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.     

Cluster ion structures formed by positive and negative chemical ionization of lactic and other hydroxyacids

Both positive ion and negative ion chemical ionization mass spectra of hydroxycarboxylic acids (hydroxyethanoic acid, 3-hydroxypropanic acid, 2-hydroxypropanoic acid and 1-phenyl-1-hydroxyethanoic acid) show intense oligomeric ions when the samples are evaporated into a chemical ionization source. The observation of oligomeric anionic and cationic species is unusual, and the parallel behavior observed between the positive and negative ion mass spectra is striking. These results are explicable in terms of evaporation of oligomers and their dehydration products from the hot probe, although gas phase clustering reactions of singly charged ions are not excluded. Hydrogen bonding and dehydration provide bonding within each cluster. The structures of the ions have been confirmed by recording the collision induced dissociations of individual cluster ions via their mass analyzed ion kinetic energy spectra. Temperature dependence of the chemical ionization mass spectra provides a method for distinguishing hydrogen bonding from covalent bonding and gives further structural information on the cluster ions.     

Analysis of amino acid mixtures by plasma desorption mass spectrometry

Plasma desorption mass spectrometry (PDMS) was investigated as a means of analysing mixtures of three, four and five amino acids in both positive- and negative-ion modes. Fifteen mixtures were tested; each mixture contained equimolar amounts of selected amino acids. The PD mass spectra exhibited MH⁺ and [M – H]^? molecular ions for all the aminoacids with different desorption–ionization yields. The spectra were more easily interpreted in the negative- than the positive-ion mode. The desorption order of the amino acids was progressively established by comparing the molecular ion desorption–ionization yields for each mixture. This desorption order was well correlated in both the positive- and negation-ion modes with the acid–base thermodynamic data for the amino acids in the gas phase. This observation gives some insight into the desorption–ionization mechanisms under PDMS conditions.     

Identification of amino acid thiohydantoin derivatives by chemical ionization mass spectrometry

The chemical ionization mass spectra of 16 amino acid thiohydantoins were examined using isobutane or ammonia as reagent gases. Except for a few cases, including some aromatic amino acids, the chemical ionization spectra were much simpler than the corresponding electron impact spectra. Therefore, the major component in the amino acid thiohydantoin mixture was easily detected by chemical ionization mass spectrometry. The combination of the chemical ionization method and thiohydantoin formation was applied successfully to the sequence analysis of model peptides.     

Rapid characterization of edible oils by direct matrix-assisted laser desorption/ionization time-of-flight mass spectrometry analysis using triacylglycerols

Direct matrix-assisted laser desorption/ionization time-of-flight mass spectrometric (MALDI-TOFMS) analysis of solutions of edible fats/oils yielded spectra useful for their rapid differentiation and classification. Results also reflected the individual fatty acid components and their degree of unsaturation. After dissolution in hexane, MALDI-MS analysis revealed spectra showing characteristic triacylglycerols (TAGs), the main fat/oil components, as sodium adduct ions. The Euclidean distances calculated using the mass and intensity values for 20 TAGs were used to evaluate and compare spectra. With cluster analysis, animal fats grouped together differently than vegetable oils and the individual oils grouped together by type. The ion abundances for the individual TAGs and their presumed compositions were used to approximate the overall fatty acid composition of canola, soybean, corn, olive and peanut oil, as well as lard. Using this approach the calculated fatty acid compositions and degree of unsaturation generally fell within about 4% of literature values. When the degree of saturation was compared with values calculated from the package labeling the differences were about 7%.     

若干气溶胶单粒子化学成分的实时测量

利用激光解吸附电离飞行时间质谱技术获得了若干已知化学成分的气溶胶粒子的飞行时间质谱,分析标识了各类气溶胶粒子的特征离子谱峰,并对一些特征峰的形成机理进行了探讨。在此基础上,对烟花火药以及纸张燃烧产生的烟气气溶胶粒子进行了实时在线测量,通过对质谱图的分析,获得了有关此两类燃烧过程产生的烟气气溶胶单粒子的化学组成信息。     

