Quantification of biogenic amines by microchip electrophoresis with chemiluminescence detection.
A highly sensitive microchip electrophoresis (MCE) method with chemiluminescence (CL) detection was developed for the determination of biogenic amines including agmatine (Agm), epinephrine (E), dopamine (DA), tyramine, and histamine in human urine samples. To achieve a high assay sensitivity, the targeted analytes were pre-column labeled by a CL tagging reagent, N-(4-aminobutyl)-N-ethylisoluminol (ABEI). ABEI-tagged biogenic amines after MCE separation reacted with hydrogen peroxide in the presence of horseradish peroxidase (HRP), producing CL emission. Since no CL reagent was added to the running buffer, the background of the CL detection was extremely low, resulting in a significant improvement in detection sensitivity. Detection limits (S/N=3) were in the range from 5.9x10(-8) to 7.7x10(-8) M for the biogenic amines tested, which were at least 10 times lower than those of the MCE-CL methods previously reported. Separation of a urine sample on a 7 cm glass/poly(dimethylsiloxane) (PDMS) microchip channel was completed within 3 min. Analysis of human urine samples found that the levels of Agm, E and DA were in the ranges of 2.61x10(-7) to 4.30x10(-7) M, 0.81x10(-7) to 1.12x10(-7) M, and 8.76x10(-7) to 11.21x10(-7) M (n=4), respectively.
Zhao
Shulin
S
College of Chemistry and Chemical Engineering, Guangxi Normal University, Guilin 51004, China.
Huang
Yong
Y
Shi
Ming
M
Liu
Yi-Ming
YM
D001679:Biogenic Amines / Q000032:analysis* / Q000652:urine; D046209:Electrophoresis, Microchip / Q000379:methods*; D006801:Humans; D008163:Luminescent Measurements / Q000379:methods*
D001679:Biogenic Amines