The effects of electron and chemical Ionization Modes on the MS Profiling of Whole Bacteria

Free fatty acid profiling of whole bacteria [Francisella tularensis, Brucella melitensis, Yersinia pestis, Bacillus anthracis (vegetative and sporulated), and Bacillus cereus] was carried out with direct probe mass spectrometry under 70-eV electron ionization (EI) and isobutane chemical ionization in both the positive (CI+) and negative modes (CI-). Electron ionization produced spectra that contained molecular ions and fragment ions from various free fatty acids. Spectra acquired with isobutane chemical ionization in the positive mode yielded molecular ions of free fatty acids as well as ions from other bacterial compounds not observed under EI conditions. Spectra obtained with negative chemical ionization did not contain as much taxonomic information as EI or CI+; however, some taxonomically significant compounds such as dipicolinic acid and poly(3-hydroxybutyrate) did produce negative ions. All ionization modes yielded spectra that could separate the bacteria by Gram-type when observed with principle components analysis (PCA). Chemical ionization in the positive ion mode produced the greatest amount of differentiation between the four genera of bacteria when the spectra where examined by PCA.     

Comparative EI,CI and FAB Mass Spectrometry of Dialkyl and Monoalkyl 2-Chloroethylphosphonates

Abstract

Comparison is made between electron ionization (EI), methane, isobutane and ammonia chemical ionization (CI) and fast atom bombardment (FAB) mass spectrometry of phosphonates related to 2-chloroethylphosphonic acid     

Structural elucidation of disinfection by-products in treated drinking water

Two unusual disinfection by-products have been detected periodically in the gas chromatography/mass spectrometry (GC/MS) characterization analyses of semi-volatile organics in treated drinking water. The electron ionization (EI) mass spectra contained mono-chlorinated and mono-brominated ions at m/z 107/109 and 151/153, respectively. Library searching techniques suggested mono-halogenated butanol structures but no matches could be found. Positive ion chemical ionization (CI) spectra contained mono-chlorinated and mono-brominated ions at m/z 105/107 and 149/151, respectively. No [M + H]+ ions were initially observed. Accurate mass measurements confirmed the empirical formulae for the significant ions in the EI spectra and the mono-halogenated butanol structures. Further CI experiments with other reagent gases and instruments revealed possible molecular weights of 139 and 183 Da, respectively. Tandem mass spectrometry (MS/MS) experiments in EI and CI were used to elucidate the fragmentation schemes. The two compounds have been tentatively identified as 1-aminoxy-1-chlorobutan-2-ol and 1-aminoxy-1-bromobutan-2-ol, respectively.     

Mass spectra of some new 3,4-dihydro-2[H]-pyridones

Electrospray ionization (ESI) and tandem mass spectrometry (MS/MS) experiments, as well as electronic impact (EI) and chemical ionization (CI) techniques, have been applied to the title compounds 1a-h. The observation of different fragmentation pathways in the three sets of spectra is in accord with different degrees of internal excitation of the investigated precursors. In ESI (methanol as solvent) and CI (methane as reagent gas) spectra, the MH(+) ion represents the most important peak, while the fragments [M - OH](+) and [M - SO](+) are either the base peak or a very abundant peak in the EI mass spectra of these compounds. ESI-MS/MS experiments on the parent ions [MH](+) show that the loss of a fragment of 140 Da corresponding to p-toluenesulfenic acid is common from all the precursors. As well as competitive pathways, the second generation ions have also been elucidated to allow some observations to be made concerning the relationships between structure type and mass spectrometric characteristics.     

HPLC,mass spectrometry,and LC/MS of gossypol and its derivatives

Conditions are reported for the reverse-phase HPLC analysis of gossypol and of some of its derivatives, including anhydrogossypol, gossypolone, gossypol hexaacetate, and Schiff's base derivatives. It is shown that field desorption (FD) and chemical ionization (CI) mass spectrometry have advantages over electron impact (EI) mass spectrometry for the characterization of these compounds and that combined high-performance liquid chromatography/mass spectrometry (LC/MS), especially when used in the CI mode, is particularly effective.     

