It is well-known that the total metal content in soils is not a good indicator of their harmful effects, leading to an overestimation of risks. Toxicological and environmental hazards depend on the chemical species and on its bioavailability to target organisms. Because a good estimation of bioavailability is difficult, a good compromise is to assess bioaccessibility, defined as the maximum amount of a pollutant which is potentially absorbable by a target organism. This study presents a comparison of different strategies to measure metal bioaccessibility in soils. Three procedures were applied to real soil samples with different levels of metal contamination: pseudo-total metal attack, selective sequential extractions and in vitro tests (deliberately developed to simulate human or mammals digestion). Considering the first step of the selective extraction procedure, which can provide the bioaccessible fraction for deposit-feeder organisms, data obtained for each metal were lower than those obtained from in vitro tests. Therefore, it is possible to highlight that this extraction tends to underestimate metal bioaccessibility in soils for humans, while in vitro tests certainly will overestimate bioaccessibility for organisms as invertebrates. If the sum of first and second step of sequential procedure is considered, results are quite similar to those obtained from in vitro tests, but this kind of procedure would require two days of work rather than a few hours required to perform an in vitro test. Results highlight the diversity among the differently defined bioaccessible fractions and the need to apply the most suitable procedure depending on the target organism.