Time domain analysis of signals for charge detection mass spectrometry.

Abstract

A charge detection mass spectrometer (CDMS) includes an electrostatic linear ion trap (ELIT), a processor, and a memory having instructions executable by the processor stored therein, the processor (a) controlling the ELIT to trap ions, (b) collecting ion measurement information as the trapped ions oscillate back and forth through the ELIT, the ion measurement information including charges induced by the ions on a charge detector of the ELIT during each passage of the ions through the ELIT and the timing of the induced charges relative to one another, (c) processing the ion measurement information in the time domain for each of a plurality of successive time windows of the ion measurement information to determine a charge magnitude of the ions during each time window, and (d) determining a charge magnitude of the trapped ions based on the charge magnitude for each of the time windows.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This international patent application claims the benefit of and priority to U.S. Provisional Patent Application No. 62 / 969,325, filed February 3, 2020, the entire disclosure of which is expressly incorporated herein by reference.

[0002] Government Rights

[0002] This invention was made with Government support under award GM1311100 from the National Institutes of Health. The United States Government has certain rights in this invention.

[0003]

[0003] This disclosure relates generally to charge detection mass spectrometry instruments, and more particularly to making mass and charge measurements with such instruments. [Background technology]

[0004]

[0004] Charge detection mass spectrometry (CDMS) is a particle analysis technique in which the mass of an ion is determined by simultaneously measuring the mass-to-charge ratio and charge of the ion, commonly referred to as "m / z". Some CDMS instruments use an electrostatic linear ion trap (ELIT) to make such measurements. Summary of the Invention [Means for solving the problem]

[0005]

[0005] The present disclosure may include one or more of the features recited in the appended claims and / or one or more of the following features and combinations of features. In one aspect, a charge detection mass spectrometer (CDMS) may comprise an electrostatic linear ion trap (ELIT), an ion source configured to provide ions to the ELIT, a charge sensitive preamplifier having an input operably coupled to the ELIT, at least one processor operably coupled to the ELIT and to an output of the amplifier, and at least one memory having instructions stored therein, which, when executed by the at least one processor, cause the at least one processor to (a) control the ELIT to trap ions provided from the ion source in the ELIT, and (b) determine whether the trapped ions are in a charge sensitive preamplifier. (c) processing the ion measurement information in the time domain for each of a plurality of successive time windows of the ion measurement information to determine a magnitude of the charge on the ion during each time window; and (d) determining a magnitude of the charge on the trapped ion based on the magnitude of the charge for each of the time windows.

[0006]

[0006] In another aspect, a method is provided for measuring the charge of an ion in an electrostatic linear ion trap including a charge detection cylinder positioned between two ion mirrors, wherein during an ion trapping event, the ion passes through the charge detection cylinder and repeatedly oscillates back and forth between the two ion mirrors each time inducing a corresponding charge in the charge detection cylinder, and during the trapping event, an ion measurement signal including the magnitude of the induced charge and the timing of the induced charge is recorded in an ion measurement file. The method may include (a) setting a time window of the ion measurement signal at the beginning of the ion measurement file; (b) generating a simulated ion measurement signal for the time window of the ion measurement signal using input parameters including estimates of signal frequency, charge magnitude, signal phase, and duty cycle; (c) iteratively processing the variance between the time window of the ion measurement signal and the simulated ion measurement signal by adjusting values ​​of the input parameters until the variance reaches convergence; (d) recording the charge magnitude value resulting from (c); (e) advancing the time window of the ion measurement signal by an incremental time; (f) repeating (b)-(d) until the time window reaches the end of the ion measurement file; and (g) determining the charge of the ion based on the charge magnitude values ​​for each of the time windows. [Brief description of the drawings]

[0007] [Figure 1] FIG. 1 is a simplified schematic of a CDMS system including an embodiment of an electrostatic linear ion trap (ELIT) with coupled control and measurement components. [Figure 2A] FIG. 2 is a close-up view of the ion mirror M1 of the ELIT shown in FIG. 1, in which the mirror electrodes of M1 are controlled to generate an ion-permeable electric field within M1. [Figure 2B] FIG. 2 is a close-up view of the ion mirror M2 of the ELIT shown in FIG. 1, in which the mirror electrodes of M2 are controlled to generate an ion-reflecting electric field within M2. [Diagram 3] 2 is a simplified diagram of an embodiment of the processor shown in FIG. 1; [Figure 4A]FIG. 4A is a simplified schematic of the ELIT of FIG. 1 showing sequential control and operation of the ion mirrors to introduce at least one ion into the ELIT, oscillate the ion back and forth between the ion mirrors and through the charge detection cylinder, and measure and record multiple charge detection events. [Figure 4B] FIG. 4B is a simplified schematic of the ELIT of FIG. 1 showing sequential control and operation of the ion mirrors to admit at least one ion into the ELIT, oscillate the ion back and forth between the ion mirrors and through the charge detection cylinder, and measure and record multiple charge detection events. [Figure 4C] FIG. 4C is a simplified schematic of the ELIT of FIG. 1 showing sequential control and operation of the ion mirrors to introduce at least one ion into the ELIT, oscillate the ion back and forth between the ion mirrors and through the charge detection cylinder, and measure and record multiple charge detection events. [Diagram 5] 1 is a simplified flow chart illustrating an embodiment of a process for analyzing signal measurements contained in an ion measurement event file in the time domain to determine the frequency and charge magnitude (z) of ions oscillating back and forth through the charge detection cylinder of an ELIT during an ion trapping event. [Figure 6] 1 is a graph of signal versus time showing one period of a simulated signal for an axial ion trajectory in an electrostatic linear ion trap. [Figure 7] 7 is an expanded graph of the simulated signal of FIG. 6 with the duty cycle varying between 40% and 60%. [Figure 8] 7 is another expanded graph of the simulated signal of FIG. 6, with the frequency varying between 10 kHz and 15 kHz. [Figure 9] 7 is an expanded graph of the simulated signal of FIG. 6 with a modified variant shown superimposed, the modified variant introducing RC damping. [Figure 10] 1 is a simplified operational flowchart illustrating an embodiment of a cross-correlation process for determining an initial estimate of the phase of a simulated ion signal. [Figure 11] 1 is a simplified operational flowchart illustrating an embodiment of an optimization algorithm for reducing variance between simulated ion signals and ion measurement signals. [Figure 12] 12 is a graph of SRS versus iteration number showing an example of convergence by the optimization algorithm of FIG. 11; DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0008]

[0020] For the purposes of promoting an understanding of the principles of the disclosure, reference will now be made to certain exemplary embodiments which are illustrated in the accompanying drawings, and specific language will be used to describe the embodiments.

[0009]

[0021] This disclosure relates to an apparatus and technique for processing a time domain ion measurement signal generated by an electrostatic linear ion trap (ELIT) of a charge detection mass spectrometer (CDMS) to simultaneously determine the ion mass-to-charge ratio and the ion charge from which the ion mass can then be determined. In this disclosure, the phrase "charge detection event" is defined as the detection of a charge induced at a charge detector of an ELIT by an ion passing the charge detector only once, and the phrase "ion measurement event" is defined as a collection of charge detection events obtained by oscillating an ion back and forth through the charge detector a selected number of times or for a selected period of time. Since the oscillation of an ion back and forth through the charge detector occurs by controlled trapping of the ion in the ELIT, as described in detail below, the phrase "ion measurement event" may alternatively be referred to herein as an "ion trapping event" or simply a "trapping event", and the phrases "ion measurement event", "ion trapping event", "trapping event", and variations thereof shall be understood to be synonymous with each other.

