Serge Pilard

Identification of Li-Based Electrolyte Degradation Products Through DEI and ESI High-Resolution Mass Spectrometry

The nature and composition of gel-like organic films forming during the cycling of Li-based cells functioning through a conversion reaction process were investigated. Besides infrared techniques, both desorption electron impact (DEI) and electrospray ionization (ESI) mass spectrometry were used to study the large amounts of films obtained after extended cycling at 55°C. We give direct evidence for the formation, depending on the type of electrolytes used that differ by the nature of either the Li-based salt (LiPF 6 , LiCF 3 SO 3 ) or solvents (dimethyl carbonate, propylene carbonate, ethylene carbonate, and their mixtures), of either phosphate-ending PEG-(polyethylene glycol) type chains, PEG chains (CH 2 -CH 2 -O) n , or polypropylene glycol chains (CH(CH 3 )-CH 2 -O) n with n values ranging from 1 to 9, and also trimethyl phosphate. The reaction schemes involving either electrochemical or chemical processes are proposed to describe the formation of such species.

The Arabidopsis sweetie mutant is affected in carbohydrate metabolism and defective in the control of growth, development and senescence

SUMMARY Sugars modulate many vital metabolic and developmental processes in plants, from seed germination to flowering, senescence and protection against diverse abiotic and biotic stresses. However, the exact mechanisms involved in morphogenesis, developmental signalling and stress tolerance remain largely unknown. Here we report the characterization of a novel Arabidopsis thaliana mutant, sweetie, with drastically altered morphogenesis, and a strongly modified carbohydrate metabolism leading to elevated levels of trehalose, trehalose-6-phosphate and starch. We additionally show that the disruption of SWEETIE causes significant growth and developmental alterations, such as severe dwarfism, lancet-shaped leaves, early senescence and flower sterility. Genes implicated in sugar metabolism, senescence, ethylene biosynthesis and abiotic stress were found to be upregulated in sweetie. Our physiological, biochemical, genetic and molecular data indicate that the mutation in sweetie was nuclear, single and recessive. The effects of metabolizable sugars and osmolytes on sweetie morphogenesis were distinct; in light, sweetie was hypersensitive to sucrose and glucose during vegetative growth and a partial phenotypic reversion took place in the presence of high sorbitol concentrations. However, SWEETIE encodes a protein that is unrelated to any known enzyme involved in sugar metabolism. We suggest that SWEETIE plays an important regulatory function that influences multiple metabolic, hormonal and stress-related pathways, leading to altered gene expression and pronounced changes in the accumulation of sugar, starch and ethylene.

Development and formulation of a 0.2% oral solution of midazolam containing γ-cyclodextrin

In absence of dedicated children formulation, intravenous formulations of midazolam, which exhibit strong bitterness, are occasionally used for oral or sublingual administration. In order to improve the quality and the acceptance by children of a midazolam anesthesia premedication, a new 0.2% (w/v) aqueous solution for oral administration has been prepared. The final formulation was obtained by the adjunction of a sweetener (sucralose), an aroma (orange aroma) and gamma-cyclodextrin to a citric acid solution of midazolam. The gamma-cyclodextrin forms an inclusion complex with the hydrophobic midazolam as evidenced using nuclear magnetic resonance spectroscopy (stoichiometry 1:1, K=283 M(-1)). A sterile filtration method was selected for the formulation microbial preservation using liquid chromatography coupled to high resolution mass spectrometry (LC-HRMS). Finally, a routine high performance liquid chromatography (HPLC) method is proposed for the quantitative determination of global midazolam amount in the pharmaceutical preparation.

