Biosynthesis of Quinoline by a Stick Insect.

The Peruvian stick insect Oreophoetes peruana is the only known animal source for unsubstituted quinoline in nature. When disturbed, these insects discharge a defensive secretion containing quinoline. Analysis of samples obtained from l-[2',4',5',6,'7'-2H5]tryptophan-fed stick insects demonstrated that the insects convert it to [5,6,7,8-2H4]quinoline by removing the 2'-CH moiety in the indole ring of tryptophan. Analogous experiments using l-[1'-15N]tryptophan and l-[1'-15N,15NH2]tryptophan showed that the indole-N atom is retained while the α-amino group is eliminated during the biosynthesis. Mass spectra recorded from quinoline derived from [2-13C1]tryptophan-fed insects indicated that the α-carbon atom of tryptophan is incorporated as the C-2 atom of the quinoline ring.

Rapid, Selective, and Sensitive Method for Semitargeted Discovery of Congeneric Natural Products by Liquid Chromatography Tandem Mass Spectrometry.

Natural product congeners serve a useful role in the understanding of natural product biosynthesis and structure-activity relationships. A minor congener with superior activity, selectivity, and modifiable functional groups could serve as a more effective lead structure and replace even the original lead molecule that was used for medicinal chemistry modifications. Currently, no effective method exists to discover targeted congeners rapidly, specifically, and selectively from producing sources. Herein, a new method based on liquid-chromatography tandem-mass spectrometry combination is evaluated for targeted discovery of congeners of platensimycin and platencin from the extracts of Streptomyces platensis. By utilizing a precursor-ion searching protocol, tandem mass spectrometry not only confirmed the presence of known congeners but also provided unambiguous detection of many previously unknown congeners of platensimycin and platencin. This high-throughput and quantitative method can be rapidly and broadly applied for dereplication and congener discovery from a variety of producing sources, even when the targeted compounds are obscured by the presence of unrelated natural products.

LC-MS analysis of p-aminosalicylic acid under electrospray ionization conditions manifests a profound solvent effect.

Selected-ion recording (SIR) or multiple-reaction monitoring (MRM) protocols are widely employed for the quantification of targeted analytes by liquid chromatography-mass spectrometry (LC-MS). After chromatographic separation, analytes are desolvated and converted to gaseous ions usually by electrospray-ionization. The chromatographic peaks generated in this way are then integrated for quantification. It is generally assumed that the chromatographic peak intensities are dependent only on the selected MRM-transition protocols and the instrumental parameters set on the mass spectrometer. Using p-aminosalicylic acid (PAS) as a model compound, we demonstrate that the nature of the LC mobile phase exerts a significant effect on the chromatographic peak intensities. Under identical mass spectrometric conditions, chromatographic peak intensities recorded with methanol as the mobile phase were drastically different from those acquired using acetonitrile as the eluent. In fact, the product-ion mass spectra recorded with protonated PAS under different solvent conditions were qualitatively different. The observed differences were attributed to the existence of different protomers of PAS in the gas phase in dissimilar ratios under different solvent-spray conditions. Results from ion-mobility mass spectrometry experiments confirmed this hypothesis. For example, when PAS was sprayed from an acetonitrile solution, the arrival-time profile recorded from the mass-selected m/z 154 ion for protonated PAS showed essentially one arrival-time peak for the N-protonated tautomer. In contrast, the profile recorded from a methanolic PAS solution showed a different arrival-time peak for a more mobile protomer, which was recognized as the carbonyl-protonated PAS. The coexistence of protomers in different and variable ratios in an ensemble of ions generated by electrospray ionization of a single pure compound wields strong ramifications on the identification and quantification of analytes by LC-MS. However, the inclusion of an ion-mobility separator before the mass-selected ions are fragmented and detected by mass spectrometry ameliorates the complications rendered by the coexistence of different protomers and deprotomers.

Biosynthetic origin of benzoquinones in the explosive discharge of the bombardier beetle Brachinus elongatulus.