[0010]

[0022] Referring to FIG. 1, a CDMS system 10 is shown that includes an embodiment of an electrostatic linear ion trap (ELIT) 14 with associated control and measurement components. In the illustrated embodiment, the CDMS system 10 includes an ion source 12 operably coupled to the inlet of the ELIT 14. The ion source 12 illustratively includes any conventional device or apparatus that generates ions from a sample, and may further include one or more devices and / or instruments that separate, collect, filter, resolve, and / or normalize or shift the charge state of the ions according to one or more molecular characteristics. The ion source 12 may include, as one illustrative example, a conventional electrospray ionization source, a matrix-assisted laser desorption ionization (MALDI) source, etc., coupled to the inlet of a conventional mass spectrometer, although this should not be considered limiting in any way. The mass spectrometer may be of any conventional design, including, for example, but not limited to, a time-of-flight (TOF) mass spectrometer, a reflectron mass spectrometer, a Fourier transform ion cyclotron resonance (FTICR) mass spectrometer, a quadrupole mass spectrometer, a triple quadrupole mass spectrometer, a magnetic sector mass spectrometer, etc. In any case, the ion outlet of the mass spectrometer is operatively coupled to the ion inlet of the ELIT 14. The sample from which ions are generated may be any biological or other material.

[0011]

[0023] In the illustrated embodiment, the ELIT 14 is illustratively surrounded by a grounded chamber or cylinder GC and includes a charge detector CD operably coupled to opposing ion mirrors M1, M2 disposed at opposite ends of the ELIT 14. The ion mirror M1 is operably disposed between the ion source 12 and one end of the charge detector CD, and the ion mirror M2 is operably disposed at the opposite end of the charge detector CD. Each ion mirror M1, M2 defines an ion mirror region R1, R2 within the ion mirror, respectively. As described in more detail below, the regions R1, R2 of the ion mirrors M1, M2, the charge detector CD, and the space between the charge detector CD and the ion mirrors M1, M2 collectively define a central longitudinal axis 20 therethrough, which illustratively represents an ideal ion travel path between the ion mirrors M1 and M2 through the ELIT 14.

[0012]

[0024] In the illustrated embodiment, voltage sources V1, V2 are electrically connected to ion mirrors M1, M2, respectively. Each voltage source V1, V2 illustratively includes one or more switchable DC voltage sources that can be controlled or programmed to selectively generate N programmable or controllable voltages, where N can be any positive integer. Illustrative examples of such voltages are described below in conjunction with FIGS. 2A and 2B and set one of two distinct operating modes of each of the ion mirrors M1, M2, as described in detail below. In either case, ions move within the ELIT 14 near a longitudinal axis 20 that extends centrally through the charge detector CD and the ion mirrors M1, M2 under the influence of the electric fields selectively formed by the voltage sources V1, V2.

[0013]

[0025] The voltage sources V1, V2 are illustratively shown electrically connected by P signal paths to a conventional processor 16 including a memory 18 having instructions stored therein that, when executed by the processor 16, cause the processor 16 to control the voltage sources V1, V2 to selectively shape the ion transmission and reflection fields, TEF, REF, respectively, in the regions R1, R2 of the respective ion mirrors M1, M2 to generate desired DC output voltages. P may be any positive integer. In some alternative embodiments, one or both of the voltage sources V1, V2 may be programmable to selectively generate one or more constant output voltages. In other alternative embodiments, one or both of the voltage sources V1, V2 may be configured to generate one or more time-varying output voltages of any desired waveform. It will be appreciated that in alternative embodiments, more or fewer voltage sources may be electrically connected to the mirrors M1, M2.

[0014]

[0026] The charge detector CD is illustratively provided in the form of a conductive cylinder electrically connected to a signal input of a charge sensitive preamplifier CP, the signal output of which is electrically connected to the processor 16. The voltage sources V1, V2 are illustratively controlled to selectively trap ions entering the ELIT 14 and oscillate the ions back and forth between the ion mirrors M1 and M2 in the ELIT 14, so that the trapped ions repeatedly pass the charge detector CD, as described in detail below. Once the ions are trapped in the ELIT 14 and oscillate back and forth between the ion mirrors M1 and M2, the charge sensitive preamplifier CP is illustratively operable in a conventional manner to detect a charge (CH) induced in the charge detection cylinder CD as the ions pass through the charge detection cylinder CD and between the ion mirrors M1 and M2, and to generate a charge detection signal (CHD) corresponding to the detected charge. The charge detection signal CHD is illustratively recorded in the form of oscillation period values, with each oscillation period value representing ion measurement information at only one respective charge detection event. A number of such oscillation period values ​​are measured and recorded for ions trapped during each ion measurement event (i.e., during an ion trapping event), and the resulting set of recorded oscillation period values, i.e., recorded ion measurement information, for an ion measurement event is processed to determine charge, mass-to-charge ratio, and / or mass values ​​of the ions, as described below. A number of ion measurement events may be processed in this manner, and a mass-to-charge ratio and / or mass spectrum of the sample may illustratively be produced by this processing.

[0015]

[0027] 2A and 2B, there are shown, respectively, an embodiment of an ion mirror M1, M2 of the ELIT 14 shown in FIG. 1. The ion mirrors M1, M2 are illustratively identical to one another in that each includes a cascaded structure of four spaced apart conductive mirror electrodes. For each of the ion mirrors M1, M2, the first mirror electrode 301 has a thickness W1 and defines a passageway of diameter P1 through the center of the mirror electrode 301. The end cap 32 is attached or otherwise coupled to the outer surface of the first mirror electrode 301 and defines an opening A1 through the center of the end cap 32 that serves as an inlet and / or outlet to and / or from the corresponding ion mirror M1, M2, respectively. In the case of the ion mirror M1, the end cap 32 is coupled to or is part of the ion outlet of the ion source 12 shown in FIG. 1. The opening A1 for each end cap 32 illustratively has a diameter P2.

[0016]

[0028] The second mirror electrode 302 of each ion mirror M1, M2 is spaced from the first mirror electrode 301 by a space of width W2. The second mirror electrode 302, like the mirror electrode 301, has a thickness W1 and defines a passage of diameter P2 through the center of the mirror electrode 302. The third mirror electrode 303 of each ion mirror M1, M2 is similarly spaced from the second mirror electrode 302 by a space of width W2. The third mirror electrode 303 has a thickness W1 and defines a passage of width P1 through the center of the mirror electrode 303.

[0017]

[0029] The fourth mirror electrode 304 is separated from the third mirror electrode 303 by a space of width W2. The fourth mirror electrode 304 is illustratively formed at each end of a grounded cylinder GC having a thickness of W1 and disposed around the charge detector CD. The fourth mirror electrode 304 defines an opening A2 through the center of the mirror electrode 304, which is illustratively conical in shape and increases linearly between the inner and outer surfaces of the grounded cylinder GC from a diameter P3 defined at the inner surface of the grounded cylinder GC to a diameter P1 at the outer surface of the grounded cylinder GC (which is also the inner surface of each ion mirror M1, M2).

[0018]

[0030] The spaces defined between the mirror electrodes 301-304 may be voids, i.e., vacuum gaps, in some embodiments, while in other embodiments, such spaces may be filled with one or more non-conductive, e.g., dielectric, materials. The mirror electrodes 301-304 and the end caps 32 are axially aligned, i.e., collinear, such that the longitudinal axis 22 passes through the center of each aligned passageway and also passes through the center of the apertures A1, A2. In embodiments in which the spaces between the mirror electrodes 301-304 include one or more non-conductive materials, such materials similarly define respective passageways, i.e., collinear, through the axially aligned spaces, the passageways being defined through the mirror electrodes 301-304 and illustratively having diameters equal to or greater than P2. Illustratively, P1>P3>P2, although other relative diameter configurations are possible in other embodiments.

