Determination Of Collision Cross Sections Research Articles

Impact of Source Conditions on Collision Cross Section Determination by Trapped Ion Mobility Spectrometry.

Collision cross section (CCS) values determined in ion mobility-mass spectrometry (IM-MS) are increasingly employed as additional descriptors in metabolomics studies. CCS values must therefore be reproducible and the causes of deviations must be carefully known and controlled. Here, we analyzed lipid standards by trapped ion mobility spectrometry-mass spectrometry (TIMS-MS) to evaluate the effects of solvent and flow rate in flow injection analysis (FIA), as well as electrospray source parameters including nebulizer gas pressure, drying gas flow rate, and temperature, on the ion mobility and CCS values. The stability of ion mobility experiments was studied over 10 h, which established the need for a delay-time of 20 min to stabilize source parameters (mostly pressure and temperature). Modifications of electrospray source parameters induced shifts of ion mobility peaks and even the occurrence of an additional peak in the ion mobility spectra. This behavior could be essentially explained by ion-solvent cluster formation. Changes in source parameters were also found to impact CCS value measurements, resulting in deviations up to 0.8%. However, internal calibration with the Tune Mix calibrant reduced the CCS deviations to 0.1%. Thus, optimization of source parameters is essential to achieve a good desolvation of lipid ions and avoid misinterpretation of peaks in ion mobility spectra due to solvent effects. This work highlights the importance of internal calibration to ensure interoperable CCS values, usable in metabolomics annotation.

Enhanced Ion Mobility Separation and Characterization of Isomeric Phosphatidylcholines Using Absorption Mode Fourier Transform Multiplexing and Ultraviolet Photodissociation Mass Spectrometry.

The structural diversity of phospholipids plays a critical role in cellular membrane dynamics, energy storage, and cellular signaling. Despite its importance, the extent of this diversity has only recently come into focus, largely owing to advances in separation science and mass spectrometry methodology and instrumentation. Characterization of glycerophospholipid (GP) isomers differing only in their acyl chain configurations and locations of carbon-carbon double bonds (C═C) remains challenging due to the need for both effective separation of isomers and advanced tandem mass spectrometry (MS/MS) technologies capable of double-bond localization. Drift tube ion mobility spectrometry (DTIMS) coupled with MS can provide both fast separation and accurate determination of collision cross section (CCS) of molecules but typically lacks the resolving power needed to separate phospholipid isomers. Ultraviolet photodissociation (UVPD) can provide unambiguous double-bond localization but is challenging to implement on the timescales of modern commercial drift tube time-of-flight mass spectrometers. Here, we present a novel method for coupling DTIMS with a UVPD-enabled Orbitrap mass spectrometer using absorption mode Fourier transform multiplexing that affords simultaneous localization of double bonds and accurate CCS measurements even when isomers cannot be fully resolved in the mobility dimension. This method is demonstrated on two- and three-component mixtures and shown to provide CCS measurements that differ from those obtained by individual analysis of each component by less than 1%.

Multi-CRAFTI: Relative Collision Cross Sections from Fourier Transform Ion Cyclotron Resonance Mass Spectrometric Line Width Measurements.

Determination of collision cross sections (CCS) using the cross-sectional areas by the Fourier transform ion cyclotron resonance (CRAFTI) technique is limited by the requirement that accurate pressures in the trapping cell of the mass spectrometer must be known. Experiments must also be performed in the energetic hard-sphere regime such that ions decohere after single collisions with neutrals; this limits application to ions that are not much more massive than the neutrals. To mitigate these problems, we have resonantly excited two (or more) ions of different m/z to the same center-of-mass kinetic energy in a single experiment, subjecting them to identical neutral pressures. We term this approach "multi-CRAFTI". This facilitates measurement of relative CCS without requiring knowledge of the pressure and enables determination of absolute CCS using internal standards. Experiments with tetraalkylammonium ions yield CCS in reasonable agreement with the one-ion-at-a-time CRAFTI approach and with ion mobility spectrometry (IMS) when differences in collision energetics are taken into account (multi-CRAFTI generally yields smaller CCS than does IMS due to the higher collision energies employed in multi-CRAFTI). Comparison of multi-CRAFTI and IMS results with CCS calculated from structures computed at the M06-2X/6-31+G* level of theory using projection approximation or trajectory method values, respectively, indicates that the computed structures have CCS increasingly smaller than the experimental CCS as m/z increases, implying the computational model overestimates interactions between the alkyl arms. For ions that undergo similar collisional decoherence processes, relative CCS reach constant values at lower collision energies than do absolute CCS values, suggesting a means of increasing the accessible upper m/z limit by employing multi-CRAFTI.

