Characterization of an improved electrodynamic ion funnel interface for electrospray ionization mass spectrometry.

An improved electrodynamic ion funnel for ion focusing at high pressure (> 1 Torr) has been developed for a triple quadrupole mass spectrometer and its performance compared with that of an earlier prototype previously reported. The ion funnel consists of a series of ring electrodes of progressively smaller internal diameters to which rf and dc electric potentials are co-applied. The new design utilizes ring electrodes possessing larger internal diameters that taper down to a relatively larger exit aperture. In the 1-10 Torr pressure range, the new design provides significant improvement in low m/z ion transmission. Additionally, the overall ion transmission range is improved by linked scanning of the ion funnel's rf voltage concomitantly with the scanning of the quadrupole mass analyzer. Transmission of a noncovalent complex through the interface demonstrated that excessive ion heating was not problematic. Computer simulations of ion transport support the ion funnel design and help explain the relative performance of both designs. Both ion simulations and experimental results are in accord and indicate close to 100% ion transmission efficiency for electrosprayed biopolymer ions through the interface and into the mass analyzer.     

Independent control of ion transmission in a jet disrupter dual-channel ion funnel electrospray ionization MS interface

A new atmospheric pressure ionization mass spectrometer (API-MS) interface has been developed to allow the control of ion transmission through the first vacuum stage of the mass spectrometer. The described interface uses a dual-heated capillary and a dual-inlet ion funnel design. Two electrosprays, aligned with the dual-capillary inlet, are used to introduce ions from different solutions independently into the MS. The initial design was specifically aimed at developing a method for the controlled introduction of calibrant ions in highly accurate mass measurements using Fourier transform ion cyclotron resonance mass spectrometer (FTICR). The dual-channel ion funnel has different inlet diameters that are aligned with the dual capillaries. The large diameter main channel of the ion funnel is used for analyte introduction to provide optimum ion transmission. The second, smaller diameter channel inlet includes a jet disrupter in the ion funnel to modulate the ion transmission through the channel. The two inlet channels converge into a single-channel ion funnel where ions from both channels are mixed, focused, and transmitted to the mass analyzer. Both theoretical simulations and experimental results show that the transmission of different m/z species in the small diameter channel of the ion funnel can be effectively modulated by varying the bias voltage on the jet disrupter. Both static and dynamic modulations of ion transmission are demonstrated experimentally by applying either a constant DC or a square waveform voltage to the jet disrupter. High ion transmission efficiency, similar to the standard single-channel ion funnel, is maintained in the main analyte channel inlet of the ion funnel over a broad m/z range with negligible "cross talk" between the two ion funnel inlet channels. Several possible applications of the new interface (e.g., for high-accuracy MS analysis of complex biological samples) are described.     

Improved ion transmission from atmospheric pressure to high vacuum using a multicapillary inlet and electrodynamic ion funnel interface

A heated multicapillary inlet and ion funnel interface was developed to couple an electrospray ionization (ESI) source to a high-vacuum stage for obtaining improved sensitivity in mass spectrometric applications. The inlet was constructed from an array of seven thin-wall stainless steel tubes soldered into a central hole of a cylindrical heating block. An electrodynamic ion funnel was used in the interface region to more effectively capture, focus, and transmit ions from the multicapillary inlet. The interface of seven capillary inlets with the ion funnel showed more than 7 times higher transmission efficiency compared to that of a single capillary inlet with the ion funnel and a 23-fold greater transmission efficiency than could be obtained using the standard orifice-skimmer interface of a triple-quadrupole MS. The multiple-capillary inlet and ion funnel interface showed an overall 10% ion transmission efficiency and approximately 3-4% overall detection efficiency of ions from solution based (i.e., prior to electrospray). The improved performance was achieved under conditions where ESI operation is robust and results in a significant increase in dynamic range.     

A multicapillary inlet jet disruption electrodynamic ion funnel interface for improved sensitivity using atmospheric pressure ion sources

