Catecholamine measurements by high-performance liquid chromatography

Abstract

The development of sensitive detectors has allowed the use of high-performance liquid chromatography (HPLC) for measurements of catecholamines in extracts of plasma, urine, and tissue samples. Separation of the catecholamines may be effected by reversed phase chromatography or cation-exchange chromatography and quantitation by electrochemical detection (EC) or by fluorometry coupled with postcolumn derivatization according to the trihydroxyindole (THI) method. EC has a somewhat lower sensitivity than the THI method for norepinephrine (NE) and epinephrine (E). The THI method is insensitive to dopamine (DA). Basal plasma E levels of 0.1 nM (20 pg/ml) or less may be measured in sample volumes of 1-2 ml with EC. Sensitivity and reproducibility of an assay is not necessarily a guarantee of accuracy. It is argued that new methods and modifications of old methods should be validated against accepted methodology. This is rarely the case. Cation exchange HPLC with EC has been adequately validated, but only one of the reversed phase methods has been compared with radioenzymatic methodology. HPLC has the advantages of economy, speed, and more stimulating laboratory work, as compared with radioenzymatic methodology.