Abstract
This paper describes the preparation of the acid fluoride, 7-dimethylaminocoumarin-3-carbonyl fluoride (MACF), and a preliminary investigation as a precolumn fluorescence derivatization reagent for primary and secondary amines in high performance liquid chromatography (HPLC). Amines were derivatized quantitatively into the corresponding fluorescent amides by treating with MACF at 20°C for 30 s in aqueous 70% acetonitrile solution in the presence of 4-dimethylaminopyridine. The derivatives were subjected to HPLC on a TSK gel ODS-80TM column (4.6 mm i.d.×250mm, 5μm) with methanol-water (10 : 3) as the mobile phase and monitored with an excitation wavelength of 415 nm and an emission wavelength of 470 nm. The detection limits of amines were 0.6-4.6 pmol for an injection volume of 10μl.
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