A new multicapillary inlet and ion funnel interface for electrospray ionization-mass spectrometry has been developed and demonstrated to achieve higher ion transmission efficiency compared to a single-capillary inlet and ion funnel interface. Even though the distance between the end of the ESI inlet capillary and the exit of the ion funnel (10 cm) is significantly longer than that of the conventional interface (typically a few millimeters), a significant part of the directed inlet gas flow persists into the first stage of pumping and results in an increased gas load to the second chamber. A jet disrupter made of a circular metal disk placed on axis in the ion funnel enhanced the dispersion of the directed gas flow from a multicapillary inlet and was also found to improve the ion transmission. The ion funnel with the jet disrupter demonstrated a 15% improvement in ion transmission (compared to that without the jet disrupter) and simultaneously reduced the pumping speed required for the first or second stage by a factor of 2-3. Compared to the sensitivity with the standard mass spectrometer interface (an API 3000, Sciex, Concord, ON, Canada) in MS/MS operation using an interface equipped with the jet disrupter and ion funnel, a 5.3-10.7-fold enhancement in signal was observed for samples with concentrations of 100-500 pg/microL and 10.2 to 14.1-fold enhancement for concentrations of 10 to 50 pg/microL. The decreased enhancement at higher concentrations is attributed to space charge effects and detector saturation.     

Zeptomole-sensitivity electrospray ionization--Fourier transform ion cyclotron resonance mass spectrometry of proteins

Methods are being developed for ultrasensitive protein characterization based upon electrospray ionization (ESI) with Fourier transform ion cyclotron resonance mass spectrometry (FTICR-MS). The sensitivity of a FTICR mass spectrometer equipped with an ESI source depends on the overall ion transmission, which combines the probability of ionization, transmission efficiency, and ion trapping in the FTICR cell. Our developments implemented in a 3.5 tesla FTICR mass spectrometer include introduction and optimization of a newly designed electrodynamic ion funnel in the ESI interface, improving the ion beam characteristics in a quadrupole-electrostatic ion guide interface, and modification of the electrostatic ion guide. These developments provide a detection limit of approximately 30 zmol (approximately 18,000 molecules) for proteins with molecular weights ranging from 8 to 20 kDa.     

Subambient pressure ionization with nanoelectrospray source and interface for improved sensitivity in mass spectrometry

A nanoelectrospray ionization mass spectrometry (ESI-MS) source and interface has been designed that enables efficient ion production and transmission in a 30 Torr pressure environment using solvents compatible with typical reversed-phase liquid chromatography (RPLC) separations. In this design, the electrospray emitter is located inside the mass spectrometer in the same region as an electrodynamic ion funnel. This avoids the use of a conductance limiting ion inlet, as required by a conventional atmospheric pressure ESI source, and allows more efficient ion transmission to the mass analyzer. The new subambient pressure ionization with nanoelectrospray (SPIN) source improves instrument sensitivity and enables new electrospray interface designs, including the use of multi-emitter approaches. Performance of the SPIN source was evaluated by electrospraying standard solutions at 300 nL/min and comparing results with those obtained from a standard atmospheric pressure ESI source that used a heated capillary inlet. This initial study demonstrated an approximately 5-fold improvement in sensitivity when the SPIN source was used compared to a standard atmospheric pressure ESI source. The importance of desolvation was also investigated by electrospraying at different flow rates, which showed that the ion funnel provided an effective desolvation region to aid the creation of gas-phase analyte ions.     

Analysis of High Mass-to-Charge Ions in a Quadrupole Ion Trap Mass Spectrometer via an End-Cap Quadrupolar Direct Current Downscan

A method for performing mass-selective instability analysis in a three-dimensional (3-D) quadrupole ion trap is described that involves scanning a direct current (dc) voltage applied to the end-cap electrodes while holding the radio frequency (rf) potential at a fixed value. Rather than eject at the ?(z) = 1 instability line by ramping the amplitude of the drive rf potential applied to the ring electrode, as with the original mass-selective instability scan, this approach effects ion ejection along the ?(z) = 0 instability line in a process identical in principle (though it varies in its method of implementation) to the previously termed "downscan" ( Todd , J. F. J. ; Penman , A. D. ; Smith , R. D. Int. J. Mass Spectrom. Ion Processes 1991 , 106 , 117 - 135 ). A linear scan of the dc amplitude results in a nonlinear mass scale, unlike the conventional resonance ejection scan with a linear scan of the rf amplitude, and the ejection of ions in the direction of high mass-to-charge (m/z) to low m/z. However, the downscan offers some advantages over the traditional rf scan for ions of high m/z values. These include a larger scannable mass range, as well as the opportunity for improved resolution at high mass. These characteristics are demonstrated with ions of m/z 10(4)-10(5).     

Preparative separation of mixtures by mass spectrometry

A specially designed mass spectrometer which allows for preparative separation of mixtures is described. This mass spectrometer allows for large ion currents, on the order of nanoamperes, to be produced by electrospray and transmitted into a high vacuum. Accumulation of nanomole quantities of collected and recovered material in several hours is demonstrated. The use of high-velocity ions reduces space charge effects at high ion currents. Separation of mass occurs simultaneously for all ions, providing a 100% duty cycle. The use of a linear dispersion magnet avoids compression at higher m/z ratios. A deceleration lens slows the ions to allow for soft landing at low kinetic energy. The ions are neutralized by ion pairing on an oxidized metal surface. Retractable landing plates allow for easy removal of the separated components.     

