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Development of a chemiluminescence detection system using bis[4‐nitro‐2‐(3,6,9‐trioxadecyloxycarbonyl)phenyl] oxalate for the sensitive determination of the fluorescent compounds separated with an acidic mobile phase
Author(s) -
Sugiura Masaki,
Kanda Susumu,
Imai Kazuhiro
Publication year - 1993
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.1130070310
Subject(s) - chemistry , trifluoroacetic acid , peroxyoxalate , chemiluminescence , reagent , detection limit , chromatography , fluorescence , oxalate , hydrogen peroxide , elution , inorganic chemistry , organic chemistry , physics , quantum mechanics
A rotating flow mixing device having three directional inlets was developed for a high performance liquid chromatographic peroxyoxalate chemiluminescence detection system for the sensitive detection of fluorescent compounds separated with acidic mobile phases, such as those containing 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 reagents (bis[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 through 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 drug) as a typical fluorescent compound. The system was applied to the detection of dipyridamole and dansylated amino acids with a mobile phase containing trifluoroacetic acid. The detection limits for dipyridamole and each dansylated amino acid were 10 amol and sub‐fmol ( S / N =2), respectively.