AUTHOR=Chen Sze Yun , Low Chi Yuan , Loh Jun Yang , Tew Wan Yin , Ouyang Li Yun , Ong Peng Shun , Yan Chong Seng , Loh Hui Wei , Chen Ying , Xu Wei , Xu Wen , Yoon Tiem Leong , Yam Mun Fei TITLE=A rapid approach for discriminating Ganoderma species using attenuated total reflectance–Fourier transform infrared (ATR-FTIR) spectroscopy integrated with chemometric analysis and convolutional neural network (CNN) JOURNAL=Frontiers in Chemistry VOLUME=Volume 13 - 2025 YEAR=2025 URL=https://www.frontiersin.org/journals/chemistry/articles/10.3389/fchem.2025.1655760 DOI=10.3389/fchem.2025.1655760 ISSN=2296-2646 ABSTRACT=The issue of adulteration and misclassification of Ganoderma species is addressed in this research. In the study, we present a novel and comprehensive framework for Ganoderma authentication by analyzing attenuated total reflectance–Fourier transform infrared (ATR-FTIR) spectra using a combined approach of a chemometric analysis and deep learning (DL) with a convolutional neural network (CNN). The three Ganoderma species involved in this study were as follows: Ganoderma lucidum, Ganoderma sinense, and Ganoderma tsugae. Among chemometric models, orthogonal partial least squares discriminant analysis (OPLS-DA) yielded a high accuracy of 98.61%, a sensitivity of 97.92%, and a specificity of 98.96%. Additionally, the root-mean-squared error of estimation (RMSEE), root-mean-squared error of prediction (RMSEP), and root-mean-squared error of cross-validation (RMSECV) values for the OPLS-DA model were <0.3, confirming its reliability. The CNN model also performed well, achieving 89.84% accuracy, 84.75% sensitivity, and 92.38% specificity, with minimal variation during random segregation testing. Additionally, the model exhibited a precision of 0.87 ± 0.02, a recall of 0.85 ± 0.03, and an F1 score of 0.86 ± 0.03 for 10 random segregation tests. As a conclusion, both chemometric and CNN models developed in this study are efficient and robust for classifying Ganoderma species. To further validate this combined approach, we aim to implement chemometric and CNN models in other medicinal herb authentication in the future.