Ion funnel trap interface for orthogonal time-of-flight mass spectrometry

A combined electrodynamic ion funnel and ion trap coupled to an orthogonal acceleration (oa)-time-of-flight mass spectrometer was developed and characterized. The ion trap was incorporated through the use of added terminal electrodynamic ion funnel electrodes enabling control over the axial dc gradient in the trap section. The ion trap operates efficiently at a pressure of approximately 1 Torr, and measurements indicate a maximum charge capacity of approximately 3 x 10(7) charges. An order of magnitude increase in sensitivity was observed in the analysis of low concentration peptides mixtures with orthogonal acceleration (oa)-time-of-flight mass spectrometry (oa-TOF MS) in the trapping mode as compared to the continuous regime. A signal increase in the trapping mode was accompanied by reduction in the chemical background, due to more efficient desolvation of, for example, solvent related clusters. Controlling the ion trap ejection time was found to result in efficient removal of singly charged species and improving signal-to-noise ratio (S/N) for the multiply charged analytes.     

Ion soft landing using a rectilinear ion trap mass spectrometer

A new ion soft landing instrument has been built for the controlled deposition of mass selected polyatomic ions. The instrument has been operated with an electrospray ionization source; its major components are an electrodynamic ion funnel to reduce ion loss, a 90-degree bent square quadrupole that prevents deposition of fast neutral molecules onto the landing surface, and a novel rectilinear ion trap (RIT) mass analyzer. The ion trap is elongated (inner dimensions: 8 mm x 10 mm x 10 cm). Three methods of mass analysis have been implemented. (i) A conventional mass-selective instability scan with radial resonance ejection can provide a complete mass spectrum. (ii) The RIT can also be operated as a continuous rf/dc mass filter for isolation and subsequent soft landing of ions of the desired m/ z value. (iii) The 90-degree bent square quadrupole can also be used as a continuous rf/dc mass filter. The mass resolution (50% definition) of the RIT in the trapping mode (radial ion ejection) is approximately 550. Ions from various test mixtures have been mass selected and collected on fluorinated self-assembled monolayers on gold substrates, as verified by analysis of the surface rinses. Desorption electrospray ionization (DESI) has been used to confirm intact deposition of [Val (5)]-Angiotensin I on a surface. Nonmass selective currents up to 1.1 nA and mass-selected currents of up to 500 pA have been collected at the landing surface using continuous rf/dc filtering with the RIT. A quantitative analysis of rinsed surfaces showed that the overall solution-to-solution soft landing yields are between 0.2 and 0.4%. Similar experiments were performed with rf/dc isolation of both arginine and lysine from a mixture using the bent square quadrupole in the rf/dc mode. The unconventional continuous mass selection methods maximize soft landing yields, while still allowing the simple acquisition of full mass spectra.     

Pulsed acceleration charge detection mass spectrometry: application to weighing electrosprayed droplets

We describe a new approach to measuring the masses of individual macroions. The method employs a pulsed acceleration tube located between two sensitive image charge detectors. The charge and velocity of the macroion are recorded with the first image charge detector. The ion is pulse accelerated through a known voltage drop, and then the charge and velocity are remeasured using the second image charge detector. The mass of the ion is deduced from its charge and its initial and final velocities. The approach has been used to measure masses in the 10(10)-10(14) Da range with z = 10(3)-10(6) and m/z = 10(6)-10(9). It should be extendable to masses of <10(6) Da. We have used the method to determine the size and charge of water droplets transmitted through a capillary interface and an aperture interface. The droplets detected from the aperture interface are approximately 1 order of magnitude smaller in mass than those detected from the capillary interface. The droplets from both interfaces have relatively low charges, particularly with the capillary interface where they are only charged to a small fraction of the Rayleigh limit. These results suggest that the aerodynamic breakup of the droplets plays a significant role in the mechanism of electrospray ionization.     

