Substrates from four and a half year old constructed wetlands built to treat wastewater from an active metal mine were analysed for elevated metal and sulphur concentrations by chemical sequential extractions and x-ray diffraction analyses. Amounts of Fe, Pb, Zn and S were quantified in substrates from the first cells of multi-celled (in-series) treatment wetland systems at three different depths. The analyses showed that the majority of metals removed from the wastewater were retained in residual immobile forms in the upper 0-5 cm of the waterlogged anaerobic substrates. Although substantial concentrations of metals and sulphur were retained in the substrates, the amounts were generally not sufficient to allow accurate mineralogical identification by x-ray diffraction. Classification of the sediments using x-ray techniques was further confounded by the highly organic nature of the wetland substrates. These results suggest that chemical analyses of wetland substrates may still provide a clearer interpretation of metal accumulation over time, especially in wastewaters characterised by relatively low metal concentrations flowing through organically rich substrates. While x-ray diffraction can provide useful interpretation of sediment crystallography and mineralogy, there are limitations in using this technology to characterise young wetland substrates.