Determination of catecholamines in human plasma by high-performance liquid chromatography: comparison between a new method with fluorescence detection and an established method with electrochemical detection

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Abstract

We report a sensitive fluorimetric method, in which catecholamines are concentrated from plasma by liquid-liquid extraction and derivatized with the selective fluorescent agent 1,2-diphenylethylenediamine prior to chromatography. Optimal conditions for extraction, derivatization and chromatography were investigated. With α-methylnorepinephrine as internal standard, the chromatographic separations are complete within 6 min. Limits of detection are 0.3 pg for norepinephrine and epinephrine and 0.5 pg for dopamine. Coefficients of variation are low (3–7%). Comparison of plasma catecholamine values determined with this method and with an established method with electrochemical detection (n=135) shows good correlation (r=0.94–1.00), and regression lines are close to lines of identity.

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