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Bark paintings represent a unique artform that originated in Australia and continues to this day. This paper presents a scientific investigation into both the pigments and binders used by Aboriginal artists from across the Northern Territory (NT) and the Kimberley region in Western Australia (WA) and represents the first major study dedicated to identifying these materials in samples taken from paintings. Laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS), Fourier transform infrared (FTIR) and Raman spectroscopy were used to identify the pigments in the four colors commonly associated with works by Aboriginal artists: red, yellow, black, and white. Iron oxides are used for red and yellow. Typically, kaolin is used for white and carbon or natural manganese ores are used for black, although the use of other pigments such as huntite (white) and battery black were seen in paintings from specific locations. Evidence of trade could not be established during this study; however, analysis shows that artists may use different pigments based on location and availability. A combination of FTIR and pyrolysis-gas chromatography-mass spectrometry (pyrolysis-GC-MS) was utilized to investigate the presence of a binder in the paint. Plant-based materials were identified as the binder in the majority of the paintings. The use of orchid juice was common, both as a preparatory layer on the bark substrate and as a paint binder. The presence of synthetic resins was more widespread than anticipated, attributed to both conservation efforts and more modern painting practices. [ABSTRACT FROM AUTHOR]