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A new fluorogenic reagent for labelling carboxylic acids in HPLC
Gikas, E.
Derventi, M.
Panderi, I.
Vavayannis, A.
Kazanis, M.
Parissi-Poulou, M.
A new fluorescent reagent for carboxylic acids, N-(4-bromomethyl-7-hydroxy-2-oxo-2H-6-chromenyl) bromoacetamide (Br-MAMC) was synthesized. The fluorescence properties of Br-MAMC were monitored for a series of polar and non-polar solvents. Its applicability as a precolumn derivatization reagent in high-performance liquid chromatography was examined using undecylenic acid as a model compound. The derivatization reaction was performed in acetone in the presence of potassium carbonate and the crown ether, 18-crown-6 at 30°C for 20 min. The reaction solution was subjected to a high-performance liquid chromatographic procedure on a reversed-phase ODS C-18 column (150 × 4.6mm i.d., 5μm) with a mobile phase of acetonitrile and water (75:25, v/v) at a flow rate of 1.0 mL/min. The derivative of undecylenic acid was detected fluorometrically; excitation and emission wavelenghts were set at 345 and 435 nm, respectively. The detection limit of the method was determined to be 12.5 pg (signal to noise ratio = 3) of the derivatized undecylenic acid on column. The derivatives of butyric, hexanoic, octanoic, decanoic, and dodecanoic acid with Br-MAMC were also well resolved with a mobile phase of acetonitrile and water (60:40, v/v).
(EN)
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