DEVELOPMENT OF A CHEMILUMINESCENCE DETECTION SYSTEM USING IS[4-NITRO-2-(3,6,9-TRIOXADECYLOXYCARBONYL)PHENYL] OXALATE FOR THE SENSITIVE DETERMINATION OF THE FLUORESCENT COMPOUNDS SEPARATED WITH AN ACIDIC MOBILEPHASE

A rotating flow mixing device having three directional inlets was deve loped for a high performance liquid chromatographic peroxyoxalate chem iluminescence detection system for the sensitive detection of fluoresc ent compounds separated with acidic mobile phases, such as those conta ining trifluoroacetic acid. The diameters of the three inlets were 0.3 , 0.3 and 0.5 mm i.d., for an acidic column eluate, chemiluminogenic r eagents is[4-nitro-2-(3,6,9-trioxadecyloxycarbonyl)phenyl] oxalate and hydrogen peroxide) and imidazole solution (for neutralization and as a catalyst), respectively. The three mixed solutions flowed out throug h an outlet (0.5 mm i.d.) at the centre of the top of the vessel. The reaction conditions to give the appropriate signal-to-noise ratio (S/N ) were investigated using dipyridamole (an antiplatelet aggregation dr ug) as a typical fluorescent compound. The system was applied to the d etection of dipyridamole and dansylated amino acids with a mobile phas e containing trifluoroacetic acid. The detection limits for dipyridamo le and each dansylated amino acid were 10 amol and sub-fmol (S/N = 2), respectively.