Molecular ion fragmentation and its effects on mass isotopomer abundances of fatty acid methyl esters ionized by electron impact

We have analyzed the isotopomer abundance ratios of an equimolar mixture of nine fatty acid methyl esters (decanoate, undecanoate, laurate, tridecanoate, myristate, pentadecanoate, palmitate, heptadecanoate, and stearate) by selected-ion monitoring gas chromatography/electron impact/mass spectrometry (GC/EI/MS). The abundance of the second lowest m/z isotopomer (IM1) increased disproportionately compared with the abundance of the lowest m/z isotopomer (IM0) as a function of: (1) increasing sample size; (2) decreasing repeller voltage; and (3) decreasing alkyl chain length. We also compared the abundance of the third lowest m/z isotopomer (IM2) and the abundance of the second lowest m/z isotopomer (IM1) of methyl palmitate and [4,4-2H2]methyl palmitate. We observed that the IM2/IM1 for methyl palmitate was significantly lower than IM2/IM1 for [4,4-2H2]methyl palmitate. From these results, as well as a consideration of basic principles of ion chemistry and ion physics, we conclude that gas-phase chemistry, specifically proton (or deuteron) transfer from fragment ions to molecules, is a major contributor to the sample size dependence observed in mass isotopomer abundance measurements of fatty acid methyl esters ionized by EI. Our results and analysis do not support hydrogen abstraction as the reaction mechanism. In addition, we calculate that rearranged molecular ions are unlikely to contribute significantly to intermolecular proton transfer because of their relatively brief lifetime. We also discuss alternative analytical techniques which might improve the precision and accuracy of isotopomer measurements by reducing molecular ion fragmentation.     

Comparison of EI and metastable atom bombardment ionization for the identification of polyurethane thermal degradation products

Polyurethanes are widely used in the manufacture of commercial products such as foams and paints. During combustion, these polymers can generate isocyanates, which induce adverse health effects. Polymer pyrolysis (Py) hyphenated with mass spectrometry (MS) allows the investigation of polymer thermal degradation over time/temperature. A diphenylmethanediisocyanate (MDI) polyurethane foam was analyzed with electron ionization (EI) and metastable atom bombardment (MAB) ionization at a pyrolysis temperature of 400 °C. The recently introduced MAB ionization source uses discrete energy stored in metastable atoms of gases to ionize the analytes. This characteristic allows modulation of the ionization energy by simply changing the ionization gas. The extensive fragmentation of molecular ions observed using EI 70 eV is not totally eliminated with EI 10 eV. However, only molecular ions are observed with MAB using N₂ as the ionization gas. Temperature gradients were used to separate the products generated during the thermal degradation of a 1,6-hexamethylenediisocyanate (HDI) polyurethane paint. The analysis of mass spectra was facilitated owing to a selective desorption of pyrolysis products. Furthermore, changing the MAB ionization gas allows elucidation of the structure of the pyrolysis products by controlling the extent of their fragmentation. During these experiments, isocyanic acid, methyleneisocyanate, ethyleneisocyanate, propylisocyanate and butylisocyanate were detected.     

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.     

A comparison of electron ionization mass spectra obtained at 70 eV,low electron energies,and with cold EI and their NIST library identification probabilities

Electron ionization (EI) mass spectra of 46 compounds from several different compound classes were measured. Their molecular ion abundances were compared as obtained with 70‐eV EI, with low eV EI (such as 14 eV), and with EI mass spectra of vibrationally cold molecules in supersonic molecular beams (Cold EI). We further compared these mass spectra in their National Institute of Standards and Technology (NIST) library identification probabilities. We found that

1. Low eV EI is not a soft ionization method, and it has little or no influence on the molecular ion relative abundances for large molecules and those with weak or no molecular ions.
2. Low eV EI for compounds with abundant or dominant molecular ions in their 70 eV mass spectra results in the reduction of low mass fragment ions abundances thereby reducing their NIST library identification probabilities thus rarely justifies its use in real‐world applications.
3. Cold EI significantly enhances the relative abundance of the molecular ions particularly for large compounds; yet, it retains the low mass fragment ions; hence, Cold EI mass spectra can be effectively identified by the NIST library.
4. Different standard EI ion sources provide different 70 eV EI mass spectra. Among the Agilent technologies ion sources, the “Extractor” exhibits relatively abundant molecular ions compared with the “Inert” ion source, while the “High efficiency source” (HES) provides mass spectra with depleted molecular ions compared with the “Inert” ion source or NIST library mass spectra.