Mass spectra of new substituted 2-amino-4H-pyrans: a retro-Diels-Alder reaction pattern

New substituted 2-amino-3-cyano-4H-pyrans have been studied by electron ionization (EI), chemical ionization (CI) and electrospray ionization (ESI) mass spectrometry. The retro-Diels-Alder reaction (RDA) is the main fragmentation pattern observed in the EI spectra forming an unsaturated ketone as the diene fragment. In contrast, a different RDA reaction takes place yielding an unsaturated amide as diene fragment together with the unsaturated ketone in the CI spectra. The MS/MS spectra obtained using an ESI source reveal that the favoured fragmentation by collision induced dissociation (CID) is the elimination of the substituent at the C4 position with formation of a stable pyrilium cation.     

An overview of the retro-Diels-Alder reaction in semiunsaturated heterocyclic rings: mass spectra of new substituted 1,4,5,6,7,8-hexahydroquinolines and their oxo-analogues 5,6,7,8-tetrahydro-4H-chromenes

Electron impact ionization (EI), chemical ionization (CI), electrospray ionization (ESI) and tandem mass spectrometry (MS/MS) were used to investigate a number of relatively large and structurally related new heterocycles such as substituted 1,4,5,6,7,8-hexahydroquinolines and their oxa-analogues 5,6,7,8-tetrahydro-4H-chromenes. In the EI spectra the hexahydroquinolines undergo the loss of the substituent attached at the C4 position, while the 4H-chromenes undergo a retro-Diels-Alder reaction (RDA) after elimination of the C4 substituent. Under chemical ionization conditions the RDA reaction is observed only for the 4H-chromenes. The ESI-MS/MS spectra reveal results similar to the EI and CI spectra, since the 4H-chromenes undergo RDA reactions while the hexahydroquinolines form a very stable even-electron pyridium ion derived from the loss of the C4 substituent.     

Cyclization of the substituted <Emphasis Type="Italic">N</Emphasis>-(<Emphasis Type="Italic">Ortho</Emphasis>-cyclopropylphenyl)-<Emphasis Type="Italic">N′</Emphasis>-aryl ureas and thioureas in the gas phase and solution

Electron ionization (EI), chemical ionization (CI), tandem mass spectrometry, high-resolution measurements, and labeling studies as well as quantum chemical calculations were used to understand the behavior of the molecular radical cations (EI) and protonated molecules (CI) of substituted N-(ortho-cyclopropylphenyl)-N'-aryl ureas and N-(ortho-cyclopropylphenyl)-N'-aryl thioureas in a mass spectrometer. Fragmentation schemes and possible mechanisms of primary isomerization were proposed. According to the fragmentation pattern, formation of the corresponding benzoxazines and benzothiazines was considered as the major process of isomerization of the original M(+.) and MH(+), although some portions of these ions definitely transformed into other structures. The treatment of N-(ortho-cyclopropylphenyl)-N'-phenyl urea and N-(ortho-cyclopropylphenyl)-N'-phenylthiourea in solution with strong acids formed predicted 4-ethyl-N-phenyl-4H-3,1-benzoxazin-2-amin and 4-ethyl-N-phenyl-4H-3,1-benzothiazin-2-amine as principal products.     

Compatibility of electron ionization and soft ionization methods in gas chromatography/orthogonal time‐of‐flight mass spectrometry

Orthogonal acceleration time‐of‐flight (oa‐TOF) mass spectrometry (MS) was coupled to gas chromatography (GC) to measure ion yields (ratio of ion counts to number of neutrals entering the ion source) and signal‐to‐noise (S/N) in the electron ionization (EI) mode (hard ionization) as well as in the soft ionization modes of chemical ionization (CI), electron capture negative ion chemical ionization (NICI) and field ionization (FI). Mass accuracies of the EI and FI modes were also investigated. Sixteen structurally diverse volatile organic compounds were chosen for this study. The oa‐TOF mass analyzer is highly suited for FI MS and provided an opportunity to compare the sensitivity of this ionization method to the more conventional ionization methods. Compared to the widely used quadrupole mass filter, the oa‐TOF platform offers significantly greater mass accuracy and therefore the possibility of determining the empirical formula of analytes. The findings of this study showed that, for the instrument used, EI generated the most ions with the exception of compounds able to form negative ions readily. Lower ion yields in the FI mode were generally observed but the chromatograms displayed greater S/N and in many cases gave spectra dominated by a molecular ion. Ion counts in CI are limited by the very small apertures required to maintain sufficiently high pressures in the ionization chamber. Mass accuracy for molecular and fragment ions was attainable at close to manufacturer's specifications, thus providing useful information on molecular ions and neutral losses. The data presented also suggests a potentially useful instrumental combination would result if EI and FI spectra could be collected simultaneously or in alternate scans during GC/MS. Copyright © 2009 John Wiley & Sons, Ltd.     

