On-line cysteine modification for protein analysis: new probes for electrochemical tagging nanospray mass spectrometry

A series of electrogenerated selective electrophiles based on substituted benzoquinones has been characterized as tags for l-cysteine and cysteine residues in proteins. The electrophiles are generated electrochemically from the corresponding hydroquinones. It is shown from mass spectrometry analysis that the electrogenerated benzoquinone can tag the biomolecules. The rate constants pertaining to the addition of l-cysteine onto the electrogenerated benzoquinones have been determined using electrochemical techniques. The substitution patterns have been unraveled leading to the assessment of site-specific rate constants. It is shown that the rate constants are primarily dependent on the electronic nature of the substituents as expressed by the Hammett substitution constant. The apparent tagging yields observed for l-cysteine in nanospray mass spectrometry experiments do not correspond to the yields expected from the electrochemical study, as the ionisation efficiencies are highly dependent on the tag. Finally, the on-line tagging has been tested using β-lactoglobulin A and myoglobin. Based on these results, it is concluded that the tagging reaction is selective towards cysteine when it takes place in the nanospray interface. The results show that the methodology presented can be used for a rapid characterization and identification of reactive sites in biomolecules.     

Polyelectrolyte-modified short microchannel for cation separation

Three alkali cations, potassium, sodium, and lithium, have been separated within 15 s in a 1 cm long polymer microchip. The separation microchannel is modified by a polycation, poly(allylammonium chloride), which makes the channel surfaces positively charged leading to a reversed electroosmotic flow (EOF) when compared to bare channels. Due to the decreased apparent mobility of the cations, the separation resolution is improved allowing the use of shorter channels.     

Development of a chiral non-aqueous capillary electrophoretic system using the partial filling technique with UV and mass spectrometric detection

A chiral non-aqueous CE system with UV and mass spectrometric detection has been developed. The enantioseparation was promoted by diastereomeric complex (ion-pair) formation between the amines (e.g. salbutamol, atenolol) and the chiral selector, (-)-2,3:4,6-di-O-isopropylidene-2-keto-L-gulonic acid [(-)-DIKGA]. Different solvent mixtures were studied, as well as different concentrations of (-)-DIKGA and ammonium acetate in the background electrolyte. A partial filling technique was developed with a selector plug composed of (-)-DIKGA and ammonium acetate in a solvent mixture of methanol and 2-propanol. The separated enantiomers of pronethalol were detected by a Q-TOF MS system equipped with a sheath-flow electrospray ionization interface.     

On the possible catalysis by single water molecules of gas-phase hydrogen abstraction reactions by OH radicals

The gas phase hydrogen abstraction reaction between OH and CY(2)XH, where X = H, F, OH, or NH(2) and Y = H, CH(3) or F, in the absence and presence of a single water molecule is investigated using both density function theory, B3LYP, and explicitly correlated coupled cluster theory, CCSD(T)-F12. We find that a single water molecule could have a catalytic effect at low temperatures possible in laboratory experiments, but does not seem to catalyze these reactions at 298 K, and will not play a role under relevant atmospheric conditions.     

Cyclic voltammetry of highly hydrophilic ions at a supported liquid membrane

Cyclic voltammetry has been used to study the coupling of ion transfer reactions at a liquid membrane. The liquids are either supported by a porous hydrophobic membrane (polyvinylidene difluoride, PVDF) when the organic solvent is non-volatile (o-nitrophenyloctylether) or are merely a free standing organic solvent layer such as 1,2-dichloroethane comprised between two hydrophilic dialysis membranes supporting the adjacent aqueous phases. The passage of current across the liquid membrane is associated with two ion transfer reactions across the two polarised liquid liquid interfaces in series. It is shown that it is possible to study the transfer of highly hydrophilic ions at one interface by limiting the mass transfer of the other ion transfer reaction at the other interface. Indeed, for systems comprising an ion M in one aqueous phase and a reference ion R partitioned between the membrane and the other aqueous phase, the observed and simulated cyclic voltammograms have a half-wave potential determined by the Gibbs energy of transfer of M transferring at one interface and by the limiting mass transfer of R at the other interface. This new methodology opens a way to measure the Gibbs energy of transfer of highly hydrophilic or hydrophobic ions, which usually limits the potential window at single liquid liquid interfaces (ITIES).     

Nonadiabatic alignment of asymmetric top molecules: rotational revivals

The rotational revival structure of asymmetric top molecules, following irradiation by an intense picosecond laser pulse, is explored theoretically and experimentally. Numerically we solve nonperturbatively for the rotational dynamics of a general asymmetric top subject to a linearly polarized intense pulse, and analyze the dependence of the dynamical alignment on the field and system parameters. Experimentally we use time-resolved photofragment imaging to measure the alignment of two molecules with different asymmetry, iodobenzene, and iodopentafluorobenzene. Our numerical results explain the experimental observations and generalize them to other molecules. The rotational revival structure of asymmetric tops differs qualitatively from the intensively studied linear top case. Potentially it provides valuable structural information about molecules.     

Equatorial contra axial polar substituents. The relation of a chemical reaction to stereochemical substituent constants

The established rates of glycoside hydrolysis reactions were analyzed using free energy relationship plots based on substituent constants that depend on whether the substituent is axial or equatorial. In all cases good correlations were found when assuming either that the transition state had a charged ring-oxygen atom or that it had a charged anomeric carbon atom. The spontaneous hydrolysis of 2,4-dinitrophenyl beta-glycopyranosides and the acidic hydrolysis of methyl beta-D-glycopyranosides were found to give a good correlation, when 100% charge at the ring-oxygen in the transition state of these reactions is assumed. The acidic hydrolysis of methyl alpha-glycopyranosides was found to give good correlations regardless of whether 100% charge at the ring-oxygen or 100% charge at the anomeric carbon was assumed. The findings clearly demonstrate how crucial the stereochemistry of even remote polar substituents is for their electronic effect on chemical reaction.     

Dioxygen-promoted regioselective oxidative heck arylations of electron-rich olefins with arylboronic acids

Arylations of electron-rich heteroatom-substituted olefins were performed with arylboronic acids. This appears to constitute the first example of palladium(II)-catalyzed internal Heck arylations. The novel protocol exploits oxygen gas for environmentally benign reoxidation and a stable 1,10-phenanthroline bidentate ligand to promote the palladium(II) regeneration and to control the regioselectivity. Internal arylation is strongly favored with electron-rich arylboronic acids. DFT calculations support a charge-driven selectivity rationale, where phenyls substituted with electron-donating groups prefer the electron-poor alpha-carbon of the olefin. Experiments, verified by calculations, confirm the cationic nature of the catalytic route. This Heck methodology provides a facile and mild access to functionalized enamides. Controlled microwave heating and increased oxygen pressure were used to further reduce the reaction time to 1 h.