Protomers: formation,separation and characterization via travelling wave ion mobility mass spectrometry

Travelling wave ion mobility mass spectrometry (TWIM‐MS) with post‐TWIM and pre‐TWIM collision‐induced dissociation (CID) experiments were used to form, separate and characterize protomers sampled directly from solutions or generated in the gas phase via CID. When in solution equilibria, these species were transferred to the gas phase via electrospray ionization, and then separated by TWIM‐MS. CID performed after TWIM separation (post‐TWIM) allowed the characterization of both protomers via structurally diagnostic fragments. Protonated aniline (1) sampled from solution was found to be constituted of a ca. 5:1 mixture of two gaseous protomers, that is, the N‐protonated (1a) and ring protonated (1b) molecules, respectively. When dissociated, 1a nearly exclusively loses NH₃, whereas 1b displays a much diverse set of fragments. When formed via CID, varying populations of 1a and 1b were detected. Two co‐existing protomers of two isomeric porphyrins were also separated and characterized via post‐TWIM CID. A deprotonated porphyrin sampled from a basic methanolic solution was found to be constituted predominantly of the protomer arising from deprotonation at the carboxyl group, which dissociates promptly by CO₂ loss, but a CID‐resistant protomer arising from deprotonation at a porphyrinic ring NH was also detected and characterized. The doubly deprotonated porphyrin was found to be constituted predominantly of a single protomer arising from deprotonation of two carboxyl groups. Copyright © 2012 John Wiley & Sons, Ltd.     

Baseline resolution of isomers by traveling wave ion mobility mass spectrometry: investigating the eff ects of polarizable drift gases and ionic charge distribution

We have studied the behavior of isomers and analogues by traveling wave ion mobility mass spectrometry (TWIM‐MS) using drift‐gases with varying masses and polarizabilities. Despite the reduced length of the cell (18 cm), a pair of constitutional isomers, N‐butylaniline and para‐butylaniline, with theoretical collision cross‐section values in helium (Ω_He) differing by as little as 1.2 Ų (1.5%) but possessing contrasting charge distribution, showed baseline peak‐to‐peak resolution (R_p‐p) for their protonated molecules, using carbon dioxide (CO₂), nitrous oxide (N₂O) and ethene (C₂H₄) as the TWIM drift‐gas. Near baseline R_p‐p was also obtained in CO₂ for a group of protonated haloanilines (para‐chloroaniline, para‐bromoaniline and para‐iodoaniline) which display contrasting masses and theoretical Ω_He, which differ by as much as 15.7 Ų (19.5%) but similar charge distributions. The deprotonated isomeric pair of trans‐oleic acid and cis‐oleic acid possessing nearly identical theoretical Ω_He and Ω_N2 as well as similar charge distributions, remained unresolved. Interestingly, an inversion of drift‐times were observed for the 1,3‐dialkylimidazolium ions when comparing He, N₂ and N₂O. Using density functional theory as a means of examining the ions electronic structure, and He and N₂‐based trajectory method algorithm, we discuss the effect of the long‐range charge induced dipole attractive and short‐range Van der Waals forces involved in the TWIM separation in drift‐gases of differing polarizabilities. We therefore propose that examining the electronic structure of the ions under investigation may potentially indicate whether the use of more polarizable drift‐gases could improve separation and the overall success of TWIM‐MS analysis. Copyright © 2013 John Wiley & Sons, Ltd.     

Metal‐Directed Self‐Assembly of a Polyoxometalate‐Based Molecular Triangle: Using Powerful Analytical Tools to Probe the Chemical Structure of Complex Supramolecular Assemblies

A polyoxometalate‐based molecular triangle has been synthesized through the metal‐driven self‐assembly of covalent organic/inorganic hybrid oxo‐clusters with remote pyridyl binding sites. The new metallomacrocycle was unambiguously characterized by using a combination of ¹H NMR spectroscopy, 2D diffusion NMR spectroscopy (DOSY), electrospray ionization travelling wave ion mobility mass spectrometry (ESI‐TWIM‐MS), small‐angle X‐ray scattering (SAXS) and molecular modelling. The collision cross‐sections obtained from TWIM‐MS and the hydrodynamic radii derived from DOSY are in good agreement with the geometry‐optimized structures obtained by using theoretical calculations. Furthermore, SAXS was successfully employed and proved to be a powerful technique for characterizing such large supramolecular assemblies.     

