Analysis of neurotransmitter catecholamines and related amines in human urine and serum by chromatography and capillary electrophoresis with 1, 3, 5, 7-tetramethyl-8-(N-hydroxysuccinimidyl propionic ester)-difluoro-boradiaza-s-indacene

Abstract

Two sensitive and effective methods were developed for the detection of catecholamines and related biogenic amines (dopamine, epinephrine, norepinephrine, serotonin, levodopa and tyramine) using high performance liquid chromatography with fluorescence detection and capillary electrophoresis with laser-induced fluorescence detection. A BODIPY fluorescent dye, 1, 3, 5, 7-tetramethyl-8-(N-hydroxysuccinimidyl propionic ester)-difluoroboradiaza- s-indacene was used as pre-column derivatization reagent. The separation and derivatization conditions were optimized in detail. In high performance liquid chromatography with fluorescence detection method, the derivatization reaction was completed at 35 °C for 20 min. At the wavelength of λex/λem = 493 nm/513 nm, dopamine, epinephrine, norepinephrine, and levodopa derivatives achieved baseline separation within 15 min. The limits of detection (S/N = 3) were 1.0, 2.0, 5.0, and 0.5 nmol/L, respectively. In capillary electrophoresis with laser-induced fluorescence detection method, the derivatization reaction was completed at 25 °C for 20 min. Serotonin, tyramine and dopamine derivatives reached baseline separation within 10 min at the wavelength of λex = 473 nm. The limits of detection (S/N = 3) for serotonin, tyramine, and dopamine were 0.3, 0.02, and 0.2 nmol/L, respectively. The amino compounds in human serum and urine samples were detected successfully, and the recoveries were 93.3%–106.7% and 91.0%–103.1%, respectively.