Electron-capture Ionization Research Articles

Salt-Bridged Peptide Anion Gaseous Structures Enable Efficient Negative Ion Electron Capture Dissociation.

We previously discovered that electron attachment to gaseous peptide anions can occur within a relatively narrow electron energy range. The resulting charge-increased radical ions undergo dissociation analogous to conventional cation electron capture/transfer dissociation (ECD/ETD), thus enabling a novel tandem mass spectrometry (MS/MS) technique that we termed negative ion electron capture dissociation (niECD). We proposed that gaseous zwitterionic structures are required for niECD with electron capture either occurring at or being directed by a positively charged site. Here, we further evaluate this zwitterion mechanism by performing niECD of peptides derivatized to alter their ability to form zwitterionic gaseous structures. Introduction of a fixed positive charge tag, a highly basic guanidino group, or a highly acidic sulfonate group to promote zwitterionic structures in singly charged anions, rescued the niECD ability of a peptide refractory to niECD in its unmodified form. We also performed a systematic study of five sets of synthetic peptides with decreasing zwitterion propensity and found that niECD efficiency decreased accordingly, further supporting the zwitterion mechanism. However, traveling-wave ion mobility-mass spectrometry experiments, performed to gain further insight into the gas-phase structures of peptides showing high niECD efficiency, exhibited an inverse correlation between the orientationally averaged collision cross sections and niECD efficiency. These results indicate that compact salt-bridged structures are also a requirement for effective niECD.

Online protein unfolding characterized by ion mobility electron capture dissociation mass spectrometry: cytochrome C from neutral and acidic solutions.

Electrospray ionization mass spectrometry (ESI-MS) experiments, including ion mobility spectrometry mass spectrometry (ESI-IMS-MS) and electron capture dissociation (ECD) of proteins ionized from aqueous solutions, have been used for the study of solution-like structures of intact proteins. By mixing aqueous proteins with denaturants online before ESI, the amount of protein unfolding can be precisely controlled and rapidly analyzed, permitting the characterization of protein folding intermediates in protein folding pathways. Herein, we mixed various pH solutions online with aqueous cytochrome C for unfolding and characterizing its unfolding intermediates with ESI-MS charge state distribution measurements, IMS, and ECD. The presence of folding intermediates and unfolded cytochrome c structures were detected from changes in charge states, arrival time distributions (ATDs), and ECD. We also compared structures from nondenaturing and denaturing solution mixtures measured under "gentle" (i.e., low energy) ion transmission conditions with structures measured under "harsh" (i.e., higher energy) transmission. This work confirms that when using "gentle" instrument conditions, the gas-phase cytochrome c ions reflect attributes of the various solution-phase structures. However, "harsh" conditions that maximize ion transmission produce extended structures that no longer correlate with changes in solution structure.

ДИССОЦИАТИВНЫЙ ЗАХВАТ ЭЛЕКТРОНОВ МОЛЕКУЛАМИ ДИГИДРОКУМАРИНА И 5,6-МЕТИЛЕНДИОКСИ-1-ИНДАНОНА

The molecules of dihydrocoumarin and 5,6-methylenedioxy-1-indanone were studied by the negative ion resonant electron capture mass spectrometry for slow (0-15 eV) electrons. Such studies of the structure of vacant molecular orbitals (VMO) and dissociative electron attachment (DEA) for isolated molecules of prospective organic semiconducting materials, which are of great interest in terms of use as elementary components of organic nanoelectronics devices, are the most promising, since a correspondence between the energies of vacant MO isolated molecules and the maxima of the density of states in the conduction band of thin films was revealed earlier. Also, there is an idea developing in modern science, that the results of studies of vacant orbitals of individual molecules in the gas phase, in particular, using the DEA method, are necessary to identify the features of the energy structure of the conduction band of molecular organic condensates, starting from energies below the vacuum level, by the amount of binding energy of molecules in the film. In particular, it was shown that the substances studied in this work cannot be used as monomeric compounds in the design of electrically conductive polymers in the future, since they do not form negative molecular ions in the gas phase at energies close to thermal.

