Separation of attogram terpenes by the capillary zone electrophoresis with fluorometric detection

• Published in: Journal of Chromatography A 2010-11, Vol.1217 (46), p.7288-7292
• Main Authors: Kubesová, Anna, Horká, Marie, Růžička, Filip, Šlais, Karel, Glatz, Zdeněk
• Format: Article
• Language: English
• Subjects: Biological and medical sciences; Candida albicans; Capillary electrophoresis; Farnesol; Fluorometric detection; Fundamental and applied biological sciences. Psychology; General aspects; Microbiology; Mycology; Poly(ethylene glycol)4-(1-pyrene)-butanoate; Terpenes
• ISSN: 0021-9673
• DOI: 10.1016/j.chroma.2010.09.038

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 expresion of a number of virulence factors, especially morphogensis 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.