This paper questions whether the presence of biosolids amendment in metal-spiked soils alters the outcome of soil-based assays of metal bioavailability. The effects of biosolids amendment on the efficacies of six soil metal bioavailability assays (total recoverable, EDTA, Ca(NO3)2, soil solution, diffusive gradient in thin films and free ion activity) were assessed against metal concentrations in wheat shoots (Triticum aestivum) germinated in three contrasting soils, each previously incubated for either 2 weeks or 6 months following treatment with Cd, Cu, Ni and Zn +/− biosolids amendment. Overall, Ca(NO3)2 was the most accurate method to predict Cd (r2 = 0.62), Ni (r2 = 0.73) and Zn (r2 = 0.55) bioavailability in soils and therefore was used to compare variations in responses between biosolids and nonbiosolids-amended soils. Comparisons between these two groups revealed no significant differences in linear relationships for all four metals and soil types assessed. These findings not only support Ca(NO3)2 as a robust and valid method for determining soil metal bioavailability across metal matrices and soil types, but also that the presence of biosolids does not compromise the predictive power of this assay or any of the others examined.