Micromachined Bradbury-Nielsen gates

Bradbury-Nielsen gates (BNGs) are a standard way for gating or steering beams of charged particles in ion mobility spectrometry and time-of-flight mass spectrometry. They consist of a pair of interleaved electrodes that when at the same potential allow ions to pass through the electrodes undeflected and, when a voltage is applied, cause the ions to be deflected from their propagation axis. Previous efforts to construct such devices have relied on mechanical assembly by winding wires across an aperture. We describe a micromachining method for making monolithic BNGs using deep reactive ion etching of silicon-on-insulator wafers. This method enables the creation of electrodes with spacings ranging from 25 to 100 microm with a thickness of 20 microm, covering a 5 mm by 5 mm active area. We characterize the performance of these micromachined BNGs by ion imaging in a pseudorandom time-of-flight mass spectrometer.     

Kendrick mass defect spectrum: a compact visual analysis for ultrahigh-resolution broadband mass spectra

At currently achievable Fourier transform ion cyclotron resonance broadband mass spectrometry resolving power (m/deltam50% > 350,000 for 200 < m/z < 1,000), it would be necessary to spread out a conventional mass spectrum over approximately 200 m in order to provide visual resolution of the most closely resolved peaks. Fortunately, there are natural gaps in a typical mass spectrum, spaced 1 Da apart, because virtually no commonly encountered elemental compositions yield masses at those values. Thus, it is possible to break a broadband mass spectrum into 1-Da segments, rotate each segment by 90 degrees, scale each segment according to its mass defect (i.e., difference between exact and nominal mass), and then compress the spacing between the segments to yield a compact display. For hydrocarbon systems, conversion from IUPAC mass to "Kendrick" mass (i.e., multiplying each mass by 14.00000/14.01565) further simplifies the display by rectilinearizing the peak patterns. The resulting display preserves not only the "coarse" spacings (e.g., approximately 1 Da between odd and even masses, corresponding to either even vs odd number of nitrogens or 12C(c) vs 12C(c-1)13C1 elemental compositions of the same molecule; approximately 2-Da separations, corresponding to a double bond or ring; approximately 14 Da separations, corresponding to one CH2 group) but also the "fine structure" (i.e., different mass defects for different elemental compositions) across each 1-Da segment. The method is illustrated for experimental electrospray ionization FTICR ultrahigh-resolution mass spectra of a petroleum crude oil. Several thousand elemental compositions may be resolved visually in a single one-page two-dimensional display, and various compound families-class (NnOoSs), type (Z in C(c)H2(c+z)NnOoSs), and alkylation series-may be identified visually as well.     

Protein tyrosine-O-sulfation analysis by exhaustive product ion scanning with minimum collision offset in a NanoESI Q-TOF tandem mass spectrometer

Tyrosine-O-sulfated peptides were studied by nanoESI Q-TOF mass spectrometry and were found to exhibit an abundant loss of SO3 in positive ion mode under the usually nonfragmenting conditions of survey spectrum acquisition. A new strategy for the detection of tyrosine-O-sulfated peptides in total protein digests was designed based on exhaustive product ion scanning at the collision offset conditions typical for the recording of survey spectra (minimum collision offset). From these data, Q-TOF neutral loss scans for loss of 80/z and Q-TOF precursor ions scans were extracted. The specificity of this approach for analysis of tyrosine-O-sulfation was tested using a tryptic digest of bovine serum albumin spiked with sulfated hirudin (1:1 and 1000:1 molar ratio of BSA to sulfated hirudin, respectively) and using an in-solution digest of the recombinant extracellular domain of thyroid stimulating hormone receptor (ECD-TSHr). For both examples, the combination of in silico neutral loss scans for 80/z and subsequent in silico precursor ion scans resulted in a specific identification of sulfated peptides. In the analysis of recombinant ECD-TSHr, a doubly sulfated peptide could be identified in this way. Surprisingly, approximately 1/4 of the product ion spectra acquired from the tryptic digest of ECD-TSHr at minimum collision offset exhibited sequence-specific ions suitable for peptide identification. Complementary ion pairs were frequently observed, which either were b2/y(max-2) pairs or were induced by cleavage N-terminal to proline. MS/MS analysis at minimum collision offset followed by extraction of neutral loss and precursor ion scans is ideally suited for highly sensitive detection of analyte ions which exhibit facile gas-phase decomposition reactions.     

Halo ion trap mass spectrometer

We describe a novel radio frequency ion trap mass analyzer based on toroidal trapping geometry and microfabrication technology. The device, called the halo ion trap, consists of two parallel ceramic plates, the facing surfaces of which are imprinted with sets of concentric ring electrodes. Radii of the imprinted rings range from 5 to 12 mm, and the spacing between the plates is 4 mm. Unlike conventional ion traps, in which hyperbolic metal electrodes establish equipotential boundary conditions, electric fields in the halo ion trap are established by applying different radio frequency potentials to each ring. The potential on each ring can be independently optimized to provide the best trapping field. The halo ion trap features an open structure, allowing easy access for in situ ionization. The toroidal geometry provides a large trapping and analyzing volume, increasing the number of ions that can be stored and reducing the effects of space-charge on mass analysis. Preliminary mass spectra show resolution (m/Deltam) of 60-75 when the trap is operated at 1.9 MHz and 500 Vp-p.     

