Validation of a high performance liquid chromatography analysis for the determination of noradrenaline and adrenaline in human urine with an on-line sample purification

A high performance liquid chromatography (HPLC) method with fluorescence de tection including an on-line purification was established for determination of catecholamines in human urine. The method was evaluated using samples o f pooled urine spiked with catecholamines and validated for measurements of catecholamines in urine of healthy individuals in a held study. The labora tory method evaluation study showed that the recovery of the method was 0.8 2 (confidence interval (CI): 0.79-0.86) and 0.92 (CI: 0.89-0.95) for noradr enaline and adrenaline, respectively. Thus, correction factors of 0.82(-1) and 0.92(-1) were applied to correct the measurement results for this syste matic effect. Furthermore, an uncertainty budget was generated for the anal ytical method using the BIPM-approach recommended by the International Orga nization for Standardization. The relative uncertainty of the method was es timated to be 10-12%, which was consistent with the observed relative varia bility found in the method evaluation. The method was evaluated in accordan ce with EURACHEM Guidance Document No 1 concerning accreditation for chemic al laboratories with respect to accuracy and precision. The field study sho wed that the standard deviation of the method was sufficiently low to disti nguish between persons working with two different processes of garbage coll ection, i.e. collection using four wheeled containers versus collection usi ng bins. (C) 1999 Elsevier Science B.V. All rights reserved.