Ion trajectories in an electrostatic ion guide for external ion source fourier transform ion cyclotron resonance mass spectrometry

An electrostatic ion guide (EIG) that consists of concentric cylinder and central wire electrodes can transport ions efficiently from an external ion source to an ion cyclotron resonance (ICR) ion trap for mass analysis, with several advantages over current injection methods. Because the electrostatic force of the EIG captures ions in a stable orbit about the wire electrode, ions with initially divergent trajectories may be redirected toward the ICR ion trap for improved ion transmission efficiency. SIMION trajectory calculations (ion kinetic energy, 1–200 eV; elevation angle, 0.30 °; azimuthal angle, 0.360°) predict that ions of m/z 1000 may be transmitted through a strong (0.01 → 3.0-T) magnetic field gradient. Judicious choice of ion source position and EIG potential minimizes the spread in ion axial kinetic energy at the ICR ion trap. Advantages of the EIG include large acceptance angle, even for ions that have large initial kinetic energy and large radial displacement with respect to the central z-axis, low ion extraction voltage (5–20 V), and efficient trapping because ions need not be accelerated to high velocity to pass through the magnetic field gradient.     

A combined linear ion trap for mass spectrometry

We propose a novel ion cyclotron resonance ion trap capable of confining ions even at high pressure. The trap consists of three capacitively coupled axial sections, each composed of four circular cross-section rods parallel to the magnetic field axis. Ion confinement along the magnetic field direction is provided by applying the same static voltage to each set of “endcap” rods. As for a two-dimensional quadrupole mass filter, a sufficiently high rf frequency (several MHz) leads to an “effective” electrostatic “pseudopotential” well with a minimum on the trap central axis. Ions are confined radially by the combination of an applied axial static magnetic field and a radially inward-directed electric field resulting from differential rf voltages applied to each set of four rods. Ion confinement properties are revealed from a Paul traplike “stability diagram,” whereas ion trajectories are analyzed in terms of Penning-type ion cyclotron rotation, magnetron rotation, and axial oscillation motional modes. Ion cyclotron frequency increases with the strength of the rf trapping field. Ion magnetron motion becomes stable if the rf voltage is high enough. Therefore, ion trajectories can be stable even in the presence of ion-neutral collisions. Adding an ac potential to a Penning trap should dramatically increase the upper mass detection limit.     

Characterization of an Electron Ionization Source Trap Operating in the Presence of a Magnetic Field Through Computer Simulation

We explore the feasibility of conducting electron ionization (EI) in a radio-frequency (rf) ion source trap for mass spectrometry applications. Electrons are radially injected into a compact linear ion trap in the presence of a magnetic field used essentially to lengthen the path of the electrons in the trap. The device can either be used as a stand-alone mass spectrometer or can be coupled to a mass analyzer. The applied parallel magnetic field and the oscillating rf electric field produced by the trap give rise to a set of coupled Mathieu equations of motion. Via numerical simulations, electron trajectories are studied under varying intensities of the magnetic field in order to determine the conditions that enhance ion production. Likewise, the dynamic behavior of the ions are investigated in the proposed EI source trap and the fast Fourier transform FFT formalism is used to obtain the frequency spectrum from the numerical simulations to study the motional frequencies of the ions which include combinations of the low-frequency secular and the high-frequency micromotion with magnetron and cyclotron frequencies. The dependence of these motional frequencies on the trapping conditions is examined and particularly, the limits of applying a radial magnetic field to the EI ion trap are characterized.

