Quantitative analysis of monofluoroacetate in biological samples by high-performance liquid chromatography using fluorescence labeling with 9-chloromethylanthracene

Abstract

A rapid and sensitive RP-HPLC method with fluorescence detection has been developed for the quantitative analysis of trace amounts of monofluoroacetate (MFA) in biological samples as serum, food and meat. 9-Chloromethylanthracene (9-CMA) is used as the fluorescence labeling reagent. Samples were extracted and reacted with 9-chloromethylanthracene together with tetrabutylammonium bromide as catalyst at 80 °C for 50 min to give a new fluorescent derivative as 9-methyleneanthracene monofluoroacetate (MA-MFA). The resulting MA-MFA was characterized with IR, 1H NMR, 13C NMR and MS. Chromatography separation is performed on an Agilent Hypersil ODS column with a fluorescent detector employed with the excitation and emission wavelengths as 256 nm and 412 nm, respectively. Optimal conditions for derivatization, fluorescence detection and chromatographic separation have been established. The novel method yields a good linear relationship when the MFA concentration in serum within 1 and 250 ng/mL ( r = 0.9988). The detection limit (signal-to-noise ratio = 3 with 2 μL injected) was 0.25 ng/mL. The practical applicability of this method was demonstrated by quantitative determination of MFA-Na in a blood sample from a person who had ingested the poison.