Identity confirmation of anthocyanins in berries by LC-DAD-IM-QTOFMS.

Rugged analytical methods for the screening and identity confirmation of anthocyanins require a dedicated sample preparation, chromatographic setup, and the reliable generation of multiple identification points to confirm identity against the wide range of phenolic compounds typically present in food, beverage, and plant material samples. To this end, combinations of spectroscopic and mass spectrometric detection are frequently employed for this application to provide higher confidence in the absence of authentic standards. In the present work, low‐field drift tube ion mobility (DTIM) separation is evaluated for this task using a LC–DAD–DTIM–QTOFMS method. DTIM‐MS allows accurate determination of collision cross sections (DTCCS) for all analysed compounds as well as a precise alignment tool for reconciling fragment and precursor ions in data independent acquisition mode. The presented approach thereby allows for an anthocyanin screening method taking true advantage of all dimensions of the analytical platform: relative retention (RPLC), UV/VIS absorption spectrum, accurate mass, DTCCSN2, and confirmed high‐resolution fragment ions. From the analysis of authentic standards and several berry samples primarily from the Vaccinium genus, Level 1 confirmation data for six anthocyanins from the cyanidin family, and Level 2 confirmation for a further 29 anthocyanins confirmed in berry samples is provided. The method and accompanying dataset provided as part of this work provides a means to develop anthocyanin screening methods using the ion mobility dimension as an additional alignment and filtering parameter in data independent analysis acquisition across any LC–IM–MS platform.

Investigating the Potential of Ion Mobility-Mass Spectrometry for Microalgae Biomass Characterization.

Algae biomass is formed by an extremely complex set of metabolites, and its molecular characterization has been very challenging. We report the characterization of microalgae extracts via traveling wave ion mobility-mass spectrometry (TWIM-MS) by two different analysis strategies. First, the extracts were analyzed by direct infusion electrospray ionization (ESI) with no previous chromatographic separation (DI-ESI-TWIM-MS). Second, the samples were screened for metabolites and lipids using an untargeted high-throughput method that employs ultrahigh-performance liquid chromatography (UHPLC) using data-independent analysis (DIA) - MSE (UHPLC-HDMSE). Sixteen different microalgae biomasses were evaluated by both strategies. DI-ESI-TWIM-MS was able, via distinct drift times, to set apart different classes of metabolites, with the differences in the profiles of each microalga readily evident. With the UHPLC-HDMSE approach, 1251 different compounds were putatively annotated across 16 samples with 210 classified as lipids. From the normalized abundance for each annotated compound category, a detailed profiling in terms of metabolites, lipids, and lipid classes of each sample was performed. The reported workflow represents a powerful tool to determine the most suitable biotechnological applications for a given alga type and may allow for real-time monitoring of the algae composition distribution as a function of growth conditions, feedstocks, and the like. The determination of collision cross section results in improved confidence in the identification of triacylglycerols in samples, highly applicable to biofuels production. The two analysis strategies explored in this work offer powerful tools for the biomass industry by aiding in the identification of ideal strains and culture conditions for a specific application, saving analysis time and facilitating identification of a large number of constituents at once.

Structural characterization of synthetic peptides using electrospray ion mobility spectrometry and molecular dynamics simulations

Electrospray ionization-ion mobility spectrometry was employed for the determination of collision cross sections (CCS) of 25 synthetically produced peptides in the mass range between 540–3310 Da. The experimental measurement of the CCS is complemented by their calculation applying two different methods. One prediction method is the intrinsic size parameter (ISP) method developed by the Clemmer group. The second new method is based on the evaluation of molecular dynamics (MD) simulation trajectories as a whole, resulting in a single, averaged collision cross-section value for a given peptide in the gas phase. A high temperature MD simulation is run in order to scan through the whole conformational space. The lower temperature conformational distribution is obtained through thermodynamic reweighting. In the first part, various correlations, e.g. CCS vs. mass and inverse mobility vs. m/z correlations, are presented. Differences in CCS between peptides are also discussed in terms of their respective mass and m/z differences, as well as their respective structures. In the second part, measured and calculated CCS are compared. The agreement between the prediction results and the experimental values is in the same range for both calculation methods. While the calculation effort of the ISP method is much lower, the MD method comprises several tools providing deeper insights into the conformations of peptides. Advantages and limitations of both methods are discussed. Based on the separation of two pairs of linear and cyclic peptides of virtually the same mass, the influence of the structure on the cross sections is discussed. The shift in cross section differences and peak shape after transition from the linear to the cyclic peptide can be well understood by applying different MD tools, e.g. the root-mean-square deviation (RMSD) and the root mean square fluctuation (RMSF).

