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.     

Differentiation of diastereomeric N-aryltetrahydropyrano/tetrahydrofuranochromenylamines under electron ionization and chemical ionization conditions

A series of diastereomeric 4S,5S,6R/S-tetrahydropyrano- and 3S,4S,5R/S-tetrahydrofuranochromenylamine derivatives (a/b isomers; 1-26) has been studied under electron ionization (EI) and chemical ionization (CI) conditions. The EI mass spectra of all diastereomeric compounds show two characteristic fragment ions, of which one is formed by retro-Diels-Alder (RDA) reaction from the molecular ion, retaining the charge on the diene fragment, and the other [M-(HNAr)]+ ion by a simple radical loss. The RDA process is more favorable in all b isomers, whereas the radical loss is dominant in all a isomers; based on these two ions it is easy to differentiate the two diastereomers. The collision-induced dissociation (CID) spectra of all the molecular ions also show the same trend, which reflects the stereoselectivity in the formation of the two characteristic fragment ions. The results of theoretical calculations performed are in accordance with the experimental observations. The CI experiments (methane and isobutane) on all the diastereomeric compounds also enabled the differentiation of the isomers.     

Mass spectral study of Boc-carbo-beta3-peptides: differentiation of two pairs of positional and diastereomeric isomers

A mass spectral study of a series of new Boc-C-linked carbo-beta(3)-peptides prepared from C-linked carbo-beta(3)-amino acids (Caa) was carried out using liquid secondary ion mass spectrometry (LSIMS), electrospray ionization (ESI) and tandem mass spectrometry. Using the nomenclature of Roepstorff and Fohlman, the positive ion high- and low energy collision-induced dissociation (CID) of [M + H - Boc + H](+) ions of the peptides produce both N- and C-terminus ions, y(n) (+) and b(n) (+) ions, with high abundance and other ions of low abundance. Further, characteristic fragment ions of carbohydrate moiety are observed. In contrast to the CID of protonated peptide acids, the CID of [M - H](-) ions of the beta(3)-peptide acids do not give b(n)(-) ions and show abundant z(n)(-) and c(n) (-) ions which are insignificant in the former. Two pairs of positionally isomeric Boc-carbo-beta(3)-dipeptides were differentiated by the CID of [M + H](+) ions in LSIMS and ESIMS. The fragment ion [M + H - C(CH(3))(3) + H](+) formed from [M + H](+) by the loss of 2-methylprop-2-ene is relatively more abundant in the dipeptide Boc-NH-beta-hGly-Caa(S)-OCH(3) (14) containing the sugar moiety at the C-terminus whereas it is insignificant in Boc-NH-Caa(S)-beta-hGly-OCH(3) (13), which has the sugar moiety at the N-terminus. Similarly, two pairs of diastereomeric dipeptides were distinguished by the high- and low-energy CID of [M + H](+) ions. The loss of 2-methylprop-2-ene is more pronounced for Boc-NH-Caa(R)-beta-hGly-OCH(3) (17) and Boc-NH-Caa(R)-Caa(S)-OCH(3) (18) isomers whereas it is insignificant for Boc-NH-Caa(S)-beta-hGly-OCH(3) (13) and Boc-NH-Caa(S)-Caa(S)-OCH(3) (2) isomers. This was attributed to a favorable configuration of the carbohydrate moiety favoring the 'H' migration involved in the loss of 2-methylprop-2-ene from the [M + H](+) ions of isomers 17 and 18 compared with the unfavorable configuration of the carbohydrate moiety in isomers 13 and 2.     

