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Rhodiola is an increasingly widely used traditional Tibetan medicine and traditional Chinese medicine in China. The composition profiles of bioactive compounds are somewhat jagged according to different species, which makes it crucial to identify authentic Rhodiola species accurately so as to ensure clinical application of Rhodiola . In this paper, a nondestructive, rapid, and efficient method in classification of Rhodiola was developed by Fourier transform near-infrared (FT-NIR) spectroscopy combined with chemometrics analysis. A total of 160 batches of raw spectra were obtained from four different species of Rhodiola by FT-NIR, such as Rhodiola crenulata , Rhodiola fastigiata , Rhodiola kirilowii , and Rhodiola brevipetiolata . After excluding the outliers, different performances of 3 sample dividing methods, 12 spectral preprocessing methods, 2 wavelength selection methods, and 2 modeling evaluation methods were compared. The results indicated that this combination was superior than others in the authenticity identification analysis, which was FT-NIR combined with sample set partitioning based on joint x-y distances (SPXY), standard normal variate transformation (SNV) + Norris-Williams (NW) + 2nd derivative, competitive adaptive reweighted sampling (CARS), and kernel extreme learning machine (KELM). The accuracy (ACCU), sensitivity (SENS), and specificity (SPEC) of the optimal model were all 1, which showed that this combination of FT-NIR and chemometrics methods had the optimal authenticity identification performance. The classification performance of the partial least squares discriminant analysis (PLS-DA) model was slightly lower than KELM model, and PLS-DA model results were ACCU = 0.97, SENS = 0.93, and SPEC = 0.98, respectively. It can be concluded that FT-NIR combined with chemometrics analysis has great potential in authenticity identification and classification of Rhodiola , which can provide a valuable reference for the safety and effectiveness of clinical application of Rhodiola . [ABSTRACT FROM AUTHOR]