Energy Charge, Redox State, and Metabolite Turnover in Single Human Hepatocytes Revealed by Capillary Microsampling Mass Spectrometry

Metabolic analysis of single cells to uncover cellular heterogeneity and metabolic noise is limited by the available tools. In this study, we demonstrate the utility of capillary microsampling electrospray ionization mass spectrometry with ion mobility separation for nontargeted analysis of single cells. On the basis of accurate mass measurements and collision cross-section determination, a large number of chemical species, 22 metabolites and 54 lipids, were identified. To assess the cellular response to metabolic modulators, the adenylate energy charge (AEC) levels for control and rotenone treated cells were evaluated. A significant reduction in the AEC values was observed for rotenone treated cells. For the cells under oxidative stress, the mean value for the [reduced glutathione (GSH)]/[oxidized glutathione (GSSG)] ratio was significantly decreased, whereas the distribution of the [uridine diphosphate N-acetylhexosamine (UDP-HexNAc)]/[uridine diphosphate hexose (UDP-hexose)] ratio exhibited dramatic tailing to higher values. Lipid turnover rates were studied by pulse-chase experiments at the single cell level.

Ion collision cross sections and transport coefficients extended to intermediate energies and reduced electric fields forHe2+ions colliding with He

This work is devoted to the calculation of transport coefficients for He(2)(+) ions in gaseous He at intermediate reduced electric fields. These swarm data are of great interest for a better understanding of the mechanisms of formation and propagation of the fast plasma bullets or ionization waves observed in dielectric barrier plasma jet devices. For transport data, the collision cross sections required are determined from several theoretical methods based on quantum, semiclassical, and hybrid approaches and a diatomics-in-molecules model for the potential energy surfaces of He(2)(+). The corresponding collision cross sections are then used in an optimized Monte Carlo code to calculate the ion transport coefficients over a wide range of reduced electric fields extending over the experimental range. Calculated transport coefficients are compared with available experimental data at low electric fields. Moreover, an extrapolation method is used in order to determine the reduced mobility for stronger fields. A critical discussion has been performed on the pertinence and the reliability of these different methods of determination of collision cross sections needed for the calculation of ion transport data. Such ion data will be used in electrohydrodynamic and chemical kinetic models of the low-temperature plasma jet to quantify and to tune the active species production for a better use in biomedical applications.

Damping Factor Links Periodic Focusing and Uniform Field Ion Mobility for Accurate Determination of Collision Cross Sections

The methodology for obtaining accurate ion-neutral collision cross section (Ω) values for peptides and proteins using periodic focusing ion mobility spectrometry (PF IMS) is presented. A mobility dampening factor (represented by the term α) is introduced to account for the relative increase in ion-neutral collisions in PF IMS compared to uniform field ion mobility spectrometry (UF IMS) for equivalent operating conditions. The results show that α may be easily quantified both theoretically and empirically for a specific PF IMS design operating at a given pressure based upon the charge state of the analyte. By simply incorporating an α term into traditional UF IMS expressions, accurate Ω values were obtained with excellent agreement (≤4% difference) compared to UF IMS measurements found in the current literature.

Collision cross-section determination and tandem mass spectrometric analysis of isomeric carotenoids using electrospray ion mobility time-of-flight mass spectrometry.

Carotenoids are natural pigments with provitamin A and antioxidant activities. Biosynthesized in plants as their all-trans isomers, carotenoids isomerize in solution and in humans to multiple cis isomers which can have different bioavailabilities and functions. Since separation and characterization of isomeric carotenoids using high-pressure liquid chromatography (HPLC) or liquid chromatography-tandem mass spectrometry (LC-MS/MS) is time-consuming, the potential for ion mobility mass spectrometry (IM-MS) to resolve and characterize carotenoid isomers rapidly without chromatography was investigated using traveling-wave ion mobility spectrometry on a quadrupole time-of-flight mass spectrometer. The all-trans isomers of lycopene and β-carotene were separated by several milliseconds from the cis-isomers which were detected as partially overlapping peaks. The collision cross-section values of these carotenoid isomers were determined using IM-MS to be 180 and 236 Å(2) for cis-lycopene and all-trans-lycopene, and 181 and 225 Å(2) for cis-β-carotene and all-trans-β-carotene, respectively. Collision-induced dissociation MS/MS of ion mobility-resolved isomers indicated that cis and all-trans carotenoid isomers can be distinguished by their fragmentation patterns. Previous MS/MS studies of cis- and all trans-carotenoids had suggested that they produced identical tandem mass spectra, but this appears to have been the result of isomerization during ionization. Introduction of specific cis or trans isomers by infusion or HPLC resulted in cis/trans isomerization in the ion source during electrospray, and the relative levels of cis carotenoids forming in the ion source compared to the all-trans isomers were temperature dependent.

