Analytical Sciences

Fluorous and Fluorogenic Derivatization for Selective Liquid Chromatographic Analysis of Cyanide in Human Plasma

Ryoko TOMITA, Tadashi HAYAMA, Nao NISHIJO, and Toshihiro FUJIOKA

Faculty of Pharmaceutical Sciences, Fukuoka University, 8-19-1 Nanakuma, Johnan, Fukuoka 814-0180, Japan

A liquid chromatographic (LC) method with fluorous derivatization for the determination of cyanide in human plasma is described. In this method, the cyanide was transformed to a fluorous and fluorogenic compound by derivatizing with 2,3-naphthalenedialdehyde and perfluoroalkylamine reagent under mild reaction conditions (a reaction time of 5 min at room temperature). The obtained derivative was successfully retained on the perfluoroalkyl-modified LC column with the use of a high concentration of organic solvent in the mobile phase, whereas non-fluorous derivative was hardly retained, followed by fluorometric detection at excitation and emission wavelengths of 420 and 490 nm, respectively. Under the optimized conditions, the limit of detection and the limit of quantification for cyanide in a 5-μL injection volume were 1.3 μg/L (S/N = 3) and 4.4 μg/L (S/N = 10), respectively. The recovery from spiked human plasma was achieved in the range of 54 – 90% within a relative standard deviation of 3.5%. The feasibility of this method was further evaluated by applying it to the analysis of human plasma samples.

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