Enforced stripping of negative muons from $\mu$He+ ions to stimulate muon-catalyzed fusion by cyclotron resonance acceleration

Abstract The addition (sticking) of a negative muon to an $\alpha$ particle to form a $\mu$He$^+$ ion limits the number of chain reactions in muon-catalyzed fusion (MuCF), making scientific break-even in MuCF difficult. The reduction in the sticking probability by boosting the negative muon stripping using resonance radio-frequency (RF) acceleration of $\mu$He$^+$ ions has been examined with multi-particle tracking simulations. It was found that the stripping efficiency was largely depressed by ion–electron capture processes that have not been considered so far. Improvement of the fusion chain cycles is rather difficult with an RF electric field strength of 50 MV m$^{-1}$ even at a small deuterium/tritium (D/T) gas pressure of approximately 5 atm. However, in the scheme of using spatially localized D/T mixture gas streams, the sticking probability could be reduced almost tenfold even at a large gas density of 100 atm with a modest RF field strength of less than 50 MV m$^{-1}$.

Dissociation Strategies to Maximize Coverage of α-Helical Domains in Top-Down Mass Spectrometry of Integral Membrane Proteins.

Transmembrane α-helical domains of membrane proteins tend to remain structured in the gas phase, presenting a challenge for efficient electron capture/transfer dissociation during top-down dissociation mass spectrometry (MS) experiments. In this study, we compare results from different dissociation modes on a modern Orbitrap platform applied to a model integral membrane protein containing two transmembrane helices, the c-subunit of the Fo domain of the chloroplast ATP synthase. Using commercially available options, we compare collisionally activated dissociation (CAD) with the related variant higher-energy collisional dissociation (HCD) and with electron transfer dissociation (ETD). HCD performed better than CAD and ETD. A combined method utilizing both ETD and HCD (EThcD) demonstrates significant synergy over HCD or ETD alone, representing a robust option analogous to activated ion electron capture dissociation, whereby an infrared laser was used to heat the protein ion alongside electron bombardment. Ultraviolet photodissociation at 213 nm displays at least three backbone dissociation mechanisms and covered nearly 100% of backbone bonds, suggesting significant potential for this technique.

Thymidine and stavudine molecules in reactions with low-energy electrons

The negative ion formation and fragmentation processes in thymidine and stavudine were studied by means of resonant electron capture mass spectrometry in the electron energy range 0–14 eV. Stavudine is a nucleoside analog of thymidine, that differs from the latter by the presence of double bond in the deoxyribose moiety. In the energy range below 5 eV transient negative ions have been found to arise via shape resonances resulting in electron attachment into the empty π*-orbitals. The main fragmentation channels for molecular ions are the H-atom loss and N-glycosidic bond cleavage between sugar and a nucleobase moieties. The dissociative electron attachment cross sections for the fragment negative ions were measured. The structures for some observed anions and neutral counterparts were suggested basing on the thermochemical approach for dissociative reactions energy balance under the support of quantum chemical calculations. The processes of ion formation in thymidine were found to be similar to those revealed previously for deoxyuridine nucleoside. In contrast, very strong dissociative channel was found for stavudine yielding [M–R]– ions (where R is a sugar moiety) in the energy range <3 eV. The efficient fragmentation found for stavudine in the low energy electron attachment reactions may have implications for the design of nucleoside radiosensitizers for cancer therapy.

LC-APCI(-)-MS Determination of 1-Chloro-2,4-dinitrobenzene, a Model Substrate for Glutathione S-Transferases.

1-Chloro-2,4-dinitrobenzene (CDNB) is widely used as a model substrate for measuring the enzyme activity of glutathione S-transferases in toxicity studies and in studies focusing on the metabolic capacity of different test systems. To allow the quantification of CDNB at low, nontoxic concentrations, we developed a sensitive liquid chromatography-mass spectrometry (LC-MS) technique, which is based on electron capture ionization using atmospheric pressure chemical ionization (APCI) in negativeion mode. Gas-phase reactions occurring under atmospheric pressure produce specific ions that allow direct CDNB quantification down to 17 ng/mL in water. Using the new technique, we were able to verify CDNB exposure concentrations applied in two typical toxicity studies with early life stages of the common model organisms, zebrafish (Danio rerio) and a zebrafish embryonic cell line (PAC2).

