Simultaneous Measurement of Plasma Catecholamine (Norepinephrine, Epinephrine, and Dopamine) and Free N—Methyl Dopamine (Epinine) Levels, by HPLC with Electrochemical Detection

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Abstract
Epinine is the active moiety of ibopamine, a cardiovascular prodrug used in congestive heart failure. This catecholic compound shows dopaminergic and adrenergic properties. Moreover the drug seems to affect plasma catecholamine levels in patients with heart failure. Here we present a method developed for the simultaneous determination of epinine and catecholamine plasma levels. Free epinine and catecholamines were extracted from human venous plasma via an alumina adsorption procedure. The extracts underwent an ion-pair reversed-phase HPLC separation with three-electrode coulometry. Quantitation was made by an internal standard method. Coefficients of variation were < 9%. The validity was assessed as the peak height—picograms correlation (r >0.997). The detection limits were ≤ 5 pg of each catechol after extraction. This method allows about 50 low cost determination to be done in a working day.