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Silica Stationary Phase (silica + stationary_phase)
Selected AbstractsCEC-ESI ion trap MS of multiple drugs of abuseELECTROPHORESIS, Issue 7 2010Zeineb Aturki Abstract This article describes a method for the separation and determination of nine drugs of abuse in human urine, including amphetamines, cocaine, codeine, heroin and morphine. This method was based on SPE on a strong cation exchange cartridge followed by CEC-MS. The CEC experiments were performed in fused silica capillaries (100,,m×30,cm) packed with a 3,,m cyano derivatized silica stationary phase. A laboratory-made liquid junction interface was used for CEC-MS coupling. The outlet capillary column was connected with an emitter tip that was positioned in front of the MS orifice. A stable electrospray was produced at nanoliter per minute flow rates applying a hydrostatic pressure (few kPa) to the interface. The coupling of packed CEC columns with mass spectrometer as detector, using a liquid junction interface, provided several advantages such as better sensitivity, low dead volume and independent control of the conditions used for CEC separation and ESI analysis. For this purpose, preliminary experiments were carried out in CEC-UV to optimize the proper mobile phase for CEC analysis. Good separation efficiency was achieved for almost all compounds, using a mixture containing ACN and 25,mM ammonium formate buffer at pH 3 (30:70, v/v), as mobile phase and applying a voltage of 12,kV. ESI ion-trap MS detection was performed in the positive ionization mode. A spray liquid, composed by methanol,water (80:20, v/v) and 1% formic acid, was delivered at a nano-flow rate of ,200,nL/min. Under optimized CEC-ESI-MS conditions, separation of the investigated drugs was performed within 13,min. CEC-MS and CEC-MS2 spectra were obtained by providing the unambiguous confirmation of these drugs in urine samples. Method precision was determined with RSDs values ,3.3% for retention times and ,16.3% for peak areas in both intra-day and day-to-day experiments. LODs were established between 0.78 and 3.12,ng/mL for all compounds. Linearity was satisfactory in the concentration range of interest for all compounds (r2,0.995). The developed CEC-MS method was then applied to the analysis of drugs of abuse in spiked urine samples, obtaining recovery data in the range 80,95%. [source] Enantiomeric separation of mirtazapine and its metabolites by nano-liquid chromatography with UV-absorption and mass spectrometric detectionJOURNAL OF SEPARATION SCIENCE, JSS, Issue 14 2005Salvatore Fanali Abstract Mirtazapine (MIR) and two of its main metabolites, namely, 8-hydroxymirtazapine and N -desmethylmirtazapine, were separated in totheir enantiomers by nanoLC in a laboratory-made fused-silica capillary column (75 ,m ID) packed with a vancomycin-modified silica stationary phase. The simultaneous separation of the three couples of the studied enantiomers was achieved in less than 33 min, using an experimentally optimized mobile phase delivered in the isocratic mode. Optimization of the mobile-phase composition was achieved by testing the influence of the buffer pH and concentration, the water concentration, the organic modifier type and concentration, and on the retention and resolution of the analytes. The optimum mobile-phase composition contained 500 mM ammonium acetate pH 4.5/water/MeOH/MeCN, 1 : 14 : 40 : 45 v/v/v/v. Using a UV detector at 205 nm, the method was validated studying several experimental parameters such as LOD and LOQ, intraday and interday repeatability, and linearity. Good results were achieved: LOD and LOQ were in the range 5,15 and 10,40 ,g/mL, respectively (the highest value was obtained for the DEMIR enantiomers); correlation coefficients, 0.9993,0.9999; the intraday and interday precision was acceptable (RSD < 2%) using an internal standard. The method was tested for the separation of the studied enantiomers in an extracted (solid-phase) serum sample spiked with standard racemic mixture of MIR and its two metabolites. Finally, the nanoLC system was connected to a mass spectrometer through a nanoelectrospray interface and the MS, MS2, and MS3 spectra were acquired showing the potential of the system used for characterization and identification of the separated analytes. [source] Direct injection of 96-well organic extracts onto a hydrophilic interaction chromatography/tandem mass spectrometry system using a silica stationary phase and an aqueous/organic mobile phaseRAPID COMMUNICATIONS IN MASS SPECTROMETRY, Issue 23 2004Weng Naidong First page of article [source] Liquid chromatographic assay for the cyclic depsipeptide aplidine, a new marine antitumor drug, in whole blood using derivatization with trans -4,-hydrazino-2-stilbazoleBIOMEDICAL CHROMATOGRAPHY, Issue 1 2004Rolf W. Sparidans Abstract A sensitive bio-analytical assay for the depsipeptide aplidine in plasma has been modi,ed and tested for human whole blood samples. The adapted method is based on reversed-phase liquid chromatography and ,uorescence detection of the trans -4,-hydrazino-2-stilbazole derivative of the analyte. Aplidine is isolated from the matrix by solid-phase extraction on an octadecyl modi,ed silica stationary phase. After evaporation of the acetone eluate, the derivatization with the hydrazino reagent is performed in a water,acetonitrile mixture at pH = 4. The reaction mixture is injected directly into the chromatograph and the analyte is quanti,ed by ,uorescence detection at 410 and 560 nm for excitation and emission, respectively. The method has been validated in the 2,100 ng/mL range, with 2 ng/mL being the lower limit of quanti,cation. Precision and accuracy both meet the current requirements for a bioanalytical assay. The stability of aplidine in whole blood at ambient temperature and at 37°C is limited; recoveries in the range 60,85% were observed after 7 h. Further, adequate stability of aplidine in plasma at ,80 and ,20°C for 35 months could now be demonstrated. Copyright © 2003 John Wiley & Sons, Ltd. [source] True and Apparent Temperature Dependence of Protein Adsorption Equilibrium in Reversed-Phase HPLCBIOTECHNOLOGY PROGRESS, Issue 6 2002Szabelski The adsorption behavior of bovine insulin on a C8 -bonded silica stationary phase was investigated at different column pressures and temperatures in isocratic reversed-phase HPLC. Changes in the molar volume of insulin (, Vm) upon adsorption were derived from the pressure dependence of the isothermal retention factor ( k,). The values of , Vm were found to be practically independent of the temperature between 25 and 50 °C at ,96 mL/mol and to increase with increasing temperature, up to ,108 mL/mol reached at 50 °C. This trend was confirmed by two separate series of measurements of the thermal dependence of ln( k,). In the first series the average column pressure was kept constant. The second series involved measurements of ln( k,) under constant mobile-phase flow rate, the average column pressure varying with the temperature. In both cases, a parabolic shape relationship was observed between ln( k,) and the temperature, but the values obtained for ln k, were higher in the first than in the second case. The relative difference in ln( k,), caused by the change in pressure drop induced by the temperature, is equivalent to a systematic error in the estimate of the Gibbs free energy of 12%. Thus, a substantial error is made in the estimates of the enthalpy and entropy of adsorption when neglecting the pressure effects associated with the change in the molar volume of insulin. This work proves that the average column pressure must be kept constant during thermodynamic measurements of protein adsorption constants, especially in RPLC and HIC. Our results show also that there is a critical temperature, Tc , 53 °C, at which ln( k,) is maximum and the insulin adsorption process changes from an exothermic to an endothermic one. This temperature determines also the transition point in the molecular mechanism of insulin adsorption that involves successive unfolding of the protein chain. [source] Effect of coverage density and structure of chemically bonded silica stationary phases on the separation of compounds with various propertiesJOURNAL OF SEPARATION SCIENCE, JSS, Issue 6 2006Bogus, aw Buszewski Abstract The chemical character, geometry, and architecture of chemically formed surface layers determine interactions between stationary phase, analyte, and mobile phase, and therefore the retention mechanisms (partitioning, adsorption, ion exchange, steric exclusion) of separated analytes. These interactions also depend on the structure and chemical character of the solutes and the composition of the mobile phase. High-molecular-weight fullerenes (C60 and C70) and water-soluble selenium-containing peptides (833 and 2607 Da) were used for the evaluation of laboratory-prepared octadecyl stationary phases with high and low coverage density before and after end-capping. The aim of this work was to study differences in surface coverage density and homogeneity and conformational changes of chemically bonded moieties and the influence of these parameters on the separation of mixtures of selenopeptides and fullerenes with significantly different molecular masses. A topographical model of the chemically modified stationary surface is presented. [source] Hydride-based silica stationary phases for HPLC: Fundamental properties and applicationsJOURNAL OF SEPARATION SCIENCE, JSS, Issue 15 2005Joseph J. Pesek Abstract Silica hydride is a recent development in chromatographic support materials for HPLC where hydride groups replace 95% of the silanols on the surface. This conversion changes many of the fundamental properties of the material as well as the bonded stationary phases that are the result of further chemical modification of the hydride surface. The general approach for fabricating the silica hydride and subsequent bonded phases is reviewed. Properties of the silica hydride surface are compared to those of the standard material obtained in the preparation of most commercial HPLC stationary phases. Some unique chromatographic properties of hydride-based phases are described as well as some general application areas where these bonded materials may be used in preference to or have advantages not available from typical stationary phases. [source] Simultaneous determination of carotenoids, tocopherols, and ,-oryzanol in crude rice bran oil by liquid chromatography coupled to diode array and mass spectrometric detection employing silica C30 stationary phasesJOURNAL OF SEPARATION SCIENCE, JSS, Issue 14 2005Wolfgang Stöggl Abstract Crude rice bran oil contains tocopherols (vitamin E), carotenoids (vitamin A), and phytosterols, which possess antioxidant activities and show promising effects as preventive and therapeutic agents. The aim of this work was to establish methods and to compare C18 and C30 silica stationary phases in order to separate and detect tocopherols, carotenoids, and ,-oryzanol in one single run. Comparing RP-LC on silica C18 and C30, higher resolution between all target compounds was obtained using the C30 stationary phase. Methanol was used as eluent and the elution strength was increased by the addition of tert -butyl methyl ether for highly hydrophobic analytes such as ,-oryzanol. Detection was accomplished by diode array detection from 200 to 500 nm. Absorbance maxima were found at 295 nm for tocopherols, 324 nm for ,-oryzanol, and 450 nm for carotenoids. Furthermore, compounds were characterized and identified on the basis of their UV-spectra. Both RP systems were coupled to MS (LC-MS) by using an atmospheric pressure chemical ionization interface. [source] | |||||||||||||