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.
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-diphenylethyl-enediamine prior to chromatography. Optimal conditions for extraction, derivatization and chromatography were investigated. With alpha-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.
van der Hoorn
F A
FA
Department of Internal Medicine I, University Hospital Dijkzigt, Erasmus University, Rotterdam, The Netherlands.
Boomsma
F
F
Man in 't Veld
A J
AJ
Schalekamp
M A
MA
D002851:Chromatography, High Pressure Liquid; D004298:Dopamine / Q000097:blood*; D004563:Electrochemistry; D004837:Epinephrine / Q000097:blood*; D006801:Humans; D009638:Norepinephrine / Q000097:blood*; D013050:Spectrometry, Fluorescence / Q000379:methods*
D004298:Dopamine; D009638:Norepinephrine; D004837:Epinephrine