The Department of Vigilance Radiologica y Radiactividad Ambiental from CIEMAT has developed an appropriate analytical methodology for Fe-55 and Ni-63 sequential determination in environmental samples based on the procedure used by RIS0 Laboratories. The experimental results obtained in the mayor and minor elements behaviour (soil and air constituents) in the different types of resins used for separating Fe-55 and Ni-63 are showed in this report. The measuring method of both isotopes by scintillation counting has been optimized with Ultima Gold liquid with different concentrations of stable element Fe and Ni. The decontamination factors of different gamma-emitters are experimentally determined in this method with the presence of soil matrix. The Fe-55 and Ni-63 activity concentrations and their associated uncertainties have been calculated from the counting data and sample preparation. A computer application has been implemented in Visual Basic in excel sheets for: (I) obtaining the counting data from spectrometer and counts in each window, (II) representing graphically the background and sample spectrums, (III) determining the activity concentration and its associated uncertainty and (IV) calculating the characteristic limits using ISO 11929 (2007) with various confidence levels. (Author) 30 refs.