Mathilde Faure

Hydrogen isotope exchanges between water and methanol in interstellar ices

The deuterium fractionation of gas-phase molecules in hot cores is believed to reflect the composition of interstellar ices. The deutera-tion of methanol is a major puzzle, however, because the isotopologue ratio [CH 2 DOH]/[CH 3 OD], which is predicted to be equal to 3 by standard grain chemistry models, is much larger (∼20) in low-mass hot corinos and significantly lower (∼1) in high-mass hot cores. This dichotomy in methanol deuteration between low-mass and massive protostars is currently not understood. In this study, we report a simplified rate equation model of the deuterium chemistry occurring in the icy mantles of interstellar grains. We apply this model to the chemistry of hot corinos and hot cores, with IRAS 16293-2422 and the Orion KL Compact Ridge as prototypes, respectively. The chemistry is based on a statistical initial deuteration at low temperature followed by a warm-up phase during which thermal hydro-gen/deuterium (H/D) exchanges occur between water and methanol. The exchange kinetics is incorporated using laboratory data. The [CH 2 DOH]/[CH 3 OD] ratio is found to scale inversely with the D/H ratio of water, owing to the H/D exchange equilibrium between the hydroxyl (-OH) functional groups of methanol and water. Our model is able to reproduce the observed [CH 2 DOH]/[CH 3 OD] ratios provided that the primitive fractionation of water ice [HDO]/[H 2 O] is ∼2% in IRAS 16293-2422 and ∼0.6% in Orion KL. We conclude that the molecular D/H ratios measured in hot cores may not be representative of the original mantles because molecules with exchangeable deuterium atoms can equilibrate with water ice during the warm-up phase.

Investigating the kinetics of paramagnetic-beads linked alkaline phosphatase enzyme through microchannel resistance measurement in dielectric microchip

Real time monitoring of electrolyte resistance changes during hydrolysis of 4-nitrophenylphosphate (pNPP) by alkaline phosphatase (ALP) bound on paramagnetic-beads was performed into a small dielectric channel. The reaction kinetic fit with a non-competitive substrate-inhibition equation. Michaelis-Menten apparent constant, KM(app), was determined as 0.33±0.06mM and the maximum apparent rate, Vmax(app) as 98±5pMs(-1). The detection limits were 15fM for ALP and 0.75mM for pNPP. This miniaturized device constitutes a powerful tool for analysis of interaction between ligands.

Investigating of labelling and detection of transthyretin synthetic peptide derivatized with naphthalene-2,3-dicarboxaldehyde

Labelling and detection of a synthetic peptide (PN) mimicking a tryptic fragment of interest for the diagnosis of familial amyloidal polyneuropathy have been investigated optically and electrochemically. We decided to covalently label naphtalene-2,3-dicarboxyaldehyde (NDA), a fluorogenic and electroactive molecule on PN. First, the optimization of the labelling chemical reaction was performed by capillary electrophoresis coupled with laser induced fluorescence detection (CE-LIF). The analytical parameters such as separation efficiency and peak area were considered to propose this optimized derivatization reaction. The results obtained allowed us to establish the pH and ionic strength of the derivatization buffer, the molar ratio between NDA and PN and the reaction time of the labelling. Optimal conditions are obtained when [NDA]/[PN]=40, buffer pH of 9, buffer ionic strength of 70 mM and reaction time of 15 min. Second, differential pulse voltammetry (DPV) and cyclic voltammetry (CV) were also used to characterize NDA-labelled PN and different electroinactive amino acids (histidine, lysine, serine, threonine) which are in the PN sequence. The electrochemical detection experiments demonstrated that the labelled biomolecules could be also easily detected at low concentration. Moreover, the derivatization reaction could be followed to describe more precisely the labelling process of these biomolecules. Optimal conditions for labelling are obtained when [NDA]total/[CN(-)] ratio =1 and [NDA]total/[amino acid or peptide]=100 with a buffer having a pH=9 on a glassy carbon electrode. In all cases, an obvious oxidation peak for the N-2-substituted-1-cyanobenz-[f]-isoindole derivative (CBI) has been observed at 0.5-0.7 V/SCE. The multi-labelling of PN and lysine were shown with DPV. We presumed this result to occur because of the shouldered shape of the DPV peak shape. These experiments confirm that NDA can be used as a derivative agent for PN, allowing for electrochemical and fluorescence detections with a limit of detection of labelled PN estimated at 0.2 µM and 5 µM, respectively.

