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After a radiological or nuclear incident, it is necessary to give a prompt response and to know the number of persons exposed to internal contamination, to evaluate the contamination levels in each person and even and to identify the radionuclides involved.In vitrolaboratories routine monitoring measurements employed to quantify 90Sr and actinides in urine require radiochemical separation and long counting time, which implies a minimum of 1 or 2 weeks to obtain the results, respectively. In this work, rapid radiochemical separation method applied directly to urine samples is presented. It is based on minimal sample preparation, without co-precipitation phase, using extraction resin columns and vacuum box technology. Pu isotopes and 241Am are isolated, electrodeposited and measured by alpha spectrometry, whereas 90Sr is measured by liquid scintillation counting. Finally, results of the participation in European Radiation Dosimetry Group intercomparison on Emergency Bioassay exercise and Bundesamt für Strahlenschutz exercise validate the accuracy of this procedure.