Peak width‐mass correlation in CID MS/MS of isomeric oligosaccharides using traveling‐wave ion mobility mass spectrometry

Isomeric oligosaccharides γ‐cyclodextrin (γ‐CD), glucosyl‐βCD (Glc₁‐βCD) and maltosyl‐αCD (Glc₂‐αCD) were analyzed by traveling‐wave ion mobility (twIM) mass spectrometry (MS). Their formation of multicharged multimers differed from each other. The ion mobility‐mass spectrometry was useful in the self‐assembling and complex formation analyses of CD isomers. The drift times of the isomers and their product ions with the same mass were almost the same in collision‐induced dissociation (CID) MS/MS. In contrast, the ion mobility peak widths were sensitive to structural differences of the isomeric product ions. The twIM peak width (ms ‐ µs) of the product ions [M ? Glc_n + H]⁺ (n = 0 ～ 6) of γ‐CD correlated linearly with their masses (Da); the large and/or long chain product ions had wider peak widths, which were much wider than those from the general diffusion effect. This was a novel and useful ‘trend line’ to discriminate between the three isomers. Plots of [M ? Glc_{2 ～ 6} + H]⁺ of Glc₁‐βCD and [M ? Glc_{3 ～ 6} + H]⁺ of Glc₂‐αCD product ions' plots were on the same trend line as γ‐CD. The plots of [M ? Glc₁ + H]⁺ of Glc₁‐βCD and [M ? Glc_{1, 2} + H]⁺ of Glc₂‐αCD strayed from the γ‐CD line; their peak widths were narrower than those of γ‐CD. These results indicated that product ions from the chemical species of Glc₁‐β CD and Glc₂‐αCD retained their CD structure. Analyses of the IM peak widths enable us to elucidate the structures of the product ions. Copyright © 2009 John Wiley & Sons, Ltd.     

Practical guidelines for characterization of O‐diglycosyl flavonoid isomers by triple quadrupole MS and their applications for identification of some fruit juices flavonoids

Fifteen flavonoid O‐diglycosides with different interglycosidic linkage isomery and glycosylation position have been studied in order to analyze their fragmentation patterns. Initial separation was carried out using high performance liquid chromatography with diode array detection (HPLC/DAD) coupled to an electrospray ionization (ESI) interface and a triple quadrupole mass spectrometer. Some useful differences in their MS spectra have been found and discussed. As it has already been reported, [Y*]⁺/[Y₀]⁺ ratio for flavanones and [Y₁]⁺/[Y₀]⁺ ratio for other flavonoids is specific for each isomeric interglycosidic linkage. In this work it has also been observed that the abundance of these ions is dependent on the position of glycosylation. On the basis of these differences, systematic guidelines for our experimental conditions have been proposed for the differentiation of not only isomeric interglycosidic linkage but also glycosylation position using collision‐induced dissociation MS/MS (CID‐MS/MS) spectra in positive mode. These results have been successfully applied for the characterization of three diglycosyl flavonoids found in Citrus fruit juices and these conclusions have also been extrapolated for characterizing two triglycosides in the same fruits. Copyright © 2009 John Wiley & Sons, Ltd.     

An RP-LC-UV-TWIMS-HRMS and Chemometric Approach to Differentiate between Momordica balsamina Chemotypes from Three Different Geographical Locations in Limpopo Province of South Africa

Momordica balsamina leaf extracts originating from three different geographical locations were analyzed using reversed-phase liquid chromatography (RP-LC) coupled to travelling wave ion mobility (TWIMS) and high-resolution mass spectrometry (HRMS) in conjunction with chemometric analysis to differentiate between potential chemotypes. Furthermore, the cytotoxicity of the three individual chemotypes was evaluated using HT-29 colon cancer cells. A total of 11 molecular species including three flavonol glycosides, five cucurbitane-type triterpenoid aglycones and three glycosidic cucurbitane-type triterpenoids were identified. The cucurbitane-type triterpenoid aglycones were detected in the positive ionization mode following dehydration [M + H − H₂O]⁺ of the parent compound, whereas the cucurbitane-type triterpenoid glycosides were primarily identified following adduct formation with ammonia [M + NH₄]⁺. The principle component analysis (PCA) loadings plot and a variable influence on projection (VIP) analysis revealed that the isomeric pair balsaminol E and/or karavilagen E was the key molecular species contributing to the distinction between geographical samples. Ultimately, based on statistical analysis, it is hypothesized that balsaminol E and/or karavilagen E are likely responsible for the cytotoxic effects in HT-29 cells.     

