカラムからの溶出液がイオン源1に導入され、イオン源1は導入された溶出液に含まれる化合物をイオン化する。イオン源1で生成された化合物由来のイオンは図示しないイオン光学素子等を経て四重極マスフィルタ2に導入される。制御部7の制御の下でQ1駆動部61は例えば、予め指定された特定の質量電荷比を有するイオンのみを通過させるような電圧を四重極マスフィルタ2に印加する。それにより、試料由来の様々なイオンの中で、特定の質量電荷比を有するイオンのみがプリカーサイオンとして選択的に四重極マスフィルタ2を通り抜ける。
例えば、少なくとも特定の種類の化合物では、コリジョンエネルギが大きいほど開裂が起こり易く、質量電荷比が小さなプロダクトイオンが生成され易いとすれば、大きなCE値と高いm/z範囲との組み合わせでは得られるプロダクトイオンが殆どないことが想定される。そこで、(CE値,m/z範囲)の全ての組み合わせについてのMS/MS分析を実行するのではなく、有意なプロダクトイオンが観測されないことが分かっている又はそうした推定が可能である(CE値,m/z範囲)の組み合わせについてのMS/MS分析を省略するようにしてもよい。
ここでは、CE値及びm/z範囲自体は図2に示したMS/MS分析モードの例と同じであるが、CE値とm/z範囲との組み合わせの数はかなり絞られている。即ち、少なくとも或る種の化合物では、CE値が大きいほど壊れ易く質量電荷比が小さなプロダクトイオンが生じ易いと推定できる。そこで、コリジョンエネルギが最も大きいCE値=-30Vに対しては質量電荷比範囲をm/z 10-600、コリジョンエネルギが次に大きいCE値=-20Vに対しては質量電荷比範囲をm/z 600-1300、コリジョンエネルギが最も小さいCE値=-10Vに対しては質量電荷比範囲をm/z 1300-2000、にそれぞれ限定し、(CE値,m/z範囲)の組み合わせを定めている。これによって、組み合わせの総数は21になり、図2に示したMS/MS分析モードにおける組み合わせ総数の1/3である。したがって、この場合には、図2に示したMS/MS分析モードとサイクル数Nを同じにすれば、測定時間は約1/3に短縮される。また、測定時間を同じにすれば、サイクル数を約3倍にすることができ、その分だけ同じプロダクトイオンの強度が加算されるので感度が高くなる。
2…四重極マスフィルタ
3…コリジョンセル
31…イオンガイド
4…イオントラップ
41…リング電極
42、43…エンドキャップ電極
5…飛行時間型質量分離器
51…押出し電極
52…グリッド電極
53…飛行空間
54…反射器
55…イオン検出器
61…Q1駆動部
62…CC駆動部
63…IT駆動部
64…TOF駆動部
7…制御部
71…分析条件決定部
72…分析制御部
8…データ処理部
81…スペクトルデータ積算部
82…マススペクトル作成部
91…入力部
92…表示部
Claims (3)
- 試料中の化合物をイオン化するイオン源と、生成された各種イオンの中で特定の質量電荷比を有するイオンをプリカーサイオンとして選別する第1質量分離部と、該プリカーサイオンを衝突誘起解離により開裂させるイオン開裂部と、その開裂により生成された各種プロダクトイオンを質量分析する第2質量分離部及び検出器と、を具備し、前記第1質量分離部、前記イオン開裂部、若しくは前記第2質量分離部のいずれかをイオン蓄積部として動作させることによって、又は、前記第1質量分離部と前記イオン開裂部との間、若しくは、前記イオン開裂部と前記第2質量分離部との間に設けられたイオン蓄積部によって、イオンを一時的に蓄積可能であるタンデム型質量分析装置において、
a)前記イオン開裂部における衝突誘起解離の際にプリカーサイオンに付与される衝突エネルギをm段階(ただしmは2以上の整数)に変更する衝突エネルギ設定部と、
b)前記イオン蓄積部に蓄積するイオンの質量電荷比範囲を質量電荷比方向にずらしたn段階(ただしnは2以上の整数)に変更する質量電荷比範囲設定部と、
c)前記衝突エネルギ設定部によりm段階の衝突エネルギのうちの一つを設定するとともに、前記質量電荷比範囲設定部によりn段階の質量電荷比範囲のうちの一つを設定し、その衝突エネルギと質量電荷比範囲とを組み合わせた条件の下での所定のプリカーサイオンに対するプロダクトイオンの質量分析を実施するように各部の動作を制御するものであって、衝突エネルギと質量電荷比範囲との組み合わせが相違するm×n個の全ての組み合わせについて、それぞれ少なくとも1回ずつプロダクトイオンの質量分析を実施するように各部の動作を制御する分析制御部と、
d)前記分析制御部による制御の下で少なくともm×n回実施されたプロダクトイオンの質量分析により得られたスペクトルデータを積算することで、所定質量電荷比範囲に亘る一つのマススペクトルを作成するデータ処理部と、
を備えることを特徴とするタンデム型質量分析装置。 - 試料中の化合物をイオン化するイオン源と、生成された各種イオンの中で特定の質量電荷比を有するイオンをプリカーサイオンとして選別する第1質量分離部と、該プリカーサイオンを衝突誘起解離により開裂させるイオン開裂部と、その開裂により生成された各種プロダクトイオンを質量分析する第2質量分離部及び検出器と、を具備し、前記第1質量分離部、前記イオン開裂部、若しくは前記第2質量分離部のいずれかをイオン蓄積部として動作させることによって、又は、前記第1質量分離部と前記イオン開裂部との間、若しくは、前記イオン開裂部と前記第2質量分離部との間に設けられたイオン蓄積部によって、イオンを一時的に蓄積可能であるタンデム型質量分析装置において、
a)前記イオン開裂部における衝突誘起解離の際にプリカーサイオンに付与される衝突エネルギをm段階(ただしmは2以上の整数)に変更する衝突エネルギ設定部と、
b)前記イオン蓄積部に蓄積するイオンの質量電荷比範囲を質量電荷比方向にずらしたn段階(ただしnは2以上の整数)に変更する質量電荷比範囲設定部と、
c)前記衝突エネルギ設定部によりm段階の衝突エネルギのうちの一つを設定するとともに、前記質量電荷比範囲設定部によりn段階の質量電荷比範囲のうちの一つを設定し、その衝突エネルギと質量電荷比範囲とを組み合わせた条件の下での所定のプリカーサイオンに対するプロダクトイオンの質量分析を実施するように各部の動作を制御するものであって、m段階の衝突エネルギのそれぞれに対しn段階の質量電荷比範囲のうちのnよりも小さい任意の整数の質量電荷比範囲を、少なくともn段階の質量電荷比範囲が少なくとも一回ずつ選択されるようにそれぞれ組み合わせ、そのm又はnのいずれか大きいほうの値以上である全ての組み合わせについて、それぞれ少なくとも1回ずつプロダクトイオンの質量分析を実施するように各部の動作を制御する分析制御部と、
d)前記分析制御部による制御の下でm又はnのいずれか大きいほうの値以上の回数だけ実施されたプロダクトイオンの質量分析により得られたスペクトルデータを積算することで、所定質量電荷比範囲に亘る一つのマススペクトルを作成するデータ処理部と、
を備えることを特徴とするタンデム型質量分析装置。 - 請求項1又は2に記載のタンデム型質量分析装置であって、
前記第2質量分離部は飛行時間型質量分析器であることを特徴とするタンデム型質量分析装置。
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