[0019]

[0031] One region R1 is defined between the openings A1 and A2 of the ion mirror M1, and another region R2 is similarly defined between the openings A1 and A2 of the ion mirror M2. Regions R1, R2 are illustratively identical to each other in shape and volume.

[0020]

[0032] As explained above, the charge detector CD is illustratively provided in the form of an elongated conductive cylinder disposed between corresponding portions of the ion mirrors M1, M2 and spaced apart by a space of width W3. In one embodiment, W1>W3>W2, and P1>P3>P2, although other relative width configurations are possible in alternative embodiments. In either case, the longitudinal axis 20 illustratively extends centrally through a passage defined through the charge detector cylinder CD, such that the longitudinal axis 20 extends centrally through the combination of the ion mirrors M1, M2 and the charge detector cylinder CD. In operation, the ground cylinder GC is illustratively controlled to a ground potential such that the fourth mirror electrode 304 of each ion mirror M1, M2 is always at ground potential. In some alternative embodiments, the fourth mirror electrode 304 of either or both of the ion mirrors M1, M2 can be set to any desired DC reference potential or to a switchable DC or other time-varying voltage source.

[0021]

[0033] In the embodiment shown in Figures 2A and 2B, each of the voltage sources V1, V2 is configured to generate four DC voltages D1-D4, respectively, and to supply the voltages D1-D4 to a respective one of the mirror electrodes 301-304 of the respective ion mirror M1, M2. In some embodiments in which one or more of the mirror electrodes 301-304 are to be held at ground potential at all times, such one or more mirror electrodes 301-304 may alternatively be electrically connected to the ground reference of the respective voltage supply V1, V2, and the corresponding one or more voltage outputs D1-D4 may be omitted. Alternatively or additionally, in embodiments in which any two or more of the mirror electrodes 301-304 are to be controlled to the same non-zero DC value, such any two or more mirror electrodes 301-304 may be electrically connected to only one of the voltage outputs D1-D4, and unnecessary ones of the output voltages D1-D4 may be omitted.

[0022]

[0034] Each ion mirror M1, M2 is illustratively controllable and switchable between an ion transmission mode (FIG. 2A) in which the voltages D1-D4 generated by the respective voltage sources V1, V2 form an ion transmission electric field (TEF) in the respective region R1, R2 of the ion mirror by selectively applying voltages D1-D4, and an ion reflection mode (FIG. 2B) in which the voltages D1-D4 generated by the respective voltage sources V1, V2 form an ion reflection electric field (REF) in the respective region R1, R2 of the ion mirror. As shown in the example of FIG. 2A, when ions from the ion source 12 fly through the entrance aperture A1 of the ion mirror M1 into the region R1 of the ion mirror M1, the ions are focused towards the longitudinal axis 20 of the ELIT 14 by the ion transmission electric field TEF formed in the region R1 of the ion mirror M1 by selective control of the voltages D1-D4 of V1. As a result of the focusing effect of the transmission field TEF in the region R1 of the ion mirror M1, ions exiting the region R1 of the ion mirror M1 through the opening A2 of the grounded chamber GC obtain a narrow trajectory into and through the charge detector CD, in other words to keep the ion's travel path close to the longitudinal axis 20 through the charge detector CD. The same ion transmission field TEF can be selectively formed in the region R2 of the ion mirror M2 by similar control of the voltages D1-D4 of the voltage source V2. In the ion transmission mode, ions entering the region R2 from the charge detector cylinder CD through the opening A2 of M2 are focused towards the longitudinal axis 20 by the ion transmission field TEF in the region R2, which causes the ions to exit the opening A1 of the ion mirror M2.

[0023]

[0035] 2B, by way of example, an ion reflecting field REF formed in region R2 of ion mirror M2 by selectively controlling voltages D1-D4 of V2 acts to slow and stop ions entering ion region R2 from charge detection cylinder CD through ion entrance aperture A2 of M2, and to focus ions toward central longitudinal axis 20 within region R2 of ion mirror M2 in order to accelerate the stopped ions in the opposite direction back through aperture A2 of M2 and into the end of charge detection cylinder CD adjacent M2, as shown by ion trajectory 42, and maintain a narrow ion trajectory through charge detector CD and back toward ion mirror M1. The same ion reflecting field REF can be selectively formed in region R1 of ion mirror M1 by similar control of voltages D1-D4 of voltage source V1. In the ion reflection mode, ions entering region R1 from the charge detection cylinder CD through the opening A2 of M1 are slowed down and stopped by the ion reflection field REF generated in region R1, then accelerated in the opposite direction back through the opening A2 of M1 into the end of the charge detection cylinder CD adjacent M1 and focused towards the central longitudinal axis 20 in region R1 of the ion mirror M1 to maintain a narrow trajectory of the ions through the charge detector CD and back towards the ion mirror M1. Ions passing through the ELIT 14 and being reflected by the ion reflection field REF in the ion regions R1, R2 are considered to be trapped in the ELIT 14, allowing the ions to continue to move back and forth between the ion mirrors M1 and M2 through the charge detection cylinder CD, just as described.

[0024]

[0036] An exemplary set of output voltages D1-D4 generated by voltage sources V1, V2, respectively, for controlling each ion mirror M1, M2 in the above-mentioned ion transmission and reflection modes is shown below in Table 1. It will be understood that the following values ​​of D1-D4 are provided by way of example only, and that other values ​​may alternatively be used for one or more of D1-D4. [Table 1]

[0025]

[0037] 1-2B as defining cylindrical passages therethrough, it will be appreciated that in alternative embodiments, either or both of the ion mirrors M1, M2 and / or the charge detection cylinder CD may define non-cylindrical passages therethrough, such that one or more of the passages through which the longitudinal axis 20 passes centrally exhibit a non-circular cross-section and profile. In still other embodiments, the cross-section of the passage defined through ion mirror M1 may be different from the passage defined through ion mirror M2, regardless of the shape of the cross-sectional profile.

[0026]

[0038] Referring now to Figure 3, there is shown an embodiment of the processor 16 shown in Figure 1. In the illustrated embodiment, the processor 16 includes a conventional amplifier circuit 40 having an input for receiving a charge detection signal CHD generated by a charge sensitive preamplifier CP and an output electrically connected to an input of a conventional analog-to-digital (A / D) converter 42. The output of the A / D converter 42 is electrically connected to a processor 50 (P1). The amplifier 40 is operable in a conventional manner to amplify the charge detection signal CHD generated by the charge sensitive preamplifier CP, and the A / D converter is then operable in a conventional manner to convert the amplified charge detection signal to a digital charge detection signal CDS.

[0027]

[0039] The processor 16 shown in FIG. 3 further includes a conventional comparator 44 having a first input receiving a charge detection signal CHD generated by the charge sensitive preamplifier CP, a second input receiving a threshold voltage CTH generated by a threshold voltage generator (TG) 46, and an output electrically connected to the processor 50. The comparator 44 is operable in a conventional manner to generate a trigger signal TR at an output of the comparator 44 that depends on the magnitude of the charge detection signal CDH compared to the magnitude of the threshold voltage CTH. In one embodiment, the comparator 44 is operable to generate an "inactive" trigger signal TR at or near a reference voltage, e.g., ground potential, for as long as CHD is less than CTH, and to generate an "active" TR signal at or near a power supply voltage of the circuits 40, 42, 44, 46, 50, or otherwise distinguishable from an inactive TR signal, if CHD is equal to or greater than CTH. In alternative embodiments, the comparator 44 may be operable to generate an "inactive" trigger signal TR at or near the power supply voltage as long as CHD is less than CTH, and to generate an "active" trigger signal TR at or near the reference potential when CHD is equal to or greater than CTH. Those skilled in the art will recognize that other trigger signal magnitudes and / or trigger signal polarities may be used, so long as such magnitudes and / or trigger signal polarities are distinguishable by the processor 50 to form the "inactive" and "active" states of the trigger signal TR. It will also be understood that any other such trigger signal magnitudes and / or trigger signal polarities are intended to fall within the scope of this disclosure. The comparator 44 may further be designed to include a desired amount of hysteresis in a conventional manner to prevent rapid switching of the output between the reference voltage and the power supply voltage in either case.

