On The Time Scale of Internal Energy Relaxation of AP-MALDI and nano-ESI Ions in a Quadrupole Ion Trap

Recently reported results (Konn et al. [14]) on the collisional cooling of atmospheric pressure matrix assisted laser desorption ionization (AP-MALDI) and nano-electrospray ionization (nano-ESI) generated ions in a quadrupole ion trap mass spectrometer (QITMS) are inconsistent with measured collisional cooling rates. The work reported here presents a re-examination of those previous results. Collision induced dissociation (CID) has been used to probe various properties of ions contained in a QITMS. It is shown experimentally that when trapping large numbers of ions, an effective dc trapping voltage is induced that varies with changes in the size of the ion cloud. A decrease in the resonant frequency for maximum CID efficiency is observed as the cool time between parent ion isolation and CID is increased. Ion trajectories in a QITMS are simulated to demonstrate how ion density changes over the course of parent ion isolation. The effect of space charge on ion motion is simulated, and Fourier transformations of ion axial motion plus simple calculations corroborate the experimentally observed transient frequency shifts. The relative stability of ions formed by AP-MALDI and nano-ESI is compared under low charge density conditions. These data show that the ions have reached equilibrium internal energy and, thus, that differences in dissociation onsets and “50% fragmentation efficiency points” between the ionization mechanisms are due to the formation of distinct ion conformations as previously shown in reference [28]. The conclusions of Konn et al. [14] are based on invalid experimental procedures as well as inappropriate comparisons of QITMS data to low-pressure FT-ICR data

High Amplitude Short Time Excitation: A Method to Form and Detect Low Mass Product Ions in a Quadrupole Ion Trap Mass Spectrometer

Collision induced dissociation (CID) in a quadrupole ion trap mass spectrometer using the conventional 30 ms activation time is compared with high amplitude short time excitation (HASTE) CID using 2 ms and 1 ms activation times. As a result of the shorter activation times, dissociation of the parent ions using the HASTE CID technique requires resonance excitation voltages greater than conventional CID. After activation, the rf trapping voltage is lowered to allow product ions below the low mass cut-off to be trapped. The HASTE CID spectra are notably different from those obtained using conventional CID and can include product ions below the low mass cut-off for the parent ions of interest. The MS/MS efficiencies of HASTE CID are not significantly different when compared with the conventional 30 ms CID. Similar results were obtained with a two-dimensional (linear) ion trap and a three-dimensional ion trap

Selective Ion Isolation/Rejection Over a Broad Mass Range in the Quadrupole Ion Trap

AbstractTechniques are presented for mass-selective ion manipulation over a wide mass range in a three-dimensional quadrupole. The methods use an auxiliary, low-amplitude radio-frequency signal applied to the endcap electrodes. This signal is either held at a single frequency as the fundamental radio-frequency trapping amplitude is ramped or swept over a frequency range while the fundamental radio-frequency trapping amplitude is held at a fixed level. Ion isolation and ejection are demonstrated for ions formed within the ion trap using electron ionization and for ions injected into the ion trap formed either by an air-sustained glow discharge or by electrospray. Mass-selective ion ejection is used to reduce matrix-ion-induced space charge during ion injection, thereby producing signal enhancement for the detection of 2,4,6-trinitrotoluene in air. Mass-selective isolation of ions with mass-to-charge ratios above the normal operating range (m/z 650) for the ion trap is also demonstrated after injection of myoglobin ions formed via electrospray

Paper spray mass spectrometry for high-throughput quantification of nicotine and cotinine

