Determination of primary and secondary aliphatic amines by N-hydroxysuccinimidyl 4,3,2 '-naphthapyrone-4-acetate and reversed-phase high-performance liquid chromatography

A simple procedure for precolumn fluorescence derivatization of low molecul ar weight primary and secondary aliphatic amines with N-hydroxysuccinimidyl 4,3,2 ' -naphthapyrone-4-acetate is described. The derivatization reaction proceeds in aqueous solution at room temperature with a sodium borate buff er of pH 8.50 and gives corresponding stable fluorescent derivatives of pri mary amines and secondary amines within 5 and 15 min, respectively. Separat ion of eight-derivatized aliphatic amines has been optimized on a C-8 colum n with complete resolution in less than 15 min by isocratic elution with 56 % aqueous methanol as the eluent. The derivatives were detected spectrofluo rimetrically at 422 urn with excitation at 353 nm. The detection limits (si gnal-to-noise ratio = 3) of primary amines and secondary amines can reach f mol level for an injection volume of 20 mul. As a preliminary application, the method was used to determine aliphatic amines in lake water. (C) 2001 E lsevier Science B.V. All rights reserved.