[0028]

[0040] The processor 50 is illustratively operable to generate a threshold voltage control signal THC and provide the THC to the threshold generator 46 to control the operation of the threshold generator 46. In some embodiments, the processor 50 is programmed or programmable to control the generation of the threshold voltage control signal THC to control the threshold voltage generator 46 to generate a CTH having a desired magnitude and / or polarity. In other embodiments, a user can, for example, through a downstream processor, for example, via a virtual control and visualization unit, give instructions in real time to the processor 50 to control the generation of the threshold voltage control signal THC to control the threshold voltage generator 46 to generate a CTH having a desired magnitude and / or polarity. In any case, the threshold voltage generator 46 is illustratively implemented in some embodiments in the form of a conventional controllable DC voltage source configured to respond to a digital form of the threshold control signal THC, for example in the form of a single serial digital signal or multiple parallel digital signals, to generate an analog threshold voltage CTH having a polarity and magnitude defined by the digital threshold control signal THC. In some alternative embodiments, the threshold voltage generator 46 may be provided in the form of a conventional digital-to-analog (D / A) converter that generates an analog threshold voltage CTH, having a magnitude, and in some embodiments, a polarity, defined by a digital threshold control signal THC, in response to a series or parallel digital threshold voltage TCH. In some such embodiments, the D / A converter may form part of the processor 50. Those skilled in the art will recognize other conventional circuits and techniques for selectively generating a threshold voltage CTH of a desired magnitude and / or polarity in response to one or more digital and / or analog forms of the control signal THC, and it will be understood that any such other conventional circuits and / or techniques are intended to fall within the scope of this disclosure.

[0029]

[0041] In addition to the aforementioned functions performed by the processor 50, the processor 50 is further operable to control the voltage sources V1, V2 to selectively generate ion transmission and reflection fields in the regions R1, R2 of the ion mirrors M1, M2, respectively, as described above in connection with FIGS. 2A, 2B. In some embodiments, the processor 50 is programmed or programmable to control the voltage sources V1, V2. In other embodiments, the voltage sources V1 and / or V2 may be programmed or otherwise controlled in real time by a user, for example, through a downstream processor 52, for example, via a virtual control and visualization unit. In any case, the processor 50 is exemplarily provided in one embodiment in the form of a field programmable gate array (FPGA) that is programmed or otherwise commanded by a user to collect and store charge detection signals CDS at charge detection events and ion measurement events, generate threshold control signals TCH from which the magnitude and / or polarity of the threshold voltage CTH is determined or derived, and control the voltage sources V1, V2. In this embodiment, the memory 18 described in relation to Fig. 1 is integrated into and forms part of the programming of the FPGA. In an alternative embodiment, the processor 50 may be provided in the form of one or more conventional microprocessors or controllers and one or more associated memory units having stored therein instructions that, when executed by the one or more microprocessors or controllers, cause the one or more microprocessors or controllers to operate exactly as described. In other alternative embodiments, the processing circuitry 50 may be implemented simply in the form of one or more conventional hardware circuits designed to operate as described above, or as a combination of one or more such hardware circuits and at least one microprocessor or controller operable to execute instructions stored in the memory and that operate as described above.

[0030]

[0042] The embodiment of the processor 16 shown in FIG. 3 further includes a second processor 52, illustratively coupled to the first processor 50 and also to at least one memory unit 54. In some embodiments, the processor 52 may include one or more peripheral devices, such as a display monitor, one or more input and / or output devices, etc., while in other embodiments, the processor 52 may not include any such peripheral devices. In either case, the processor 52 is illustratively configured, i.e., programmed, to execute at least one process for analyzing ion measurement events. Data received by the processor 50 through the charge detection signal CDS in the form of charge magnitude and charge timing data (i.e., detection of the timing of the charge mutually induced by the ions in the charge detection cylinder) is illustratively transferred directly from the processor 50 to the processor 52 for processing and analysis upon completion of each ion measurement event.

[0031]

[0043] In some embodiments, the processor 52 is exemplarily provided in the form of a high-speed server operable to perform both collection / storage and analysis of such data. In such embodiments, one or more high-speed memory units 54 may be coupled to the processor 52 and operable to store data received and analyzed by the processor 52. In one embodiment, the one or more memory units 54 exemplarily include at least one local memory unit that stores data being used or to be used by the processor 52, and at least one persistent storage memory unit that stores data long term. In such an embodiment, the processor 52 is exemplarily provided in the form of a Linux server (e.g., OpenSuse Leap42.1) with four Intel® Xeon™ processors (e.g., E5-465L v2 12 cores, 2.4 GHz). In this embodiment, an improvement of more than 100 times in the average analysis time of a single ion measurement event file is realized compared to a conventional Windows® PC (e.g., i5-2500K, 4 cores, 3.3 GHz). Similarly, the processor 52 of this embodiment, together with the high speed / performance memory unit 54, illustratively allows for over 100 times improvement in data storage speed. Those skilled in the art will recognize one or more other high speed data processing and analysis systems that may be implemented as the processor 52, and it will be understood that any such one or more other high speed data processing and analysis systems are intended to fall within the scope of this disclosure. In alternative embodiments, the processor 52 may be provided in the form of one or more conventional microprocessors or controllers and one or more associated memory units having instructions stored therein that, when executed by the one or more microprocessors or controllers, cause the one or more microprocessors or controllers to operate as described herein.

[0032]

[0044] In the illustrated embodiment, the memory unit 54 illustratively has stored therein instructions executable by the processor 52 for analyzing ion measurement event data generated by the ELIT 14 and determining ion mass spectral information of the sample being analyzed. In one embodiment, the processor 52 is operable to receive ion measurement event data from the processor 50 in the form of charge magnitude and charge detection timing information measured during each of a plurality of "charge detection events" (as that term is defined above) that constitute an "ion measurement event" (as that term is defined above), process such charge detection events that constitute such ion measurement event, determine ion charge and mass-to-charge data, and then determine ion mass data from the ion charge and mass-to-charge data. A plurality of ion measurement events can be processed in a similar manner to produce mass spectral information of the sample being analyzed.

[0033]

[0045] As briefly described above in connection with Figures 2A and 2B, the voltage sources V1, V2 are illustratively controlled by the processor 50, e.g., via the processor 52, to selectively form ion transmission and ion reflection electric fields in the region R1 of the ion mirror M1 and in the region R2 of the ion mirror M2 to guide ions introduced into the ELIT 14 from the ion source 12 through the ELIT 14 and then selectively trap and confine a single ion in the ELIT 14, whereby the trapped ion repeatedly passes through the charge detector CD as it oscillates back and forth between M1 and M2. With reference to Figures 4A-4C, simplified diagrams of the ELIT 14 of Figure 1 are shown illustrating examples of such sequential control and operation of the ion mirrors M1, M2 of the ELIT 14. In the following examples, the processor 52 is described as controlling the operation of the voltage sources V1, V2 in accordance with the programming of the processor 52, although it will be understood that the operation of the voltage source V1 and / or the operation of the voltage source V2 may be virtually controlled, at least in part, by the processor 50.

