Determination of catecholamines and serotonin by micellar electrokinetic chromatography with laser-induced fluorescence detection.
6-Oxy-(N-succinimidyl acetate)-9-(2'-methoxycarbonyl) fluorescein, a new synthesized fluorescent reagent, was established for the first time as a label for the sensitive analysis of catecholamines (CAs) and serotonin (5-HT) by micellar electrokinetic chromatography (MEKC) with laser-induced fluorescence detection. After careful study on the derivatization conditions such as pH value, reagent concentration, temperature and reaction time, the labeling reaction was accomplished as quickly as 7 min with stable yield. The separation parameters for the CAs and 5-HT were also optimized in detail. The derivatives were baseline separated in a running buffer containing 30 mM boric acid and 15 mM sodium dodeculsulfate at pH 9.0. The detection limits ranged from 5 x 10(-10) to 2 x 10(-9) M (signal-to-noise ratio = 3). The rapid and sensitive method was also applied to the determination of the CAs and 5-HT of urine samples.
Cao
Li-Wei
LW
Department of Chemistry, Jinan University, Guangzhou 510632, People's Republic of China. lwcao@126.com
D002138:Calibration; D002395:Catecholamines / Q000652:urine*; D020374:Chromatography, Micellar Electrokinetic Capillary / Q000379:methods*; D006801:Humans; D007834:Lasers; D012701:Serotonin / Q000652:urine*; D013050:Spectrometry, Fluorescence / Q000379:methods*
D002395:Catecholamines; D012701:Serotonin