Controlled ion fragmentation in a 2-D quadrupole ion trap for external ion accumulation in ESI FTICR mass spectrometry

Undesired fragmentation of electrospray generated ions in an rf multipole traps can be problematic in many applications. Of special interest here is ion dissociation in a 2-D quadrupole ion trap external to a Fourier transform ion cyclotron resonance mass spectrometer (FTICR MS) used in proteomic studies. In this work, we identified the experimental parameters that determine the efficiency of ion fragmentation. We have found that under the pressure conditions used in this study there is a specific combination of the radial and axial potential well depths that determines the fragmentation threshold. This combination of rf and dc fields appears to be universal for ions of different mass-to-charge ratios, molecular weights, and charge states. Such universality allows the fragmentation efficiency of the trapped ions in the course of capillary liquid chromatography (LC) separation studied to be controlled and can increase the useful duty cycle and dynamic range of a FTICR mass spectrometer equipped with an external rf only 2-D quadrupole ion trap.     

Stacked-ring electrostatic ion guide

In 1969 Bahr, Gerlich, and Teloy introduced an rf device that consisted of a stack of ring electrodes, with charge sign alternation between neighboring rings, to store or transport ions. Here we propose to operate such a device with electrostatic potentials rather than rf potentials: ions that move axially along the center of the guide are thereby subjected to an oscillating electrical potential similar to the sinusoidal rf potential in familiar rf-only multipole ion guides. The oscillating potential of the stacked-ring static ion guide focuses ions by exerting a field gradient force on the ions so as to push ions toward the central axis where the field is weakest. The stacked-ring ion guide produces an effectively static “pseudopotential” that is much steeper at the edge (potential varies as e ^r) compared to a quadrupole or octupole guide (for which the potential varies as r ² or r ⁶, where r is radial position) and that is much flatter near the center of the guide (for potentially higher ion flux). Advantages of the new ion guide include static rather than rf potential, low electrical noise, a large field-free region near the central axis of the guide, and simple mechanical construction. A disadvantage of the stacked-ring ion guide is that high ion axial kinetic energy is required; ions with axial kinetic energy that is too low may be trapped in the shallow pseudopotential well between adjacent ring electrodes.     

Computer simulation of the gap-tripole ion trap with linear injection, 3D ion accumulation,and versatile packet ejection

The behavior of a completely new ion trap is shown with SIMION 7.0 simulations. The simulated trap, which was a mix of a linear and a 3D trap, was made by axially setting two ion guides with a gap between them. Each guide consisted of three rods with three symmetrically delayed radio frequency (rf) voltages (tripole). The "injected" ions were linearly contained by pulsed potentials on the entrance and exit plates. Then the three-dimensional (3D) rf field in the gap, which was created by the tripole special rod arrangement, could trap the ions when the translational energy was dampened by collisions with low-pressure nitrogen. Because the injected ions were trapped in the small gap, the trapping cycle could be repeated many times before ion ejection, so a high concentrated ion cloud could be obtained. This trapping and accumulation methodology is not possible in most conventional multipole linear traps with even number of poles. Compared with quadrupole linear trap at the same rf amplitude, tripole lost more ions due to strong charge repulsion in the ion cloud. However, tripole could catch up the ions at higher voltage. Radial and axial mass-independent ejection of the ions localized in the tripole gap was very simple, compared with conventional linear ion traps that need extra and complicated electrodes for effective axial ejection.     

Electrospray ionization—Fourier transform ion cyclotron resonance mass spectrometry at 11.5 tesla: Instrument design and initial results

Initial results obtained using a new electrospray ionization (ESI) Fourier transform ion cyclotron resonance (FTICR) mass spectrometer operated at a magnetic field 11.5 tesla are presented. The new instrument utilized an electrostatic ion guide between the ESI source and FTICR trap that provided up to 5% overall transmission efficiency for light ions and up to 30% efficiency for heavier biomolecules. The higher magnetic field in combination with an enlarged FTICR ion trap made it possible to substantially improve resolving power and operate in a more robust fashion for large biopolymers compared to lower field instruments. Mass resolution up to 10⁶ has been achieved for intermediate size biopolymers such as bovine ubiquitin (8.6 kDa) and bovine cytochrome c (12.4 kDa) without the use of frequency drift correction methods. A mass resolution of 370,000 has been demonstrated for isotopically resolved molecular ions of bovine serum albumin (66.5 kDa). Comparative measurements were made with the same spectrometer using a lower field 3.5-tesla magnet allowing the performance gains to be more readily quantified. Further improvements in pumping capacity of the vacuum system and efficiency of ion transmission from the source are expected to lead to further substantial sensitivity gains.     