Evaluating electron ionization mass spectral library search results

As the size of various collections of electron ionization mass spectra gets larger, there is a continuing and increasing propensity to rely on the results of computerized library searches. The results of these computerized searches do not necessarily account for a spectrum that is produced by the mixture of two or more different compounds. Sometimes the submitted spectrum is not that of a compound whose spectrum is in the library. The quality of the spectrum submitted to the library is often such that the numerical confidence level reported for the search result is so low that it will cause the result to be disregarded. When the sample spectrum is matched against library spectra that have been condensed, the search result can be misleading. Three different examples of mass spectral search results are examined: one, with a high confidence level that the unknown has been identified, but the results are incorrect; one, where the spectrum of the unknown compound is not in the library; one, where consideration of the numerical search results would cause a positive identity not to be confirmed.     

Study of two diastereomeric pairs of regiosomeric 2-isoxazolines by tandem mass spectrometry with electron impact ionization

On electron impact (EI) ionization, two cis/trans pairs of 4-methyl-5-phenyl and 4-phenyl-5-methyl regioisomeric 3-carbethoxy-2-isoxazolines showed normal mass spectra and mass-analysed ion kinetic energy (MIKE) spectra of metastable (MI) and collision-activated (CA) molecule ions, allowing unequivocal differentiation of the regioisomers. The cis/trans stereoisomers of each regioisomer showed very similar normal mass spectra. Very interestingly, the cis- and trans-4-phenyl-5-methyl stereoisomers appeared reasonably differentiated by the molecule ion MIKE spectra, whereas the 4-methyl-5-phenyl regioisomeric pair of stereoisomers did not. The influence of the phenyl substituent to the fragmentation processes was notable. Some fragments of interest were studied by comparison of their MIKE spectra with those of model ions, generated by EI from suitable substrates, including (i) the isomeric α,β-unsaturated oxime, namely ethyl (Z)-2-(hydroxymino)-3-methyl-4-phenylbut-3-enoate, a by-product of importance for the mechanism(s) of the addition/cycloaddition reactions of nitrile oxides to alkenes and (ii) trans-β-methylstyrene, a dipolarophilic reactant in the same reactions. The favoured heterocyclic C(5)–O(1) bond cleavage occurred only for the ionized 4-methyl-5-phenyl 2-isoxazoline pair, leading to a distonic ion of relevance, as it can represent either a reasonable precursor for both the isomerization to the ionized α,β-unsaturated oxime and the EI-induced cycloreversion yielding ionized β-methylstyrene, or the ionized form of a zwitterionic intermediate, which had been proposed previously for the addition/cycloaddition mechanism(s) in the solution phase, currently under study.     

Mass spectral fingerprints of detergents in gasolines using electrospray ionization

Mass spectral fingerprints of detergents in Austrian, Hungarian, Uruguayan, and Chilean gasolines have been obtained using electrospray ionization mass spectrometry. Polymers or copolymers were observed based on ether motifs (inter-peak spacings of 44, 58 and 72 u) in all samples. Austrian gasoline was found to also contain polymers based on isobutene-amine.     

Mass spectrometric characterization of different norandrosterone derivatives by low-cost mass spectrometric detectors using electron ionization and chemical ionization

The abuse of nortestosterone in sport is an important problem in doping-control analysis. In order to detect the main urinary metabolite of nortestosterone, norandrosterone (NA), sensitive and specific methodology is necessary. In this context the use of a low-cost mass spectrometric detector such as the Finnigan MAT ion-trap detector (ITD) was studied. The electron ionization (EI) and positive-ion chemical ionization (PICI) mass spectra of the methoxime-trimethylsilyl, trimethylsilyl-enol trimethylsilyl ether and pentafluoropropionic ester derivatives of NA are described. The limits of detection of these derivatives are compared with those obtained by the Hewlett-Packard mass selective detector (MSD), another low-cost mass spectrometric detector and operating only in the EI mode. For the derivatives of the reference standard of NA the ITD has in the EI mode the same limit of detection, in the range of 0.5 to 1 ng injected on the column, as the MSD. However, under these conditions the ITD provides more spectrometric information, because it gives full scan data. Moreover, with the same or even improved limits of detection the ITD can operate in the PICI mode. On the other hand, for the analysis of NA isolated from urine samples, the performance of the MSD was better than that of the ITD. The ion trapping technique is probably limited when the chemical background is high.     