Gas chromatography/Fourier transform infrared/mass spectrometry coupling: a tool for Li-ion battery safety field investigation

As electric vehicles may have a positive impact on global warming, worldwide endeavour is devoted to improving the performance, durability and safety of Li-ion batteries considered as the most promising technology. To help the characterisation and identification of volatile compounds released upon batteries ageing or during a system malfunction-induced thermal event, we implemented the coupling of the gas chromatography (GC) technique with mass spectrometry (MS) and Fourier transform infrared (FTIR) analytical tools. Through two detailed examples related to the thermal runaway phenomenon and the battery swelling, this paper provides evidence that the complementarities of these techniques allow us to detect and then accurately identify a vast array of volatile molecules ensuing from electrochemically/chemically driven electrolyte degradation. Hence, this GC/FTIR/MS equipment will be powerful in studying the impact of new electrolyte molecules on the battery functioning or safety and in assessing its degradation state after long-term or unexpected premature capacity loss.

Identification of an N-(hydroxysulfonyl)oxy metabolite using in vitro microorganism screening, high-resolution and tandem electrospray ionization mass spectrometry

Preliminary metabolic profiling of a drug under pre-clinical development revealed the presence of a minor unknown metabolite with a positive ion electrospray ionization (ESI) mass spectrum identical to that of the unchanged compound. Since the low concentration of the compound did not allow any additional experiments, preparative bioconversion using fungi was used to obtain a substantial amount of the molecule. Negative ion ESI-MS and tandem mass spectrometry (MS/MS) in combination with accurate mass measurements obtained on a quadrupole/time-of-flight instrument (Q-TOF) led to the positive identification of a hydroxylamide sulfoconjugated metabolite. Copyright

Benefits of Methylated Cyclodextrins in the Development of Midazolam Pharmaceutical Formulations

Midazolam (MDZ) is a benzodiazepine commonly administered in preanesthesia of children by oral or by sublingual routes. To mask its bitter taste and enhance its aqueous solubility, we already developed a 0.2% (w/v) MDZ oral solution containing γ-cyclodextrin (γ-CD), which proves to be better accepted by children in pediatrics at University Hospital of Amiens. To improve the MDZ solubility, its closed form proportion in acidobasic equilibrium and its chemical stability, nuclear magnetic resonance, liquid chromatography-electrospray-high-resolution mass spectrometry, and tandem mass spectrometry methods were used to highlight the advantages of using partially methylated CD (2,6 di-O-methyl-β-cyclodextrin) and randomized methylated-β-cyclodextrin (RAMEB). The formation of 1:1 inclusion complex offered an improvement of the MDZ solubility and an increase of the closed and pharmacologically active form with a 33% gain when compared with the aqueous solution without CD. It was also demonstrated that RAMEB had a protecting effect on the MDZ degradation because it was found in almost 95% of remaining MDZ solution after 3 months at 40°C.

Tropane alkaloid profiling of hydroponic Datura innoxia Mill. Plants inoculated with Agrobacterium rhizogenes.

INTRODUCTION Hydroponics has been shown as a possible way to produce high quality plant biomass with improved phytochemical levels. Nevertheless, effects of plant biotic and abiotic environment can lead to drastic changes and plant growth conditions must be optimised. OBJECTIVE To evaluate how much microbes and Agrobacterium rhizogenes TR7 wild strain may affect the tropane alkaloid profile in Datura innoxia Mill. plants cultivated in hydroponic conditions. METHODOLOGY Datura innoxia Mill. plants were cultivated in hydroponic with sterile or non-sterile conditions. For half of the non-sterile plants, Agrobacterium rhizogenes TR7 strain was added to the nutrient solution for hydroponics. The tropane alkaloid content of leaves and roots was analysed by UFLC/ESI-HRMS and MS/MS. The metabolite profiles were compared using partial least square-discriminant analysis. RESULTS In sterile conditions, aerial parts contained more scopolamine than the roots. However, the diversity of tropane alkaloids was greater in roots. Furthermore, 21 known compounds and four non-elucidated tropane alkaloids were found. The tropane alkaloid profile was shown to be statistically different between sterile and non-sterile hydroponic conditions. The levels of 3-acetoxy-6-hydroxytropane and 3-hydroxylittorine were higher in plants inoculated with A. rhizogenes. Five other tropane compounds were found in higher amounts in non-axenic control plants. Hyoscyamine and scopolamine total contents were much higher in the whole plant co-cultivated with A. rhizogenes TR7 than in controls. Furthermore, the leaves and roots of axenic plants contained more alkaloids than non-sterile ones. CONCLUSION In hydroponic conditions, microbes induced variations of the phytochemical levels. Addition of A. rhizogenes TR7 into the nutrient solutions improved the total hyoscyamine and scopolamine production.