Bombardier beetles are well-known for their remarkable defensive mechanism. Their defensive apparatus consists of two compartments known as the reservoir and the reaction chamber. When challenged, muscles surrounding the reservoir contract sending chemical precursors into the reaction chamber where they mix with enzymes resulting in an explosive discharge of a hot noxious chemical spray containing two major quinones: 1,4-benzoquinone and 2-methyl-1,4-benzoquinone (toluquinone). Previously, it has been speculated that the biosynthesis of all benzoquinones originates from one core precursor, 1,4-hydroquinone. Careful ligation of the base of the reservoir chamber enabled us to prevent the explosive reaction and sample untransformed reservoir fluid, which showed that it accumulates significant quantities of 1,4-hydroquinone and 2-methyl-1,4-hydroquinone. We investigated the biosynthetic mechanisms leading to quinone formation by injecting or feeding Brachinus elongatulus beetles with stable-isotope-labeled precursors. Chemical analysis of defensive secretion samples obtained from 1,4-hydroquinone-d6-administered beetles demonstrated that it underwent conversion specifically to 1,4-benzoquinone. Analogously, results from m-cresol-d8 injected or fed beetles confirmed that m-cresol is metabolized to 2-methyl-1,4-hydroquinone, which is then oxidized to 2-methyl-1,4-benzoquinone in the hot spray. Our results refute the previous claim that 1,4-hydroquinone is the precursor of all substituted benzoquinones in bombardier beetles and reveal that they are biosynthetic products of two independent pathways. Most likely, the aforementioned biosynthetic channel of hydroxylation of appropriate phenolic precursors and subsequent oxidation is not restricted to bombardier beetles; it could well be a general pathway that leads to the formation of all congeners of benzoquinones, one of the most widely distributed groups of defensive compounds in arthropods. Graphical abstract.

HCN emission by a Polydesmid Millipede Detected Remotely by Reactive Adsorption on Gold Nanoparticles Followed by Laser Desorption/Ionization Mass Spectrometry (LDI-MS).

Hydrocyanic acid (HCN) is a well-known defensive allomone in the chemical arsenal of millipedes in the order Polydesmida. The presence of HCN in the headspace vapor of adult Xystocheir dissecta (Wood, 1867), a common millipede from the San Francisco Bay Area, was traced by laser desorption/ionization-mass spectrometry (LDI-MS). To accomplish this, the headspace vapor surrounding caged, live millipedes was allowed to diffuse passively over gold-nanoparticle (AuNP) deposits placed at various distances from the emitting source. The stainless steel plates with AuNP deposits were removed and irradiated by a 355-nm laser. The gaseous ions generated in this way were detected by time-of-flight mass spectrometry. The intensity of the mass spectrometric peak detected at m/z 249 for the Au(CN)2- complex anion was compared to that of the residual Au- signal (m/z 197). Using this procedure, HCN vapors produced by the live millipedes could be detected up to 50 cm away from the source. Furthermore, the addition of H2O2, as an internal oxygen source for the gold cyanidation reaction that takes place in the AuNP deposits, significantly increased the detection sensitivity. Using the modified H2O2 addition procedure, HCN could now be detected at 80 cm from the source. Moreover, we found a decreasing intensity ratio of the Au(CN)2-/Au- signals as the distance from the emitting source increased, following an exponential-decay distribution as predicted by Fick's law of diffusion. Graphical abstract.

Alkyl-Dimethylpyrazines in Mandibular Gland Secretions of Four Odontomachus Ant Species (Formicidae: Ponerinae).

Ponerine ants are known to contain mixtures of pyrazines in their mandibular glands. We analyzed the mandibular gland contents of four ponerine species (Odontomachus chelifer, O. erythrocephalus, O. ruginodis, and O. bauri) by gas chromatography coupled with mass spectrometry, and found that each species contains specific mixtures of trisubstituted alkylpyrazines among other volatiles. Attempts to identify alkylpyrazines solely by mass spectral interpretation is unrealistic because spectra of positional isomers are indistinguishable. To avoid misidentifications, we synthesized a large number of reference compounds and compared their mass spectral and gas chromatographic properties with those present in the Odontomachus species under investigation. Most of the compounds identified were 2-alkyl-3,5-dimethylpyrazines. Interestingly, when the third substituent was an isopentyl group, the two methyl groups were found to be located at the 2 and 5 ring positions. Using our data, we recognized several misidentifications in previous publications.