Micro-synthesis and electron ionization mass spectral analysis of O-alkyl N,N-dialkylphosphoramidocyanidates

This communication describes microsynthesis and GC/MS analysis of O-alkyl N,N-dialkylphosphoramidocyanidates (ADAPCs), which are analogues of chemical warfare agent, Tabun. The study was undertaken with a view to develop spectral data base of ADAPCs for verification purpose of Chemical Weapons Convention (CWC). Reported microsynthetic approach has advantages over traditional synthesis in terms of efficiency, synthetic waste, and exposure to toxic chemicals. GC/MS analysis of variety of these compounds (ADAPCs) was performed. Based on the obtained mass spectra of structurally diverse ADAPCs, the fragmentation routes are proposed, which explains most of the characteristic ions.     

Mass spectrometric behaviour of simple oxazolidines under electron impact and chemical ionization

The 70 eV electron impact mass spectra of 33 differently substituted oxazolidines were studied to determine the effect of substituents and the existence of the ring—chain tautomeric equilibrium on the decomposition of the molecular ions. Most of the fragmentations can be rationalized to start as the cleavage initiated by the radical site at nitrogen. Isomeric compounds showed different spectra and were easily differentiated. The position of the ring—chain equilibrium could be located only roughly. The chemical ionization mass spectra of the compounds were also recorded, with ammonia, isobutane, acetone or methane as reagent gas. Methane was the only reagent gas that promoted extensive fragmentation of the protonated molecules. However, no information about the position of ring-chain tautomerism was obtained under these conditions. Analogously to other related five-membered heterocycles, the oxazolidines reacted under acetone chemical ionization conditions to afford [M + CH₃CO]⁺ adduct ions. These adducts were stable, however, and unlike those of 1,3-dioxolanes and 1,3-oxathiolanes, they did not decompose and form stable oxonium ions.     

(E)-4-alkoxycarbonylalkylthiochalcones: differentiation of isomeric derivatives by electron ionization mass spectrometry

The principal fragmentation pathways of the molecular ions of 18 new (E)-4-alkoxycarbonylalkylthiochalcones have been investigated. It has been shown that the data derived from electron ionization mass spectra (the relative abundance of the fragment ions and values of coefficients micro) can be used to differentiate the isomers. The fragmentation rules deduced here could help in the characterization of other chalcones of these types.     

Mass spectral analysis of N-oxides of Chemical Weapons Convention related aminoethanols under electrospray ionization conditions

N,N'-Dialkylaminoethanols are the hydrolyzed products or precursors of chemical warfare agents such as V-agents and nitrogen mustards, and they are prone to undergo oxidation in environmental matrices or during decontamination processes. Consequently, screening of the oxidized products of aminoethanols in aqueous samples is an important task in the verification of chemical weapons convention-related chemicals. Here we report the successful characterization of the N-oxides of N,N'-dialkylaminoethanols, alkyl diethanolamines, and triethanolamine using positive ion electrospray ionization mass spectrometry. The collision-induced dissociation (CID) spectra of the [M+H](+) and [M+Na](+) ions show diagnostic product ions that enable the unambiguous identification of the studied N-oxides, including those of isomeric compounds. The proposed fragmentation pathways are supported by high-resolution mass spectrometry data and product/precursor ion spectra. The CID spectra of [M+H](+) ions included [MH-CH(4)O(2)](+) as the key product ion, in addition to a distinctive alkene loss that allowed us to recognize the alkyl group attached to the nitrogen. The [M+Na](+) ions show characteristic product ions due to the loss of groups (R) attached to nitrogen either as a radical (R) or as a molecule [R+H or (R-H)] after hydrogen migration.     

