Biomedical subjects

Sebastiaan Eeltink

Publications and source records attributed to Sebastiaan Eeltink.

12 recordsLinked to original sources

PubMed2007

Recent advances in the control of morphology and surface chemistry of porous polymer-based monolithic stationary phases and their application in CEC.

This review focuses on developments in the field of polymer-based monolithic columns for CEC published in the literature since the beginning of the year 2005. The possibility of in-situ preparation as well as easy control over their porous properties and surface chemistries clearly make monolithic separation media an attractive alternative to capillary columns packed with particles. Different variables such as polymerization conditions, morphology, and surface chemistry are shown to directly affect performance of monolithic capillary columns in terms of efficiency, analysis time, and retention.

Capillary Electrochromatography↗

PubMed2006

Stability and repeatability of capillary columns based on porous monoliths of poly(butyl methacrylate-co-ethylene dimethacrylate).

Monolithic poly(butyl methacrylate-co-ethylene dimethacrylate) capillary columns have been prepared via either thermally or photochemically initiated polymerization of the corresponding monomers and the repeatability of their preparation has been explored. Three separate batches of 5 columns each were prepared using thermal and photochemical initiation for a total of 30 columns. All 30 capillary columns were tested in liquid chromatography-electrospray ionisation mass spectrometry mode for the separation of a model mixture of three proteins--ribonuclease A, cytochrome c and myoglobin. Excellent repeatability of retention times was observed for the proteins as evidenced by relative standard deviation (RSD) values of less than 1.5%. Somewhat broader variations with RSD values of up to 10% were observed for the pressure drop in the columns. The stability of retention times was also monitored using a single monolithic column and no significant shifts in either retention times or back pressure was observed in a series of almost 2200 consecutive protein separations.

Chromatography, High Pressure Liquid↗

PubMed2006

Selection of comparison criteria and experimental conditions to evaluate the kinetic performance of monolithic and packed-bed columns.

The present study concerns the problem of finding appropriate experimental conditions and comparison criteria to assess the kinetic performance of LC supports with different sizes or morphologies. A general procedure, based on evaluating each support for its own optimal mobile-phase composition, is proposed. The practical elaboration of the procedure is illustrated using the specific case of a capillary LC separation of a series of polycyclic aromatic test compounds employing silica-monolith capillary columns and capillary columns packed with 6-microm porous particles. To compare the systems for their ability to yield the fastest critical-pair separation, plate-height measurements are transformed into an effective plate number kinetic plot, i.e., a plot of the extrapolated retention time divided by the square of the extrapolated effective plate number (t(R)/N(eff)(2)) versus N(eff). This type of data representation provides a direct and universal basis to compare the kinetic performance of different LC supports and it corrects for differences in retention strength arising from different phase ratios.

Chromatography, Liquid↗

PubMed2006

Recent applications in capillary electrochromatography.

In this review research papers on the application of CEC are summarized that have been published between May 2003 and May 2005. First, a short overview is given of trends and developments in CEC that may increase the applicability of the separation technique. Next, application-oriented research using CEC is described in biochemical studies, including proteomics and genomics, in the analysis of food and natural products, and in pharmaceutical, industrial, and environmental analysis.

Animals↗

PubMed2006

Open-tubular capillary columns with a porous layer of monolithic polymer for highly efficient and fast separations in electrochromatography.

Open-tubular columns for CEC separations having inner-wall coated with a thin layer of porous monolithic polymer have been studied. A two-step process including (i) UV-initiated polymerization leading to a layer of porous poly(butyl methacrylate-co-ethylene dimethacrylate), and (ii) UV-initiated grafting of ionizable monomers appear to be well suited for the preparation of these columns. The thickness of the porous polymer layer is controlled by the percentage of monomers in the polymerization mixture and/or length of the irradiation time. The layer thickness significantly affects retention, efficiency, and resolution in open-tubular CEC. Under optimized conditions, column efficiencies up to 400,000 plates/m can be achieved. Use of higher temperature and application of pressure enables a significant acceleration of the open-tubular CEC separations.

Capillary Electrochromatography↗

PubMed2006

Determination of major carotenoids in vegetables by capillary electrochromatography.

A simple and rapid method for the isocratic separation and determination of carotenoids (carotenes and xanthophylls) in vegetables by CEC is described. The capillary column (100 microm ID, 25 cm effective length) was packed with 3 microm Hypersil ODS particles. The optimized mobile phase contained 60% ACN, 35% THF and 5% of a 5 mM Tris aqueous buffer of pH 8. beta-Carotene, lycopene and lutein were separated with efficiencies of 66 000-128 000 plates/m within a short time (less than 12 min for the last peak eluted, 13/13'-cis-beta-carotene). An excellent resolution of the three carotenoids, as well as partial resolution of their geometrical isomers, was achieved. Application to the determination of the analytes in carrot, tomato, spinach and corn was demonstrated.

Carotenoids↗

PubMed2005

Performance limits of monolithic and packed capillary columns in high-performance liquid chromatography and capillary electrochromatography.

A method is proposed for the comprehensive characterization and comparison of columns in the high-performance liquid chromatographic (HPLC) and capillary electrochromatographic (CEC) modes. Using this approach, column parameters such as the number of plates, the eddy-diffusion and mass-transfer contributions to peak broadening, the permeability, and the analysis time are incorporated in a single graph and a comparison in terms of efficiency and speed is obtained. The chromatographic performance of silica-based and polymer-based monolithic capillary columns is discussed and a comparison is made with the performance of packed columns. Also, the potential of ultra-high-pressure liquid chromatography is discussed in this context. In the HPLC mode, the best results were obtained with silica monoliths; in the CEC mode, the low-density methacrylate-ester-based monoliths showed the best performance.