Effect of Electrospray Ionization Source Conditions on the Tautomer Distribution of Deprotonated p-Hydroxybenzoic Acid in the Gas Phase.

The deprotonation site of p-hydroxybenzoic acid upon electrospray ionization has been a subject of fervent debate in several articles in the Journal of the American Chemical Society and elsewhere. General consensus is that electrospray ionization mass spectrometry (ESI-MS) experimental results reflect the situation in solution to a considerable extent. Our research, using ion-mobility mass spectrometry, challenges the notion that ESI-MS results directly reflect solution-phase structures and demonstrates that the relative populations of the thermodynamically less favored gaseous carboxylate tautomer or the thermodynamically more favored gaseous phenoxide tautomer, generated from the same aqueous solution of p-hydroxybenzoic acid by ESI, can be varied back and forth by changing the probe position, capillary voltage, desolvation-gas temperature, sample infusion flow rate, and cone voltage. In other words, solvent effects are not the primary criteria that determine the relative population distributions of tautomeric carboxylate (C(-)) and phenoxide (P(-)) ions (m/z 137) generated by electrospray ionization of p-hydroxybenzoic acid. In addition, we propose that the observed ratio of the P(-) and C(-) forms indirectly reflects the relative contribution of the charge-residue or ion-evaporation process that occurs during the electrospray ion generation process.

Ambulation of Incipient Proton during Gas-Phase Dissociation of Protonated Alkyl Dihydrocinnamates.

Upon activation in the gas phase, protonated alkyl dihydrocinnamates undergo an alcohol loss. However, the mechanism followed is not a simple removal of an alkanol molecule after a protonation on the alkoxy group. The mass spectrum of the m/z 166 ion for deuteron-charged methyl dihydrocinnamate showed two peaks of 1:5 intensity ratio at m/z 133 and 134 to confirm that the incipient proton is mobile. The proton initially attached to the carbonyl group migrates to the ring and randomizes before a subsequent transfer of one of the ring protons to the alkoxy group for the concomitant alcohol elimination. Moreover, protonated methyl dihydrocinnamate undergoes more than one H/D exchange. The spectra recorded from m/z 167 and 168 ions obtained for di- and tri-deuterio isotopologues showed peak pairs at m/z 134, 135 and 135, 136, at 1:2 and 1:1 intensity ratios, respectively, confirming the benzenium ion intermediate achieves complete randomization before the proton transfer. Additionally, protonated higher esters of alkyl dihydrocinnamates undergo a cleavage of the O-CH2 bond to form an ion/neutral complex, which, upon activation, dissociates generating a carbenium ion and dihydrocinnamic acid, or rearranges to generate protonated dihydrocinnamic acid and an alkene by a nonspecific proton transfer.

Real-time monitoring of in situ gas-phase H/D exchange reactions of cations by atmospheric pressure helium plasma ionization mass spectrometry (HePI-MS).

An enclosed atmospheric-pressure helium-plasma ionization (HePI-MS) source avoids, or minimizes, undesired back-exchange reactions usually encountered during deuterium incorporation experiments under ambient-pressure open-source conditions. A simple adaptation of an ESI source provides an economical way of conducting gas phase hydrogen/deuterium (H/D) exchange reactions (HDX) in real time without the need for complicated hardware modifications. For example, the spectrum of [(2)H8]toluene recorded under exposed ambient conditions showed the base peak at m/z 96 due to fast leaching of ring hydrogens because of interactions with H2O vapor present in the open source. Such D/H exchanges are rapidly reversed if the deuterium-depleted [(2)H8]toluene is exposed to D2O vapor. In addition to the enumeration of labile protons, our procedure enables the identification of protonation sites in molecules unambiguously, by the number of H/D exchanges observed in real time. For example, molecules such as tetrahydrofuran and pyridine protonate at the heteroatom and consequently undergo only one H/D exchange, whereas ethylbenzene, which protonates at a ring position of the aromatic ring, undergoes six H/D exchanges. In addition, carbocations generated in situ by in-source fragmentation of precursor protonated species, such as benzyl alcohol, do not undergo any rapid H/D exchanges. Because radical cations, second-generation cations (ions formed by losing a small molecule from a precursor ion), or those formed by hydride abstraction do not undergo rapid H/D exchanges, our technique provides a way to distinguish these ions from protonated molecules.

