Soil organic matter can be intimately associated with mineral particles of various sizes. For structural studies, soil organic matter can be isolated in particle size fractions after complete dispersion of the aggregates by ultrasonication. The ultrasonic dispersion energy necessary for complete dispersion was investigated in three A and two B horizons originating from four soils differing in pedogenesis (Gleysol, Phaeozem, Podzol, Alisol), organic C (4.2–34.5 g kg–1) and clay content (24–294 g kg–1). Calorimetric calibration of five probe-type ultrasonifiers revealed that the actual energy output from an instrument can depart widely from its nominal output, and that this discrepancy varies from instrument to instrument. Calorimetric calibration is therefore essential for consistency and comparisons between laboratories. Between 450 and 500 J ml–1 of ultrasonic dispersion energy was enough to disperse completely all samples investigated. The particle size distributions obtained were close to those from standard analysis, except for smaller yields (–20 to –80 g kg–1) of sand size fractions, which suggests that dispersion by ultrasound is more effective. Based on total C, C:N ratio and distribution of dissolved C, no detachment of soil organic matter from primary organomineral complexes and no redistribution between particle size fractions could be detected in the range 30–590 J ml–1 of dispersion energy.