This study is concerned with the reconstruction of the palaeoenvironmental history of southeastern Australia for the last ∼300 ka by establishing a luminescence chronology of dune sand deposition in the western Murray Basin (South Australia). In the study area, vast fields of palaeodunes, stabilised by vegetation, provide evidence of past environmental change. In total 98 samples were collected from dune sand layers at 13 different dune sections. The time of their deposition was determined using optically stimulated luminescence dating of single quartz grains, accounting for the impact of post-depositional mixing by the use of a finite mixture model. The oldest depositional phase demonstrates that dune sand layers of great antiquity are preserved in the western Murray Basin, ranging up to at least 380 ka. Phases of substantial dune sand deposition were identified for the periods 18–38 ka and 63–72 ka. Older depositional phases also exist, but are poorly resolved due to relatively large errors of the luminescence ages. Aeolian deposition during the last termination and the Holocene is relatively limited, with a slight clustering of ages at the time of the Antarctic Cold Reversal and from 5–8 ka. Two modern ages give evidence of very recent dune sand deposition. Comparison with other palaeoclimate records from the region suggests that phases with high aeolian sedimentation coincide with more arid conditions and breaks in the dune record with more humid phases. Thus, although dune records are often discontinuous and their interpretation in palaeoclimatic terms is not always straightforward, the palaeodunes of the western Murray Basin show a good preservation of phases of aeolian activity and provide useful information for palaeoenvironmental reconstruction.