A study on the solution and gas-phase chemistry of Mn(III) and Fe(III) tetraarylporphyrin complexes by fast-atom bombardment mass spectrometry

Fast-atom bombardment mass spectrometry has been used to investigate the chemical behavior of Fe(III) and Mn(III) tetraarylporphyrins (TAP) in both the condensed and gas phases and to clarify the mechanisms responsible for the production of positive and negative ions. The differences in the behavior of Fe(III) and Mn(III) complexes in the positive ion mode could be correlated with those in their electronic structures and knowledge of the mechanism for the generation of negatively charged species was applied to characterize the counterion coordinated to the Mn(III)-TAP. Thus, the unprecedented, complete characterization of even complex Mn(III)-TAP was made possible.     

Structural study of Mn(III)-tetraarylporphyrin complexes by fast atom bombardment mass spectrometry

A series of 27 Mn(III)-tetraarylporphyrins bearing heterogeneous substituents on the phenyl rings at the meso positions were subjected to positive ion fast atom bombardment mass spectrometry The source spectra yielded the molecular ion and a few peaks confirmative of the chemical structure. Major processes were hydrogenolytic loss of the aryl rings or of their substituents. Molecules carrying a side-chain underwent loss of the chain either as a solution process (amide linkage) or as a gas-phase process (phenolic ether linkage). Collisionally activated dissociation mass-analysed ion kinetic energy spectroscopy allowed discrimination between the two types of fragmentation processes and some other previously unreported gas-phase reaction channels to be recognized.     

Redox processes of ruthenium (II) polypyridine complexes induced by fast-atom bombardment mass spectrometry

Fast-atom bombardment (FAB) mass spectrometry in the negative ionization mode enables the sputtering into the gas phase of the ruthenium complexes [Ru(2,2′-bipyridine[bpy])₂(2,5-bis) (pyrydil)pyrazine[dpp])](PF₆)₂; [Ru(bpy)₂,(2,3dpp)](PF₆)₂;[Ru(bpy)₂,(2,3-dpp-Me)]( PF₆)₃; and [Ru(bpy)₂(?-2,3-dpp)]₂ RuCl₂(PF₆)₄ as intact radical anions. These data, combined with those avaiiable from the positive FAB spectra allow a full characterization of the analytes.     

Quantitative analysis of the molecular species of monosialogangliosides by continuous-flow fast-atom bombardment mass spectrometry.

Negative-ion continuous-flow fast-atom bombardment mass spectrometry was evaluated as a means for the quantitative analysis of N-acetylneuraminyl-galactosyl-glucosyl-ceramide (NeuAc-GM3) and N-acetylgalactosaminyl-(N-acetylneuraminyl)galactosyl-glucosyl-ceramide (NeuAc-GM2). This study was carried out on a 7070-EQ mass spectrometer (VG Analytical, Manchester, UK) using a home-made continuous-flow fast-atom bombardment probe with a mixture of methanol + water + triethanolamine (70:27:3, v/v/v) as the mobile phase. Utilizing 100 ng of acetyl-lysogalactosyl-N-acetylgalactosaminyl-(N-acetylneuraminyl)g alactosyl-glucosyl-ceramide (acetyl-lysoGM1) as an internal standard, standard curves for NeuAc-GM3 d18:1-16:0, NeuAc-GM3 d18:1-18:0 and Neuac-GM2 d18:1-18:0 were found to be linear over the range 5-250 ng, with associated correlation coefficients of 0.990-0.997. The lower limit of detection was found to be 2.5 ng. Satisfactory results could also be obtained when the calibration curves were derived from the deprotonated molecular ions of a mixture of the NeuAc-GM2 and NeuAc-GM3 classes. Using this approach, quantitative determination of NeuAc-GM3 d18:1-16:0 from rat adrenal gland was performed using N-acetylneuraminic acid assay as a test control. We found 278 +/- 36 ng of this species in 1 mg of tissue (three replicate experiments). The procedure represents a sensitive method for the quantitation of monosialogangliosides and its capability to give molecular species information.     