The rapid release of new tobacco products requires high-throughput quantitative methods to support tobacco research. Sample preparation for LC-MS and GC-MS is time consuming and limits throughput. Paper spray tandem mass spectrometry (PS-MS/MS) is proposed and validated as a simple and rapid method for quantification of nicotine and cotinine in complex matrices to support tobacco-related research. Air liquid interface (ALI) human tracheobronchial epithelial cell (HTBEC) cultures were exposed to tobacco smoke using a Vitrocell VC-10 smoking machine. Apical culture washes (phosphate buffered saline, PBS) and basolateral media were analyzed with the PS-MS/MS method. GC-MS/MS was used as a comparative quantitative technique. The PS-MS/MS approach allowed for direct spotting of samples on the paper substrate, whereas the GC-MS/MS method required additional sample preparation in the form of solvent-solvent extraction. Limits of quantitation (LOQs) were higher with the PS-MS/MS approach than GC-MS/MS, but still below the relevant concentrations found in HTBEC smoke exposure experiments as well as most clinical applications. PS-MS/MS is readily achieved on mass spectrometers that include atmospheric pressure inlets, and allows for convenient quantification from complex matrices that would otherwise require additional sample preparation and chromatographic separation. © 2017 The Royal Society of Chemistry

Enhancement of mass resolution in the quadrupole ion trap via resonance ejection

The Finnigan MAT ion trap mass spectrometer (ITMS) has unit mass resolution and a nominal upper mass/charge limit of only 650 when operated in the standard mass-selective instability method for acquiring mass spectra. Mass resolution up to {approx}2500 can be achieved with the use of the axial modulation technique. Ions can also be ejected at a q{sup z} value other than that corresponding to the mass-selective instability boundary by application of an auxiliary signal to the end-cap electrodes. When the frequency of this signal is resonant with the secular frequency for a particular m/z, such ions can become kinetically excited and ejected from the trap via a process known as resonance ejection. We demonstrate the capability to mass-selectively isolate or eject ions over a wide mass range and to generate mass spectra of electrospray-generated ions by scanning the frequency of the resonance ejection signal applied to the end-cap electrodes. A block diagram of the experimental modifications to the ITMS required for frequency-swept resonance ejection is shown. 2 refs., 5 figs

Optimization of Peptide Separations by Differential Ion Mobility Spectrometry

Differential ion mobility spectrometry (DIMS) has the ability to separate gas phase ions based on their difference in ion mobility in low and high electric fields. DIMS can be used to separate mixtures of isobaric and isomeric species indistinguishable by mass spectrometry (MS). DIMS can also be used as a filter to improve the signal-to-background of analytes in complex samples. The resolving power of DIMS separations can be improved several ways, including increasing the dispersion field and increasing the amount of helium in the nitrogen carrier gas. It has been previously demonstrated that the addition of helium to the DIMS carrier gas provides improves separations when the dispersion field is the kept constant as helium content is varied. However, helium has a lower breakdown voltage than nitrogen. Therefore, as the percent helium content in the nitrogen carrier gas is increased, the highest dispersion field accessible decreases. This work presents the trade-offs between increasing dispersion fields and using helium in the carrier gas by comparing the separation of a mixture of isobaric peptides. The maximum resolution for a separation of a mixture of three peptides with the same nominal molar mass was achieved by using a high dispersion field (~72 kV/cm) with pure nitrogen as the carrier gas within the DIMS assembly. The conditions used to achieve the maximum resolution also exhibit the lowest ion transmission through the assembly, suggesting that it is necessary to consider the trade-off between sensitivity and resolution when optimizing DIMS conditions for a given application

Simultaneous Collision Induced Dissociation of the Charge Reduced Parent Ion during Electron Capture Dissociation

A method of performing collision induced dissociation (CID) on the charge-reduced parent ion as it is formed during electron capture dissociation (ECD), called ECD+CID, is described. In ECD+CID, the charge-reduced parent ion is selectively activated using resonant excitation and collisions with the helium bath gas inside a linear quadrupole ion trap ECD device (ECDLIT). It has been observed that ECD+CID can improve the sequence coverage for melittin over performing ECD alone (i.e., from 76 % to 88 %). Perhaps just as important, ECD+CID can be used to reduce the extent of multiple electron capture events observed when performing ECD in the ECDLIT. Consequently, the abundance of mass-to-charge ratios corresponding to ECD product ions that contain neutralized protons is decreased, simplifying the interpretation of the product ion spectrum