A method for accurate on-line ultra-trace analysis of palladium by inductively coupled plasma mass spectrometry has been developed. After separation of interfering cations by cation exchange chromatography, palladium was selectively adsorbed on to a C18 micro-column on-line which had been reversibly loaded with the complexing agent N,N-diethyl-N′-benzoylthiourea (DEBT). The palladium complex formed was eluted with methanol and introduced into an ICP-SFMS via microconcentric nebulization with membrane desolvation. Quantification of palladium was carried out by isotope dilution analysis. The results obtained via the isotope ratios of 105Pd/108Pd and 106Pd/108Pd agreed within their measurement uncertainty. For solid samples (total intake 100 mg, final sample volume of 10 mL after closed vessel microwave digestion) the limit of detection was 0.24 ng g−1 palladium (2 mL of digested sample). The necessity of combining matrix separation and pre-concentration for elimination of spectral interferences was demonstrated using soil samples. The method was validated by analysis of the certified reference material BCR-723 (road dust).