红霉素类抗生素的自身化学电离质谱研究

采用自身化学电离离子化(SCI)/四极质谱法,测定了5个红霉素类抗生素,不仅得到了丰度较高的准分子离子(MH)^+,而且也得到了丰富的碎片离子,并对其裂解规律进行了总结。结果表明对于在电离过程中不稳定的化合物,SCI/四极质谱法既可确定其分子量,也可提供丰富的分子结构信息。此方法简便、快速,且图谱简单有助于结构分析。     

Mass spectrometry of partially methylated alditol acetates derived from hydroxyethyl starch

The degradation and derivatization of hydroxyethyl starch to partially methylated alditol acetates (PMAAs) allows its detection by gas chromatography/mass spectrometry. The derivatization was performed by permethylation of the carbohydrate, hydrolysis of the permethylated polysaccharide, reduction of the resulting monosaccharides to alditoles and finally acetylation. A close similarity in the fragmentation of the PMAAs obtained was observed in both electron ionization (EI) and chemical ionization (CI) mass spectra owing to the comparable structures of the derivatives. CI measurements permitted the recognition of introduced hydroxyethyl groups in the glucose residues by detection of [M(+)+1]-60 signals. Investigations concerning the EI fragmentation schemes allowed secure determinations of monohydroxyethyl monosaccharides and differentiations between the possible positions (C-2, C-3 and C-6) of the substituted hydroxyethyl groups. Proposed generations of the main fragment ions are presented.     

Cluster chemical ionization for improved confidence level in sample identification by gas chromatography/mass spectrometry

Upon the supersonic expansion of helium mixed with vapor from an organic solvent (e.g. methanol), various clusters of the solvent with the sample molecules can be formed. As a result of 70 eV electron ionization of these clusters, cluster chemical ionization (cluster CI) mass spectra are obtained. These spectra are characterized by the combination of EI mass spectra of vibrationally cold molecules in the supersonic molecular beam (cold EI) with CI-like appearance of abundant protonated molecules, together with satellite peaks corresponding to protonated or non-protonated clusters of sample compounds with 1-3 solvent molecules. Like CI, cluster CI preferably occurs for polar compounds with high proton affinity. However, in contrast to conventional CI, for non-polar compounds or those with reduced proton affinity the cluster CI mass spectrum converges to that of cold EI. The appearance of a protonated molecule and its solvent cluster peaks, plus the lack of protonation and cluster satellites for prominent EI fragments, enable the unambiguous identification of the molecular ion. In turn, the insertion of the proper molecular ion into the NIST library search of the cold EI mass spectra eliminates those candidates with incorrect molecular mass and thus significantly increases the confidence level in sample identification. Furthermore, molecular mass identification is of prime importance for the analysis of unknown compounds that are absent in the library. Examples are given with emphasis on the cluster CI analysis of carbamate pesticides, high explosives and unknown samples, to demonstrate the usefulness of Supersonic GC/MS (GC/MS with supersonic molecular beam) in the analysis of these thermally labile compounds. Cluster CI is shown to be a practical ionization method, due to its ease-of-use and fast instrumental conversion between EI and cluster CI, which involves the opening of only one valve located at the make-up gas path. The ease-of-use of cluster CI is analogous to that of liquid CI in ion traps with internal ionization, and is in marked contrast to that of CI with most other standard GC/MS systems that require a change of the ion source.     