Mass and Collision Cross-Section Determination Using a Low-Vacuum Mass Spectrometer

A simple low-vacuum mass spectrometer (LVMS) operating in the milliTorr pressure range was developed. The instrument resolves masses by time-of-flight measurements and employs a high-gain, fast-response detector that can operate at these pressures. This instrument allows simultaneous determination of mass and collision cross sections of the ions with the bath gas. Here we demonstrate the LVMS's abilities to determine total collision cross sections for the collisions of organic ions with three background gases, He, N(2), and SF(6). As a demonstration of the system capabilities, the unimolecular interconversion of photochemically produced C(7)H(7)(+) to the tropylium ion structure is investigated.

Determination of collision cross section by dynamical absorption saturation in OsO 4

We investigated the saturation dynamics of the 192OsO 4-absorption line P(46)A 1 2 in the infrared ν 3-band, induced by excitation with CO 2-laser light, and determined the decay constant T 1 of the saturation as a function of the OsO 4 gas pressure. From the linear dependence of the relaxation rate T 1 −1 on the pressure we additionally determined the collision cross section σ = 6.6 × 10 −18m 2 which is substantial for the evaluation of the pressure broadening of saturation dips in the saturation spectroscopy of OsO. The method allows to measure the influence of collisions on homogeneous line widths, measured in saturation exeriments.

The determination of collision cross-sections

Collision cross-sections of ions, σ0t, can be accurately obtained by extrapolating σt to zero target gas pressure, thus eliminating the error in the collision path length, l(n), caused by target gas leaking out of the collision cell. σ0t values for various ions have been determined and contain structural information. For normal alkanes (n = 1–10) σOt steadily increases from 3.4 to 14.2Å2 by 1.2Å2 per methylene unit. The observation that ions with cyclic structures have smaller total collision cross-sections than corresponding non-cyclic species could be general, as observed for cyclohexane, cyclohexanone and the C2H3O+ ions from oxirane with respect to acetyl cations. The benzene and pyridine dications cannot be considered as pure six-membered ring species since their σ0t values are significantly larger than those of the singly charged ions.

On the determination of collision cross sections by nuclear Doppler shift

We report on measurements of the energy loss of ions in matter by “Inverted Doppler Shift Attenuation” (IDSA). This new method is an inversion of the “Doppler Shift Attenuation” (DSA) method for the determination of lifetimes of nuclear states. While in DSA the exact knowledge of the velocity dependent energy loss functiondE/ds is required, it is shown that in IDSA this function or the absolute collision cross section, respectively, can be determined from the Doppler spectrum of an excited nuclear fragment recoiling in matter, whose lifetime is known. No corrections or assumptions concerning the collision processes are necessary. 7Li* fragments (E γ=478 keV) from the10B(n, α)7Li* reaction produce an easily observable and analysable Doppler spectrum. Here boron must be a constituent or an implanted impurity of the material to be investigated. The experimental set-up is described. An exact relativistic analysis of the Doppler spectrum is given. The measured collision cross sections turn out (a) to be proportional tov within the range 1.5 · 108≦v≦4.8 · 108 cm/sec, (b) are thus only due to pure “electronic” collisions, and (c) sensitively dependent on the charge distributions of the target atoms.

Ion Cyclotron Resonance and the Determination of Collision Cross Sections

It is shown that the ion cyclotron resonance absorption line in weakly ionized gases may be interpreted to obtain $\frac{e}{m}$ ratio, the number, and the collision frequency of the ions. From the observed linewidths, the collision frequencies of several ions are found as a function of pressure and electric field strength. From these data the ion-molecule collision cross section for ${\mathrm{N}}_{2}^{+}$ and ${\mathrm{Ar}}^{+}$ in their parent gases are found to be 125 and 140 ${\mathrm{\AA{}}}^{2}$, respectively, at high $\frac{E}{p}$ and 185 and 215 ${\mathrm{\AA{}}}^{2}$, respectively, at low $\frac{E}{p}$. Observed absorption and dispersion line shapes fit closely the Lorentz line shape derived theoretically with the assumption of constant mean free time. The measurement chamber consists of an ion source and drift region in a strong magnetic field within which a transverse rf electric field may be applied. A narrow band, derivative-type system is used to detect the power absorption as a function of frequency.