Simultaneous determination of 10 kinds of biogenic amines in rat plasma using high-performance liquid chromatography...

We describe a simple, rapid, selective and sensitive HPLC method coupled with fluorescence detection for simultaneous determination of 10 kinds of biogenic amines (BAs: tryptamine, 2-phenethylamine, putrescine, cadaverine, histamine, 5-hydroxytryptamine, tyramine, spermidine, dopamine and spermine). BAs and IS were derivated with dansyl chloride. Fluorescence detection (λ_ex /λ_em = 340/510 nm) was used. A satisfactory result for method validation was obtained. The assay was shown to be linear over the ranges 0.005-1.0 μg/mL for tryptamine, 2-phenethylamine and spermidine, 0.025-1.0 μg/mL for putrescine, 0.001-1.0 μg/mL for cadaverine, 0.25-20 μg/mL for histamine, 0.25-10 μg/mL for 5-hydroxytryptamine and dopamine, and 0.01-1.0 μg/mL for tyramine and spermine. The limits of detection and the limits of quantification were 0.3-75.0 ng/mL and 1.0-250.0 ng/mL, respectively. Relative standard deviations were ≤5.14% for intra-day and ≤6.58% for inter-day precision. The recoveries of BAs ranged from 79.11 to 114.26% after spiking standard solutions of BAs into a sample at three levels. Seven kinds of BAs were found in rat plasma, and the mean values of tryptamine, 2-phenethylamine, putrescine, cadaverine, histamine, spermidine and spermine determined were 52.72 ± 7.34, 11.45 ± 1.56, 162.56 ± 6.26, 312.75 ± 18.11, 1306.50 ± 116.16, 273.89 ± 26.41 and 41.51 ± 2.07 ng/mL, respectively.