Circumambulatory movement of negative charge ("ring walk") during gas-phase dissociation of 2,3,4-trimethoxybenzoate anion.

A dramatic "ortho effect" was observed during gas-phase dissociation of ortho-, meta-, and para-methoxybenzoate anions. Upon activation under mass spectrometric collisional activation conditions, anions generated from all three isomers undergo a CO2 loss. Of the m/z 107 ions generated in this way, only the 1-dehydro-2-methoxybenzene anion from the ortho isomer underwent an exclusive formaldehyde loss. A peak for a formaldehyde loss in the spectra of 2,4-, 2,5-, and 2,6-dimethoxybenzoates and the absence of an analogous peak from 3,4- and 3,5-dimethoxy derivatives confirmed that this is a diagnostically useful ortho-isomer-specific phenomenon. Moreover, the spectrum from 2,3-dimethoxybenzoic acid showed peaks for two consecutive formaldehyde losses. The 1-dehydro-2,3,4-trimethoxybenzene anion (m/z 167) generated from 2,3,4-trimethoxybenzoate in this way endures three consecutive eliminations of formaldehyde units. For this, the negative charge, initially located on position 1, circumambulates to position 2, then to position 3, and finally to position 4 to form the final phenyl anion. The proposed stepwise fragmentation pathway, which resembles the well-known E1cB-elimination mechanism, is supported by tandem mass spectrometric observations made with 2-[(13)C(2)H3]methoxy-3-[(13)C]methoxy-4-methoxybenzoic acid, and ab initio calculations. In addition, the spectra of ions such as 1-dehydro-3,4-dimethoxybenzene anion show peaks for consecutive methyl radical losses, a feature that establishes the 1,2-relationship between the two methoxy groups.

Determination of low levels of 2H-labeling using high-resolution mass spectrometry: application in studies of lipid flux and beyond.

RATIONALE: The ability to measure low levels of (2)H-labeling is important in studies of metabolic flux, e.g. one can estimate lipid synthesis by administering (2)H2O and then measuring the incorporation of (2)H into fatty acids. Unfortunately, the analyses are complicated by the presence of more abundant naturally occurring stable isotopes, e.g. (13)C. Conventional approaches rely on coupling gas chromatographic separation of lipids with either quadrupole-mass spectrometry (q-MS) and/or pyrolysis-isotope ratio mass spectrometry (IRMS). The former is limited by high background labeling (primarily from (13)C) whereas the latter is not suitable for routine high-throughput analyses. METHODS: We have contrasted the use of continuous flow-pyrolysis-IRMS against high-resolution mass spectrometry (i.e. Qq-FT-ICR MS) for measuring the (2)H-enrichment of fatty acids and peptides. RESULTS: In contrast to IRMS, which requires ~30 min per analysis, it is possible to measure the (2)H-enrichment of palmitate via direct infusion high-resolution mass spectrometry (HRMS) in ~3 min per sample. In addition, Qq-FT-ICR MS enabled measurements of the (2)H-enrichment of peptides (which is not possible using IRMS). CONCLUSIONS: High-resolution mass spectrometry can be used to measure low levels of (2)H-labeling so we expect that this approach will enhance studies of metabolic flux that rely on (2)H-labeled tracers, e.g. (2)H2O. However, since the high-resolution analyses require greater amounts of a given analyte one potential limitation centers on the overall sensitivity. Presumably, future advances can overcome this barrier.

Manipulating Non-Dissociative Transformations of Gaseous Ion Ensembles Prior to Ion-Mobility Separation.