Trehalose determination in linseed subjected to osmotic stress. HPAEC-PAD analysis: an inappropriate method

Trehalose is a non-reducing disaccharide involved in stress tolerance in plants. To understand better the role of trehalose in the osmotic stress response in linseed (Linum usitatissimum), trehalose content in leaves was studied. First, the method commonly used for sugar determination, high performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD), gave unsatisfactory results and the separation efficiency could not be improved by varying the elution conditions. The same problem was also found in the model plant: Arabidopsis thaliana. After clearly highlighting a co-elution of trehalose in these two species by a trehalase assay and liquid chromatography-high resolution mass spectrometry analysis, gas chromatography-mass spectrometry (GC-MS) was used as the analytical method instead. These results confirmed that trehalose content is currently overestimated by HPAEC-PAD analysis, approximately 7 and 13 times for A. thaliana and linseed respectively. Thus GC-MS gave more satisfactory results for trehalose quantification in plants. With this method, trehalose accumulation was observed in linseed during an osmotic stress (-0.30 MPa), the quantity (31.49 nmol g(-1) dry weight after 48 h) appears too low to assign an osmoprotector or osmoregulator role to trehalose in stressed linseed.

The effect of pymetrozine (Plenum WG-50®) on proboscis extension conditioning in honey bees (Apis mellifera: Hybrid var. Buckfast)

Experiments are designed to examine the effects of pymetrozine (Plenum WG-50), a recent systemic pesticide of the pyridine-azomethin family, on Pavlovian conditioning of harnessed foragers. In one set of experiments bees learned a task in which they associated a conditioned stimulus with feeding. A second set of experiments required the bees to learn a discrimination task. Within each experiment, bees received 5 μl of sucrose only, the recommended field dose of Plenum (5 μL of .3 gL(-1), .16 gL(-1) of pymetrozine measured), or 100 times the field dose of Plenum WG 50 thirty minutes prior to training (5 μL of 30 gL(-1), 14 gL(-1) of pymetrozine measured). The Plenum WG 50 was diluted with .88 M sucrose to facilitate the drinking of the pesticide. In addition to varying the concentration, we also investigated the effect of Plenum WG 50 on bees confined to an observation hive and a hive located outside. The results indicated that prior exposure to Plenum WG 50 affected Pavlovian conditioning only when bees were exposed to 100 times the recommended dosage.

Structural investigation of an exopolysaccharide substituted with a lactyl ether group produced by Raoultella terrigena Ez-555-6 isolated in the Chernobyl exclusion zone

Raoultella terrigena strain Ez-555-6, isolated from a root nodule of Medicago sativa harvested in the Chernobyl exclusion zone, produces a non-referenced high-molecular-mass exopolysaccharide (EPS). The structure of this EPS was determined using a combination approach including monosaccharide composition (GLC-FID, HPAEC-PAD), determination of glycosylation sites (GLC-EIMS) and 1D/2D NMR ((1)H, (13)C) and ESIMS (HR, MS/MS) studies of oligosaccharides obtained from mild acid hydrolysis. The EPS was found to be a charged pentasaccharide with a repeating unit composed of D-galactose, D-glucose, D-mannose and D-glucuronic acid (1:2:1:1). Lactic acid and O-acetyl substituents were localized on galactose and glucose residues, respectively, as presented in the following structure: