The influence of metal concentration, solution pH and exposure time on the phytoextraction (i.e. separation using vascular plants) of Au was investigated for the known metallophytes Brassica juncea (BJ) and Medicago sativa (MS). Metal uptake was inferred using Inductively Coupled Plasma Optical Emission Spectroscopy (ICP-OES) and in vivo localisation and distribution using proton induced X-ray emission spectroscopy (mu-PIXE). MS roots accumulated a maximum of 287 mg Au g(-1) (dry biomass) and BJ roots a maximum of 227 mg Au g-1 (dry biomass), both when exposed to a 10,000 ppm aqueous solution of KAuCl4. MS was found to accumulate comparatively greater quantities of Au than BJ across higher substrate concentrations (40-10,000 ppm Au) whereas BJ was found to be a better accumulator of Au at lower concentrations (5-20 ppm Au). In general MS showed an increase in Au uptake with an increase in Au substrate concentration and the time exposed, whereas for BJ the maximum uptake was observed after 48 h of exposure at higher concentrations (100-10,000 ppm), and then decreased at longer exposure times. The uptake ratio (UR), defined as the ratio of Au concentration in plant tissues to the concentration in the substrate, increased with increasing concentration and exposure time, to a maximum of 995 for MS roots after 72 h exposure. Metal translocation from roots to shoots in BJ increased with increasing substrate concentration, however in the shoots, metal uptake increased from 24 to 48 h and then decreased at 72 h, indicating some threshold level had been reached and metal was then being excluded from the cells, possibly through the phloem to the Au solution. Elemental distribution maps of plant tissues measured using mu-PIXE, show Au present across the entire sample, ranging from the epidermis and cortex, with the greatest concentration occurring within the central stele. This result is suggestive of xylem loading. These results collectively suggest that the separation of Au using vascular plants for applications in mining (phytomining) and remediation (phytoremediation) are viable technologies. (C) 2009 Elsevier B.V. All rights reserved.