Journal
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
Volume 406, Issue 23, Pages 5669-5676Publisher
SPRINGER HEIDELBERG
DOI: 10.1007/s00216-013-7514-9
Keywords
Chemiluminescence; Neurotransmitters; Neurotransmitter metabolites; Brain; Urine; Two-dimensional high-performance liquid chromatography
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High-performance liquid chromatography with chemiluminescence detection based on the reaction with acidic potassium permanganate and formaldehyde was explored for the determination of neurotransmitters and their metabolites. The neurotransmitters norepinephrine and dopamine were quantified in the left and right hemispheres of rat hippocampus, nucleus accumbens and prefrontal cortex, and the metabolites vanillylmandelic acid, 3,4-dihydrophenylacetic acid, 5-hydroxyindole-3-acetic acid and homovanillic acid were identified in human urine. Under optimised chemiluminescence reagent conditions, the limits of detection for these analytes ranged from 2.5 x 10(-8) to 2.5 x 10(-7) M. For the determination of neurotransmitter metabolites in urine, a two-dimensional high-performance liquid chromatography (2D-HPLC) separation operated in heart-cutting mode was developed to overcome the peak capacity limitations of the one-dimensional separation. This approach provided the greater separation power of 2D-HPLC with analysis times comparable to conventional one-dimensional separations.
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