Liquid chromatography of aromatic amines with photochemical derivatization and tris(bipyridine)ruthenium(III) chemiluminescence detection

We have shown by flow injection that tris(bipyridyl)ruthenium(III) [Ru(bpy)(3)(3+)] chemiluminescence (CL) detection of some aromatic amines can be enhanced by on-line photochemical derivatization. Two of the aromatic amino acids, tryptophan, and tyrosine as well as the peptide phenylalanine-alanine and other primary aromatic amines such as L-dopa, phentermine, and tryptamine upon irradiation with UV light are found to give an increased CL signal on the order of 4-9 times that for nonirradiated compounds. For benzylamine, phenethylamine, and phenylalanine, the improved CL detectability upon photolysis is about 15-16 times better. Chemiluminescence detection limits of the photolyzed compounds are generally 2-20 pmol, significantly better than those by UV-Vis detection at 254 nm. GC-MS work has been done to identify the products of some of the photolysis reactions and explain the enhanced CL detectability. The fact that other aromatic amines without a one or two carbon spacer from the aromatic ring to the amine group such as aniline, m- and p-phenylenediamine, and N,N'-dimethylaniline did not show any CL signal improvement upon irradiation with UV light suggests that there is some selectivity in the reaction. CL detection of aromatic amino acids after on-line photochemical derivatization and HPLC has been shown. (C) 1998 Elsevier Science B.V. All rights reserved.