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Separation of attogram terpenes by the capillary zone electrophoresis with fluorometric detection
Authors:Kubesová Anna  Horká Marie  Růžička Filip  Slais Karel  Glatz Zdeněk
Affiliation:1. Institute of Analytical Chemistry of the ASCR, v.v.i., Veve?í 97, 602 00 Brno, Czech Republic;2. Department of Biochemistry, Faculty of Science, Masaryk University Brno, Czech Republic;3. Department of Microbiology, Faculty of Medicine, Masaryk University Brno, Czech Republic
Abstract:An original method based on capillary zone electrophoresis with fluorimetric detection has been developed for the determination of terpenic compounds. The method is based on the separation of a terpenes dynamically labeled by the non-ionogenic tenside poly(ethylene glycol) pyrenebutanoate, which was used previously for the labeling of biopolymers. The background electrolytes were composed of taurine-Tris buffer (pH 8.4). In addition to the non-ionogenic tenside aceton and poly(ethylene glycol) were used as the additives. The capillary zone electrophoresis with fluorometric detection at the excitation wavelength 335 nm and the emission wavelength 463 nm was successfully applied to the analysis of tonalid, cholesterol, vitamin A, ergosterol, estrone and farnesol at level of 10(-17) mol L(-1). Farnesol, is produced by Candida albicans as an extracellular quorum-sensing molecule that influences expression of a number of virulence factors, especially morphogenesis and biofilm formation. It enables this yeast to cause serious nosocomial infections. The sensitivity of this method was demonstrated on the separation of farnesol directly from the cultivation medium.
Keywords:Capillary electrophoresis   Fluorometric detection   Poly(ethylene glycol)4-(1-pyrene)-butanoate   Terpenes   Farnesol   Candida albicans
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