Integrated derivatization-chemiluminescence detection system for the determination of β-carboline alkaloids by high-performance liquid chromatography

An integrated derivatization-chemiluminescence detection system was used for the high-performance liquid chromatographic determination of β-carboline alkaloids such as harmol, harmalol, norharmane, harmane, harmine and harmaline by the chemiluminescence produced in their reaction with the bis(2,4-dinitrophenyl)oxalate (DNPO)-hydrogen peroxide system. The ability of the proposed detection system based on its zero-dead-volume to overcome the problems associated with the use of this oxalate ester in peroxyoxalate chemiluminescence detection in HPLC was exploited to develop a sensitive chromatographic method for the determination of the above-mentioned hallucinogens. The method excels over other alternatives based on different detection techniques, such as UV and fluorescence spectroscopy, in terms of limits of detection. The proposed method was validated by determining the alkaloids (the identification and quantification of some are reported for the first time) in Heliconiini butterfly specimens with good results.