Controlled Ion Ejection from an External Trap for Extended m/z Range in FT-ICR Mass Spectrometry

An auxiliary rf waveform of the same amplitude and phase applied to all the rods of an ion accumulation multipole creates an m/z-dependent axial pseudo potential. Controlled decrease of the auxiliary rf amplitude releases ions from the accumulation multipole sequentially from high to low m/z. The slope of the auxiliary rf voltage ramp is adjusted so that ions of different m/z reach the center of the ICR cell at the same time point, which mitigates the typical time dispersion observed in external source FT-ICR and extends the observable mass range for a single data acquisition by 2- to 3-fold. For complex mixture analysis, twice the number of elemental compositions are assigned when the auxiliary rf ejection is applied compared with the standard gated trapping.

A novel fourier transform ion cyclotron resonance mass spectrometer with improved ion trapping and detection capabilities

A novel Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometer has been developed for improved biomolecule analysis. A flared metal capillary and an electrodynamic ion funnel were installed in the source region of the instrument for improved ion transmission. The transfer quadrupole is divided into 19 segments, with the capacity for independent control of DC voltage biases for each segment. Restrained ion population transfer (RIPT) is used to transfer ions from the ion accumulation region to the ICR cell. The RIPT ion guide reduces mass discrimination that occurs as a result of time-of-flight effects associated with gated trapping. Increasing the number of applied DC bias voltages from 8 to 18 increases the number of ions that are effectively trapped in the ICR cell. The RIPT ion guide with a novel voltage profile applied during ion transfer provides a 3- to 4-fold increase in the number of ions that are trapped in the ICR cell compared with gated trapping for the same ion accumulation time period. A novel ICR cell was incorporated in the instrument to reduce radial electric field variation for ions with different z-axis oscillation amplitudes. With the ICR cell, called trapping ring electrode cell (TREC), we can tailor the shape of the trapping electric fields to reduce dephasing of coherent cyclotron motion of an excited ion packet. With TREC, nearly an order of magnitude increase in sensitivity is observed. The performance of the instrument with the combination of RIPT, TREC, flared inlet, and ion funnel is presented.     

A new technique for unbiased external ion accumulation in a quadrupole two-dimensional ion trap for electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry.

External ion accumulation in a two-dimensional (2D) multipole trap has been shown to increase the sensitivity, dynamic range and duty cycle of a Fourier transform ion cyclotron resonance (FTICR) mass spectrometer. However, it is important that trapped ions be detected without significant bias at longer accumulation times in the external 2D multipole trap. With increasing ion accumulation time pronounced m/z discrimination was observed when trapping ions in an accumulation quadrupole. In this work we show that superimposing lower rf-amplitude dipolar excitation over the main rf-field in the accumulation quadrupole results in disruption of the m/z discrimination and can potentially be used to achieve unbiased external ion accumulation with FTICR.     

Fourier Transform Ion Cyclotron Resonance Mass Resolution and Dynamic Range Limits Calculated by Computer Modeling of Ion Cloud Motion

