Ion mobility spectrometry (IMS) can delineate gas-phase ions and probe their geometries. Coupling with electrospray ionization and MS has brought IMS to structural biology, revealing the macromolecular folding and subunit connectivity. However, the orientational averaging of ion–molecule collision cross sections (Ω) in the linear and field asymmetric waveform IMS (FAIMS) diminishes the resolution and structural specificity. In the novel low-field differential (LOD) IMS, a field too weak for ion heating (and thus FAIMS) aligns strong macrodipoles, capturing their magnitudes and directional Ω across the dipole (Ω⊥). However, the bisinusoidal waveforms (from FAIMS) have compromised the resolution, measurement accuracy, and correlation to the ion properties. Large ions amenable to LODIMS have low mobility and diffuse slowly, allowing the waveform frequencies down to ∼10 kHz. The low field and frequency permit generating the ideal rectangular waveforms with a flexible frequency and duty cycle by direct switching (impractical for FAIMS) in a miniature low-power format. This new IMS stage is evaluated for the exemplary large protein albumin (66 kDa) previously studied using the bisinusoidal waveform. The flat voltages and greater form factor initiate the differential IMS effect at lower fields, expand the separation space, and enable the quantification of Ω⊥ values by varying the duty cycle.
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Experimental Evaluation of Higher Order Stability Zones Using a Digitally Operated Quadrupole Mass Filter
Recent improvements to the comparison-based method of digital waveform generation increased the reproducibility of the waveforms so that the higher-order Mathieu stability zones can be accessed reliably. Digitally driven quadrupole mass filters access these zones using a fixed AC voltage and rectangular waveforms that are defined by a duty cycle. In this context, the duty cycle is the fraction of the waveform period where the waveform remains in the high state. Because digitally driven quadrupoles navigate stability using a duty cycle, there is no need to apply a resolving DC offset between electrode pairs. Accessing the higher stability zones using a conventional resonantly-tuned RF requires the use of thousands of AC and DC voltages making the mode of operation less accessible with these devices. Stability zones higher than (1,1) and (2,1) have theoretical resolving powers that are on the order 1,140 and 3,447 at FWHM which drives efforts to practically access these operational conditions. Accessing these zones digitally requires the use of extremely precise waveforms. In a previous effort, waveform generation produced waveforms to reliably access the (1,1) and (2,1) zones without impacting performance. However, recent work found more improvement was needed to reliably access neighboring higher stability zones. Derived from that work, it was determined that a waveform resolution of ~10 ppm or less was needed to reliably access the (3,1) and (3,2) zones. The present work utilized digital waveforms that achieve this level of precision to experimentally access and characterize attributes of the (3,1) and (3,2) zones. This work dives into the investigation of different beam energies to overcome the destabilizing fringing fields, improve transmission, and their overall effect on the experimental resolving power and signal-to-noise. In addition, the AC voltage of the driving RF was varied to understand the effects on the initial ion beam energy that is needed to achieve balanced separation and how the overall signal-to-noise is affected. Lastly, an assessment was made on the effects of the temporal parameters of a digital mass scan on peak sensitivity, peak fidelity, and overall duration for a scan.
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- Award ID(s):
- 2103645
- PAR ID:
- 10529698
- Publisher / Repository:
- American Chemical Society
- Date Published:
- Journal Name:
- Journal of the American Society for Mass Spectrometry
- ISSN:
- 1044-0305
- Subject(s) / Keyword(s):
- mass spectrometry digital waveform technology quadrupole
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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