Temperature-programmed pyrolysis hyphenated with metastable atom bombardment ionization mass spectrometry (TPPy/MAB-MS) for the identification of additives in polymers

Thermoanalytical techniques are currently used for the analysis of additives contained in polymers that cannot be easily dissolved, extracted, or hydrolyzed. With these techniques, the polymers are heated to liberate the additives trapped in the polymer matrix. If the polymer is heated slowly, up to its thermal degradation, the technique is called temperature-programmed pyrolysis (TPPy). For TPPy experiments, mass spectrometry is generally used as the detection method. The ionization sources commonly used in mass spectrometry, such as CI and EI, can cause fragmentation during the ionization process. Fragmentation decreases the sensitivity of the molecular ions and increases the risks of interferences with the compounds coming from the matrix. An energy-tunable ionization technique, called metastable atom bombardment (MAB), is proposed for TPPy/MS experiments. With this ionization source, the energy of ionization depends on the metastable gas used. With low-energy metastable gases such as Xe or N(2), fragmentation is reduced compared to CI, whereas with medium-energy metastable gases such as Ar or Kr, the fragmentation is similar to that observed with CI. TPPy/MAB-MS was performed on an unknown polyurethane-based car paint. The detection of molecular ions and characteristic fragments with MAB(N(2)) led to the identification of two light stabilizers: Bis(1,2,2,6,6-pentamethyl-4-piperidinyl)sebacate (BPPS) and 2-(2H-benzotriazol-2-yl)-4,6- di-tert-pentylphenol (PTPP). Using MAB(Ar) to simulate CI, the molecular ion and one of the two characteristic fragments of BPPS were not detected, thus confirming the advantage of using MAB(N(2)) ionization for TPPy/MS experiments.     

Influence of fluorine atoms on the electron impact and chemical ionization mass spectrometric fragmentation of methyl x-deoxy-x-fluoro-per-O-methyl-β-D-galactopyranosides

Fully methylated methyl x-deoxy-x-fluoro-β-D -galactopyranosides were studied using electron impact (EI) and chemical ionization (CI) mass spectrometry and by gas chromatography (GC)/mass spectrometry. Metastable daughter- and parent-ion measurements and high-resolution measurements were used to evaluate the fragmentation schemes. Both the presence and the position of the electronegative fluorine atom influences the fragmentation pathways of the permethylated compounds. The individual methyl x-deoxy-x-fluoro-per-O-methyl-β-D - galactopy-ranosides have different GC retention times. This, together with the characteristic differences present in the EI or CI (methane or isobutane) mass spectra, allows the location of fluorine in these substances to be unambiguously determined.     

Fast-atom bombardment of the cyclic acetals: Evidence indicating the predominant involvement of condensed-phase processes in ionization

A series of cyclic acetals, the 2-phenyl-l,3 dioxolanes, and their deuterated analogues were studied by electron ionization (EI), chemical ionization (CI), and fast-atom bombardment (FAB) mass spectrometry to gain insight into the primary ionization processes for these compounds in FAB/liquid secondary ion mass spectrometry. Comparison of EI and CI data with that of FAB led to the conclusion that the predominant [M - H]+ ion observed in FAB for the nondeuterated cyclic acetals cannot to a large extent be rationalized in thermodynamic terms by known gas-phase ion-molecule reactions. Instead, a condensed-phase model in which the multicharged transition state for hydride abstraction is better solvated than the transition state for proton transfer appears to be a plausible explanation for the FAB data obtained for the nonlabeled cyclic acetals; however, this explanation is not entirely sufficient to rationalize the FAB data for the deuterated cyclic acetals. For these compounds, a dramatic time dependence of protonation versus hydride abstraction is observed that suggests that beam-induced reactive species are responsible for hydride abstraction in the condensed phase. This time dependence can be interpreted in terms of a buildup of highly reactive beam-induced species in the bulk of solution. Comparison of the results obtained for deuterated acetals with different surface activities support this hypothesis. (J Am Sot Mass     

Characteristic features of mass-spectra of some nitroxide radicals

Some compounds containing mono- and polynitroxide radicals have been investigated and characterized by electron ionization (EI), chemical ionization (CI), and matrix-assisted laser desorption-ionization (MALDI) mass spectrometry. Some characteristics of the mass spectra of these radicals are demonstrated. It was found that, under CI and MALDI conditions, ions [M+nH]⁺ are formed along with monoprotonated molecules [M+H]⁺, depending on the type of the radical n = 2–5.     

