Effect of the pH and the importance of the internal standard on the measurement of the urinary catecholamines by high-performance liquid chromatography.

Clinical manifestations of phaeochromocytoma are not always sufficient for its early diagnosis. It is therefore necessary to confirm hypersecretion of catecholamines. Current methods for the measurement of catecholamines are based on their oxidative properties, and the majority of the laboratories often use HPLC methods for catecholamine testing. However, the extraction procedures used for the biogenic amines differ. We use a method of ion-exchange chromatography which is performed at pH 6.5. In order to avoid the spontaneous oxidation of the catecholamines, the urine samples has to be collected on HCl, which gives a pH of approximately 2. Occasionally, the acidified urine sample has a pH less than 1 requiring the addition of NaOH to reach a pH of 6.5, necessary for the adsorption of catecholamines on cation exchanger resins. This phenomenon produces a decrease in the peak areas but the use of an internal standard allows the final results to be corrected.