Separation and detection of small amounts of catecholamines by high-performance liquid chromatography.

The effects of mobile phase pH, flow-rate, column temperature, etc., on the separation of catecholamines by high-performance liquid chromatography (HPLC) on Zipax SCX and Zorbax ODS columns were investigated. Fluorimetric detection methods based on post-column reaction using trihydroxyindole and o-phthalaldehyde methods were employed and the results are compared. Optimal reaction conditions ensuring a high sensitivity of the fluorimetric detection were established. Based on the results, a new HPLC system utilizing the trihydroxyindole method has been produced on a trial basis. The favourable detection limits (ca. 20--30 pg) achieved permit highly accurate and selective assays for noradrenaline and/or adrenaline in tissues and urine.