[0034]

[0046] As shown in FIG. 4A, the ELIT control sequence begins with the processor 52 controlling the ion mirror M1 in an ion transmission mode of operation (T) by controlling the voltage source V1 to form an ion transmission field in the region R1 of the ion mirror M1, and also controlling the voltage source V2 to control the ion mirror M2 in an ion transmission mode of operation (T) by similarly forming an ion transmission field in the region R2 of the ion mirror M2. As a result, ions generated by the ion source 12 travel into the ion mirror M1 and are focused toward the longitudinal axis 20 by the ion transmission field formed in the region R1 as they travel into the charge detection cylinder CD. The ions then travel through the charge detection cylinder CD into the ion mirror M2 where the ion transmission field formed in the region R2 of M2 focuses the ions toward the longitudinal axis 20, causing them to pass through the exit aperture A1 of M2, as shown by the ion trajectory 60 shown in FIG. 4A.

[0035]

[0047] 4B, after both ion mirrors M1, M2 have operated in the ion transmission mode of operation for a selected period of time and / or until ion transmission through the ion transmission mode of operation has been successfully achieved, the processor 52 is illustratively operable to control the ion mirror M2 to an ion reflection mode of operation (R) by controlling the voltage source V2 to form an ion reflection field in a region R2 of the ion mirror M2 while maintaining the ion mirror M1 in the ion transmission mode of operation (T) as shown. As a result, at least one ion generated by the ion source 12 enters the ion mirror M1 and is focused towards the longitudinal axis 20 by the ion transmission field formed in the region R1, which causes the at least one ion to travel through the ion mirror M1 into the charge detection cylinder CD, just as described in connection with FIG. 4A. The ion then travels through the charge detection cylinder CD into the ion mirror M2, where the ion reflection field formed in the region R2 of M2 reflects the ion to move in the opposite direction back into the charge detection cylinder CD, as shown by the ion's trajectory 62 in FIG. 4B.

[0036]

[0048] 4C, after an ion reflecting electric field is formed in region R2 of ion mirror M2, processor 52 is operable to control ion mirror M1 in an ion reflecting mode of operation (R) by controlling voltage source V1 to form an ion reflecting field in region R1 of ion mirror M1 while maintaining ion mirror M2 in the ion reflecting mode of operation (R) to trap ions in ELIT 14. In some embodiments, processor 52 is illustratively operable, i.e. programmed, to control ELIT 14 in a "random trapping mode" or "continuous trapping mode" in which processor 52 is operable to control ion mirror M1 in a reflecting mode of operation (R) after ELIT 14 has operated in the state shown in FIG. 4B, i.e., with M1 in an ion transmitting mode and M2 in an ion reflecting mode, for a selected period of time. ELIT 14 is controlled to operate in the state shown in FIG. 4B until the selected period has elapsed. In other embodiments, the processor 52 is illustratively operable, i.e. programmed, to control the ELIT 14 in a "triggered trapping mode" in which there is a substantially higher probability of trapping a single ion in the ELIT 14 as compared to a random trapping mode. In the "triggered trapping mode," the processor 52 is operable to control the ion mirror M1 to a reflective operating mode (R) after an ion is detected passing through the charge detection cylinder CD.

[0037]

[0049] In either case, once both ion mirrors M1, M2 are controlled to the ion-reflecting mode of operation (R) and trapping an ion in the ELIT 14, the ion is oscillated back and forth between the ion mirrors M1 and M2 by the opposing ion-reflecting fields formed in the regions R1 and R2 of the ion mirrors M1 and M2, respectively, passing each time through the charge detection cylinder CD, as shown by the ion's trajectory 64 shown in FIG. 4C and as described above. In one embodiment, the processor 50 is operable to maintain the operating state shown in FIG. 4C until the ion has passed through the charge detection cylinder CD a selected number of times. In an alternative embodiment, the processor 50 is operable to maintain the operating state shown in FIG. 4C for a selected period of time after controlling M1 (and in some embodiments M2) to the ion-reflecting mode of operation (R). In either embodiment, the number of cycles or time spent in the state shown in Figure 4C can illustratively be programmed, for example, by instructions stored in memory 54 or controlled through a user interface, and in either case, the ion detection event information obtained for each pass of an ion through the charge detection cylinder is temporarily stored in processor 50, for example in the form of an ion measurement file, which can illustratively have a predefined data or sample length. When an ion has passed through charge detection cylinder CD a selected number of times or oscillated back and forth between ion mirrors M1 and M2 for a selected time, the total number of charge detection events stored in processor 50 defines an ion measurement event, and once an ion measurement event is completed, the stored ion detection events defining the ion measurement event, for example an ion measurement event file, are passed to or retrieved by processor 52. 4A-4C then reverts to the sequence shown in Figure 4A where voltage sources V1, V2 are controlled as described above to control ion mirrors M1, M2, respectively, into an ion transmission mode of operation (T) by creating ion transmission fields within regions R1, R2 of ion mirrors M1, M2, respectively. The illustrated sequence is then repeated as many times as necessary.

[0038]

[0050] Previously, ion measurement event files were analyzed in the frequency domain using a Fast Fourier Transform (FFT) algorithm. In such an embodiment, the mass-to-charge ratio (m / z) of an ion was calculated from the fundamental oscillation frequency (f0) of the signal using a calibration constant (C) (Equation 1), and the charge of the ion was determined by the magnitude of the fundamental frequency peak in the FFT.

[0039]

number

[0040]

[0051] Since only the fundamental frequency was used in determining the charge of the ions, the signal can be thought of as being represented as just one sine wave. However, since the higher harmonics are ignored, a significant amount of information about the signal is not used by the FFT. This means that the signal needs to be measured for a longer period of time to obtain charge state resolution. A more complete representation of the waveform reduces the uncertainty in the amplitude, which improves charge accuracy and shortens the acquisition time required to reach charge state resolution. Furthermore, while the magnitude of the peaks in the FFT analysis depends on factors such as the duty cycle of the signal, the signal amplitude in the time domain is constant for a given charge, and amplitude measurements in the time domain are duty cycle independent. These properties make analysis in the time domain advantageous in applications with time-varying signal transients, such as those found in CDMS, where the ion oscillation frequency and signal duty cycle change as ions lose energy through collisions with background gas and electrostatic interactions with the detection cylinder.

[0041]

[0052] In the following, a process is described for analyzing signal measurements contained in a time domain ion measurement event file in combination with an FFT that incorporates information contained in higher harmonics by fitting the signal measurements to a simulated waveform in order to measure ion charge more accurately. In the following description, the ELIT is designed such that the time domain charge detection signal CHD stored in the ion measurement event file is a square wave signal (i.e., with a 50% duty cycle), but it will be understood that in alternative implementations, the ELIT may be designed such that the duty cycle of the time domain charge detection signal CHD is greater than or less than 50%. The following algorithm improves the accuracy of charge magnitude determination by 15% to 20% compared to an FFT by measuring a 50% duty cycle signal contained in the ion measurement file, which reaches the statistical lower limit of the uncertainty of the amplitude of a square wave corrupted by Gaussian noise. The best charge standard deviation that can be achieved with a square wave is determined by the standard deviation of the noise (σ) according to the following relationship: noise ), and the number of points that the waveform spends in a HI state compared to the points spent in a LO state (N HI and N LO )

[0042]

number

[0043]

[0053] 5, there is shown a simplified flow chart of an embodiment of a process 100 that analyzes signal measurements contained in a time domain ion measurement event file to determine the frequency and charge magnitude (z) of an ion oscillating back and forth through the charge detection cylinder of the ELIT during an ion trapping event. From this frequency determination, the mass to charge ratio (m / z) of the ion is determined from Equation 1, and the mass of the ion is determined as the product of m / z and z. Process 100 is illustratively stored in the memory of processor 16 in the form of instructions executable by processor 16 to perform the functions of process 100.