Mass spectral characterization of tetracyclines by electrospray ionization,H/D exchange,and multiple stage mass spectrometry

Electrospray ionization (ESI) and collisionally induced dissociation (CID) mass spectra were obtained for five tetracyclines and the corresponding compounds in which the labile hydrogens were replaced by deuterium by either gas phase or liquid phase exchange. The number of labile hydrogens, x, could easily be determined from a comparison of ESI spectra obtained with N2 and with ND3 as the nebulizer gas. CID mass spectra were obtained for [M + H]+ and [M - H]- ions and the exchanged analogs, [M(Dx) + D]+ and [M(Dx) - D]- , and produced by ESI using a Sciex API-III(plus) and a Finnigan LCQ ion trap mass spectrometer. Compositions of product ions and mechanisms of decomposition were determined by comparison of the MS(N) spectra of the un-deuterated and deuterated species. Protonated tetracyclines dissociate initially by loss of H2O (D2O) and NH3 (ND3) if there is a tertiary OH at C-6. The loss of H2O (D2O) is the lower energy process. Tetracyclines without the tertiary OH at C-6 lose only NH3 (ND3) initially. MSN experiments showed easily understandable losses of HDO, HN(CH3)2, CH3 - N=CH2, and CO from fragment ions. The major fragment ions do not come from cleavage reactions of the species protonated at the most basic site. Deprotonated tetracyclines had similar CID spectra, with less fragmentation than those observed for the protonated tetracyclines. The lowest energy decomposition paths for the deprotonated tetracyclines are the competitive loss of NH3 (ND3) or HNCO (DNCO). Product ions appear to be formed by charge remote decompositions of species de-protonated at the C-10 phenol.     

Mass spectral hydrogen rearrangements of 1,1′-bicycloalkyldiols under electron impact

1,1′-Bicycloalkyl-2,2′-diols (mixtures of stereoisomers) and 1,1′-diols on electron impact give rise to strong peaks equivalent to [M-1]⁺ions, corresponding to cycloalkenone fragments and obviously involving hydrogen atom migrations. Deuterium labelling and substitution techniques reveal the operation of different rearrangement mechanisms for the two series of isomeric compounds. Thus, in the 1,1′-bicycloalkyl-2,2′-diols a ring hydrogen atom is involved in the rearrangement, whereas in the 1,1′-bicycloalkyl-1,1′-diols (pinacols) the hydroxylic hydrogen atoms migrate.     

Differentiation of the diastereomeric synthetic precursors of isofebrifugine and febrifugine by electrospray ionization tandem mass spectrometry

Febrifugine is an alkaloid with potent antimalarial activity isolated from Dichroa febrifuga and Hydrangea umbellate, and it exists naturally with its diastereomeric component, isofebrifugine. Here we report the differentiation of diastereomeric synthetic precursors of isofebrifugine (1, cis) and febrifugine (2, trans) and a structurally similar model diastereomeric pair without a halogen substituent (3 and 4) by electrospray ionization (ESI) tandem mass spectrometry. Compounds 1-4 contain a tert-butoxycarbonyl (BOC) substituent, and the collision-induced dissociation (CID) spectra of the [M+H](+), [M+Na](+) and [M+Li](+) ions of 1-4 include the expected product ions corresponding to the loss of C(4)H(8) (isobutene) and of C(5)H(8)O(2) (BOC-H). Loss of C(5)H(8)O(2) is dominant in cis isomers (1 and 3) and/or loss of C(4)H(8) ions is dominant in trans isomers (2 and 4). The decomposition of [M+H](+) ions shows stereoselectivity in the formation of the [M+H-(BOC-H)-C(3)H(5)OBr](+) and [M+H-(BOC-H)-C(6)H(5)CH(2)OH](+) ions. The [M+Cat](+) ions (where Cat = Na or Li) additionally show loss of NaBr and HBr from [M+Cat-(BOC-H)](+), and these product ions are constantly more abundant in cis isomers than in trans isomers. The stereoselectivity for the product ion corresponding to the loss of [(BOC-H)+C(3)H(5)OBr] from [M+H](+) ions differs from that from [M+Cat](+) ions.