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K Gaudin

Publications and source records attributed to K Gaudin.

8 recordsLinked to original sources
PubMed2006

Liquid chromatography on porous graphitic carbon with atmospheric pressure photoionization mass spectrometry and tandem mass spectrometry for the analysis of glycosphingolipids.

The study of several structural variations (the length, the degree of unsaturation and hydroxylation of the alkyl chains, the number and nature of osidic residues) helped understand the behaviour of neutral glycosphingolipids (GSLs) on porous graphitic carbon stationary phase (PGC). Atmospheric pressure photoionization mass spectrometry (APPI) and tandem mass spectrometry were used to perform the detection and the identification of molecular species in positive mode where [M+H](+) and [M-H(2)O+H](+) ions provided structural information on the fatty acid and the sphingoid base. The retention of GSLs increased with the hydrocarboneous volume of their alkyl chains and with the number of osidic residues in agreement with hydrophobic properties and polar retention effect of graphite, respectively. The presence of polar groups, such as OH-group or double bond within alkyl chains, decreased their retention. The coupling of chromatography on PGC with APPI tandem mass spectrometry detection appeared a powerful technique to discriminate isobaric molecules. Isobaric solutes differing by the position of two double bonds or by the repartition of hydrocarboneous skeleton were discriminated according to their chromatographic comportment or their mass spectrum, respectively. Among isobaric molecules, only few structures differing by the nature of osidic residue were not discriminated (i.e. glucosylceramide and galactosylceramide with similar ceramide skeleton were co-eluted and no difference in mass spectra was observed).

Atmospheric Pressure↗
PubMed2005

Modelling of ceramide interactions with porous graphite carbon in non-aqueous liquid chromatography.

Interactions of solutes on porous graphitic carbon (PGC) with non-aqueous mobile phases are studied by the linear solvation energy relationship (LSER). Studies have been carried out with eight binary mixtures composed of a weak solvent (acetonitrile or methanol) and a strong solvent (tetrahydrofuran, n-butanol, CH2Cl2, 1,1,2-trichloro-2,2,1-trifluoroethane). The systematic analysis of a set of test compounds was performed for each solvent mixture in isocratic mode (50:50). The results were compared to those obtained on PGC with hydro-organic liquids and supercritical fluids. They were then correlated with the observed retention behaviour of lipid compounds, more particularly ceramides.

Ceramides↗
PubMed2005

Direct determination of niflumic acid in a pharmaceutical gel by ATR/FTIR spectroscopy and PLS calibration.

A simple, rapid and convenient analytical method without sample handling procedure is proposed for the determination of niflumic acid in a pharmaceutical gel with attenuated total reflectance/Fourier transform infrared spectroscopy (ATR/FTIR). A partial least square (PLS) calibration model for the prediction of niflumic acid contents was developed using 81 and 27 spectra of standard gels as training and validation sets, respectively. The used spectral range of niflumic acid for the establishment of this model was 2300-1100 cm(-1). All spectra were obtained in the transmittance mode, then normalized and first derivative transformed. The model yielded a regression coefficient R2 equal to 1 for the training set and a root mean square error of prediction (RMSEP) equal to 0.2 for the validation set. The percentage recoveries of the method for the analysis of Niflugel ranged from 96.60 to 101.02%.

Anti-Inflammatory Agents, Non-Steroidal↗
PubMed2004

Optimisation of the separation of four major neutral glycosphingolipids: application to a rapid and simple detection of urinary globotriaosylceramide in Fabry disease.