Differentiation of diastereomeric conduramine derivatives under electron ionization and chemical ionization mass spectral conditions

Diastereomeric conduramine derivatives, i.e., (1R,2S,3R/S,6S)-6-(N-carbomethoxyamino) 1,2-O-isopropylidenecyclohex-4-ene-1,2,3-triol (1 and 2) and their O-acetyl derivatives (3 and 4), were studied using gas chromatography (GC) with electron ionization (EI) and chemical ionization (CI). The EI mass spectra of diastereomeric pairs show consistent differences in the relative abundances of characteristic ions. The EI fragmentation patterns are based on precursor/product ion spectra, high-resolution mass spectrometry (HRMS) and deuterium labelling. The CI spectra show differences from the EI spectra, and the isobutane/CI spectra are much simpler than the methane/CI spectra. The differences shown in the CI spectra are similar to those shown in the product ion spectra of [M+H](+) ions generated under electrospray ionization (ESI) conditions. Theoretical calculations are performed to understand the observed differences. The differences in the relative stabilities of molecular ions, or protonated molecules at different sites, can explain the observed differences in the spectra.     

Identification of fatty acids in fishes collected from the Ohio River using gas chromatography-mass spectrometry in chemical ionization and electron impact modes

Analyses of fatty acid (FA) composition in freshwater fishes promote understanding of the potential relationship between fish health or human nutrition and specific FAs. Therefore, the chemical identity of FAs in endemic fishes must be established. Paddlefish, sauger, and white bass were collected from the Ohio River. The structural identification of esterified FAs from fish-fillet lipids was conducted using gas chromatography-mass spectrometry (GC-MS). The same 13 FAs, composing more than 90% of the mass of FAs extracted by techniques used in this research, were found in all three species examined. Carbon chain length and degree and position of unsaturation were determined from the characteristic ionization and fragmentation of FA methyl esters (FAMEs) resulting from GC-MS electron impact (EI) and chemical ionization (CI) modes. Assignment of structure to the extracted FAs required complementary interpretation of both EI and CI MS. The EI spectra observed substantiate findings reported in the literature. The novelty of this research is in the thorough interpretation of CI spectra for which less data are available. The observations reported for analyses of fishes will be useful to all researchers studying FAs regardless of sample media.     

Development of a new electron ionization/field ionization ion source for gas chromatography/time‐of‐flight mass spectrometry

We have developed a combined EI/FI source for gas chromatography/orthogonal acceleration time‐of‐flight mass spectrometry (GC/oaTOFMS). In general, EI (electron ionization) and FI (field ionization) mass spectra are complementary: the EI mass spectrum contains information about fragment ions, while the FI mass spectrum contains information about molecular ions. Thus, the comparative study of EI and FI mass spectra is useful for GC/MS analyses. Unlike the conventional ion sources for FI and EI measurements, the newly developed source can be used for both measurements without breaking the ion source vacuum or changing the ion source. Therefore, the combined EI/FI source is more preferable than the conventional EI or FI ion source from the viewpoint of the reliability of measurements and facility of operation. Using the combined EI/FI source, the complementarity between EI and FI mass spectra is demonstrated experimentally with n‐hexadecane (100 pg): characteristic fragment ions for the n‐alkane such as m/z 43, 57, 71, and 85 are obtained in the EI mass spectrum, while only the parent peak of m/z 226 (M⁺) without any fragment ions is observed in the FI mass spectrum. Moreover, the field desorption (FD) measurement is also demonstrated with poly(ethylene glycol)s M600 (10 ng) and M1000 (15 ng). Signals of [M+H]⁺, [M+Na]⁺ and [M+K]⁺ are clearly detected in the FD mass spectra. Copyright © 2009 John Wiley & Sons, Ltd.     

Mass-spectrometric studies of new 6-nitroquipazines—serotonin transporter inhibitors

Six synthesized 6-nitroquipazine derivatives were examined by electron ionization (EI) and electrospray ionization (ESI) mass spectrometry in positive and negative ion mode. The compounds exhibit high affinity for the serotonin transporter (SERT) and belong to a new class of SERT inhibitors. The EI mass spectra registered in negative ion mode showed prominent molecular ions for all the compounds studied. All EI mass spectra and all ESI mass spectra showed similar fragmentation pathways of molecular ions, but the pathways differed between EI and ESI. The differences were explained with the aid of theoretical evaluation of the stability of the respective radical ions (EI MS) and protonated ions (ESI MS).