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We introduce a new atmospheric pressure charge stripping (AP-CS) method for the electrospray ionization mass spectrometry (ESI-MS) analysis of heterogeneous mixtures, utilizing ion/ion proton transfer reactions within an experimental ion source to remove excess charge from sample ions and thereby reduce spectral congestion. The new method enables the extent of charge stripping to be easily controlled, independent of primary ionization, and there are no complications due to adduct formation. Here, we demonstrate AP-CS with a Xevo G2-S Q/TOF from Waters-Micromass using an ion source originally designed for atmospheric pressure-electron capture dissociation (AP-ECD) experiments; repurposing the AP-ECD ion source for AP-CS requires only adding a supplemental reagent (e.g., a perfluorocompound) to scavenge the electrons and generate anions for the charge-stripping reactions. Results from model peptides are first presented to demonstrate the basic method, including differences between the AP-CS and AP-ECD operating modes, and how the extent of charge stripping may be controlled. This is followed by a demonstration of AP-CS for the ESI-MS analysis of several large poly(ethylene glycol)s (PEGs), up to 40 kDa, typical of those used in biopharmaceutical development. [ABSTRACT FROM AUTHOR]

Abstract: This review presents our perspective on the state-of-the-art of atmospheric pressure photoionization (APPI) for LC/MS. Its focus is on APPI''s capabilities and how to utilize them fully. The introduction includes a brief recounting of the history of APPI''s development, as well as a summary of its operating principles and current position in the field. The primary ionization mechanisms in APPI are then addressed, including direct analyte photoionization (PI), dopant/solvent PI, and thermospray. Next a summary of the ion–molecule reaction pathways available for analyte ionization is presented, along with the conditions required for activating them. APPI''s performance characteristics are then examined. In effect, this review is an interim report on progress made since Rafaelli and Saba concluded that “The ability…to direct the preferential ion formation towards one particular type…can be extremely useful for qualitative and quantitative determinations. For this purpose, a better insight in the processes involved in the ionization step is strongly needed” [A. Raffaelli, A. Saba, Mass Spectrom. Rev. 22 (2003) 318]. In the conclusion, we focus on areas of APPI technology identified as being either unoptimized or largely unexplored, and having the potential to be improved upon—the crux being that with further research and development improvements in the performance, capabilities, and ease-of-use of APPI may reasonably be anticipated. [Copyright &y& Elsevier]

The sensitivity of dopant-assisted atmospheric pressure photoionization (DA-APPI) for LC/MS is generally reduced at higher solvent flow rates. Theory suggests that quenching of excited-state precursors to the dopant ions, via collisions with vaporized solvent molecules, may be one mechanism responsible for this trend. To ascertain if the primary rate of ionization is affected by quenching, experiments were performed utilizing an ionization detector to determine the primary ion current generated by irradiating vaporized mixtures of toluene dopant and methanol solvent. The results indicate that no loss of primary ion current occurs as the solvent flow is increased, provided the dopant-to-solvent ratio is held constant. Additional primary ion current can always be generated by increasing the dopant flow rate and/or the lamp power. Thus, quenching of excited-state precursors to the dopant ions, leading to a reduction in the primary rate of ionization, is not the mechanism responsible for the observed loss of sensitivity at higher liquid solvent flow rates. [Copyright &y& Elsevier]

Atmospheric pressure photoionization can provide high ionization efficiency simultaneously to both polar and nonpolar compounds delivered in reversed-phase solvent. The method to achieve this utilizes toluene as a dopant and simply requires that the solvent flow be limited so that reactions between toluene photoions and the organic component of the solvent are not driven to completion. Under these conditions, toluene photoions remain in the source for ionizing nonpolar compounds via charge exchange (electron transfer), while protonated solvent ions are available for proton-transfer reactions with polar molecules. The reagent ion mixture is then suitable for ionizing a wide range of both polar and nonpolar compounds. The critical effect of solvent flow rate is demonstrated here with results for a test analyte, 9-methylanthracene, which may be ionized by either charge exchange or proton transfer. For a solvent of 50:50 methanol/water (v/v), lowering the flow from 200 to 50 μL min-1 results in a 10x increase in charge exchange ionization efficiency-further flow reductions provide even greater enhancements. This method is compatible with sample delivery by direct infusion and micro- and narrow-bore LC, as well as conventional LC using a flow splitter. [ABSTRACT FROM AUTHOR]

It is well documented since the early days of the development of atmospheric pressure ionization methods, which operate in the gas phase, that cluster ions are ubiquitous. This holds true for atmospheric pressure chemical ionization, as well as for more recent techniques, such as atmospheric pressure photoionization, direct analysis in real time, and many more. In fact, it is well established that cluster ions are the primary carriers of the net charge generated. Nevertheless, cluster ion chemistry has only been sporadically included in the numerous proposed ionization mechanisms leading to charged target analytes, which are often protonated molecules. This paper series, consisting of two parts, attempts to highlight the role of cluster ion chemistry with regard to the generation of analyte ions. In addition, the impact of the changing reaction matrix and the non-thermal collisions of ions en route from the atmospheric pressure ion source to the high vacuum analyzer region are discussed. This work addresses such issues as extent of protonation versus deuteration, the extent of analyte fragmentation, as well as highly variable ionization efficiencies, among others. In Part 1, the nature of the reagent ion generation is examined, as well as the extent of thermodynamic versus kinetic control of the resulting ion population entering the analyzer region. [Figure not available: see fulltext.] [ABSTRACT FROM AUTHOR]