Structural characterization of steroidal saponins by electrospray ionization and fast-atom bombardment tandem mass spectrometry

The structural characterization of four steroidal saponin compounds involving two and three sugar groups, namely spirostanol saponins and furostanol saponins, were investigated by positive ion fast-atom bombardment (FAB), electrospray ionization mass spectrometry (ESI-MS) and tandem mass spectrometry (MS/MS) techniques. Important structural information was obtained from collision-induced dissociation (CID) and FAB-MS spectra with different liquid matrices. It was found that a characteristic fragmentation involving the loss of 144 Da arising from the cleavage of the E-ring was observed when there was no sugar chain at the C-26 position. When a glucoside group was substituted at the C-26 position, this C-26 sugar moiety was preferentially eliminated. All of these compounds produced a major product ion with a stable skeleton structure at m/z 255. The results of this paper can assist structural analysis of mixtures of steroidal saponins.     

Electron ionisation and fast-atom bombardment mass spectrometry study of diaryl carbonates

Electron ionisation mass spectrometry studies were performed previously for p-diphenyl carbonate and some monosubstituted diphenyl carbonates. In this work, p-diphenyl carbonate and p-methoxyphenylphenyl carbonate are re-examined, and p-chlorophenyl phenyl carbonate and two disubstituted diphenyl carbonates, bis (p-chlorophenyl) carbonate and p-methoxyphenyl-p-fluorophenyl carbonate, are studied for the first time. The previously established fragmentation routes were observed for all compounds investigated. Some other different sequences were observed, and a fragmentation path, other than decarboxylation, of the molecular ion is proposed. In the fast-atom bombardment study it was observed that the M(+*)/[MH](+) ion abundance ratio increased from 0.44 for compound 1 to 2.95 for compound 5. [MH](+) is not a dominant ion in most of compounds studied, in spite of the presence of a carbonyl group, a strong proton acceptor. The presence of two oxygen atoms bonded to the carbonyl group appears to induce delocalisation of the electron pairs, thus deactivating the carbonyl site for protonation. In addition, m-nitrobenzyl alcohol (NBA) being a relatively aprotic/hydrophobic matrix reinforces the deactivation for protonation. Because the carbonate group and NBA are common features to the study, the contributions of the substituents were taken into account to explain the different behaviour of the five compounds with respect to protonation.     

Analysis of saxitoxin in urine by continuous-flow fast-atom bombardment mass spectrometry.

An improved method of saxitoxin analysis in urine using continuous-flow fast-atom bombardment mass spectrometry was developed. Parameters studied were matrix composition, matrix flow, temperature of probe tip, probe-tip design and sample extraction. Optimal detection was obtained using the following matrix composition: 5% glycerol, 0.5% acetic acid, 0.025% sodium dodecylsulfate, 0.1% polyethylene glycol (PEG) 400 and 0.5% PEG 300; probe-tip temperature: (approximately 55 degrees C); flow rate: 5 or 8 microL per min.; probe tip: Olson-Hogge design. The STX standard was detected at 200 pg with signal-to-noise ratio of 11. The percent recovery of saxitoxin from human urine after clean-up on a weak cation exchange column was 75%.     

A coordinated approach towards the molecular weight determination of peptides by gel electrophoresis and fast-atom bombardment mass spectrometry

The molecular weight of peptides can be determined by fast-atom bombardment mass spectrometry (FAB-MS) following polyacrylamide gel electrophoresis (PAGE). Initial results combining the fast and efficient separation of peptides by PAGE and the unambiguous determination of molecular weights by FAB-MS have been demonstrated for the three peptides bradykinin, neurotensin and gramicidin S. This method has also been applied to the determination of the molecular weights of two fragments from the tryptic digest of horse-heart cytochrome c.     

Analysis of bovine beta-casein tryptic digest by continuous-flow fast-atom bombardment mass spectrometry.

Liquid chromatography/fast-atom bombardment mass spectrometry was used to partially confirm the amino-acid sequence of the protein, beta-casein. The study demonstrates that the technique is capable of the rapid and accurate identification of peptide fragments from tryptic digests and that chromatographic integrity is maintained during the analysis. The power of the technique derives from the ability to determine both the retention time and the molecular weight of the eluting components. Each of the components yields a prominent pseudo-molecular ion (MH+), the majority exhibiting sufficient fragmentation to confirm their structure unambiguously.     

Analysis of arachidonic-acid-containing molecular species in glycerophospholipid classes from rat kidney by fast-atom bombardment mass spectrometry

The qualitative analysis of the molecular species of phosphatidylinositol, phosphatidylethanolamine, phosphatidylcholine and phosphatidylserine from a rat kidney, by fast-atom bombardment mass spectrometry after separation and purification by liquid chromatography is described. The present results indicate that a high content of arachidonic-acid-containing molecular species within major glycerophospholipid classes is present in this biological sample.     

The structure characterization of dinucleoside and nucleoside glucopyranosides lipophilic phosphotriesters by fast-atom bombardment tandem mass spectrometry

Rules for the gas-phase fragmentation mechanism of the negative ions of lipophilic phosphotriester molecules of biological interest have been established by fast-atom bombardment mass spectrometry/mass spectrometry. The mass-analyzed ion kinetic energy spectra of the [M ? H]^? of dinucleoside (1–4) and nucleoside glucopyranoside (5–9) phosphotriesters show that in the absence of charges on the phosphate bridge, the availability of acidic protons on the 5′-end nucleobase drives a preferred reaction path which leads to 5′-O-nucleotide or 6-O-glucopyranoside monophosphate anions.     

Direct analysis of drugs by continuous-flow fast-atom bombardment and tandem mass spectrometry

The techniques of continuous-flow fast-atom bombardment (CF-FAB) and tandem mass spectrometry (MS/MS) are combined and applied to the analysis of small molecular mass drugs (mol.wt less than 500 Da). The approach involves the interfacing of a CF-FAB inlet with a triple-stage quadrupole mass spectrometer, enabling the acquisition of collision-activated decomposition mass spectra of the drugs after FAB ionization. The relationship between a stable sample surface on the CF-FAB probe tip and the quality of the mass spectrum is discussed, as are practical methods for obtaining and maintaining surface stability. CF-FAB MS/MS spectra for several drugs are presented, including penicillin G, phentolamine, cocaine and benzoylecgonine. Minimum detection limits range from 50-500 pg injected, depending on the compound. The reproducibility of the integrated areas of peaks from repetitive injections is approximately five per cent. Data are also presented for the direct CF-FAB MS/MS analysis of cocaine and benzoylecgonine in spiked urine samples.     

A study of metal chelation of dinucleotide analogs in the gas phase by fast-atom bombardment mass spectrometry

Fast-atom bombardment (FAB) mass spectrometry was used to investigate the interaction of proton and alkali metal ions with dinucleotide analogs such as T-n-T (T = thymine moiety, n = polyether chain, e.g., triethylene, tetraethylene, pentaethylene, and hexaethylene ether 1–4), A-n-T (A = adenine unit 5–8), and T-n-OMe (9–12) in 3-nitrobenzyl alcohol matrix. The [M + H]⁺ ion is the most abundant ion for the A-n-T series, whereas in 1–4 and 9–12 the (TC₂H₄)⁺ ion is the most abundant. Formation of [M + H -C₂H₄O]⁺ ions, a characteristic fragmentation of crown ethers under electron ionization, is observed for compounds 1–12 and is more pronounced in 6 and 7. An abundant [M ? H]^? ion is observed for all the compounds studied under negative ion FAB due to the presence of the (-CO-NH-CO-) group of thymine, an indication of existence of intramolecular H bonding. The FAB mass spectra of 1–12 with alkali metal ions (Li⁺, Na⁺, K⁺, Rb⁺, and Cs⁺) showed formation of abundant metal-coordinated ions ([M + Met]⁺ and [TC₂H₄ + Met]⁺). Compounds 3, 4, 6, 7, and 10–12 showed ions due to the substitution of the thymine moiety by a hydroxyl group ([M + Met ? 108]⁺, Met = metal ion). For compound 3 alone, substitution of two thymine groups ([M + Met - 216]⁺) was observed. Metastable ion studies were used to elucidate the structures of these potentially significant ions, and the ion formule were confirmed with high resolution measurements. Selectivity toward metal complexation with ligand size was seen in the T-n-T and A-n-T series and was even more pronounced in A-n-T series. These dinucleotide analogs fall in the following order of chelation of alkali metal ions, acyclic glymes < dinucleotide analogs (acyclic glymes substituted with nitrogen bases) < crown ethers, which places them in perspective as receptor models.     