Fragmentation and slow autoneutralization of isolated negative molecular ions of phthalocyanine and tetraphenylporphyrin.

Macrocyclic tetrapyrrolic compounds, such as naturally occurring or artificial porphyrins and phthalocyanines, have unique and highly attractive properties for applications in medicine and technology. The interaction of free-base phthalocyanine (H2Pc) and tetraphenylporphyrin (H2TPP) molecules with low-energy (0-15 eV) electrons was studied in vacuo by means of negative ion resonant electron capture mass spectrometry. Close similarities in formation and decay of negative ions of these compounds were revealed. Efficient formation of long-lived molecular negative ions (MNIs) was observed in the incident electron energy range of 0-8 eV, unprecedentedly wide for organic compounds and comparable to the range characteristic to carbon atomic clusters, fullerenes. Experiments testify to the strong persistence of MNIs of both compounds to dissociative decay, isomerization, and electron autodetachment. Lifetimes of MNIs as a function of incident electron energy were measured and it was concluded that the isolated anions may retain additional electrons in a time scale of up to hundreds of seconds at standard temperature due to the high adiabatic electron affinity of these large molecules. For the representatives of dyes and photochromic compounds comprehensively studied in terms of interaction with light, the present work highlights yet another unique property of these molecules, namely the capability to attach and durably retain an additional electron of low, pre-ionization energy.

Supercritical fluid extraction of pesticides in sediment from the Okavango Delta, Botswana, and determination by gas chromatography with electron capture detection (GC-ECD) and mass spectrometry (GC-MS)

The study reports supercritical fluid extraction (SFE) of pesticides in sediment samples from the Okavango Delta, the headwaters of which originate from Angola and traverse through Namibia into Botswana. An acetone modifier and fractional extraction by pressure ramping gave recoveries ranging from 55 to 86% for the 15 pesticides studied. Hexachlorobenzene (HCB), aldrin and 4, 4-dichlorodiphenyltrichloroethane (4, 4'-DDT) were identified at concentration ranges of 1.1 to 30.3, 0.5 to 15.2 and 1.4 to 55.4 µg/g, respectively by gas chromatography with electron capture detection (GC-ECD) and were unequivocally confirmed by gas chromatography time-of-flight mass spectrometry (GC-ToF-MS). The study indicated an increase of pesticide concentrations in the direction of water flow from the Panhandle (point of entry) to the lower delta. The results show that there are ramifications associated with activities either upstream or downstream; hence close monitoring is required for the long-term preservation of the delta.

Ion mobility spectrometers and electron capture detector – A comparison of detection capabilities

Electron capture detectors (ECDs) and detectors used in ion mobility spectrometry (IMS) have been successfully used for the detection of numerous compounds including hazardous substances. The general principles of their operations are similar and based on sample component ionization and measurement of the signal using the differences in the mobility of electric charge carriers. Differences in sensitivity result from various parameters of these instruments. Value of electric field intensity in ionic reactors have an influence on ionization process. The main goal of the performed tests was to compare the analytical properties of ECD and two types of IMS detectors: a drift tube spectrometer (DT IMS) and a differential mobility spectrometer (DMS).In the work performed, the efficiency of ionization and the response of detectors to selected analytes were compared. ECD, DT IMS and DMS were equipped with 63-Ni radioactive sources. Analytes have been ionized via electron capture process or dissociative electron transfer. Results obtained for oxygen and chloro-substituted organic compounds (carbon tetrachloride, benzyl chloride, chloroform, 2-chloroethyl ethyl sulfide) were used to calculate the relative signal and to compare the ionization efficiency for three detectors. The phenomena observed experimentally were related to energy dependencies and electron capture cross-sections of analytes. The efficiency of ionization in DT IMS was also compared for electron capture when nitrogen was used as the carrier gas, and when the ionization process was based on the collisions of the analyte molecules with the O2- with the use of air.