Direct mass spectrometric analysis of Bacillus spores

Spores from the Bacillus species, B. cereus, B. anthracis, B. thuringensis, B. lichenformis, B. globigi, and B. subtilis, were examined by direct probe mass spectrometry using electron ionization (EI) and positive and negative chemical ionization (CI). Molecular ions from free fatty acids and nucleic acids were observed in the 70eV spectra as were fragments from glycerides. Spectra obtained with isobutane positive chemical ionization (CI(+)) were dominated by ions associated with pyranose compounds such as N-acetylglucosamine (NAG). Unlike the positive ion spectra, the negative ion spectra of the spores were very simple and contained few peaks. The M(-.) ion from dipicolinic acid (DPA) was the base peak in the negative ion spectra of all spore species except those from B. lichenformis. The negative ion of DPA produced such a strong signal that 10(8) colony forming units (CFUs) of B. cereus spores could be detected directly in 0.5 g of ground rice. Principal component analysis (PCA) of the spectra revealed that only CI(+) spectra contained differences that could be used to identify the spectra by species. Differentiation of the CI(+) spectra by PCA was attributed to variances in the peaks associated with the bacterial polymer poly(3-hydroxybutyrate) (PHB) and NAG. Similar differences in PHB and NAG peaks were detected in the CI(+) spectra of a suite of vegetative Bacillus stains grown with various media.     

Equilibrium conditions in laser-desorbed plumes: thermodynamic properties of alpha-cyano-4-hydroxycinnamic acid and protonation of amino acids

The equilibrium nature of a plume of laser desorbed material is explored through the application of a simple equilibrium model to the ion signals observed in 355 nm laser desorption/ionization mass spectra of mixtures of the MALDI matrix alpha-cyano-4-hydroxycinnamic acid (alphaCHCA) with the amino acids glycine, alanine, valine, isoleucine, and phenylalanine. In these studies it is found that there are systematic and predictable increases in the relative yield of protonated amino acid with increases in amino acid gas-phase basicity. In addition, the thermodynamic values extracted from the equilibrium plot are shown to be in good agreement with values obtained from computational investigation of plausible alphaCHCA proton donor species. These results are supportive of a picture wherein the laser-desorbed material is viewed as a dense plume in which facile charge transfer occurs leading, ultimately, to a thermodynamically equilibrated distribution of proton donor and proton acceptor species.     

High-resolution triacylglycerol mixture analysis using high-temperature gas chromatography / mass spectrometry with a polarizable stationary phase,negative ion chemical ionization,and mass-resolved chromatography

High-temperature gas chromatography/mass spectrometry (HT-GC/MS) has been employed to study the behavior of mixtures of triacylglycerol molecular species on a polarizable stationary phase (immobilized 65% phenyl methyl silicone). The use of negative ion chemical ionization (NICI) at an ion source block temperature of 300 °C overcomes problems with interpretation of electron ionization (EI) mass spectra produced during the HT-GC/MS. The NICI spectra of triacylglycerols produced under these conditions contain abundant [RCO₂]^?, [RCO₂ ? 18]^?, and [RCO₂ ? 19]^? ions, believed to be produced by nucleophilic gas-phase ammonolysis, that are used to identify the individual fatty acid moieties associated with peaks in triacylglycerol total ion chromatograms. The polarizable stationary phase produces significantly enhanced resolution of triacylglycerol molecular species compared to hightemperature stable apolar stationary phases, such as immobilized dimethyl polysiloxanes. The resolution of complex natural mixtures of triacylglycerols can be further improved by use of the Biller-Biemann enhancement technique to produce mass-resolved chromatograms. Investigation of the mass-resolved chromatograms provides important information with regard to the factors that affect elution orders of individual triacylglycerol molecular species. The analysis of mixtures of authentic triacylglycerols by HT-GC/MS via NICI provides data that relate to the analytical limits of the technique for the analysis of triacylglycerols that bear both saturated and polyunsaturated fatty acyl moieties.     

Chemical ionization mass spectrometry of complex molecules—VI: Peptides

The chemical ionization mass spectra of a series of simple peptides containing six or fewer amino acids have been studied. Using methane as the reactant gas we found cleavage of the peptide bond occurs in two ways, yielding either the acyl carbonium ion or the complementary ammonium ion. The observation of both types of fragments permits the determination of the amino acid sequence of the peptide. The ammonium ions provide an additional sequence determining route compared to that available from electron-impact spectra. ‘Sequence-determing ions,’ especially the quasimolecular ion at m/e [M+1] are usually more intense than in the electron-impact mass spectra.     

The mass spectrometry of polychlorinated dibenzo-p-dioxins

The negative ion chemical ionization mass spectra of polychlorinated dibenzo-p-dioxins using oxygen, methane and methane/oxygen are reported together with their methane positive ion chemical ionization mass spectra and conventional electron impact spectra. The methane/oxygen negative ion chemical ionization mass spectra proved to be the most useful of the negative ion spectra for structure determination.     

A semi‐quantitative approach for the rapid screening and mass profiling of naphthenic acids directly in contaminated aqueous samples

We report the use of a direct sampling, online analytical approach for the determination of acid extractable naphthenic acids in complex aqueous samples, known as condensed phase membrane introduction mass spectrometry (CP‐MIMS). The technique employs a capillary hollow fibre semi‐permeable membrane probe configured for immersion into a pH adjusted sample. A continuously flowing methanol acceptor phase transfers naphthenic acids to an electrospray ionization source, operated in negative ion mode, whereupon they are analysed by mass spectrometry as [M–H]^? ions. High‐resolution mass spectrometry is used to characterize the influence of sample pH on membrane transport of multiple components of complex naphthenic acid mixtures. We demonstrate the use of CP‐MIMS for semi‐quantitative analysis of real‐world samples using selected ion monitoring and full scan mass spectra at unit mass resolution. The technique has also been employed to continuously monitor the temporal evolution in the mass profile and concentrations of individual naphthenic acid isomer classes in heterogeneous solutions during adsorption processes. Copyright © 2015 John Wiley & Sons, Ltd.     

Sites of protonation in phenothiazines: Methane and ammonia chemical ionization mass spectra

Positive ion methane and ammonia chemical ionization mass spectra for ten phenothiazine derivatives are reported. The fragmentations observed in the chemical ionization mass spectra are rationalized in terms of the location of the added proton. High-resolution measurements are used to confirm empirical formulae of the ions in the mass spectra. Changes in the mass spectra with a change in the chemical ionization reagent gas from methane to ammonia are described. A comparison with positive ion secondary ion mass spectra of the same compounds show that the amount of fragmentation is higher in the secondary ion mass spectra, but the same types of ions are observed in spectra produced by both ionization methods.     

Definitive identification of lead(II)-amino acid adducts and the solid state structure of a lead-valine complex

Electrospray ionization mass spectra of lead(II) nitrate-amino acid mixtures enable unequivocal identification of lead complexes for each of the essential amino acids and a valine complex is reported as the first crystallographically characterized lead-amino acid complex.     

The novel use of gas chromatography-ion mobility-time of flight mass spectrometry with secondary electrospray ionization for complex mixture analysis

Increasing the dimensionality of an analysis enables more detailed and comprehensive investigations of complex mixtures. One dimensional separation techniques like gas chromatography (GC) and ion mobility spectrometry (IMS) provide limited chemical information about complex mixtures. The combination of GC, ion mobility spectrometry, and time-of-flight mass spectrometry (GC-IM-TOFMS) provides three-dimensional separation of complex mixtures. In this work, a hybrid GC-IM-TOFMS with a secondary electrospray ionization (SESI) source provided four types of analytical information: GC retention time, ion mobility drift time, mass-to-charge ratios, and ion intensity. The use of secondary electrospray ionization enables efficient and soft ionization of gaseous sample vapors at atmospheric pressure. Several complex mixtures, including lavender and peppermint essential oils, were analyzed by GC-SESI-IM-TOFMS. The resulting 3D data from these mixtures, each containing greater than 50 components, were plotted as 3D projections. In particular, post-processed data plotted in three dimensions showed that many mass selected GC peaks were resolved into different ion mobility peaks. This technique shows clear promise for further in-depth analyses of complex chemical and biological mixtures.     

The application of a new sampling technique using an atomizer for chemical ionization mass spectrometry to free amino acids,drug components,higher phthalates and oligomers of styrene and ethyleneglycol

A new vacuum atomizing sample introduction device was developed and applied to take stable and reproducible chemical ionization mass spectra of various involatile compounds. As examples, chemical ionization spectra of some free amino acids, drug components in a cold medicine, higher phthalates, oligomers of styrene and ethyleneglycol, etc., are presented.     

Mass spectra of synthesized mixed bromochloroacetic acids

Mass spectra of methyl esters of three common drinking water chlorination by-products, bromodichloroacetic acid, dibromochloroacetic acid and tribromoacetic acid, are presented. The spectra are obtained in electron impact and chemical ionization modes, using a quadrupole mass spectrometer. Proposed fragment ion assignments are presented in detail. Fragment clusters were identified from their m/z values and confirmed with their relative isotope ratios. The detailed mass spectra presented are especially useful because standards for these drinking water contaminants are not yet commercially available.     

Vibrational and photoionization spectroscopy of biomolecules: aliphatic amino acid structures

The aliphatic amino acids glycine, valine, leucine, and isoleucine are thermally placed into the gas phase and expanded into a vacuum system for access by time of flight mass spectroscopy and infrared (IR) spectroscopy in the energy range of 2500-4000 cm(-1) (CH, NH, OH, and stretching vibrations). The isolated neutral amino acids are ionized by a single photon of 10.5 eV energy (118 nm), which exceeds by less than 2 eV their reported ionization thresholds. As has been reported for many hydrogen bonded acid-base systems (e.g., water, ammonia, alcohol, acid clusters, and acid molecules), the amino acids undergo a structural rearrangement in the ion state (e.g., in simplest form, a proton transfer) that imparts sufficient excess vibrational energy to the ion to completely fragment it. No parent ions are observed. If the neutral ground state amino acids are exposed to IR radiation prior to ionization, an IR spectrum of the individual isomers for each amino acid can be determined by observation of the ion intensity of the different fragment mass channels. Both the IR spectrum and fragmentation patterns for individual isomers can be qualitatively identified and related to a particular isomer in each instance. Thus, each fragment ion detected presents an IR spectrum of its particular parent amino acid isomer. In some instances, the absorption of IR radiation by the neutral amino acid parent isomer increases a particular fragmentation mass channel intensity, while other fragmentation mass channel intensities decrease. This phenomenon can be rationalized by considering that with added energy in the molecule, the fragmentation channel populations can be modulated by the added vibrational energy in the rearranged ions. This observation also suggests that the IR absorption does not induce isomerization in the ground electronic state of these amino acids. These data are consistent with theoretical predictions for isolated amino acid secondary structures and can be related to previous IR spectra of amino acid conformers.     

Analysis of tetrapyrrolic pigments and derivatives in sediments by high performance liquid chromatography/atmospheric pressure chemical ionization mass spectrometry

A new application is described of the on-line high-performance liquid chromatography/positive and negative ion atmospheric pressure chemical ionization mass spectrometry (APCI(+/–)-MS) to the identification of tetrapyrrolic compounds in sediment samples containing complex mixtures of natural pigments and their diagenetic products. The method relies on the production of radical anions in order to determine the molecular masses of the molecules which yield abundant fragment ions when analyzed in the positive mode. This approach allows to verify the occurrence of coeluting components by examining their negative ion spectra and to complete their identification on the basis of their fragment ions produced in the positive ion spectra. Received: 24 September 1998 / Revised: 23 December 1998 / Accepted: 29 December 1998     

Analysis of tetrapyrrolic pigments and derivatives in sediments by high performance liquid chromatography/atmospheric pressure chemical ionization mass spectrometry

A new application is described of the on-line high-performance liquid chromatography/positive and negative ion atmospheric pressure chemical ionization mass spectrometry (APCI(+/–)-MS) to the identification of tetrapyrrolic compounds in sediment samples containing complex mixtures of natural pigments and their diagenetic products. The method relies on the production of radical anions in order to determine the molecular masses of the molecules which yield abundant fragment ions when analyzed in the positive mode. This approach allows to verify the occurrence of coeluting components by examining their negative ion spectra and to complete their identification on the basis of their fragment ions produced in the positive ion spectra. Received: 24 September 1998 / Revised: 23 December 1998 / Accepted: 29 December 1998