Particle-in-Cell (PIC) ion trajectory calculations provide the most realistic simulation of Fourier transform ion cyclotron resonance (FT-ICR) experiments by efficient and accurate calculation of the forces acting on each ion in an ensemble (cloud), including Coulomb interactions (space charge), the electric field of the ICR trap electrodes, image charges on the trap electrodes, the magnetic field, and collisions with neutral gas molecules. It has been shown recently that ion cloud collective behavior is required to generate an FT-ICR signal and that two main phenomena influence mass resolution and dynamic range. The first is formation of an ellipsoidal ion cloud (termed “condensation”) at a critical ion number (density), which facilitates signal generation in an FT-ICR cell of arbitrary geometry because the condensed cloud behaves as a quasi-ion. The second phenomenon is peak coalescence. Ion resonances that are closely spaced in m/z coalesce into one resonance if the ion number (density) exceeds a threshold that depends on magnetic field strength, ion cyclotron radius, ion masses and mass difference, and ion initial spatial distribution. These two phenomena decrease dynamic range by rapid cloud dephasing at small ion density and by cloud coalescence at high ion density. Here, we use PIC simulations to quantitate the dependence of coalescence on each critical parameter. Transitions between independent and coalesced motion were observed in a series of the experiments that systematically varied ion number, magnetic field strength, ion radius, ion m/z, ion m/z difference, and ion initial spatial distribution (the present simulations begin from elliptically-shaped ion clouds with constant ion density distribution). Our simulations show that mass resolution is constant at a given magnetic field strength with increasing ion number until a critical value (N) is reached. N dependence on magnetic field strength, cyclotron radius, ion mass, and difference between ion masses was determined for two ion ensembles of different m/z, equal abundance, and equal cyclotron radius. We find that N and dynamic range depend quadratically on magnetic field strength in the range 1–21 Tesla. Dependences on cyclotron radius and Δm/z are linear. N depends on m/z as (m/z)^–2. Empirical expressions for mass resolution as a function of each of the experimental parameters are presented. Here, we provide the first exposition of the origin and extent of trade-off between FT-ICR MS dynamic range and mass resolution (defined not as line width, but as the separation between the most closely resolved masses).     

A new hybrid electrospray Fourier transform mass spectrometer: design and performance characteristics

A new hybrid electrospray quadrupole Fourier transform mass spectrometry (FTMS) instrument design is shown and characterized. This instrument involves coupling an electrospray source and mass-resolving quadrupole, ion accumulation, and collision cell linear ion trap system developed by MDS Sciex with a home-built ion guide and ion cyclotron resonance (ICR) cell. The iterative progression of this design is shown. The final design involves a set of hexapole ion guides to transfer the ions from the accumulation/collision trap through the magnetic field gradient and into the cell. These hexapole ion guides are separated by a thin gate valve and two conduction limits to maintain the required <10(-9) mbar vacuum for FTICR. Low-attomole detection limits for a pure peptide are shown, 220 000 resolving power in broadband mode and 820 000 resolving power in narrow-band mode are demonstrated, and mass accuracy in the <2 ppm range is routinely available provided the signal is abundant, cleanly resolved, and internally calibrated. This instrument design provides high experimental flexibility, allowing Q2 CAD, SORI-CAD, IRMPD, and ECD experiments with selected ion accumulation as well as experiments such as nozzle skimmer dissociation. Initial top-down mass spectrometry experiments on a protein is shown using ECD.     

Dynamically Harmonized FT-ICR Cell with Specially Shaped Electrodes for Compensation of Inhomogeneity of the Magnetic Field. Computer Simulations of the Electric Field and Ion Motion Dynamics

The recently introduced ion trap for FT-ICR mass spectrometers with dynamic harmonization showed the highest resolving power ever achieved both for ions with moderate masses 500?C1000?Da (peptides) as well as ions with very high masses of up to 200?kDa (proteins). Such results were obtained for superconducting magnets of very high homogeneity of the magnetic field. For magnets with lower homogeneity, the time of transient duration would be smaller. In superconducting magnets used in FT-ICR mass spectrometry the inhomogeneity of the magnetic field in its axial direction prevails over the inhomogeneity in other directions and should be considered as the main factor influencing the synchronic motion of the ion cloud. The inhomogeneity leads to a dependence of the cyclotron frequency from the amplitude of axial oscillation in the potential well of the ion trap. As a consequence, ions in an ion cloud become dephased, which leads to signal attenuation and decrease in the resolving power. Ion cyclotron frequency is also affected by the radial component of the electric field. Hence, by appropriately adjusting the electric field one can compensate the inhomogeneity of the magnetic field and align the cyclotron frequency in the whole range of amplitudes of z-oscillations. A method of magnetic field inhomogeneity compensation in a dynamically harmonized FT-ICR cell is presented, based on adding of extra electrodes into the cell shaped in such a way that the averaged electric field created by these electrodes produces a counter force to the forces caused by the inhomogeneous magnetic field.     