[0044]

[0054] Process 100 begins at step 102 where a time window counter N is initialized to 1 (or some other constant value). Process 100 is illustratively designed to analyze signal measurements contained in an ion measurement event file by analyzing signal measurements in each of a number of consecutive time windows in the measurement event file. This file windowing technique advantageously reduces the effect of time-varying frequency and duty cycle on the measured amplitude, so long as the frequency and duty cycle of the signal measurements do not change substantially within each time window, thereby allowing approximations of these parameters to be constant over the duration of each window. In one illustrative implementation in which the ion measurement event file is approximately 100 milliseconds long and contains approximately 1,000 cycles of signal measurements, the time windows are illustratively selected to be 10 milliseconds long each, with each of the 10 time windows containing 100 cycles of signal measurements.

[0045]

[0055] The process 100 continues at step 102 with the processor 16 performing an FFT analysis of a first time window of measurements of a signal contained in an ion measurement event file (hereinafter ion measurement signal IMS) to determine the fundamental oscillation frequency (F) of the first time window of the IMS signal in the conventional manner as described above. FFT ) and the magnitude of the charge (CH FFT ) for later use in process 100. FFT Multiply by 2.955 ADC bits / e to get the time-domain signal amplitude in ADC bits.

[0046]

[0056] Following step 104, the process 100 proceeds to step 106 where the processor 16 is operable to generate a simulated ion signal (SIS) for an Nth time window using input parameters F, CH, PH, and DC, where F is frequency, CH is charge magnitude, PH is phase, and DC is duty cycle.

[0047]

[0057] In one exemplary implementation, the SIS was generated by simulating the trajectory of one 130 eV / z ion with m / z of 25,600TH in the ELIT using the Beeman algorithm in Fortran (modified Velocity Verlet algorithm) at 10.02306 kHz with the electric field calculated using SIMION8.1. The signal of the ion was generated by superimposing the trajectory of the ion on a potential array where the charge detection cylinder has +1 and all other electrodes are held at ground. This generates a signal 160, normalized to +1 according to Green's reciprocity theorem, as shown in the example of FIG. 6. The signal 160 is divided into two sections, one negative-going transition 160A and one positive-going transition 160B. One period of the signal was fitted in OriginPro2018 using two bi-dose sigmoidal curves. One curve was used for the positive going transition 160B (for when the ion enters the detection cylinder) and another curve was used for the negative going transition 160A (for when the ion leaves the cylinder) according to the following formula:

[0048]

number

[0049] where t is time, p is a generic fitting parameter for a sigmoid curve, and I1 and I2 represent the times at which the SIS waveform 160 rises or falls. The values ​​of I1 and I2 have been further adjusted by ΔDC to change the duty cycle of the positive-going transitions while the negative-going transitions remain constant, as shown in the example of FIG. 7 where the duty cycle of the generated waveform 160 varies from 40% (trace 170A) to 60% (trace 170C), with a nominal 50% duty cycle (trace 170B) shown for comparison.

[0050]

[0058] In formula 3, f scaling is the desired frequency divided by the nominal frequency used in initially creating the analytic function for this waveform (e.g., 10.02306 kHz). Such an exemplary analytic function is shown in FIG. 8, where the signal 160 varies between a lower frequency 180A, e.g., 10 kHz, and a higher frequency 180B, e.g., 15 kHz. T is the wave period time, the time elapsed since the last wave period began is t-t0, and A is the amplitude. The phase is adjusted by adding a phase time to t0 which shifts the waveform by a specified amount of time. The variables h1 and h2 represent the speed at which the wave transitions from LO to HI or HI to LO states. Smaller values ​​of h produce a more rounded waveform, while larger values ​​of h produce a waveform with sharper transitions. The variable h scaling adjusts h multiplicatively to adjust the transition slope. The minimum and maximum values ​​of these curves were constrained to 0 and +1, respectively, so that the curves could be joined end to end to produce a periodic waveform. The SIS waveform was then scaled to an amplitude of 1500 ADC bits and centered at zero.

[0051]

[0059] As shown in the example of FIG. 9, to accommodate the RC damping 190 noted in existing mass spectrometers, a discrete-time first order recursive implementation of the high-pass filter (τ=7.89320623×10 -5seconds) was applied to the SIS waveform 160. The RC decay point i of the SIS waveform function was generated by multiplying the decay constant by the symmetric numerical derivative of the waveform according to the following equation:

[0052]

number

[0053]

[0060] The τ constant was determined by applying a square wave generated by a function generator to an antenna close to the charge detection cylinder of the spectrometer and fitting various RC values ​​to the square wave in the time domain to find the best fit. The variable Δt represents the time of a single ADC sample (400 ns).

[0054]

[0061] F=F FFT , CH=CH FFT 5, step 106 of process 100 for the first pass of the first time window (N=1) of the simulated ion signal SIS with PH=zero and an estimate of DC=49.2%, corresponding to the duty cycle of ions moving in an axial trajectory at 130 eV / z. Cross-correlation is further exemplarily performed in step 106 between the first time window of the IMS and the SIS, as illustrated by the example of process 200 shown in FIG. 10. In this process 200, the initial SIS (PH=0) is cross-correlated with the IMS by shifting the SIS by one sampling point (e.g., 400 ns) and then calculating the variance between the IMS and the SIS at each phase, e.g., using a conventional sum of residual squares (SRS). The minimum of the resulting correlation function (where the phases of the IMS and SIS coincide with the closest acquisition point) then exemplarily serves as an initial non-zero estimate of the phase of the PH of the SIS signal.

[0055]

[0062] From step 106, once a simulated ion signal (SIS) has been generated and initial input parameter values ​​entered as just described, the process 100 proceeds to step 108 where the processor 16 is operable to determine the variance between the IMS and the SIS. In one embodiment, the variance of the signal is determined using a conventional sum of squared residuals (SRS) according to the following formula, where in one implementation M=25,000 acquisition points (the number of points in a 10 millisecond window of the IMS file), although in alternative implementations M can be any positive integer.

[0056]

number

[0057] In alternative embodiments, other conventional variance determination equations and / or processes may be used.

[0063] In any case, following step 108, process 100 proceeds to step 110 where processor 16 is operable to determine whether the variance process performed in step 108 has converged. Convergence in step 110 is illustratively performed by comparing the result of Equation 5 with the result of a previous run of Equation 5. Since in the first run of step 110, Equation 5 has only been run once, process 100 follows the NO branch of step 110 and proceeds to step 112 where processor 16 is operable to run an optimization algorithm configured to reduce the variance between the IMS and the SIS.

[0058]

[0064] The variance between the IMS and the SIS determined in step 108 for each combination of input parameters generates a cost function that can be minimized, illustratively using any of a variety of conventional optimization algorithms, in step 112. In one exemplary implementation, a conventional gradient descent method is illustratively used as the optimization algorithm. This particular optimization method is beneficial in the current situation because a significant increase in throughput can be realized by using a fast first-order approximation algorithm. The gradient descent method allows the analysis method to be accelerated without significantly increasing computational costs to keep up with real-time data acquisition. In alternative embodiments, one or more other conventional optimization algorithms can be used.

[0059]

[0065] In gradient descent optimization, the IMS and SIS are compared by calculating the SRS between the IMS and SIS for a particular set of input parameters. The input parameters are then varied by relatively small amounts to determine the numerical partial derivatives of the SRS with respect to each of the input parameters. Following the calculation of the partial derivatives, in step 114, the input parameters are adjusted by multiplying each partial derivative of the SRS by a unique learning rate (γ) for each input parameter based on the individual convergence rate of the variance. X n If u is a vector of parameters at n iterations and γ is a vector of learning rates, the equation for gradient descent at step n+1 can be written as follows (Equation 6): where F, DC, PH, CH, and S represent the frequency, duty cycle, phase, amplitude, and transition slope parameters used in the synthesis of the noiseless waveform, respectively.