These conclusions are demonstrated and supported by experimental data in nine figures and two tables.     

Electron ionization mass spectra of some norbornane/ene-fused 2-N-phenyliminoperhydro-1,3-oxazines

The 70 eV electron ionization (EI) mass spectra were recorded for eight norbornane/ene-fused 2-N-phenyl-iminoperhydro-1,3-oxazines, and the fragmentation patterns were studied by metastable ion analysis and exact mass measurement. Whereas the stereoisomeric unsaturated compounds could not be distinguished on the basis of their EI mass spectra, the stereoisomeric saturated compounds gave rise to clearly different spectra. The ionized unsaturated compounds decomposed mainly by two consecutive retro-Diels-Alder (RDA) reactions. A methyl substituent on the ring nitrogen strongly influenced the charge distribution on the RDA fragments. The ionized saturated compounds fragmented through several pathways. Loss of cyclopentadiene from the molecular ion was the energetically favoured fragmentation reaction for the saturated di-endo-fused compounds but was unimportant for the di-exo-fused compounds.     

Determination of elemental compositions by gas chromatography/time‐of‐flight mass spectrometry using chemical and electron ionization

Many metabolomic applications use gas chromatography/mass spectrometry (GC/MS) under standard 70 eV electron ionization (EI) parameters. However, the abundance of molecular ions is often extremely low, impeding the calculation of elemental compositions for the identification of unknown compounds. On changing the beam‐steering voltage of the ion source, the relative abundances of molecular ions at 70 eV EI were increased up to ten‐fold for alkanes, fatty acid methyl esters and trimethylsilylated metabolites, concomitant with 2‐fold absolute increases in ion intensities. We have compared the abundance, mass accuracy and isotope ratio accuracy of molecular species in EI with those in chemical ionization (CI) with methane as reagent gas under high‐mass tuning. Thirty‐three peaks of a diverse set of trimethylsilylated metabolites were analyzed in triplicate, resulting in 342 ion species ([M+H]⁺, [M–CH₃]⁺ for CI and [M]^{+ .} , [M–CH₃]^{+ .} for EI). On average, CI yielded 8‐fold more intense molecular species than EI. Using internal recalibration, average mass errors of 1.8 ± 1.6 mm/z units and isotope ratio errors of 2.3 ± 2.0% (A+1/A ratio) and 1.7 ± 1.8% (A+2/A ratio) were obtained. When constraining lists of calculated elemental compositions by chemical and heuristic rules using the Seven Golden Rules algorithm and PubChem queries, the correct formula was retrieved as top hit in 60% of the cases and within the top‐3 hits in 80% of the cases. Copyright © 2010 John Wiley & Sons, Ltd.     

Classical electron ionization mass spectra in gas chromatography/mass spectrometry with supersonic molecular beams