Application of sustained off-resonance irradiation: The beat frequency measurement and radial separation of mass-selected ions

Newer applications of sustained off-resonance irradiation (SORI) in Fourier-transform ion cyclotron resonance (FT-ICR) spectrometry are described. SORI induced circular gyrations of ions with their cyclotron radii varying sinusoidally at the corresponding beat frequency. The beat is observed in the temporal variation of FT-ICR peak height as a function of the duration of the SORI burst, and it allows the direct measurement of the beat frequency. The effective cyclotron frequency is determined from the measured beat frequency and the applied burst frequency. The frequency uncertainty is greatly reduced by nearly 3 orders of magnitude compared with the uncertainty in FT-ICR peak frequency that fluctuates with the rf burst duration. In addition, the oscillating cyclotron radii of the irradiated ions allow the radial separation of mass-selected ions so that the mass-selective photodissociation can be carried out with m-bromotoluene ions.     

Matrix-assisted laser desorption/ionization fourier transform ion cyclotron resonance mass spectrometry with pulsed in-source collision gas and in-source ion accumulation

A new matrix-assisted laser desorption/ionization (MALDI) source for Fourier transform ion cyclotron resonance mass spectrometry (FTMS) has been developed. The new source is equipped with a hexapole ion guide. The sample on the laser target is one millimeter from the hexapole ion guide, so that ions are desorbed directly into the guide. A device for pulsing collision gas in direct proximity to the laser target makes it possible to cool the ions, which have a kinetic energy spread of several electron volts when produced by the MALDI process. These ions are trapped in the hexapole where positive potentials at the laser target and at an extraction plate help trap ions along the longitudinal axis. After a pre-defined trapping time the voltage of the extraction plate is reversed and the trapped ions are extracted for transmission to the ion cyclotron resonance cell. Accumulation of ions from multiple laser shots in the hexapole before mass spectrometric analysis increases sensitivity. Preliminary sensitivity studies with substance P show that 10 attomoles of analyte applied on the target can be detected with a signal-to-noise (S/N) ratio >15.     

Reduction of axial kinetic energy induced perturbations on observed cyclotron frequency

With Fourier transform ion cyclotron resonance (FTICR) mass spectrometry one determines the mass-to-charge ratio of an ion by measuring its cyclotron frequency. However, the need to confine ions to the trapping region of the ion cyclotron resonance (ICR) cell with electric fields induces deviations from the unperturbed cyclotron frequency. Additional perturbations to the observed cyclotron frequency are often attributed to changes in space charge conditions. This study presents a detailed investigation of the observed ion cyclotron frequency as a function of ion z-axis kinetic energy. In a perfect three-dimensional quadrupolar field, cyclotron frequency is independent of position within the trap. However, in most ICR cell designs, this ideality is approximated only near the trap center and deviations arise from this ideal quadrupolar field as the ion moves both radially and axially from the center of the trap. To allow differentiation between deviations in observed cyclotron frequency caused from changes in space charge conditions or differences in oscillation amplitude, ions with identical molecular weights but different axial kinetic energy, and thus amplitude of z-axis motion, were simultaneously trapped within the ICR cell. This allows one to attribute deviations in observed cyclotron frequency to differences in the average force from the radial electric field experienced by ions of different axial amplitude. Experimentally derived magnetron frequency is compared with the magnetron frequency calculated using SIMION 7.0 for ions of different axial amplitude. Electron promoted ion coherence, or EPIC, is used to reduce the differences in radial electric fields at different axial positions. Thus with the application of EPIC, the differences in observed cyclotron frequencies are minimized for ions of different axial oscillation amplitudes.     

