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Biomedical subjects

A Baillet

Publications and source records attributed to A Baillet.

At least 19 recordsLinked to original sources

The benefit of the retrofitting of a conventional LC system to micro LC: a practical evaluation in the field of bioanalysis with fluorimetric detection.

The interests in liquid micro-chromatography (higher column efficiencies, increase in sensitivity) are now well established. The enhancement of fluorimetric response induced by the reduction of the inner diameter of columns (4.6, 3.0, 1.0 and 0.3 mm respectively) coupled with adapted detection cells to control the loss of efficiency (8 micro L for the two first columns and 100 nL for the two smaller ones) has been studied in the bioanalytical field, using the plasma determination of native fluorescent antibacterial agents: fluoroquinolones. Ten-fold enhancement of the signal can easily be obtained when substituting a 0.3 mm i.d. column and 100 nL detection cell for a 4.6 mm i.d. column, and 8 micro L detection cell. In addition to inner diameter reduction, the detection cell geometry appears to be an essential parameter to obtain the best enhancement of the recorded signal. Hence, the enhancement of signal with micro-chromatography with fluorimetric detection appears to be a compromise between column inner diameter and flow cell volume reduction.

Chromatography, Liquid↗

Assessment of the retention properties of poly(vinyl alcohol) stationary phase for lipid class profiling in liquid chromatography.

Potentialities of polymerized vinyl alcohol on silica gel were assessed for class separation of simple lipids, sphingolipids, glyceroglycolipids and phospholipids by high-performance liquid chromatography. A screening of pure solvents in binary gradient elution and a chemometric approach was used to define a rugged two segment linear gradient formed from four solvents for total lipid class separation. Triethylamine and formic acid were added in all mobile phase components for acidic phospholipid separation and evaporative light scattering response enhancement. Simple analytical procedures are described for the analysis of complex lipid materials.

Chromatography, High Pressure Liquid↗

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↗

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↗

Modulated release of triterpenic compounds from a O/W/O multiple emulsion formulated with dimethicones: infrared spectrophotometric and differential calorimetric approaches.

From the use of silicones within O/W/O multiple emulsions, we can expect, two principal advantages: (1) the silicones with the lowest molecular weight decrease the oily touch; (2) due to the large range of viscosity, this excipient should influence the skin distribution of actives after topical application. The purpose of our work is to highlight these advantages. Multiple emulsions were formulated with several dimethicones and with drug model. Firstly, the effects of different dimethicones incorporated within multiple emulsions were studied, through in vitro penetration results. Secondly, we investigated the residual film on the skin by Differential Scanning Calorimetry (DSC) and by Fourier Transform Infrared (FTIR) to determine its structure. Correlations were established between the silicone structure and the distribution of drugs in different skin levels or between the silicone structure and the percutaneous penetration. The incorporation of silicones within O/W/O multiple emulsions seems to be an efficient means of modulating the penetration and the distribution of drugs in the skin.

Animals↗

Quantitative HPLC analysis of sunscreens and caffeine during in vitro percutaneous penetration studies.

This report describes rapid analytical HPLC for the quantification of five UV filters (octyl methoxycinnamate, benzophenone-3, benzophenone-4, octyl triazone and octocrylene) and of caffeine in various skin layers (stratum corneum, dermis, epidermis and receptor fluid) and in cosmetic preparations. The predominant purpose of the study was to establish standard operating procedures for rapid analysis of the compounds in various skin samples. Particular attention was paid to the preparation of biological samples whose natural constitution could interfere with the quantitative analysis. Our methods used the isocratic chromatographic mode in an RP-HPLC with UV detection and did not involve centrifugation or evaporation. Our results were validated in terms of specificity, linearity, precision, accuracy and limits of detection and quantification. The first results, obtained after in vitro experiments, are presented in this report.

Adolescent↗

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↗

[Epidemiology of suicide attempts in Reunion Island].

In order to specify the epidemiological characteristics of attempted suicides on Reunion island, a quantitative study was carried out, using Emergency Room registers of all the hospitals on the island. This enabled collection of 9526 files. The results revealed a high proportion of suicide attempts (335/100,000/year), particularly prevalent in rural areas to the south of the island. There was a tendency towards over representation of women, though less distinct than in Europe, as well as an abnormally frequent use of pesticides or herbicides. Medication and violent means were not used as often as in France. These outcomes, in particular the very high frequency (especially in the rural areas of the south), along with the significance of medico-psychological factors, should be taken into account by anthropologists, linguists, clinical specialists, in order to help determine preventive measures.

Adolescent↗

Fosfomycin determination in serum by capillary zone electrophoresis with indirect ultraviolet detection.