Molecules with multiple sites capable of accepting protons form ensembles of protomers. The manifested protomer ratios in such ensembles are influenced by many experimental conditions. In a Synapt G2 ion mobility (IM)-enabled mass spectrometry system, there are several physical locations where ion population changes can be manifested. Using APCI-generated protomers of aminonaphthalenes, we investigated its intramolecular proton transfers from the N-protomer to the C-protomer. This lossless transformation of the N-protomer to the thermodynamically favored C-protomer can take place in the ion source itself. Initially, we learned that the cone gas slows down the transformation to the C-protomer. Gaseous ions are then accelerated in the first vacuum region, where ions undergo collisional activation (heating), which facilitates the transformation to the C-protomer. Afterward, the ions are mass selected and transferred to the pre-IM (Trap)-collision cell, where ions can also be transformed to the thermodynamically favored protomers. Trap accumulated ions are then released to the IM separator via a helium-filled entry cell. The role of helium is to minimize ion activation and scattering taking place upon entry to the high-pressure T-Wave IM separator (TWIMS). The helium cell is known to increase the IM peak resolution. However, we found that significant changes occur depending on the presence or absence of helium. Without helium, source-generated protomers rapidly changed to a predominantly thermodynamically favorable ensemble protomers. Apparently, the introduction of helium into the precell induced a dramatic decrease in collisional "heating" effect, which effectively slowed down the conversion rate of the amino-protomer into the more favorable ring-protomer. The final message is that mobilograms should not be considered as direct real-time, or intrinsic, representations of the protomer ratios in the ion source.

Direct detection of inorganic nitrate salts by ambient pressure helium-plasma ionization mass spectrometry.

Inorganic nitrates in solid deposits were detected directly by ambient-pressure helium-plasma ionization-mass spectrometry (HePI-MS), without the need for extensive sample preparation. Nitrates were detected even from complex matrices such as meats and fruit juices. Any electrospray-ionization mass spectrometer can be modified to perform ambient-pressure HePI-MS by simply passing helium through the metal capillary intended for liquid-sample delivery. Nitrates on paper strips, glass slides, or cotton swabs (sometimes wetted with a mineral acid) were inserted directly into the ambient-pressure HePI source. The spectra acquired under negative-ion generating conditions showed a peak at m/z 62 for the nitrate ion, along with a lower-intensity peak at m/z 125 for the nitrate adduct of nitric acid. Apparently, it is nitric acid that is initially transferred to the gas phase, forming an ion-molecule complex with hydroxyl anions present in the plasma. The ion-neutral complex then dissociates by eliminating water to produce gaseous NO(3)(-) ions. This hypothesis was supported by the observation that certain solid nitrate salts, which were not readily amenable to HePI (notably the alkali nitrates), were immediately detected as m/z 62 and 125 ions upon acidification by a strong acid. Quantitative evaluations showed that the nitrate-signal response versus the deposited mass is linear for over 3 orders of magnitude. With the use of (15)N-labeled nitrate (m/z 63), the limit of detection was determined to be as low as 200 fg.

Enhancement of laser desorption ionization mass spectrometric signals of cesium iodide by elemental sulfur.

RATIONALE: The utility of elemental sulfur as a matrix for inorganic salts such as CsI, AgI, and KI was investigated because the conventional matrices deployed to generate gaseous ions from organic compounds, upon irradiation with a laser beam, are not suitable for inorganic salts. METHODS: Sulfur and inorganic salts were admixed and irradiated with a 337-nm UV laser. Laser desorption ionization (LDI) mass spectra were recorded in both positive and negative ion mode on a time-of-flight mass spectrometer. RESULTS: The positive ion laser desorption ionization mass spectrum of CsI showed peaks at m/z 133, 393, etc. for [(CsI)(n)Cs](+) ions. Similarly, negative ion spectra showed peaks at m/z 387, 647, etc. for [(CsI)(n)I](-) ions. However, for n >2 ion clusters, the intensities of peaks were negligibly small in both ionization modes. In contrast, spectra recorded from CsI admixed with elemental sulfur showed peaks up to n = 13 for (CsI)(n)Cs(+), and n = 9 for (CsI)(n)I(-). A similar enhancement of ion abundances by sulfur was observed for the cluster ions generated from KI and AgI. CONCLUSIONS: The dramatic increase in intensities of the higher-mass CsI cluster peaks suggests that sulfur acts as a laser-absorbing matrix for inorganic salts far superior to conventional matrices such as 2,5-dihydroxybenzoic acid and α-cyano-4-hydroxycinnamic acid.

Impact of Ambient Vapors on Spectra of 4-Nitroaniline Recorded under Atmospheric Solids Analysis Probe (ASAP) Mass Spectrometric Conditions.