ФИПРОНИЛ: ХИМИКО-АНАЛИТИЧЕСКОЕ ОБЕСПЕЧЕНИЕ КОНТРОЛЯ БЕЗОПАСНОСТИ ПИЩЕВОЙ ПРОДУКЦИИ

This work describes the development of the analytical methodology for determining fipronil and metabolite of a fipronil-sulfone residues in eggs, products egg dry food (eggs power), meat, offal of a poultry by gas chromatography with two detecting systems: electron-capture detection (GC-ECD) and mass spectrometry (GC-MS). Two approaches of sample preparation are presented: it agrees to the first - substances extract mix acetone acetonitrile, clear an aliquot of extract the adsorptive chromatography on silica gel and the florisil. The second approach includes extraction of analytes the acetonitrile containing 1% of acetic acid in the presence of MgSO4 and NaCl, cleaning with dispersive solid-phase extraction with use of mix of sorbents (C18, PSA, GCB). LOQ: 0,002–0,005 mg/kg, recovery – 82–104 %, RSD repeatability – 5,9–13,0 %.

Replacing H+ by Na+ or K+ in phosphopeptide anions and cations prevents electron capture dissociation.

By successively replacing H+ by Na+ or K+ in phosphopeptide anions and cations, we show that the efficiency of fragmentation into c and z˙ or c˙ and z fragments from N-Cα backbone bond cleavage by negative ion electron capture dissociation (niECD) and electron capture dissociation (ECD) substantially decreases with increasing number of alkali ions attached. In proton-deficient phosphopeptide ions with a net charge of 2-, we observed an exponential decrease in electron capture efficiency with increasing number of Na+ or K+ ions attached, suggesting that electrons are preferentially captured at protonated sites. In proton-abundant phosphopeptide ions with a net charge of 3+, the electron capture efficiency was not affected by replacing up to four H+ ions with Na+ or K+ ions, but the yield of c, z˙ and c˙, z fragments from N-Cα backbone bond cleavage generally decreased next to Na+ or K+ binding sites. We interpret the site-specific decrease in fragmentation efficiency as Na+ or K+ binding to backbone amide oxygen in competition with interactions of protonated sites that would otherwise lead to backbone cleavage into c, z˙ or c˙, z fragments. Our findings seriously challenge the hypothesis that the positive charge responsible for ECD into c, z˙ or c˙, z fragments can generally be a sodium or other metal ion instead of a proton.

Characterization of the iron-binding properties of pyoverdine using electron-capture dissociation-tandem mass spectrometry.

Pyoverdines (PVD) are a group of siderophores produced by fluorescent Pseudomonads. Identification of PVD variants mostly relies on liquid chromatography-tandem mass spectrometry (LC-MS/MS) using collision-induced dissociation (CID). Here, both CID and the novel dissociation technique electron-capture dissociation (ECD) were applied to characterize PVD succinamide and its Fe(III)-chelated complex. The results clearly showed that ECD produced diagnostic side chain fragmentation of the PVD peptide chain and preserved the labile Fe(III) binding to the chromophore in contrast to CID. The ECD technique is therefore expected to support the understanding of strain-specific Fe(III) transport processes of PVDs.

On the use of electron capture rate constants to describe electron capture dissociation mass spectrometry of peptides.

Electron capture dissociation (ECD) tandem mass spectrometry (MS/MS) is a powerful analytical tool for peptide and protein structure analysis. The product ion abundance (PIA) distribution in ECD MS/MS is known to vary as a function of electron irradiation period. This variation complicates the development of a method of peptide identification by correlation of ECD MS/MS data with experimental and theoretical mass spectra. Here, we first develop a kinetic model to describe primary electron capture by peptide dications leading to product ion formation and secondary electron capture resulting in product ion neutralization. We apply the developed kinetic model to calculate product ion formation rate constants and electron capture rate constants of product ions from ECD mass spectra acquired using various electron irradiation periods. Contrary to ECD PIA distributions, the product ion formation rate constants are shown to be independent of electron irradiation period and, thus, may be employed to characterize ECD product ion formation more universally. The electron capture rate constants of product ions in ECD Fourier transform ion cyclotron resonance MS were found to correlate (with a correlation factor, R(2), of ca 0.9) with ion mobility cross sections of product ions in electron transfer dissociation. Finally, we demonstrate that the electron irradiation period influences the ratio of radical and even-electron c and z product ions.

