Pre-analytical factors of variation need to be carefully considered and investigated in efforts to harmonize all aspects of the total testing process. This study aimed to evaluate contamination and stability in copper (Cu) analysis of serum and urine by flame atomic absorption spectroscopy (FAAS) and to compare the stability of urine Cu in controls and in D-penicillamine (D-PA) administration. Cu was measured by AAnalyst 400, Perkin Elmer, USA. Blood was collected in BD Vacutainer®SSTTM II Advance tubes and BD Vacutainer® Trace Element tubes. Sterile polyethylene and polypropylene vessels for collection, transportation, storage and preliminary preparation of samples were used in urinalysis. Stability in serum and 24 h urine was evaluated in two temperature regimens: 15-25°C and 2-8°C, for particular time of storage. No significant differences (p = 0.20) in Cu concentration was found between the two types of tested tubes with patient`s sera. The stability of the samples (serum and urine) was better at refrigeration temperature. In urine the stability was better in D-PA administration.Standardization of Cu analysis could be achieved by assessing the aspects of pre-analytical factors of variations.