A rapid tool to discriminate rhino horn and ivory samples from different mammalian species based on the combination of near-infrared reflection (NIR) spectroscopy and chemometrics was evaluated. In this study, samples from the Australian Museum mammalogy collection were scanned between 950 nm and 1650 nm using a handheld spectrophotometer and analyzed using principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA). An overall correct classification rate of 73.5% was obtained for the classification of all samples. This study demonstrates the potential of NIR spectroscopy coupled with chemometrics as a means of a rapid, nondestructive classification technique of horn and ivory samples sourced from a museum. Near-infrared spectroscopy can be used as an alternative or complementary method in the detection of horn and ivory assisting in the combat of illegal trade and aiding the preservation of at-risk species.