Cross sections of electron capture and electron capture ionization versus the impact parameter in collisions of a proton with multielectron atoms

The absolute differential cross sections of scattering of hydrogen atoms resulting from an electron capture and an electron capture ionization are measured for collisions of 4.5- and 11-keV protons with argon and xenon atoms. The range of scattering angles is 0°–2°. From the scattering differential cross section found experimentally, the probabilities of single-electron capture and electron capture ionization as a function of the impact parameter are calculated. The dependences of the incident particle scattering angle on the impact parameter (deviation function) for interactions with Ar and Xe atoms are calculated in terms of classical mechanics using the Moliere—Yukawa potential to describe the interaction of atomic particles. Analysis is given to the probabilities of electron capture and electron capture ionization versus the impact parameter and to the distribution of the electron density on different electron shells in a target atom versus a distance to the core. It is concluded that only electrons from the outer shell of the target atom are involved in the process of electron capture ionization. The cross section of electron capture ionization is calculated in the proton energy range 5–20 keV.

Negative electrospray ionization on porous supporting tips for mass spectrometric analysis: electrostatic charging effect on detection sensitivity and its application to explosive detection

The simplicity and easy manipulation of a porous substrate-based ESI-MS technique have been widely applied to the direct analysis of different types of samples in positive ion mode. However, the study and application of this technique in negative ion mode are sparse. A key challenge could be due to the ease of electrical discharge on supporting tips upon the application of negative voltage. The aim of this study is to investigate the effect of supporting materials, including polyester, polyethylene and wood, on the detection sensitivity of a porous substrate-based negative ESI-MS technique. By using nitrobenzene derivatives and nitrophenol derivatives as the target analytes, it was found that the hydrophobic materials (i.e., polyethylene and polyester) with a higher tendency to accumulate negative charge could enhance the detection sensitivity towards nitrobenzene derivatives via electron-capture ionization; whereas, compounds with electron affinities lower than the cut-off value (1.13 eV) were not detected. Nitrophenol derivatives with pKa smaller than 9.0 could be detected in the form of deprotonated ions; whereas polar materials (i.e., wood), which might undergo competitive deprotonation with the analytes, could suppress the detection sensitivity. With the investigation of the material effects on the detection sensitivity, the porous substrate-based negative ESI-MS method was developed and applied to the direct detection of two commonly encountered explosives in complex samples.

Capabilities and limitations of direct analysis in real time orbitrap mass spectrometry and tandem mass spectrometry for the analysis of synthetic and natural polymers

Despite the widespread use of direct analysis in real time mass spectrometry (DART-MS), its capabilities in terms of accessible mass range and the types of polymers that can be analysed are not well known. The goal of this work was to evaluate the capabilities and limitations of this ionization technique combined with orbitrap mass spectrometry and tandem mass spectrometry, for the characterization (structural and polydispersity metrics) of various synthetic and natural polymers. The capabilities and limitations of DART-MS (and -MS(2)), using an orbitrap mass spectrometer, for polymer analysis were evaluated using various industrial synthetic polymers and biopolymers. Stainless steel mesh screens secured on a movable rail were used as the sampling surface, onto which 5 μL of various polymers dissolved in tetrahydrofuran were added. Assignment of spectral features and calculation of molecular weight and polydispersity metrics were performed using Polymerix™ software and the results were compared with those obtained by gel-permeation chromatography (GPC). Protonated oligomers and ammonium adducts were instantaneously detected as the major ionisation products in positive ion mode. Only perfluoropolyethers (PFPEs) were ionised in negative mode and detected as [M](-·) ions. Only singly charged molecular species were observed for all oligomers under study, allowing for a rapid determination of the molecular weight and polydispersity metrics of polymers. At elevated DART gas temperatures (400-500°C) the molecular weight and polydispersity metrics compared fairly well with those obtained by GPC, with polymers whose masses ranged from 200 g x mol(-1) to 4000 g x mol(-1). DART-MS allowed the direct and rapid analysis (mass spectra and tandem mass spectra of all the polymers were acquired in seconds) based on the exact masses of their [M+H](+) and [M+NH4](+) ions (in the positive mode) or [M](-·) ions (for polymers having a high sensitivity toward electron-capture ionisation such as PFPEs), as well as the exact masses of their product ions, for both synthetic and natural polymers under ambient conditions without any sample pre-treatment.