Effect of external electromagnetic waves on a confocal elliptical multiconductor transmission line structure

The influence of external electromagnetic waves on a new kind of transmission line called a confocal elliptical multiconductor transmission line structure is introduced. The capacitance and inductance matrices are obtained to attain transmission line equations. Then a uniform plane wave, TE/sup z/ or TM/sup z/mode, is applied to the structure (normal incidence) to find the induced voltages and currents along the conductors using Bernardi?s method. Finally, the results of this analytical method are verified using the fullwave analysis software.     

Phases of the SAW reflection and transmission coefficients for short reflectors on 128 degree LiNbO3

We study numerically the phase of surface acoustic waves reflected by or transmitted through short reflectors comprising only 1-3 aluminium electrodes on 128 degree YX-cut lithium niobate (LiNbO3). The electrodes have a finite thickness, and they are either open-circuited or grounded. The center-to-center distance between adjacent electrodes d corresponds roughly either to half of the characteristic wavelength d proportional to lambda0/2 or to d proportional to lambda0, for the reflectors operating at the fundamental and second harmonic modes, respectively. We use software based on the finite-element and boundary-element methods (FEM/BEM) for numerical experiments with a tailored test structure having 3 interdigital transducers (IDTs), simulating experimental conditions with an incident wave and reflected and transmitted surface acoustic wave (SAW). Using artificial enhancement of time resolution in conjunction with the fast Fourier transform (FFT) and time-gating, calculation of the Y-parameters in a relatively wide frequency range allows us to determine the phase of the reflection and transmission coefficients.     

Limitation of time-of-flight resolution in the ultra high mass range

In this work, we have examined the reason for the deterioration of resolution and mass accuracy of time-of-flight mass analyzers with increasing mass after the expansion-induced kinetic energy has been eliminated by collisional cooling in an ion guide. Theoretically, removing the expansion-induced kinetic energy by collisional cooling permits the ions to travel along the ion guide axes without significant deviation so that they can be injected into the analyzer in a well-collimated ion beam with well-defined kinetic energy. If the ions can be injected into an orthogonal acceleration time-of-flight mass analyzer (oa-TOF) in this manner, high-resolution mass analysis can be obtained regardless of mass or m/z. Unfortunately, high resolution did not result. It is our contention that the effusive expansion out of the first ion guide yields dispersive axial ejection that reduces TOF resolving power with increasing mass not m/z.     

Electrospray ionization tandem mass spectrometry for structural elucidation of protonated brevetoxins in red tide algae

Brevetoxins, the toxic components of "red tide" algae, all share one of two robust polycyclic ether backbone structures, but they are distinguished by differing side-chain substituents. Electrospray ionization mass spectrometry analyses of brevetoxins have shown that the polyether structure invariably has a very high affinity for sodium cations that results in the production of abundant (M + Na)+ ions even when sodium cations are only present as impurities. Because the ionic charge tends to remain localized on the sodium atom and because at least two bonds must be broken in order to produce polycyclic backbone fragmentation, it is extremely difficult to obtain abundant product ions (other than Na+) from (M + Na)+ brevetoxin precursor ions in low-energy collision-induced dissociation (CID) MS/MS experiments. This report establishes that acid additives (oxalic acid, trifluoroacetic acid, and particularly hydrochloric acid) in aqueous methanol solutions can promote high yields of protonated brevetoxin molecules (MH+ ions) for Btx-1, -2, and -9 brevetoxins. Most importantly, unlike their (M + Na)+ counterparts, MH+ precursor ions offer readily detectable product ions in CID MS/MS experiments, even under low-energy collisions. This direct structural characterization approach has provided decomposition information from brevetoxins that was previously inaccessible, including the identification of diagnostic product ions for "type A" brevetoxins (m/z 611) and "type B" brevetoxins (m/z 779, 473, 179) and characteristic ions for Btx-1 (m/z 221, 139), Btx-2 (m/z 153), and Btx-9 (m/z 157, 85). Precursor ion scans and constant neutral loss scans are proposed to enable screening of individual type A or type B brevetoxins present in naturally occurring mixtures.