Capillary zone electrophoresis with indirect ultraviolet detection was used for the determination of fosfomycin in serum. Running buffer consisted of a mixture of 200 mM sodium borate with 10 mM phenylphosphonic acid used as ultraviolet absorbing background electrolyte. Relationships between the pH of the buffer and the efficiency of the separation (migration times and selectivities) or the sensitivity of detection were investigated. The method was then validated over a 10-100 micrograms ml-1 concentration range to be applied to further therapeutic drug monitoring. The choice of ethylphosphonic acid as internal standard is discussed. The specificity and the linearity of the technique are demonstrated. The inter-day precision was satisfactory with a relative standard deviation of less than 2%. Accuracy was calculated with a standard error near 0.5 and 18% for 100 and 10 micrograms ml-1, respectively.

Electrolytes↗

Analysis of carbohydrate-mediated heterogeneity and characterization of N-linked oligosaccharides of glycoproteins by high performance capillary electrophoresis.

Capillary zone electrophoresis (CZE) has been investigated as an alternative method to analyze the carbohydrate moieties of glycoproteins. Carbohydrate-mediated microheterogeneity of the recombinant plasminogen activator (rt-PA) was examined. The glycoprotein was resolved in multiple electrophoretic species using CZE but the separation was complicated by adsorption of the molecules to the wall of the capillary. The influence of several parameters, such as pH, molarity of the buffer and addition of a cationic additive, on the separation of glycopeptides was investigated. High resolution and reproducible separations of rt-PA glycopeptides carrying hybrid and complex type chains were obtained using either a 100 mM phosphate buffer, pH 6.6, or a 100 mM Tricine buffer, pH 8.2, containing 1.25 mM of putrescine. N-Oligosaccharides from fetuin, t-PA and alpha 1-acid glycoprotein were separated within 20 min on the basis of both their sialic acid content and their structure. The use of an oligosaccharide fingerprinting technique, such as the present one, could have many applications in biotechnology to assess, for example, the consistency of production of a glycoprotein or for analytical glycoprotein chemistry.

Animals↗

Liquid chromatographic analysis of monosaccharides with phenylisocyanate derivatization.

Because of the lack of sensitivity in carbohydrate analysis, HPLC pre-column derivatization techniques which give strongly UV absorbing compounds have been reported. These techniques have the disadvantage of leading to several chromatographic peaks from each reducing sugar. To enhance both the sensitivity and the selectivity of such specific separation problems, a simplex procedure was applied to optimize the phenylisocyanate derivatization of monosaccharides: a major and stable derivative was formed under the optimal conditions established. The method was extended to deoxysugars and methylglycosides. The limit of detection was 0.2-1 ng for all sugars tested.

Chromatography, High Pressure Liquid↗

[Contribution of capillary electrophoresis in the analysis of molecules with pharmaceutical and biotechnological value].

Capillary electrophoresis entails a variety of separation modes which are: capillary zone electrophoresis, capillary gel electrophoresis, micellar electrokinetic capillary chromatography, isoelectric focusing and isotachophoresis. Many applications are already reported. The fundamentals of the various modes are discussed along and typical separations in the pharmaceutical and biotechnologic fields are presented. The advantages and disadvantages in comparison to other analytical techniques are also discussed.

Biotechnology↗

Simultaneous determination of gluconolactone, galactonolactone and galactitol in urine by reversed-phase liquid chromatography: application to galactosemia.

A reversed-phase liquid chromatographic assay was developed for the specific evaluation of metabolic by-products in the urine of galactosemic patients and based on the simultaneous determination of gluconolactone, galactonolactone and galactitol. The procedure involved a lyophilization step and the formation of phenylisocyanate derivatives, followed by injection directly into the chromatograph. Analytical results showed good selectivity, linearity, precision and accuracy. The method enabled the detection of levels as low as 0.05-0.1 ng, and compared favourably with other published techniques for the estimation of aldonic acids in biological fluids.

Chromatography, High Pressure Liquid↗

Liquid chromatographic method for the determination of the carbohydrate moiety of glycoproteins. Application to alpha 1-acid glycoprotein and tissue plasminogen activator.

A rapid procedure is described for the qualitative and quantitative analysis of the carbohydrate composition of glycoproteins by liquid chromatography with light-scattering detection. The analysis was carried out in three steps. First, the glycoprotein samples were purified by a two-step purification on a Sephadex G-25 column with a 90% yield. Second, the selectivity of the separation and the sensitivity of detection of monosaccharides, as methyl glycosides obtained by direct methanolysis of glycoproteins, were improved by modified simplex optimization of the methanolysis parameters (temperature, methanolic hydrochloric acid strength and reaction time) determined at 66 degrees C, 1.2 M and 8.1 h for alpha 1-acid glycoprotein (alpha-AGP) and 73 degrees C, 1.5 M and 12.5 h for tissue plasminogen activator (tPA). Finally, the method was applied to the determination of the carbohydrate moiety of the two N-glycosylated glycoproteins alpha-AGP and tPA.

Acetylglucosamine↗