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Increasing accounts of fraud and persistent labeling problems have brought the authenticity of leather products into question. In this study, we developed an extremely simplified workflow for real-time, in situ , and unambiguous authentication of leather samples using rapid evaporative ionization mass spectrometry (REIMS) coupled with an electric soldering iron. Initially, authentic leather samples from cattle, sheep, pig, deer, ostrich, crocodile, and snake were used to create a chemometric model based on principal component analysis and linear discriminant analysis algorithms. The validated multivariate statistical model was then used to analyze and generate live classifications of commercial leather samples. In addition to REIMS analysis, the microstructures of leathers were characterized by scanning electron microscopy to provide complementary information. The current study is expected to provide a high-throughput tool with superior efficiency and accuracy for authenticating the identity of leathers and other consumer products of biogenic origin. Image 1 • A REIMS methodology was developed for real-time, in situ , and unambiguous authentication of leather samples. • A multivariate statistical model was built based on chemometric analysis of authentic leather samples. • The chemometric model was validated and used to analyze commercial leather samples. • Leather microstructures were characterized by SEM to provide complimentary information. [ABSTRACT FROM AUTHOR]