Top‐down structural analysis of posttranslationally modified proteins by Fourier transform ion cyclotron resonance‐MS with hydrogen/deuterium exchange and electron capture dissociation

High-resolution structural characterization of posttranslationally modified proteins represents a challenge for traditional structural biology methods such as crystallography and NMR. In this study, we have used top-down hydrogen/deuterium exchange MS (HDX-MS) with precursor ion selection and electron capture dissociation to determine the impact of oxidative modification on calmodulin (CaM) at an average resolution of 2.5 residues, with complete sequence coverage. The amide deuteration status of native CaM determined by this method correlates well with previously reported crystallographic and NMR data. In contrast, methionine oxidation caused almost complete deuteration of all residues in the protein in 10 s. The oxidative-modification-induced secondary and tertiary structure loss can be largely recovered upon calcium ligation, which also resulted in a substantial increase of amide protection in three of the four calcium-binding loops in oxidatively modified CaM (CaMox ). However, the structure of α-helix VI is not restored by cofactor binding. These results are discussed in terms of different target binding and activation capabilities displayed by CaM and CaMox . The isoform-specific top-down HDX structural analysis strategy demonstrated in this study should be readily applicable to other oxidatively modified proteins and other types of PTMs, and may help decipher the structure and function of specific protein isoforms.

Formation of doubly charged negative ions under the conditions of the resonant electron capture by fluorofullerenes

Doubly charged negative ions formed when electrons with controlled energies interact with isolated fluorinated fullerene molecules C60F n (n = 36, 48) have been detected and investigated by resonant electron capture mass spectrometry. The dependence of the intensity of the formation of doubly charged negative ions of fluorofullerenes on the energy of attached electrons has been measured. An original method, which is based on the experimental data and does not require additional calibration quantities, has been developed for estimating the absolute cross section for the formation of doubly charged negative ions. The absolute cross sections for the formation of the most intensely formed ions C60F 36 2− and C60F 48 2− are estimated to be about 1.1 × 10−24 and 1.5 × 10−24 m2 at their maximum-yield energies of 2.0 and 1.6 eV, respectively.

Analytical Methodology for the Determination of Organochlorine Pesticides in Vegetation

Due to the extensive use of organochlorine pesticides (OCPs) for agricultural purposes and their high persistence and low biodegradability, they have become an important group of contaminants. Detection and quantification of pesticide residues in food, particularly fruits and vegetables, is of growing concern for producers, consumers, and governments. The most widely used pretreatment for the extraction of pesticides in plants is based on solvent extraction liquid-solid extraction (LSE). LSE can be carried out using Soxhlet, shake-flask, homogenization, sonication, and, more recently, microwave-assisted extraction, pressurized liquid extraction, and supercritical fluid extraction. Furthermore, new analytical procedures using the extraction with sorbents, such as solid-phase microextraction, stir bar sorptive extraction, and matrix solid-phase dispersion, have also been used. On the other hand, a wide range of cleanup methods (liquid-liquid extraction, solid-phase extraction, gel permeation chromatography, and dispersive solid-phase extraction; and chromatographic techniques with electron capture detector and mass spectrometry detector; and HPLC with a ultraviolet detector are reported in the literature. This article reviews the applicability, advantages, and disadvantages of various sample preparation techniques (traditional and new techniques) for the analysis of OCPs in different plants and plant materials. It covers more than 15 years of published methods in which pesticide residues have been determined in a wide range of vegetation samples (fruits, horticultural samples, medicinal plants, tree leaves, etc.) by the use of chromatographic techniques after various sample preparation steps. A great number of applications in different plant material are provided. To the best of the authors' knowledge, previously published reviews have not covered as wide and exhaustive range of vegetation matrixes as presented here. A summary of pesticide levels cited in the literature is included.