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BACKGROUND: Traditional methods of evaluating herbs were mainly based on chromatographic techniques. They usually included tedious sample preparation procedures, taking tens of minutes to hours, and consume solvents as well as standards for external calibration. In this paper, the feasibility of employing a fluorescence fingerprint coupled with multi-way chemometrics analysis for quality evaluation and traceability of Bletilla striata were investigated.<br>RESULTS: Relative concentrations of four markers presented in B. striata were determined by using a four-component self-weighted alternating trilinear decomposition (SWATLD) model. These markers could be applied to accurate classification and quality control of B. striata samples from different regions. Furthermore, multiway principal component analysis, multilinear partial least squares discriminant analysis (PLS-DA), unfolded PLS-DA, and SWATLD-PLS-DA models were applied to classify the B. striata samples according to their geographic origins. Consistent results were obtained showing that B. striata samples could be successfully grouped based on their geographical origins and quality.<br>CONCLUSION: Our results revealed that the method developed can be used for quality evaluation and traceability of B. striata. Compared with the chromatographic methods, the method employed in this study was more convenient, simpler, and more sensitive. © 2018 Society of Chemical Industry