Thermally desorbed 4-nitroaniline (4-NA), upon atmospheric pressure chemical ionization (APCI), generates gaseous ions for its protonated species. The APCI mass spectrum recorded under mild in-source ion-activating conditions from 4-NA showed a peak at m/z 139, whereas that acquired under high ion-activating conditions showed two additional peaks at m/z 122 (•OH loss) and 92 (•NO loss). The spectrum changed instantaneously when acetonitrile vapor was introduced to the source. In the new spectrum, both m/z 122 and 92 peaks were absent, while a new peak appeared at m/z 93. Ion-mobility separation carried out with the m/z 139 ion revealed that the initial ion represented the thermodynamically favored nitro-protonated tautomer. The ion population changed to an ensemble dominated by the less-favored amino-protomer when acetonitrile vapor was introduced to the ion source. The amino-protomer, upon collisional activation, loses •NO2 to generate an m/z 93 ion, which was confirmed to be the 4-dehydroanilinium ion. Ion mobility provided a practical way to monitor the changes secured by acetonitrile vapor because the two protomers showed different arrival times. Under spray-ionization conditions, the formation of the thermodynamically less favored protomer has been attributed to kinetic trapping. Our study demonstrated that the less favored amino-protomer could be generated by introducing acetonitrile vapor under nonspray conditions. Apparently, under APCI conditions, protonated water vapor attaches to the nitro group to generate a proton-bound heterodimer, which upon activation dissociates to yield the nitro-protomer. In contrast, protonated acetonitrile makes a tighter complex preferentially with the amino group, which upon activation breaks to the amino-protomer.

Dependence of Collision-Induced Mass Spectra of Protonated Michler's Ketone on the Nature of LC-MS Mobile Phase.

Michler's ketone (MK) is a dimethylamino ketone that undergoes facile protonation under electrospray-ionization conditions to produce an ion of m/z 269. Initial LC-MS results showed that the collision-induced dissociation (CID) spectra of the m/z 269 ion depend heavily on the composition of the chromatographic mobile phase. Subsequent ion-mobility separation of the mass-selected m/z 269 ion revealed that protonated MK exists as two tautomeric forms. Moreover, the relative population of the two protomeric forms in the ion ensemble depends on the nature of the ambient molecules present in the atmospheric pressure ion source. For example, the ion-mobility arrival-time profile acquired from the mass-selected m/z 269 ion generated from an acetonitrile solution showed two peaks of near equal intensity. The peak with the shorter arrival time represented the O-protomer and that with the longer arrival time represented the N-protomer. However, when methanol or ammonia vapors were introduced to the ambient-pressure ion source, the intensity of the N-protomer peak decreased rapidly and that of the O-protomer signal soared until it became the dominant peak. When the introduction of methanol (or ammonia) vapors was stopped, the mobilogram signals gradually reverted back to their initial intensities. To rationalize this observation, we propose that the N-protomer of MK in the presence of methanol vapor undergoes transformation to the O-protomer by a Grotthuss-type mechanism via a methanol-based solvent bridge.

Monoisotopic Mass?

The current IUPAC-recommended definition of the term "monoisotopic mass" of a chemical species is based on the most abundant isotopes of the constituent elements. It has even been proposed to constrain the definition to be based only on the atomic masses of the most abundant stable isotopes. Such an approach is flawed because in this way several elements and their compounds, in addition to isotopically enriched species, would not merit to be assigned a monoisotopic mass. Furthermore, for large molecules, such as proteins, the monoisotopic mass as currently defined loses its significance. Therefore, we propose to eliminate using the current definition altogether. Instead, the term isotopologue mass should be applied uniformly to every species denoted by a specific chemical formula.

Fragmentation pathways of deprotonated ortho-hydroxybenzyl alcohol.

The ortho, meta, and para isomers of hydroxybenzyl alcohol can be unequivocally distinguished by the collision-induced dissociation mass spectra of their anions. The presence of a prominent peak at m/z 121 for an elimination of a dihydrogen molecule renders the ortho-isomer spectrum markedly different from those of its meta and para congeners. Investigations carried out with deuterium-labeled isotopologues of the ortho isomer verified that the labile hydrogen atom on the hydroxyl group and one of the benzylic hydrogen atoms are specifically removed in the formation of the m/z 121 ion. The ortho-isomer spectrum also showed a prominent peak at m/z 93. Experimental data indicated that the m/z 93 product ion originates either from a two-step H2 and CO elimination mechanism or from a direct loss of a HCHO molecule from the precursor anion. The intensity ratio of the m/z 93 and 94 peaks in the spectrum recorded from the m/z 124 ion generated from a sample of o-hydroxybenzyl alcohol dissolved in D2 O supported the notion that the direct HCHO loss is the more dominant pathway for the generation of the phenolate ion under low activation conditions. In contrast, the two-step mechanism becomes the more dominant pathway under high collisional activation conditions. The spectrum also showed a weak peak at m/z 105 for a water loss. Based on computational data, the m/z 105 ion generated in this way appears to be a composite generated from a common ion-neutral complex intermediate in which a hydroxyl anion is positioned equidistantly between one of the benzylic hydrogens and a nearby hydrogen atom of the benzene ring. Upon activation, the complex dissociates to form either a phenide or a quinone methide anion. The reaction forming a carbon dioxide adduct under ion-mobility conditions was used to support the proposed water-loss mechanism.

Evidence of Gas-Phase Attachment of Molecular Oxygen to Deprotonated Hydroquinone During Ion-Mobility Mass Spectrometry.

Gas-phase addition of dioxygen to certain ions is a well-known phenomenon in mass spectrometry. For this reaction to occur, the presence of a distonic radical site on the precursor ion is thought to be a prerequisite. Herein, we report that oxygen adduct formation can take place also with deprotonated hydroquinone, which in fact is an even-electron species without a radical site. When the product-ion spectrum of the m/z 109 ion, generated by electrospray ionization from a solution of hydroquinone in acetonitrile, was recorded under ion-mobility conditions, a new peak was observed at m/z 141. However, an analogous peak was not visible in the spectrum acquired under nonmobility conditions (i.e., without any gas introduced to the mobility cell). Presumably, traces of oxygen present in the collision gas instigate an ion-molecule reaction to produce an adduct of m/z 141, which upon activation results in CO and H2O loss to form a product ion of m/z 95. Isotope-labeling studies confirmed that one of the hydrogen atoms from the hydroxy group and another from the aromatic ring contribute to the water loss instigated from the m/z 141 adduct. Furthermore, computational methods indicated the three-dimensional structure of the ground-state deprotonated hydroquinone to be distinctly different from those of its 1,2- and 1,3-isomers. Calculations predicted that all atoms in the two m/z 109 ions generated from catechol and resorcinol lie on one plane. In contrast, the structure of the m/z 109 ion from hydroquinone was significantly different. Computations predicted that the hydrogen atom on the intact hydroxyl group of deprotonated hydroquinone protrudes out of plane from rest of the atoms. Consequently, the exposed OH group can interact with an incoming dioxygen molecule. Computations conducted at the CAM-B3LYP/6-311++g(2d,2p) level of theory detected a minimum energy crossing point (MECP) at -4.3 kJ mol-1 below the separated O2 + deprotonated hydroquinone triplet threshold. In contrast, similar calculations conducted for catechol and resorcinol yielded MECPs of +116.9 and +69.1 kJ mol-1, respectively, above the associated triplet thresholds. These results indicated that the curve crossing required to form singlet products upon reaction with triplet O2 is favorable in the case of hydroquinone and unfavorable in the cases of catechol and resorcinol. In practical terms, the selective oxygen addition appears to be a diagnostically useful reaction to differentiate hydroquinone from its ring isomers.

Mild route to generate gaseous metal anions.

Gaseous metal anions such as Na(-), K(-), Cs(-), and Ag(-) can be generated at ambient temperatures by the collision-induced dissociation of the anions of several dicarboxylic acid salts, including oxalate, maleate, fumarate, succinate, and glutamate salts. The formation of gaseous metal anions in this way is unprecedented because the metal is initially present in its cationic form. The mild process described here could facilitate novel applications of metal anions as selective reagents for gas-phase ion-molecule and ion-ion reactions. Ab initio calculations were used to describe the dissociation process for anions of the oxalate salts. The formation of alkalides occurs via production of a metal-carbon dioxide anion intermediate with a bidentate three-center two-electron bond to the metal. The metal atom acquires a partial negative charge in the intermediate structure.