High-performance liquid chromatography of long chain aliphatic aldehydes with peroxyoxalate chemiluminescence detection utilizing a fluorogenic reagent, 4-(N,N-dimethylaminosulphonyl)-7-hydrazino-2,1,3-benzoxadiazole.

A peroxyoxalate chemiluminescence detection in high-performance liquid chromatography was examined for determination of long chain aliphatic aldehydes using 4-(N,N-dimethylaminosulphonyl)-7-hydrazino-2,1,3-benzoxadiazole (DBD-H) as a fluorogenic labelling reagent. A mixture of 0.6 mM bis(2,4,6-trichlorophenyl)oxalate and 100 mM H2O2 in acetonitrile was used as a post-column chemiluminescent reagent. DBD-hydrazones of five long chain aliphatic aldehydes (C11-C18:1) were eluted and well separated within 16 min with a mobile phase of 5 mM imidazole in acetonitrile. The detection limits were 14-18 fmol at a signal-to-noise ratio (S/N) of 2. DBD-hydrazones of four aliphatic aldehydes (C6-C11) were also eluted and well separated within 30 min with a mobile phase of 5mM imidazole buffer (pH 7.5)-acetonitrile (20/80, v/v). The detection limits were 86-152 fmol (S/N = 2).