| Summary: | <i>Rhodiola</i>, especially <i>Rhodiola crenulate</i> and <i>Rhodiola rosea</i>, is an increasingly widely used traditional medicine or dietary supplement in Asian and western countries. Because of the phytochemical diversity and difference of therapeutic efficacy among <i>Rhodiola</i> species, it is crucial to accurately identify them. In this study, a simple and efficient method of the classification of <i>Rhodiola crenulate</i>, <i>Rhodiola rosea</i>, and their confusable species (<i>Rhodiola serrata</i>, <i>Rhodiola yunnanensis</i>, <i>Rhodiola kirilowii</i> and <i>Rhodiola fastigiate</i>) was established by UHPLC fingerprints combined with chemical pattern recognition analysis. The results showed that similarity analysis and principal component analysis (PCA) could not achieve accurate classification among the six <i>Rhodiola</i> species. Linear discriminant analysis (LDA) combined with stepwise feature selection exhibited effective discrimination. Seven characteristic peaks that are responsible for accurate classification were selected, and their distinguishing ability was successfully verified by partial least-squares discriminant analysis (PLS-DA) and orthogonal partial least-squares discriminant analysis (OPLS-DA), respectively. Finally, the components of these seven characteristic peaks were identified as 1-(2-Hydroxy-2-methylbutanoate) β-D-glucopyranose, 4-O-glucosyl-p-coumaric acid, salidroside, epigallocatechin, 1,2,3,4,6-pentagalloyglucose, epigallocatechin gallate, and (+)-isolarisiresinol-4′-O-β-D-glucopyranoside or (+)-isolarisiresinol-4-O-β-D-glucopyranoside, respectively. The results obtained in our study provided useful information for authenticity identification and classification of <i>Rhodiola</i> species.
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