A polydimethylsiloxane/glass capillary electrophoresis microchip for the analysis of biogenic amines using indirect fluorescence detection.
A polydimethylsiloxane-glass capillary microchip is fabricated for the rapid analysis of a mixture of common biogenic amines using indirect fluorescence detection. Using a running buffer of phosphate and 2-propanol, and Rhodamine 110 as a background fluorophore, both co-ionic and counter-ionic systems are explored. Studies demonstrate the separation and analysis of cations using indirect fluorescence detection for the first time in a chip-based system. Resulting electrophoretic separations are achieved within a few tens of seconds with detection limits of approximately 6 microM. The reduced sample handling and rapid separations afforded by the coupling of indirect fluorescence detection with chip-based capillary electrophoresis provide a highly efficient method for the analysis and detection of molecules not possessing a chromophore or fluorophore. Furthermore, limits of detection are on a par with reported chip-based protocols that incorporate precolumn derivatisation with fluorescence detection. The current device circumvents lengthy sample preparation stages and therefore provides an attractive alternative technique for the analysis biogenic amines.
AstraZeneca/SmithKline Beecham Centre for Analytical Sciences, Imperial College of Science, Technology and Medicine, Department of Chemistry, South Kensington, London, UK.
Nigel P
NP
Beard
Andrew J
AJ
de Mello
D000818:Animals; D001679:Biogenic Amines / Q000032:analysis / Q000302:isolation & purification*; D004129:Dimethylpolysiloxanes; D019075:Electrophoresis, Capillary / Q000295:instrumentation* / Q000379:methods / Q000592:standards; D005453:Fluorescence; D005456:Fluorescent Dyes; D006801:Humans; D008904:Miniaturization; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012828:Silicones
D001679:Biogenic Amines; D004129:Dimethylpolysiloxanes; D005456:Fluorescent Dyes; D012828:Silicones; C013830:baysilon