A simple method for the separation of the four major neutral glycosphingolipids, present in all human tissue, was developed. This gradient normal phase-HPLC method utilises a polyvinyl alcohol bonded stationary phase and an evaporative light-scattering detection (ELSD). Screening pure solvents in a binary gradient elution mode allowed, in a first step, to assess the behaviour of the studied solutes and to select the solvents for further mobile phase optimisation. The proportion of the remaining solvents was defined to reach a maximal resolution. The reduction of the analysis time and the enhancement of the signal were obtained by optimising the gradient slope and the flow-rate. Optimal levels of triethylamine and formic acid (TEA-FA) for the enhancement of the evaporative light scattering detector response were established at 0.1% (v/v). Thus, the optimal conditions for the separation of the four glycosphingolipids was obtained with a gradient elution from a 100% chloroform to a 100% acetone:methanol (90:10 (v/v)) mobile phase at 0.2 ml min-1, using a 10% min-1 gradient slope. Finally, this method was applied to detect the excess of one of the neutral sphingolipids, namely globotriaosylceramide (Gb3) in the urine of patients affected with Fabry disease. A liquid-liquid extraction of the sediments obtained from an aliquot of only ten ml of urine proved sufficient to detect the excess of Gb3 present in both hemizygote and heterozygote patients. In all, the ability of our method to detect abnormal amounts of Gb3 in urinary sediments could allow the diagnosis of weakly symptomatic Fabry patients in large screening programs

Chromatography, High Pressure Liquid↗
PubMed2001

Structural analysis of commercial ceramides by gas chromatography-mass spectrometry.

A simple method using gas chromatography-mass spectrometry was applied to analyse structures of ceramides. Identification of trimethylsilylated ceramides were obtained in short analysis times (derivatization of ceramides in 30 min at room temperature and 20 min gas chromatography mass spectrometry run) even for complex mixtures. For example in ceramide Type III, 18 peaks were observed which represent 27 various structures. The coeluted compounds were ceramides containing the same functional groups and the same carbon number but with a different distribution on the two alkyl chains of the molecule. They were accurately differentiated by mass spectrometry. Therefore, 83 structures of trimethylsilylated ceramides were identified in 11 different commercial mixtures. For 52 structures of these, mass spectral data were not described in the literature, neither full mass spectra nor characteristic fragments.

Ceramides↗
PubMed2000

Retention behaviour of ceramides in sub-critical fluid chromatography in comparison with non-aqueous reversed-phase liquid chromatography.

This study was devoted to the development of an analytical method for ceramide analysis in packed subcritical fluid chromatography (pSubFC). Monofunctional grafted silica support was found to be more suitable for ceramide analysis. Five Kromasil columns were coupled and the parameters, temperature, pressure and percentage of organic modifier in CO2 were optimised, considering selectivity and analysis time. The final conditions were 31 degrees C, 6% of methanol (MeOH) and 13 MPa. In these conditions the selectivity for structural differences (methylene group, unsaturation or two different bases) were studied. As classically observed, the methylene selectivity decreased with the increase of the eluotropic strength. Moreover, unlike in non-aqueous reversed-phase liquid chromatography (NARP-LC), adding a further unsaturation and two further methylene groups on ceramide results to an increase of retention in pSubFC. Moreover, this last technique allowed to separate ceramides with the same total number of carbons containing unsaturated fatty acids, when the distribution of carbon number of the two chain is very different. These results had enabled to plot retention chart in order to predict ceramide structure in view to identify additional ceramide. This retention chart was finally compared with the one already obtained in NARP-LC.

Ceramides↗
PubMed1999

Postcolumn fluorescence as an alternative to evaporative light scattering detection for ceramide analysis with gradient elution in non-aqueous reversed-phase liquid chromatography.

Ceramide analysis was developed with gradient elution in non-aqueous reversed-phase liquid chromatography with evaporative light scattering detection (ELSD) or postcolumn fluorescence detection. Fluorescence detection (excitation, 360 nm; emission, 425 nm) after postcolumn formation of mixed assemblies between eluted ceramides and 1,6-diphenyl-1,3,5-hexatriene was developed. In comparison with ELSD, fluorescence detection allows a better detection of the minor species ceramide from ceramide type III (commercial mixture of non-hydroxy fatty acid-sphingosine) and appears to be more sensitive for quantitation of ceramides at low concentrations. The fluorescence response is linear over a wide range of injected amount of ceramide III (expressed as stearoyl-phytosphingosine): 10 ng to 1000 ng. The response of ELSD is non linear but can be linearized in double logarithmic coordinates for calculations over a narrow range, e.g. between 10 to 350 ng ceramide III injected. The lower quantitation limits of these two detectors are similar: 5 ng ceramide III was injected.

Ceramides↗