Studies of peptide a- and b-type fragment ions using stable isotope labeling and integrated ion mobility/tandem mass spectrometry

The structures of peptide a- and b-type fragment ions were studied using synthetic peptides including a set of isomeric peptides, differing in the sequence location of an alanine residue labeled with ¹⁵N and uniformly with ¹³C. The pattern of isotope labeling of second-generation fragment ions derived via a _n and b _n ions (where n=4 or 5) suggested that these intermediates existed in part as macrocyclic structures, where alternative sites of ring opening gave rise to different linear forms whose simple cleavage might give rise to the observed final products. Similar conclusions were derived from combined ion mobility/tandem MS analyses where different fragmentation patterns were observed for isomeric a- or b-type ions that display different ion mobilities. These analyses were facilitated by a new approach to the processing of ion mobility/tandem MS data, from which distinct and separate product ion spectra are derived from ions that are incompletely separated by ion mobility. Finally, an example is provided of evidence for a macrocyclic structure for b _n ions where n=8 or 9.     

Traveling-wave ion mobility mass spectrometry analysis of isomeric modified peptides arising from chemical cross-linking

Traveling-wave ion mobility (TWIM) coupled to mass spectrometry (MS) has emerged as a powerful tool for structural and conformational analysis of proteins and peptides, allowing the analysis of isomeric peptides (or proteins) with the same sequence but modified at different residues. This work demonstrates the use of the novel TWIM-MS technique to separate isomeric peptide ions derived from chemical cross-linking experiments, which enables the acquisition of distinct product ion spectra for each isomer, clearly indicating modification on different sites. Experiments were performed with four synthetic peptides, for which variable degrees of mobility separation were achieved. In cases of partially overlapping mobility arrival time distributions (ATDs), extracting the ATDs of fragment ions belonging to each individual isomer allowed their separation into two distinct ATDs. Accumulation over regions from the specific ATDs generates the product ion spectrum of each isomer, or a spectrum highly enriched in their fragments. The population of both modified peptide isomers was correlated with the intrinsic reactivities of different Lys residues from reactions conducted at different pH conditions.     

Identification of common glycosyl groups of flavonoid O‐glycosides by serial mass spectrometry of sodiated species

Flavonoid O‐glycosides are a ubiquitous and important group of plant natural products in which a wide variety of sugars are O‐linked to an aglycone. Determining the identity of the sugars, and the manner in which they are linked, by mass spectrometry alone is challenging. To improve the identification of common O‐linked di‐ and trisaccharides when analysing mixtures of flavonoid O‐glycosides by liquid chromatography/mass spectrometry (LC/MS), the fragmentation of electrosprayed sodium adducts in an ion trap mass spectrometer was investigated. The sodium adducts [M + Na]⁺ of kaempferol 3‐O‐glycosides generated sodiated glycosyl groups by the neutral loss of kaempferol. The product ion spectra of these sodiated glycosyl groups differed between four isomeric kaempferol 3‐O‐rhamnosylhexosides and four isomeric kaempferol 3‐O‐glucosylhexosides in which the primary hexose was either glucose or galactose and bore the terminal glucose or rhamnose at either C‐2 or C‐6. Fragmentation of sodiated glycosyl groups from linear O‐triglucosides and branched O‐glucosyl‐(1 → 2)‐[rhamnosyl‐(1 → 6)]‐hexosides produced sodiated disaccharide residues, and the product ion spectra of these ions assisted the identification of the complete sugar. The product ion spectra of the sodiated glycosyl groups were consistent among flavonoid O‐glycosides differing in the position at which the sugar was O‐linked to the aglycone, and the nature of the aglycone. The abundance of sodiated species was enhanced by application of a pre‐trap collision voltage, without the need to dope with salt, allowing automated LC/MS methods to be used to identify the glycosyl groups of common flavonoid O‐glycosides, such as rutinosides, robinobiosides, neohesperidosides, gentiobiosides and sophorosides. Copyright © 2011 John Wiley & Sons, Ltd.     