[0060]

number

[0061]

[0066] Note that the transition slope S does not apply to square waves because the transition is instantaneous, and in cases such as the process shown in FIG. 5, the transition slope is omitted. In either case, the process loops back to step 106 where the processor 16 is operable to generate a new simulated ion signal (SIS) using the adjusted parameters obtained by execution of step 114. This iterative process of steps 106-114 is also shown in alternative form by process 210 shown in FIG. 11 and continues until the processor 16 determines that a convergence limit has been reached in step 110. In one embodiment, this convergence limit is determined by the SRS (SRS n ) and the SRS in the previous iteration (SRS n-1 ) is set by the ratio of SRS n / SRS n-1 If x becomes sufficiently close to 1, e.g., between 0.99999999 and 1, for more than a predetermined number of iterations, e.g., 50 iterations, the processor 16 is operable to determine that the fit has converged. An example of a portion of an optimally fitted waveform 240 upon convergence is shown in FIG. 13, superimposed on a noise corrupted IMS signal 230.

[0062]

[0067] Following the YES branch of step 110, the process 100 proceeds to step 116 where the processor is operable to determine the frequency F(N), charge magnitude CH(N), and duty cycle DC(N) of the Nth time window of the IMS fitted to the SMS. The frequency F(N) of the Nth time window of the ion measurement signal IMS is illustratively calculated directly from the time-based transitions of the signal cycle (e.g., about 100 cycles in the exemplary embodiment described above). The charge magnitude CH(N) of the Nth time window of the ion measurement signal IMS is illustratively calculated as the average value of the amplitude of the cycles that make up the Nth time window, and DC(N) is the final value of DC at convergence.

[0063]

[0068] Following step 116, the process 100 proceeds to step 118 where the processor 16 is operable to determine whether the last time window of the ion measurement signal IMS has been processed. If NO, the process 100 proceeds to step 120 where the processor 16 is operable to advance the time window by a period ΔT, e.g., 10 milliseconds. The processor 16 is then operable to advance a time window counter N by 1 and set initial values ​​of the input parameters F, CH, and DC in step 122. After the first time window (N=1) is analyzed, the first guess for the next window consists of the frequency, duty cycle, and charge amplitude that were best fitted in the previous window. For every N≧2 windows, illustratively the first 50 iterations of the iterative process of steps 106-114 are reserved to find the phase PH of the next window, and then in subsequent iterations, all parameters are optimized until convergence is reached. This is shown graphically in FIG. 12 as a graph of SRS versus iteration number, where waveform 220 shows that the first 50 iterations in which the phase PH is found are relatively flat, after which the waveform 220 moves towards convergence.

[0064]

[0069] If, in step 116, the processor 16 determines that the last time window of the IMS has been processed, the process 100 continues to calculate the fundamental frequency F of the ion measurement signal. IMS The process proceeds to step 124 where the frequency values ​​F(N) for multiple time windows are processed by the processor 16 to determine the fundamental frequency F(N). In some embodiments, measurements of the oscillation of the ions in the ELIT are not recorded immediately to allow transients due to the switching voltages of the ion mirrors M1, M2 to subside. The ions then typically lose energy as they oscillate back and forth between the ion mirrors M1 and M2 due to collisions with background gas and electrostatic interactions with the charge detection cylinder. Such loss of energy results in an increase in frequency as the ions continue to oscillate back and forth between the ion mirrors M1 and M2, as shown graphically in FIG. 11. In such embodiments, the fundamental frequency F IMScan illustratively be calculated by fitting a line to the frequency F(N) for all time windows as a function of time, and then finding the frequency F before the ion loses any energy. IMS This is determined by extrapolating to the beginning of the capture event to determine the fundamental frequency F IMS is denoted as f0. The mass-to-charge ratio of the ion is then F IMS In other embodiments with shorter trapping times and / or improved ELIT structures, the ions do not appreciably lose energy during the trapping event, and in such embodiments, the fundamental frequency F IMS may be calculated as the average of F(N) over N windows.

[0065]

[0070] The processor 16 further processes the charge magnitude values ​​CH(N) for the multiple time windows to obtain the ion charge magnitude CH at step 124. IMS Since the charge is constant throughout the IMS file, the charge CH IMS is illustratively determined by averaging the charge magnitude values ​​CH(N) over all N windows.

[0066] example

[0071] FFT analysis of 1000 files containing a square wave signal of 100 ms duration corrupted by 1000 ADC bit RMSD of Gaussian noise gave a charge RMSD of 1.65 elementary electrons (e). Time domain analysis of the same files using the techniques described herein gave an RMSD of 1.35e. In addition, the amplitude reported in the time domain analysis is independent of RC decay, resulting in a 1% increase in signal-to-noise ratio. This represents a total improvement in charge precision of 19% compared to the FFT. The theoretical optimum charge RMSD was achieved using time domain analysis with a 50% duty cycle square wave (σ per Eq. 1). noise = 1000 ADC bits, N at 50% duty cycle HI =N LO = 125,000 points, σ best= 4 ADC bits or 1.35 elementary charges. An identical analysis performed on a file containing simulated ion signals corrupted by 1000 ADC bits RMSD of Gaussian noise gave an RMSD of 1.65e for the FFT analysis and 1.45e for the time domain analysis, indicating a 13% charge accuracy improvement.

[0067]

[0072] The reduced improvement in charge accuracy of the simulated ion signal compared to a square wave can be understood by considering the Hessian of the second partial derivatives for each of the parameters fitted by this algorithm.

[0068]

number

[0069] If the Hessian is diagonally dominant, the optimization problem is well-posed with a clear global minimum and the uncertainties due to each of the parameters do not couple with each other. In this situation, the parameters are linearly independent and a first-order gradient descent algorithm can quickly solve such a problem. This is realized with square wave signals, where the transitions between the HI and LO states of the signal are instantaneous (at least within the time resolution provided by the sampling frequency of 2.5 MHz). This means that the height of each transition and the time at which the transition occurs are independent of parameters such as the amplitude, frequency, duty cycle, and phase of the signal. On the other hand, the Hessian of an ion signal that transitions stepwise between HI and LO states is not diagonally dominant and has a significant contribution due to mixed partial derivatives that link the parameters and their respective uncertainties with each other. This means that the rise and fall times of the transitions are functions of frequency that couple the uncertainty of the frequency to the uncertainty of all other parameters (i.e., not knowing when the transition occurs means that the duty cycle cannot be assigned with confidence and is compensated for by erroneous amplitude measurements). There is no unique solution to this ill-posed optimization problem, and it is difficult to converge towards a minimum of the cost function when the signal is obscured by noise. Parameter interdependencies are minimized by designing a detection system that produces a signal that transitions sharply between LO and HI states. For example, this can be achieved by minimizing the inner diameter of the detection cylinder to give fast rise and fall times for the ion signal.

[0070]

[0073] In an alternative embodiment, the bidos sigmoid equation to which the ion signal was best fitted may be modified to fit the signal generated by the mass spectrometer, taking into account distortions of the signal shape resulting from geometric imperfections and / or other design features of the ELIT. The more accurate the actual instrument ion signal is known, the more accurately the waveform synthesis function may be applied to fit the instrument signal. Note that in the waveform synthesis subroutine, any function may be used to fit any signal, but the theoretical accuracy of the parameters depends on the characteristics of the waveform. Finally, a faster optimization algorithm, or an algorithm more suitable for nonlinear optimization problems, such as the simplex optimizer, may be used to fit a noise-free waveform to the signal. In addition, a significant increase in throughput may be realized by designing a system that generates a signal that may be fitted with a fast first-order gradient descent algorithm with momentum, such as AMS Grad. Alternatively, the number of steps required to reach convergence may be minimized by using a quadratic optimization scheme, such as Newton's method. These improvements make it possible to perform time domain analysis of files in combination with real-time FFT analysis.

