The use of depleted uranium in military ordinance has led to an increasing need to determine isotope-specific uranium concentrations in environmental matrices. To this end, gamma-ray spectrometry, ICP-MS and INAA methods have been validated, in accordance with the ISO 17025 standard. Reporting limits of 0.21 (U-235) and 0.91 (U-238) ng/L were obtained by ICP-MS analysis of water. Higher reporting limits were obtained for INAA (U-238 only) validations of water and gamma-ray spectrometric validations of soil and water. Accredited methods have been used to determine uranium concentration and isotope ratio of samples obtained from the Defence Research and Development Canada Valcartier, Quebec.