Liquid chromatography coupled with ultraviolet absorbance detection,electrospray ionization,collision‐induced dissociation and tandem mass spectrometry on a triple quadrupole for the on‐line characterization of polyphenols and methylxanthines in green coffee beans

Liquid chromatography coupled with a photodiode array detector, electrospray ionization, collision‐induced dissociation and tandem mass spectrometry (LC‐DAD/ESI‐CID‐MS/MS) on a triple quadrupole (QqQ) has been used to detect and characterize polyphenols and methylxanthines in green coffee beans: three phenolic acids (caffeic acid, ferulic acid and dimethoxycinnamic acid), three isomeric caffeoylquinic acids (M_r 354), three feruloylquinic acids (M_r 368), one p‐coumaroylquinic acid (M_r 338), three dicaffeoylquinic acids (M_r 516), three feruloyl‐caffeoylquinic acids (M_r 530), four p‐coumaroyl‐caffeoylquinic acids (M_r 500), three diferuloylquinic acids (M_r 544), six dimethoxycinnamoyl‐caffeoylquinic acids (M_r 544), three dimethoxycinnamoyl‐feruloylquinic acids (M_r 558), six cinnamoyl‐amino acid conjugates, three cinnamoyl glycosides, and three methylxanthines (caffeine, theobromine and theophylline). Dimethoxycinnamic acid, three isomers of dimethoxycinnamoyl‐caffeoylquinic acids and another three of dimethoxycinnamoyl‐feruloylquinic acids, as well as the three cinnamoyl glycosides, had not previously been reported in coffee beans. Structures have been assigned on the basis of the complementary information obtained from UV‐visible spectra, relative hydrophobicity, scan mode MS spectra, and fragmentation patterns in MS² spectra (both in the positive and negative ion modes) obtained using a QqQ at different collision energies. A structure diagnosis scheme is provided for the identification of different isomers of polyphenols and methylxanthines. Copyright © 2009 John Wiley & Sons, Ltd.     

ESI‐MS/MS analysis of protonated N‐methyl amino acids and their immonium ions

Methylation is one of the important posttranslational modifications of biological systems. At the metabolite level, the methylation process is expected to convert bioactive compounds such as amino acids, fatty acids, lipids, sugars, and other organic acids into their methylated forms. A few of the methylated amino acids are identified and have been proved as potential biomarkers for several metabolic disorders by using mass spectrometry–based metabolomics workstation. As it is possible to encounter all the N‐methyl forms of the proteinogenic amino acids in plant/biological systems, it is essential to have analytical data of all N‐methyl amino acids for their detection and identification. In earlier studies, we have reported the ESI‐MS/MS data of all methylated proteinogenic amino acids, except that of mono‐N‐methyl amino acids. In this study, the N‐methyl amino acids of all the amino acids ( 1 ‐ 21 ; including one isomeric pair) were synthesized and characterized by ESI‐MS/MS, LC/MS/MS, and HRMS. These data could be useful for detection and identification of N‐methyl amino acids in biological systems for future metabolomics studies. The MS/MS spectra of [M + H]⁺ ions of most N‐methyl amino acids showed respective immonium ions by the loss of (H₂O, CO). The other most common product ions detected were [MH‐(NH₂CH₃]⁺, [MH‐(RH)]⁺ (where R = side chain group) ions, and the selective structure indicative product ions due to side chain and N‐methyl group. The isomeric/isobaric N‐methyl amino acids could easily be differentiated by their distinct MS/MS spectra. Further, the MS/MS of immonium ions inferred side chain structure and methyl group on α‐nitrogen of the N‐methyl amino acids.     

Design, synthesis, and traveling wave ion mobility mass spectrometry characterization of iron(II)- and ruthenium(II)-terpyridine metallomacrocycles

New metallomacrocycles composed of 2,2':6',2″-terpyridine (tpy) ligands and Ru(II) or Fe(II) transition metal ions were prepared by stepwise directed assembly and characterized by 2D diffusion NMR spectroscopy (DOSY), electrospray ionization traveling wave ion mobility mass spectrometry (ESI TWIM MS), and molecular modeling. The supramolecular polymers synthesized include a homonuclear all-Ru hexamer as well as heteronuclear hexamer and nonamer with alternating Ru/Ru/Fe metal centers. ESI MS yields several charge states from each supramacromolecule. If ESI is interfaced with TWIM MS, overlapping charge states and the isomeric components of an individual charge state are separated based on their unique drift times through the TWIM region. From experimentally measured drift times, collision cross-sections can be deduced. The collision cross-sections obtained for the synthesized supramacromolecules are in good agreement with those predicted by molecular modeling for macrocyclic structures. Similarly, the hydrodynamic radii of the synthesized complexes derived from 2D DOSY NMR experiments agree excellently with the radii calculated for macrocyclic architectures, confirming the ESI TWIM MS finding. ESI TWIM MS and 2D DOSY NMR spectroscopy provide an alternative approach for the structural analysis of supramolecules that are difficult or impossible to crystallize, such as the large macrocyclic assemblies investigated. ESI TWIM MS will be particularly valuable for the characterization of supramolecular assemblies not available in the quantity or purity required for NMR studies.     

Quantitative analysis of positional isomers of triacylglycerols via electrospray ionization tandem mass spectrometry of sodiated adducts

Herein we report a reversed‐phase high‐performance liquid chromatography tandem mass spectrometry (RP‐HPLC/MS/MS) method for the analysis of positional isomers of triacylglycerols (TAGs) in vegetable oils. The fragmentation behavior of [M + X]⁺ ions (X = NH₄, Li, Na or Ag) was studied on a quadrupole‐time‐of‐flight (Q‐TOF) mass spectrometer under low‐energy collision‐induced dissociation (CID) conditions. Mass spectra that were dependent on the X⁺ ion and the nature and position of the acyl substituents were observed for four pairs of 'AAB/ABA'‐type TAGs, namely PPO/POP, OOP/OPO, LLO/LOL and OOL/OLO (where P is 16:0, palmitic acid; O is 18:1, oleic acid; and L is 18:2, linoleic acid). For the majority of [M + X]⁺ adducts, the loss of the fatty acid in the outer positions (sn‐1 or sn‐3) was favored over the loss in the central position (sn‐2), which enabled the determination of the fractional abundance of the isomers. Ratios of the intensity of fragment ions at various AAB/ABA compositions produced linear calibration curves with positive slopes, comparable to those obtained traditionally by ESI‐MS/MS of [M + NH₄]⁺ adducts. The only exceptions were the [M + Ag]⁺ adducts of the PPO/POP system, which produced calibration curves with negative slopes. Sodium adducts provided the most consistent level of isomeric discrimination for the TAGs studied and also offered the most convenience in that they required no additive to the mobile phase. Therefore, calibration curve data derived from [M + Na]⁺ adducts were applied to the quantification of TAG regioisomers in sunflower and olive oils. The regiospecific analysis showed that palmitic acid was typically located at positions sn‐1 or sn‐3, whereas unsaturated fatty acids, oleic and linoleic acids were mostly found at the sn‐2 position. Copyright © 2010 Crown in the right of Canada. Published by John Wiley & Sons, Ltd.     

Differentiation of Boc‐protected α,δ‐/δ,α‐ and β,δ‐/δ,β‐hybrid peptide positional isomers by electrospray ionization tandem mass spectrometry

Two new series of Boc‐N‐α,δ‐/δ,α‐ and β,δ‐/δ,β‐hybrid peptides containing repeats of L ‐Ala‐δ⁵‐Caa/δ⁵‐Caa‐L ‐Ala and β³‐Caa‐δ⁵‐Caa/δ⁵‐Caa‐β³‐Caa (L ‐Ala = L ‐alanine, Caa = C‐linked carbo amino acid derived from D ‐xylose) have been differentiated by both positive and negative ion electrospray ionization (ESI) ion trap tandem mass spectrometry (MS/MS). MSⁿ spectra of protonated isomeric peptides produce characteristic fragmentation involving the peptide backbone, the Boc‐group, and the side chain. The dipeptide positional isomers are differentiated by the collision‐induced dissociation (CID) of the protonated peptides. The loss of 2‐methylprop‐1‐ene is more pronounced for Boc‐NH‐L ‐Ala‐δ‐Caa‐OCH₃ (1), whereas it is totally absent for its positional isomer Boc‐NH‐δ‐Caa‐L ‐Ala‐OCH₃ (7), instead it shows significant loss of t‐butanol. On the other hand, second isomeric pair shows significant loss of t‐butanol and loss of acetone for Boc‐NH‐δ‐Caa‐β‐Caa‐OCH₃ (18), whereas these are insignificant for its positional isomer Boc‐NH‐β‐Caa‐δ‐Caa‐OCH₃ (13). The tetra‐ and hexapeptide positional isomers also show significant differences in MS² and MS³ CID spectra. It is observed that ‘b’ ions are abundant when oxazolone structures are formed through five‐membered cyclic transition state and cyclization process for larger ‘b’ ions led to its insignificant abundance. However, b₁⁺ ion is formed in case of δ,α‐dipeptide that may have a six‐membered substituted piperidone ion structure. Furthermore, ESI negative ion MS/MS has also been found to be useful for differentiating these isomeric peptide acids. Thus, the results of MS/MS of pairs of di‐, tetra‐, and hexapeptide positional isomers provide peptide sequencing information and distinguish the positional isomers. Copyright © 2010 John Wiley & Sons, Ltd.     

Approach to the study of flavone di‐C‐glycosides by high performance liquid chromatography‐tandem ion trap mass spectrometry and its application to characterization of flavonoid composition in Viola yedoensis

The mass spectrometric (MS) analysis of flavone di‐C‐glycosides has been a difficult task due to pure standards being unavailable commercially and to that the reported relative intensities of some diagnostic ions varied with MS instruments. In this study, five flavone di‐C‐glycoside standards from Viola yedoensis have been systematically studied by high performance liquid chromatography‐electrospray ionization‐tandem ion trap mass spectrometry (HPLC‐ESI‐IT‐MSⁿ) in the negative ion mode to analyze their fragmentation patterns. A new MS² and MS³ hierarchical fragmentation for the identification of the sugar nature (hexoses or pentoses) at C‐6 and C‐8 is presented based on previously established rules of fragmentation. Here, for the first time, we report that the MS² and MS³ structure‐diagnostic fragments about the glycosylation types and positions are highly dependent on the configuration of the sugars at C‐6 and C‐8. The base peak (^0,2X₁^0,2X₂^? ion) in MS³ spectra of di‐C‐glycosides could be used as a diagnostic ion for flavone aglycones. These newly proposed fragmentation behaviors have been successfully applied to the characterization of flavone di‐C‐glycosides found in V. yedoensis. A total of 35 flavonoid glycosides, including 1 flavone mono‐C‐hexoside, 2 flavone 6,8‐di‐C‐hexosides, 11 flavone 6,8‐di‐C‐pentosides, 13 flavone 6,8‐C‐hexosyl‐C‐pentosides, 5 acetylated flavone C‐glycosides and 3 flavonol O‐glycosides, were identified or tentatively identified on the base of their UV profiles, MS and MSⁿ (n = 5) data, or by comparing with reference substances. Among these, the acetylated flavone C‐glycosides were reported from V. yedoensis for the first time. Copyright © 2014 John Wiley & Sons, Ltd.     

Dereplication of known pregnane glycosides and structural characterization of novel pregnanes in Marsdenia tenacissima by high‐performance liquid chromatography and electrospray ionization‐tandem mass spectrometry

In the search for novel natural products in plants, particularly those with potential bioactivity, it is important to efficiently distinguish novel compounds from previously isolated, known compounds, a process known as dereplication. In this study, electrospray ionization‐multiple stage tandem mass spectrometry (ESI‐MSⁿ) was used to study the behaviour of 12 pregnane glycosides and genins previously isolated from Marsdenia tenacissima, a traditional Chinese medicinal plant, as a basis for dereplication of compounds in a plant extract. In addition to [M + Na]⁺ and [M + NH₄]⁺ ions, a characteristic [M‐glycosyl + H]⁺ ion was observed in full‐scan mode with in‐source fragmentation. Sequential in‐trap collision‐induced dissociation of [M + Na]⁺ ions from 11,12‐diesters revealed consistent preferred losses of substituents first from C‐12, then from C‐11, followed by losses of monosaccharide fragments from the C‐3 tri‐ and tetrasaccharide substituents. A crude methanol extract of M. tenacissima stems was analysed using high‐performance liquid chromatography coupled to ESI‐MS. Several previously isolated pregnane glycosides were dereplicated, and the presence of an additional nine novel pregnane glycosides is predicted on the basis of the primary and fragment ions observed, including two with a previously unreported C₄H₇O C‐11/C‐12 substituent of pregnane glycosides. This study is the first report of prediction of the structures of novel pregnane glycosides in a crude plant extract by a combination of in‐source fragmentation and in‐trap collision‐induced dissociation and supports the usefulness of LC‐ESI‐MSⁿ not only for dereplication of active compounds in extracts of medicinal plants but also for detecting the presence of novel related compounds. Copyright © 2012 John Wiley & Sons, Ltd.     

