The objective of this research was to develop a rapid spectroscopic technique as an alternative method for the differentiation and authentication of gelatin sources in food products by using attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectra combined with chemometrics. Clear discrimination and classification of all the studied gelatin sources (bovine, porcine, and fish) were achieved by hierarchical cluster and principle component analysis (PCA). Amide-I (1700-1600 cm(-1)) and Amide-II (1565-1520 cm(-1)) spectral bands were used in a chemometric method. Moreover, ATR-FTIR spectral data successfully discriminated pure bovine gelatin from mixture of bovine and porcine gelatins, which is very important for the food industry. The method that we adopted could be beneficial for rapid, simple and economic determination of both gelatin presence and its origin from food products such as yogurt, ice cream, milk dessert or other gelatin containing products such as pharmaceuticals and cosmetics. (C) 2015 Elsevier Ltd. All rights reserved.