Improved ion extraction from a linear octopole ion trap: SIMION analysis and experimental demonstration

Externally generated ions are accumulated in a linear octopole ion trap before injection into our 9.4 T Fourier transform ion cyclotron resonance (FT-ICR) mass analyzer. Such instrumental configuration has previously been shown to provide improved sensitivity, scan rate, and duty cycle relative to accumulated trapping in the ICR cell. However, inefficient ion ejection from the octopole currently limits both detection limit and scan rate. SIMION 7.0 analysis predicts that a dc axial electric field inside the linear octopole ion trap expedites and synchronizes the efficient extraction of the octopole-accumulated ions. Further SIMION analysis optimizes the ion ejection properties of each of three electrode configurations designed to produce a near-linear axial potential gradient. More efficient extraction and transfer of accumulated ions spanning a wide m/z range promises to reduce detection limit and increase front-end sampling rate (e.g., to increase front-end resolution for separation techniques coupled with FT-ICR mass analysis). Addition of the axial field improves experimental signal-to-noise ratio by more than an order of magnitude.     

Successive Resonances for Ion Ejection at Arbitrary Frequencies in an Ion Trap

The use of successive resonances for ion ejection is demonstrated here as a method of scanning quadrupole ion traps with improvement in both resolution and sensitivity compared with single frequency resonance ejection. The conventional single frequency resonance ejection waveform is replaced with a dual-frequency waveform. The two included frequencies are spaced very closely and their relative amplitudes are adjusted so that the first frequency that ions encounter excites them to higher amplitudes where space charge effects are less prominent, thereby giving faster and more efficient ejection when the ions come into resonance with the second frequency. The method is applicable at any arbitrary frequency, unlike double and triple resonance methods. However, like double and triple resonance ejection, ejection using successive resonances requires the rf and AC waveforms to be phase-locked in order to retain mass accuracy and mass precision. The improved performance is seen in mass spectra acquired by rf amplitude scans (resonance ejection) as well as by secular frequency scans.

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Graphical Abstract ?

     

Space charge effects on relative peak heights in fourier transform-ion cyclotron resonance spectra

Ion trajectory calculations have confirmed that space charge interactions can be a source for mass discrimination seen in Fourier transform-ion cyclotron resonance (FT-ICR) spectra. As compared with the previously recognized mechanism of z-axis excitation, ion-ion repulsion is a mechanism which specifically affects relative peak heights of ions close in mass, and is most severe for low excitation radiofrequency (rf) amplitudes. In this mechanism, Coulomb repulsion significantly perturbs the motion of the ion clouds during excitation and alters the final cyclotron orbital radii. Under these conditions peak heights do not accurately reflect the true ion abundances in the FT-ICR spectrometer. Mass discrimination can be minimized by using low numbers of ions, low ion densities, and a short, high amplitude rf excitation waveform. Experimental observation of the relative peak heights of the m/z 91, 92, and 134 ions in n-butylbenzene gives quantitative confirmation of the results of the trajectory calculations. Chirp, SWIFT, and impulse excitation were modeled: impulse excitation was found to be most effective in minimizing the effects of space charge interactions.     

Ion Behavior in an Electrically Compensated Ion Cyclotron Resonance Trap

We recently described a new electrically compensated trap in FT ion cyclotron resonance mass spectrometry and developed a means of tuning traps of this general design. Here, we describe a continuation of that research by comparing the ion transient lifetimes and the resulting mass resolving powers and signal-to-noise (S/N) ratios that are achievable in the compensated vs. uncompensated modes of this trap. Transient lifetimes are ten times longer under the same conditions of pressure, providing improved mass resolving power and S/N ratios. The mass resolving power as a function of m/z is linear (log-log plot) and nearly equal to the theoretical maximum. Importantly, the ion cyclotron frequency as a function of ion number decreases linearly in accord with theory, unlike its behavior in the uncompensated mode. This linearity should lead to better control in mass calibration and increased mass accuracy than achievable in the uncompensated mode.