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Determination of catecholamines and serotonin by micellar electrokinetic chromatography with laser‐induced fluorescence detection
Author(s) -
Cao LiWei
Publication year - 2007
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.809
Subject(s) - chemistry , chromatography , reagent , micellar electrokinetic chromatography , derivatization , fluorescence , electrokinetic phenomena , detection limit , yield (engineering) , high performance liquid chromatography , organic chemistry , physics , materials science , quantum mechanics , metallurgy
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 m m boric acid and 15 m m sodium dodeculsulfate at pH 9.0. The detection limits ranged from 5 × 10 −10 to 2 × 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. Copyright © 2007 John Wiley & Sons, Ltd.