[0071]

[0074] While this disclosure has been illustrated and described in detail in the foregoing drawings and description, it should be understood that this disclosure should be considered as illustrative and not limiting in character, and that only exemplary embodiments of this disclosure have been shown and described, and that all changes and modifications within the spirit of this disclosure are desired to be protected. For example, it will be understood that the ELIT 14 shown in the accompanying figures and described herein is presented by way of example only, and that the concepts, structures, and techniques described above may be directly implemented in ELITs of various alternative designs. Any such alternative ELIT designs may include, for example, any one or combination of multiple ELIT regions, more, fewer, and / or differently shaped ion mirror electrodes, more or fewer voltage sources, more or fewer DC or time-varying signals generated by one or more of the voltage sources, one or more ion mirrors defining additional electric field regions, and the like. As another example, in some alternative embodiments, the process illustrated in FIG. 5 may be implemented by determining the magnitude of the charge of the ion at the capture event, CH IMS (i.e., z), and the conventional FFT approach described above may be used to determine the mass-to-charge ratio (m / z). As yet another example, the process shown in Figure 5 may be modified to take into account possible variations in frequency measurements within one or more time windows, and / or the entire ion measurement file may be processed to take into account any such possible variations in frequency measurements.

Claims

1. 1. A method of measuring charge on an ion in an electrostatic linear ion trap including a charge detection cylinder disposed between two ion mirrors, wherein the ion repeatedly oscillates back and forth between the two ion mirrors each time the ion passes through the charge detection cylinder during an ion trapping event and induces a corresponding charge on the charge detection cylinder, and an ion measurement signal including a magnitude of the induced charge and a timing of the induced charge is recorded in an ion measurement file during the trapping event, the method comprising: (a) setting a time window for the ion measurement signal at the beginning of the ion measurement file; (b) generating a simulated ion measurement signal for the time window of the ion measurement signal using input parameters including estimates of signal frequency, charge magnitude, signal phase, and duty cycle; (c) iteratively processing the variance between the ion measurement signal and the simulated ion measurement signal for the time window by adjusting values ​​of the input parameters until the variance reaches convergence; (d) recording the charge magnitude value resulting from (c); (e) advancing the time window of the ion measurement signal by an incremental time; (f) repeating (b) through (d) until the time window reaches the end of the ion measurement file; (g) determining the charge of the ion based on the charge magnitude values ​​for each of the time windows; A method comprising:

2. 2. The method of claim 1, wherein (b) comprises processing the time window of the ion measurement signal to determine the estimate of signal frequency and charge magnitude.

3. 3. The method of claim 2, wherein processing the time window of the ion measurement signal comprises: calculating a Fast Fourier Transform (FFT) of the time window of the ion measurement signal; and determining the estimates of signal frequency and charge magnitude based on the (FFT).

4. for a first run of (b) for the time window of the ion measurement signal at the beginning of the ion measurement file, the initial estimate of the signal phase is set to zero; 4. The method of claim 1, wherein (b) further comprises the steps of cross-correlating the simulated ion measurement signal with the ion measurement signal for the time window, and updating the estimate of the signal phase to a minimum value obtained from the cross-correlation.

5. (c) (1) determining a variance between the ion measurement signal and the simulated ion measurement signal for the time window; (2) performing an optimization process to reduce the variance between the ion measurement signal and the simulated ion measurement signal for the time window; and (3) adjusting the values ​​of the input parameters based on the results of the optimization process. The method of any one of claims 1 to 4, comprising:

6. (c) further comprises recording the adjusted charge magnitude value upon convergence of the dispersion; (g) comprises determining the charge magnitude of the ions based on the adjusted charge magnitude values ​​for each of the time windows. The method according to claim 5.

7. The method of claim 1 , wherein (e) further comprises the step of setting the input parameters to the adjusted input parameter values ​​obtained from (c).

8. (d) further comprises, after step (c), recording a value of a frequency of the ion measurement signal for the time window; determining a frequency of oscillation of the ion during the ion trapping event based on the frequency values ​​for each of the time windows.

8. The method according to any one of claims 1 to 7.

9. computing a Fast Fourier Transform (FFT) of the ion measurement file; determining a frequency of oscillation of the ion during the ion capture event based on the FFT; The method of claim 1 , further comprising:

10. determining a mass to charge ratio of the ion based on the determined frequency of oscillation of the ion during the trapping event; determining a mass of the ion based on the determined mass to charge ratio of the ion and the determined charge of the ion; 10. The method of claim 8 or 9, further comprising:

11. an electrostatic linear ion trap (ELIT); an ion source configured to provide ions to the ELIT; a charge sensitive preamplifier having an input operably coupled to the ELIT; at least one processor operatively coupled to the ELIT and to an output of the charge sensitive preamplifier; At least one memory 13. A charge detection mass spectrometer (CDMS), comprising: instructions stored in the memory that, when executed by the at least one processor, cause the at least one processor to: (a) control the ELIT to trap ions provided from the ion source in the ELIT; (b) collect ion measurement information based on an output signal generated by the charge sensitive preamplifier as the trapped ions oscillate back and forth through the ELIT, the ion measurement information including a charge induced by the ion at a charge detector of the ELIT during each pass of the ion through the ELIT and timing of the induced charges relative to one another; and (c) process the ion measurement information in the time domain for each of a plurality of successive time windows of the ion measurement information to determine a magnitude of charge of the ion during each time window and determine the magnitude of charge of the trapped ion based on the magnitude of charge for each of the time windows. the ELIT comprises a charge detection cylinder disposed between two ion mirrors, the ions repeatedly oscillate back and forth between the two ion mirrors each time the ions pass through the charge detection cylinder during an ion capture event and induce a corresponding charge in the charge detection cylinder, and an ion measurement signal including a magnitude of the induced charge and a timing of the induced charge is recorded in an ion measurement file during the capture event; (c) (i) setting a time window for the ion measurement signal at the beginning of the ion measurement file; (ii) generating a simulated ion measurement signal for the time window of the ion measurement signal using input parameters including estimates of signal frequency, charge magnitude, signal phase, and duty cycle; (iii) iteratively processing the variance between the ion measurement signal and the simulated ion measurement signal for the time window by adjusting values ​​of the input parameters until the variance reaches convergence; (iv) recording the charge magnitude value resulting from (iii); and (v) advancing the time window of the ion measurement signal by an incremental time; (vi) repeating steps (ii)-(iv) until the time window reaches the end of the ion measurement file; (vii) determining the charge of the ion based on the charge magnitude values ​​for each of the time windows; and A charge detection mass spectrometer (CDMS).

12. (iv) further comprises, after (iii), recording a value of a frequency of the ion measurement signal for the time window; 12. The CDMS of claim 11 , wherein the instructions stored in the at least one memory further comprise instructions executable by the processor for determining a frequency of the oscillation of the ion during the ion trapping event based on the frequency value in each of the time windows.

13. 12. The CDMS of claim 11 , wherein the instructions stored in the at least one memory further comprise instructions executable by the processor to calculate a Fast Fourier Transform (FFT) of the ion measurement file and determine a frequency of the oscillation of the ions based on the FFT during the ion capture event.

14. 14. The CDMS of claim 12 or 13, wherein the instructions stored in the at least one memory include instructions executable by the processor to determine a mass to charge ratio of the ion during the capture event based on the determined frequency of oscillation of the ion, and to determine a mass of the ion based on the determined mass to charge ratio of the ion and the determined charge of the ion.

Images