A major benefit of gas chromatography/mass spectrometry (GC/MS) with a supersonic molecular beam (SMB) interface and its fly-through ion source is the ability to obtain electron ionization of vibrationally cold molecules (cold EI), which show enhanced molecular ions. However, GC/MS with an SMB also has the flexibility to perform 'classical EI' mode of operation which provides mass spectra to mimic those in commercial 70 eV electron ionization MS libraries. Classical EI in SMB is obtained through simple reduction of the helium make-up gas flow rate, which reduces the SMB cooling efficiency; hence the vibrational temperatures of the molecules are similar to those in traditional EI ion sources. In classical EI-SMB mode, the relative abundance of the molecular ion can be tuned and, as a result, excellent identification probabilities and very good matching factors to the NIST MS library are obtained. Classical EI-SMB with the fly-through dual cage ion source has analyte sensitivity similar to that of the standard EI ion source of a basic GC/MS system. The fly-through EI ion source in combination with the SMB interface can serve for cold EI, classical EI-SMB, and cluster chemical ionization (CCI) modes of operation, all easily exchangeable through a simple and quick change (not involving hardware). Furthermore, the fly-through ion source eliminates sample scattering from the walls of the ion source, and thus it offers full sample inertness, tailing-free operation, and no ion-molecule reaction interferences. It is also robust and enables increased column flow rate capability without affecting the sensitivity.     

Relative molecular mass information from aliphatic nitro compounds: A chemical ionization study

The mass spectra of a number of aliphatic nitro compounds have been studied using electron Ionization (EI) and a variety of chemical Ionization (CI) techniques in attempts to obtain relative molecular mass information. The use of positive ion ammonia chemical Ionization techniques gave very satisfactory results, providing abundant [M + NH4]⁺ ions, not only from both primary and secondary nitro compounds, but also from the much more labile tertiary nitro compounds. However, the use of methane and isobutane positive ion CI or EI conditions resulted in facile fragmentation with little relative molecular mass information being made available. Negative ion CI using methane, isobutane or ammonia as moderating gases all gave abundant [M ? 1]^? ions with primary and secondary nitro compounds but at much reduced sensitivity.     

Assessment of metastable atom bombardment (MAB) ionization mass spectrometry for the fast determination of heterocyclic aromatic amines in cooked meat

An investigation of metastable atom bombardment (MAB) ionization mass spectrometry for the fast characterization of mutagenic/carcinogenic heterocyclic aromatic amines (HAAs) formed during heating processes of meats is presented. The aim of our study was to use the selective ionization of MAB to develop a detection method for HAAs in non-purified meat extracts, thus avoiding purification and concentration steps and reducing analysis time. Sample introduction into the MAB ion source was achieved by pyrolysis, allowing the direct and fast insertion of complex food extracts into the mass spectrometer. Analysis conditions were optimized on standard HAAs by using different ionization gases for the MAB process. Metastable nitrogen was selected as the best MAB gas for the analysis of HAAs. Ionization selectivity is shown by the detection of heterocyclic amines in non-purified chicken meat extracts spiked with HAAs. A quantitative approach is also presented by using pyrograms as chromatograms for quantification purposes. HAAs determination using Py-MAB-ToF was finally performed on cooked chicken breast extracts and compared to an LC-APCI-MS/MS method. Although Py-MAB-ToF sensitivity remains to be improved in the present state of development of our prototype device, only 2 h from the cooking were required to obtain quantitative results in good agreement with HAAs concentrations measured by LC-MS/MS in 36 h. Figure Experimental set-up for pyrolysis-MAB-ToF mass spectrometry experiments.     

Electron impact mass spectral interpretation for some thiophosphoryl-p-acetylaminobenzenesulfonamides

This work discusses the synthesis and the fragmentation patterns for 2-(p-acetylaminosulfonamido)-2-thiono-(5,5-dimethyl-1,3,2-dioxaphosphorinane)(1) and for the p-acetylaminosulfonylamides of O,O-diethylthiophosphoric acid (2), O,O-diphenylthiophosphoric acid (3), dimethylaminocyclohexylthiophosphoric acid (4), and diethylaminophenylthiophosphoric acid (5). A thionamidic-thiolimidic structure was attributed to compounds 1-5, consistent with their IR and NMR spectra. EI mass spectra at 70 eV, high resolution (HR) mass measurements and metastable ion spectra were used to elucidate the fragmentation processes and to determine the kinetic energy release values associated with the metastable ion dissociations. HR accurate mass measurements were used to confirm the compositions of the more abundant ions.     

