Objective To establish a fingerprinting method and multi-component quantification approach for Rehmannia glutinosa leaves from different geographical origins, and to evaluate the quality of these samples using chemometric analysis. Methods The HPLC fingerprint of R. glutinosa leaves was developed and analyzed for similarity using the Traditional Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System (Version 2012.130723). Chemical constituents were identified and quantified by comparison with reference standards. Hierarchical clustering analysis (HCA), principal component analysis (PCA), and orthogonal partial least squares-discriminant analysis (OPLS-DA) were performed using Origin 2025, IBM SPSS Statistics 27.0, and SIMCA 14.1 software to evaluate the quality of R. glutinosa leaves samples and screen for differential markers. Results A characteristic HPLC fingerprint of R. glutinosa leaves from different origins was developed. In this fingerprint, 19 common peaks were calibrated, and 13 components were unequivocally identified using reference standards. The similarity evaluation of the 23 batches yielded results ranging from 0.879 to 0.997. The cluster heatmap analysis demonstrated a clear classification, dividing the samples into two groups. Principal component analysis (PCA) yielded five principal components with a cumulative variance contribution rate of 83.545%, and the score plot revealed that samples originating from Henan province clustered separately, suggesting their superior quality. Subsequently, OPLS-DA was employed, which successfully screened four compounds-catalpol, acteoside, luteolin, and apigenin-as critical markers for quality discrimination. The quantitative analysis showed that the contents of these markers were 25.474—61.784 mg/g, 11.633—47.462 mg/g, 0.096—0.848 mg/g, and 0.100—1.268 mg/g, respectively. Conclusion The developed method, integrating HPLC fingerprinting and multi-component assay, proved to be simple, accurate, and reproducible. It can systematically reflect the differences among samples of R. glutinosa leaves from different producing areas, thereby providing a reference for their quality control.

Rehmannia glutinosa Libosch.  /  HPLC  /  fingerprint  /  catalpol  /  acteoside  /  luteolin  /  apigenin  /  chemometric analysis  /  quality evaluation  /  chemometrics