Micellar electrokinetic capillary chromatography (MECC) separations and analyses of biological samples on a planar glass microchip capillary electrophoresis device with laser-induced fluorescence solute detection are discussed. A cyclic channel system which permits dead volume free repeated column switching and thus the use of various channel lengths together with a relatively low applied separation voltage is described. It features an unbiased, dead volume free electrokinetic sample inlet system of approximately 12 pL. Because of the small cross section and favorable heat dissipation in glass microstructures, MECC separations with an electric field strength of up to 2000 V/cm achieving efficiencies of submicrometer plate heights can be performed. After a separation length of 2 cm, six fluorescein isothiocyanate labeled amino acids are shown to be separable within a few seconds and with an imprecision for peak areas (or heights) and detection times of < 2% and < 0.5%, respectively. Without application of electrokinetic solute stacking, the detection limit of fluorescein isothiocyanate labeled arginine is 3.3 nM, corresponding to approximately 40 zmol injected. Furthermore, the feasibility of directly applying human urine and serum samples onto the uncoated channel system is demonstrated and first data of the successful performance of a chip-based MECC immunoassay for serum theophylline are presented. Compared to MECC in conventional fused-silica capillaries, MECC analyses on microchips can be performed 1-2 orders of magnitude faster, with higher efficiency and at no expense of accuracy and precision. Furthermore, versatility is shown to be much increased with the use of a cyclic rather than a single-path channel system. The MECC separation efficiency of fluorescein isothiocyanate labeled amino acids is shown to be comparable to that obtained by gel electrophoresis performed in the same chip layout.
Amines can easily be derivatized with fluorescein isothiocyanate isomer I (FITC) and analyzed by capillary electrophoresis (CE) using alkaline buffers with or without dodecyl sulfate micelles. This paper reports the CE analysis of FITC-derivatized amphetamine, methamphetamine, 3,4-methylenedioxymethamphetamine and beta-phenylethylamine in human urine using chip-based and fused-silica capillary instrumentation with laser-induced fluorescence detection. Data obtained via direct labeling of fortified urine are compared to those generated after FITC labeling of urinary extracts that were prepared by solid-phase extraction using a copolymer phase. For a urine volume of 5 mL with a "spiked amine": FITC ratio of 1:250, the latter approach was found to provide a sensitivity that is relevant for toxicological drug screening and confirmation (about 200 ng/mL urine). With direct labeling of 10 microL urine that was alkalinized and diluted for derivatization, the limit of identification was determined to be about 10 microg/mL, a value that is too high for practical purposes. Compared to fused-silica capillaries, electrophoresis in microstructures is shown to provide faster separations and higher efficiencies without loss of accuracy and precision.
The electrokinetic separation behavior of positively chargeable substances with various hydrophobicities over the pH range 2-11 is described, using plain aqueous buffers, binary media with organic solvents, and systems with surfactants at concentrations between 0.0005 and 75 mM or other buffer additives. Examples studied include dextromethorphan, methadone, ketoconazole, itraconazole, amiodarone, and desethyiamiodarone, compounds
During the past decade, capillary electrophoresis (CE) emerged as a promising, effective and economic approach for separation of a large variety of substances, including those encountered in clinical and forensic analysis. Reliable and automated CE instruments became commercially available and promoted the exploration of an increasing number of CE methods and fields of application. The widespread applicability of CE, its enormous separation power and high-sensitivity detection schemes make this technology an attractive and promising tool. This review discusses the principles and important aspects of CE-based assays and provides an overview of the key achievements encountered with CE in clinical and forensic analysis, including those associated with the analysis of serum proteins, hemoglobin variants, drugs and nucleic acids. Validated assays, interesting applications and future trends in clinical and forensic analysis are also discussed.
The data presented in this paper demonstrate that it is possible to fill a cyclic micrometer‐dimensional channel system formed in untreated glass with an acrylamide solution, to produce a gel matrix consisting of linear, noncrosslinked polyacrylamide (LPA) and to perform highly reproducible capillary gel electrophoretic separations with submicrometer plate heights within a few seconds. In such a system, no fluid flows are present and mass transport is based upon electrophoresis and diffusion only. Using a video system, an 8% LPA matrix and fluorescein isothiocyanate labelled amino acids as test compounds, sample introduction and separation could be visualized and volume‐defined, unbiased electroinjection from a 12 pL intersection of two channels via application of counter voltages could be optimized. Using laser‐induced fluorescence solute detection, the cyclic gel‐filled microstructure is shown to permit stable cyclic operation, an effective approach which simulates longer capillaries with improved separations at relatively low voltages. Furthermore, employing fluorescent phosphorothioate anti‐sense oligonucleotides, a 10% LPA matrix is demonstrated to be able to sieve and thereby separate oligonucleotides comprising more than five bases. © 1996 John Wiley & Sons, Inc.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.