Comparative studies on photo-ionization and electron-impact ionization of peptide derivatives

Photo-ionization (PI) mass spectrometry performed with a monochromatic photon beam was applied to a series of peptide derivatives. PI mass spectra of ten N-acylpeptide methyl esters containing two to four residues of glycine, alanine, valine, leucine, proline, tryptophan, tyrosine, phenylalanine, methionine, carboxymethylcysteine, lysine and ornithine were studied. Comparative analysis of PI (10.2 eV) and electron-impact ionization (EI) (70 eV) mass spectra shows the total number of peaks on PI to be much less than that obtained with EI, especially in the low m/e region (< 250 to 300). At the same time the relative abundance of ‘heavy’ ions, including molecular ions, is much higher in PI. The amino acid fragmentation pattern followed by N-acylpeptide esters in PI was found to be the same as for EI.     

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。     

Supersonic gas chromatography/mass spectrometry

A new gas chromatography/mass spectrometry (GC/MS) system was designed and evaluated which we have named 'Supersonic GC/MS'. It is based on a modification of a commercially available GC/MS system to include a supersonic molecular beam (SMB) MS interface. In this system the standard electron ionization (EI) ion source was replaced with a fly-through EI ion source mounted in the path of the SMB. A hyperthermal surface ionization (HSI) ion source combined with a 90 degrees ion mirror (for the EI-produced ions) was also added, and placed inside the quadrupole mass analyzer in place of its original EI ion source. The 'Supersonic GC/MS' system requires 18 cm added bench space plus the addition of an air-cooled 60 L/s diffusion pump and a 537 L/min rotary pump. The system is user friendly since all the gas flow rates, heated zones, sampling and data analysis are performed the same way as the original system and are computer-controlled via the original software. Similar EI sensitivity was obtained as with the original system for hexachlorobenzene and octafluoronaphthalene, while improved EI detection limits were demonstrated for methyl stearate and eicosane due to the significant enhancement of their molecular ion abundances. A GC/MS detection limit of 500 ag for pyrene was demonstrated using HSI. Good supersonic expansion cooling was achieved with large alkanes, despite the use of a rotary pump at the nozzle chamber instead of a diffusion pump. High temperature GC/MS analysis was demonstrated for large polycyclic aromatic hydrocarbons (PAHs) including ovalene and decacyclene (ten rings). Library searches with EI mass spectra are demonstrated, and it is explained why the enhancement of the molecular ion actually improves the library search in most cases. The analysis of large phthalate esters is also described, and the improvement obtained is shown to originate from their enhanced molecular and high mass fragment ions.     

Atmospheric-pressure Penning ionization of aliphatic hydrocarbons

A study has been made of the atmospheric-pressure Penning ionization (APPeI) of aliphatic hydrocarbons (pentane, hexane, heptane, and octane) with long-lived rare gas atoms (Rg*). The metastable rare gas atoms (He*, Ne*, Ar* and Kr*) were generated by the negative-mode corona discharge of atmospheric-pressure rare gases. In the Rg*APPeI mass spectra for aliphatic hyrocarbons, the relative abundances of fragment ions were found to increase in the order of He* --> Ne* --> Ar* --> Kr*. The order is in the opposite direction to the internal energies of the Rg*. The less fragmentation observed for He* may be because the nascent molecular ions [M(+.)]* formed by Penning ionization have lifetimes long enough for them to be collisionally deactivated in the atmospheric-pressure ion source. It was found that the relative abundances of fragment ions in Ar*APPeI increased when the sample pressure in the ion source was reduced. This is attributed to the collision of Ar* with molecular ions followed by fragmentation.     

