Abstract: Rhinoceros horn is a scarce organic gem. Because of its high value, various imitations have emerged.Ox horn and donkey hooves are the most similar imitations in appearance to those of rhinoceros horn. In order to distinguish them effectively, samples of rhinoceros born, ox horn and donkey hooves were compared from the aspects of conventional instrument tests (density, refractive index and fluorescence detection), magnification observation, infrared spectrum analysis and scanning electron microscope analysis.The results show that the magnification observation is an important test method to identify the three kinds of samples, which shows different characteristics in the direction of cross section and longitudinal surface.