Electrochemiluminescence on-a-chip: Towards a hand-held electrically powered optofluidic source

We report a microfluidic platform that integrates several parallel optical sources based on electrochemiluminescence (ECL) of 9,10-diphenylanthracene (DPA) as luminophore agent. The annihilation of DPA radicals provides a low wavelength emission at λ=430 nm in the blue-visible range. By varying the distance between electrodes for each ECL integrated source, this glass/PDMS/glass platform enabled a systematic investigation of the main electrochemical parameters involved in ECL. These parameters have been studied either in a static mode or in a dynamic one. Even at slow flow rate (~2 µl s(-1)), the renewal of electroactive species could be easily promoted inside the microfluidic channel which gives rise to a stable optical intensity for several minutes. Compared with traditional optically pumped dye sources, this microfluidic system demonstrates that ECL can be easily implemented on chip for producing much compact optofluidic sources. Such simply electrically powered system-on-chip would surely encourage the future of hand-held µTAS devices with integrated fast detection and embedded electronics.

Investigating the Kinetics of Antibody Adsorption onto Polyethylene Terephthalate (PET) Modified with Gold Nanoparticles in Flow Microchannel

The real-time monitoring of the adsorption kinetic of antibody onto polyethylene terephthalate (PET) modified with gold nanoparticles (NPs) is performed into a dielectric flow microchannel. The principle is based on the microelectrode-dielectric interface excitation by a modulated voltage excitation through the dielectric layer with high frequency range. The set-up configuration using a homemade current-to-voltage converter has been developed for fast monitoring of biomolecule adsorption without direct electrical contact of microelectrodes with the microchannel flow. The change in the interfacial admittance, related to the output voltage measured through the microchip, during antibody incubation, gives information on the kinetics involved while antibodies are adsorbed at the interface. An example of adsorption on PET modified with gold NPS was taken. Experimental data were fit with the Langmuir equation and a good correlation was obtained with this latter, where the equilibrium constant, K, was found as 1.55 × 108 M−1 with a limit of detection for antibody concentration without depletion equal to 8.25 nM.

Determination of the isomeric forms proportion of fluorogenic naphthalene-2,3-dicarboxaldehyde in a binary mixture of water:methanol using electrochemical methods

The electrochemical response of the fluorogenic label naphthalene-2,3-dicarboxyaldehyde (NDA) in a binary mixture of water/methanol was characterized with cyclic voltammetry (CV) and differential pulse voltammetry (DPV) electrochemical techniques. Naphthalene-2,3-dicarboxyaldehyde does exist in three isomeric forms in aqueous solution: the unhydrated dialdehyde (DA), the acyclic monohydrated (MA) and the cyclic hemiacetal (HAC). The study underlines that the proportion of each of them varies according to the working pH. At low and high pH, the dialdehyde form is in larger proportion than the acyclic monohydrated form. Conversely at intermediate pH, the concentration of the acyclic form is in greater proportion than the dialdehyde form. These results allowed us to determine the optimal pH of 9 for which the labeling of biomolecules could be more efficient due to the base catalyzed regeneration of the unhydrated form. At this pH, the data processing from the analysis of measured currents and estimation of diffusion coefficients of each form according to the semi-empirical models of Wilke-Chang, Scheibel, Reddy-Doraiswamy and Lusis-Ratcliff allowed us to obtain the concentration of dialdehyde (0.28 mM), acyclic monohydrated (0.57 mM) and cyclic hemiacetal monohydrated (0.15 mM) forms starting from 1mM naphthalene-2,3-dicarboxyaldehyde.