Molecular dynamics simulation of the electron ionization mass spectrum of tabun

Tabun (ethyl N,N‐dimethylphosphoramidocyanidate), or GA, is a chemical warfare nerve agent produced during the World War II. The synthesis of its analogs is rather simple; thus, it is a significant threat. Furthermore, experiments with tabun and other nerve agents are greatly limited by the involved life risks and the severe restrictions imposed by the Chemical Weapons Convention. For these reasons, accurate theoretical assignment of fragmentation pathways can be especially important. In this work, we employ the Quantum Chemistry Electron Ionization Mass Spectra method, which combines molecular dynamics, quantum chemistry methods, and stochastic approaches, to accurately investigate the electron ionization/mass spectrometry (EI/MS) fragmentation spectrum and pathways of the tabun molecule. We found that different rearrangement reactions occur including a McLafferty involving the nitrile group. An essential and characteristic pathway for identification of tabun and analogs, a two‐step fragmentation producing the m/z 70 ion, was confirmed. The present results will be also useful to predict EI/MS spectrum and fragmentation pathways of other members of the tabun family, namely, the O‐alkyl/cycloalkyl N,N‐dialkyl (methyl, ethyl, isopropyl, or propyl) phosphoramidocyanidates.     

Differentiation of 2-alkylthioorotic acids, methyl and ethyl 2-alkylthioorotates and hydrazides of 2-alkylthioorotic acids by using electron ionization mass spectra

Electron ionization mass spectra of 12 derivatives of 2-thioorotic acid have been discussed and general fragmentation routes of their molecular ions have been proposed. The compounds under discussion were three groups of four chemical species (2-alkylthioorotic acid, methyl 2-alkylthioorotate, ethyl 2-alkylthioorotate and 2- alkylthioorotic hydrazide) each with the same relative molecular mass. The comparison of selected ions relative abundances and their correlation with the abundance of molecular ions enable differentiation between isomeric or isobaric species in this class of compounds.     

Electron and chemical ionization mass spectrometry in stereochemical differentiation of some 1,3-amino alcohols

Mass spectral fragmentations of two cyclopentane, eight cyclohexane and four norbornane/one 1,3-amino alcohols were studied under electron ionization (EI) by low-resolution, high-resolution, metastable ion analysis and collision-induced dissociation (CID) techniques. All stereoisomeric compounds gave rise to identical 70 eV EI mass spectra. However, the spectra of positional isomers clearly differed. The main fragmentation pathway for the saturated compounds began as an α-cleavage reaction with respect to the nitrogen atom. For the norbornene compounds a retro-Diels—Alder reaction was favoured. Relative to the aminomethyl-substituted compounds the fragmentation patterns for the compounds having the amino group connected directly to the ring were more complicated. The chemical ionization (CI) mass spectra were recorded using ammonia, isobutane, methane, dichloromethane and acetone as reagent gas. From the norbornane/One compounds the di-exo isomers decomposed more easily than the di-endo isomers with most of the reagent gases used. Differences between stereoisomers were observed directly only under methane CI. The decomposition products of the [M + H]⁺ ions generated under ammonia and isobutane CI were studies by recording their CID mass spectra. These spectra allowed the differentiation of the stereoisomers, at least to some extent.     

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.     

Chromatographic and mass spectral studies of perfluorooctanesulfonate and three perfluorooctanesulfonamides

The chromatographic and mass spectral characteristics of perfluorooctanesulfonate (PFOS) and three nitrogen-substituted perfluorooctanesulfonamides have been obtained. A methyl/phenyl mixed-phase fused-silica capillary column was used for gas chromatographic (GC) analyses, while a C18 reversed-phase microbore column was used for liquid chromatographic (LC) analyses. Mass (MS) and tandem mass (MS/MS) spectra were generated using electron ionization (EI), argon CE, methane positive and negative ion CI, and ES ionization modes. EI spectra of the amides showed ions characteristic of both the fluorinated hydrocarbon and the sulfonamide portion of the molecules. The fragmentation pathway was studied using hydrogen/deuterium exchange, and was thought to involve a cyclic intermediate ion. Formation of molecular ions by CE and protonated molecule ions by CI to obtain molecular weight information was only partially successful. Negative ion ES-MS spectra provided intense [M-H]- anions for the amides, and an [M-K]- anion for PFOS from which molecular weight information could be obtained, while ES-MS/MS produced product ions that could be used to detect the presence of these compounds in biological or environmental samples.