On the Hydrogen Oxalate Binding Motifs onto Dinuclear Cu and Ag Metal Phosphine Complexes

We report the binding geometries of the isomers that are formed when the hydrogen oxalate ((CO₂)₂H=HOx) anion attaches to dinuclear coinage metal phosphine complexes of the form [M₁M₂dcpm₂(HOx)]⁺ with M=Cu, Ag and dcpm=bis(dicyclohexylphosphino)methane, abbreviated [MM]⁺ . These structures are established by comparison of isomer-selective experimental vibrational band patterns displayed by the cryogenically cooled and N₂-tagged cations with DFT calculations of the predicted spectra for various local minima. Two isomeric classes are identified that feature either attachment of the carboxylate oxygen atoms to the two metal centers (end-on docking) or attachment of oxygen atoms on different carbon atoms asymmetrically to the metal ions (side-on docking). Within each class, there are additional isomeric variations according to the orientation of the OH group. This behavior indicates that HOx undergoes strong and directional coordination to [CuCu]⁺ but adopts a more flexible coordination to [AgAg]⁺ . Infrared spectra of the bare ions, fragmentation thresholds and ion mobility measurements are reported to explore the behaviors of the complexes at ambient temperature.     

Distinguishing two isomeric mephedrone substitutes with selective reagent ionisation mass spectrometry (SRI‐MS)

The isomers 4‐methylethcathinone and N‐ethylbuphedrone are substitutes for the recently banned drug mephedrone. We find that with conventional proton transfer reaction mass spectrometry (PTR‐MS), it is not possible to distinguish between these two isomers, because essentially for both substances, only the protonated molecules are observed at a mass‐to‐charge ratio of 192 (C₁₂H₁₈NO⁺). However, when utilising an advanced PTR‐MS instrument that allows us to switch the reagent ions (selective reagent ionisation) from H₃O⁺ (which is commonly used in PTR‐MS) to NO⁺, O₂⁺ and Kr⁺, characteristic product (fragment) ions are detected: C₄H₁₀N⁺ (72 Da) for 4‐methylethcathinone and C₅H₁₂N⁺ (86 Da) for N‐ethylbuphedrone; thus, selective reagent ionisation MS proves to be a powerful tool for fast detection and identification of these compounds. Copyright © 2013 John Wiley & Sons, Ltd.     

Redox Non‐Innocence and Isomer‐Specific Oxidative Functionalization of Ruthenium‐Coordinated β‐Ketoiminate

This article deals with isomeric ruthenium complexes [Ru^III(L^R)₂(acac)] (S=1/2) involving unsymmetric β‐ketoiminates (AcNac) (L^R=R‐AcNac, R=H ( 1 ), Cl ( 2 ), OMe ( 3 ); acac=acetylacetonate) [R=para‐substituents (H, Cl, OMe) of N‐bearing aryl group]. The isomeric identities of the complexes, cct (cis‐cis‐trans, blue, a ), ctc (cis‐trans‐cis, green, b ) and ccc (cis‐cis‐cis_, pink, c ) with respect to oxygen (acac), oxygen (L) and nitrogen (L) donors, respectively, were authenticated by their single‐crystal X‐ray structures and spectroscopic/electrochemical features. One‐electron reversible oxidation and reduction processes of 1 – 3 led to the electronic formulations of [Ru^III(L)(L ^? )(acac)]⁺ and [Ru^II(L)₂(acac)]^? for 1 ⁺‐ 3 ⁺ (S=1) and 1^? – 3^? (S=0), respectively. The triplet state of 1 ⁺‐ 3 ⁺ was corroborated by its forbidden weak half‐field signal near g≈4.0 at 4 K, revealing the non‐innocent feature of L. Interestingly, among the three isomeric forms ( a – c in 1 – 3 ), the ctc ( b in 2 b or 3 b ) isomer selectively underwent oxidative functionalization at the central β‐carbon (C?H→C=O) of one of the L ligands in air, leading to the formation of diamagnetic [Ru^II(L)(L ′ )(acac)] (L ′ =diketoimine) in 4 / 4′ . Mechanistic aspects of the oxygenation process of AcNac in 2 b were also explored via kinetic and theoretical studies.