•First time a sequential method for Sr-90 and Po-210 was developed.•A low quantification limit was obtained combining lyophilisation and acid leaching.•The method was applied to seafood samples.•The method was fully validated through certified reference materials and intercomparison exercise.•Good chemical yields were obtained. The latest EU Council Regulation 2016/52/Euratom updates the emergency limits on radionuclides in foods including 210Po and 90Sr, two of the most important radionuclides for radiological dose from the ingestion pathway. A novel and straightforward method has been developed for sequential determination of 90Sr and 210Po in food samples using ultra low-level liquid scintillation counting and alpha-particle spectrometry. For 90Sr analysis, the method makes use of stable strontium as yield tracer, and 210Po is determined through self-deposition using 209Po as a yield tracer. The quantification limit for this method is 25.0 and 2.0Bqkg−1 for 90Sr and 210Po, respectively. The proposed radiochemical separation can be completed within 2days for a batch of 12 samples. The radiochemical procedure was validated by its application for the measurement of IAEA certified reference materials, and through participation in a national intercomparison exercise. Results are also presented in seafood from the Mediterranean coast.