Chromatography, High Pressure Liquid↗

PubMed2005

Tailoring the morphology of methacrylate ester-based monoliths for optimum efficiency in liquid chromatography.

Methacrylate ester-based monolithic stationary phases were prepared in situ in fused-silica capillaries and simultaneously in vials. The influence of the composition of the polymerization mixture on the morphology was studied with mercury intrusion porosimetry, scanning electron microscopy, and nitrogen adsorption measurements. A high-density porous polymeric material with a unimodal pore-size distribution was prepared with 40 wt % monomers and 60 wt % solvent in the mixture. A low-density material, prepared with a 20:80 ratio of monomers versus pore-forming solvent, showed a bimodal pore-size distribution and a much finer structure than the high-density monolith. The characteristic pore size could be controlled by changing the ratio of pore-forming solvents. With increasing solvent polarity, both the pore size and the dimension of the globules increased. The best efficiency in the CEC mode was obtained with an average pore size of 600 nm. Low-density monoliths exhibited lower A- and C-terms than high-density monoliths. With the optimal monolithic material, a minimum plate height of 5 mum could be obtained. The low-density monolith also performed better in the HPLC mode, giving a minimum plate height of 15 mum and a much higher flow permeability than that of the high-density material.

Journal Article↗

PubMed2005

Practical aspects of using methacrylate-ester-based monolithic columns in capillary electrochromatography.

Methacrylate-ester-based monoliths containing quaternary ammonium groups were prepared in situ in capillary columns and in simultaneous experiments in vials, employing thermal initiation. The chromatographic properties of the monoliths were determined with capillary electrochromatography (CEC), and their morphology was studied with mercury-intrusion porosimetry on the bulk materials. Materials with different, well repeatable pore-size distributions could be prepared. A satisfactory column-to-column and run-to-run repeatability was obtained for the electro-osmotic mobility, the retention characteristics (k-values) and the efficiency on the columns prepared and tested in the CEC mode. A relatively high electro-osmotic flow was observed in the direction of the positive electrode. The electro-osmotic mobility was found to be influenced only marginally by mobile-phase parameters such as the pH, ionic strength, and acetonitrile content. The retention behavior of the monolithic columns was similar to that of columns packed with C18-modified silica particles. Columns could be prepared with optimum plate heights ranging from 6 microm for unretained compounds to 20 microm for well retained (k=2.5) polyaromatic hydrocarbons. However, for specific analytes a - still unexplained - lower chromatographic column efficiency was observed.

Chromatography, Micellar Electrokinetic Capillary↗

PubMed2004

Study of the influence of the aspect ratio on efficiency, flow resistance and retention factors of packed capillary columns in pressure- and electrically-driven liquid chromatography.

The influence of the aspect ratio, rho (rho = column diameter/particle diameter), on column parameters such as efficiency, retention factors and flow resistance was studied in both high-performance liquid chromatography and capillary electrochromatography with packed capillary columns. In order to compare the true efficiencies of different columns, a procedure to account for external band broadening was applied. High efficiencies (reduced plate height h approximately 2) were obtained with capillary columns with internal diameters of 150-, 100-, and 75-microm, packed with 10-microm particles. In contrast to previous reports in the literature, no significant improvements in efficiency or flow resistance were observed when the aspect ratio of such columns was decreased. Our observations suggest that the wall effect in these types of columns is not significant. When the aspect ratio was decreased by increasing the particle size, a decrease in reduced plate height was observed. However, the results of flow resistance measurements showed that the latter effect should be attributed to differences in packing and particle batch quality rather than to differences in the aspect ratio.

Chromatography, High Pressure Liquid↗

PubMed2004

Comparison of the efficiency of microparticulate and monolithic capillary columns.

A comparison is made between the efficiency of microparticulate capillary columns and silica and polymer-based monolithic capillary columns in the pressure-driven (high-performance liquid chromatography) and electro-driven (capillary electrochromatography) modes. With packed capillary columns similar plate heights are possible as with conventional packed columns. However, a large variation is observed in the plate heights for individual columns. This can only be explained by differences in the quality of the packed bed. The minimum plate height obtained with silica monolithic capillary columns in the HPLC mode is approximately 10 microm, which is comparable to that of columns packed with 5-microm particles. The permeability of wide-pore silica monoliths was found to be much higher than that of comparable microparticulate columns, which leads to much lower pressure drops for the same eluent at the same linear mobile phase velocity. For polymer-based monolithic columns (acrylamide, styrene/divinyl benzene, methacrylate, acrylate) high efficiencies have been found in the CEC mode with minimum plate heights between 2 and 10 microm. However, in the HPLC mode minimum plate heights in the range of 10 to 25 microm have been reported.

Chromatography, High Pressure Liquid↗

PubMed2003

Recent applications in capillary electrochromatography.

The most recent and important applications in capillary electrochromatography (CEC) are summarized, covering literature published since May 2001. A selection of new developments in stationary phases for CEC is highlighted, and enantiomeric separations and chiral stationary phases are discussed. Also, CEC applications of biological molecules, pharmaceuticals, and applications in the field of industrial and environmental analysis are summarized. For this review three modes of CEC were taken into account, i.e., packed-column CEC, CEC using monolith technology, and open-tubular CEC.

Amino Acids↗