Electrospray ionization mass spectrometry of a cerium(III) phosphomolybdate complex: Condensed and gas-phase cluster chemistry

Electrospray ionization quadrupole ion trap mass spectrometry (ESI-QIT/MS) of the ammonium cerium(III) phosphomolybdate complex (NH₄)₁₁[Ce(III)(PMo₁₁O₃₉)₂] in aqueous media has revealed a concentration-dependent behavior. Under fixed instrumental parameters, the Ce-containing polyoxomolybdate complexes H₂Ce(III)P₂Mo₂₂O₇₅^3? and Ce(III)PMo₁₁O₃₈^2? are the primary species present at 11 mM (pH = 4.3); at 0.7 mM (pH = 3.6), Ce(III)PMo₁₀O₃₅^2? is the predominant species, Ce(III)PMo₁₁O₃₈^2? is quite diminished, and H₂Ce(III)P₂Mo₂₂O₇₅^3? is absent. As a result of the complex isotopic fingerprints from multiple molybdenums, compositions of such ions are difficult to assign—successive collision induced dissociation (CID) of large ions produced smaller ions for which calculated and experimental isotopic patterns could be compared. The oxidation state of Ce and the number of counter cations on negative complexes was discerned from spectra of ions containing ¹H⁺ and ⁷Li⁺. The overall result is an ESI method applicable to phosphomolybdate complexes containing redox sensitive f-block metal ions such as Ce(IV) and Pu(III/IV). Dissociation studies also gave insight into favored fragmentation pathways, and generated gas ions with empirical formulae similar to known condensed-phase ions. Deconvolution of concentration- and pH-dependent solution behavior via ESI/MS and ³¹P NMR spectroscopy showed speciation dependent on solution concentration, not on pH.     

Direct evidence for degradation of esperamicin A1 with thiol confirmed by fast-atom bombardment mass spectrometry.

A degradation of esperamicin A1, which is a potent antitumor antibiotic having DNA-cleaving activity, was studied by fast-atom bombardment (FAB) mass spectrometry. It was proved that the degradation easily occurs via the thiol group(s) in the matrix solution and not by a FAB-induced reaction. It was concluded that from the FAB mass spectra of all non-volatile compounds, the molecular weight should be determined by using at least two chemically different matrices.     

Matrix effects in fast atom bombardment mass spectrometry of cationic iridium(III) and rhodium(III) coordination complexes

Fast atom bombardment mass spectra of cationic iridium(III) and rhodium(III) coordination complexes (M⁺Cl₂L₂, X^?; where the ligand L is a dinitrogenous aromatic system) have been obtained with thioglycerol, glycerol or tetraglyme as a matrix. Two kinds of reactions, initiated by particle bombardment, have been discovered between these complexes and the matrix. First, with thioglycerol one or two chlorine atoms are substituted by a thioglycerol radical, more rapidly for rhodium compounds; secondly, when the ligand L possesses a diazo function, this function is hydrogenated depending on the ability of the matrix to generate hydrogen radicals by bombardment.     

Analysis of human pituitary growth hormone and its charge variants by fast-atom bombardment mass spectrometry.

There is evidence that even highly purified preparations of human growth hormone are not homogenous, but contain charge as well as size variants. The charge heterogeneity was suggested to be due to deamidation of the native hormone. To verify this we have applied peptide mapping followed by fast-atom bombardment mass spectrometry (FAB-MS), in order to identify fragments containing the altered amino acids. Growth hormone was purified from human pituitaries and the differently charged forms were separated by column electrophoresis in agarose suspension. The isolated components were treated with trypsin and analysed directly by FAB-MS without prior separation by reversed-phase high-performance liquid chromatography (RP-HPLC). Using this technique, approximately 80% of the hormone structure was recovered and two deamidation sites were found in the fragment T15 (FDTNSHNDDALLK). The results clearly elucidated the potential use of FAB-MS for the fast screening of other variants of the growth hormone which are known to exist.