Chemical Fingerprints Combined with Chemometric Analysis to Evaluate and Distinguish Between Plantago Asiatica L. and Plantago Depressa Willd

BACKGROUND: Plantago, also known as plantain, has been used as a traditional herb in Asia for centuries to treat urinary tract inflammation and kidney disease. Recently, there has been increasing interest in its potential applications in the food industry. In China, there are two common species, namely Plantago asiatica L. (PAL) and P. depressa Willd. (PDW), which are not distinguished in folk and clinical practices, leading to safety and efficacy risks in plantago applications. OBJECTIVES: The objective of this study is to establish a reliable method for differentiating and evaluating the quality of PAL and PDW by integrating fingerprint analysis with multivariate chemometrics. METHODS: HPLC was used to collect characteristic chemical information from the whole plant in 13 PAL and 18 PDW samples, establishing fingerprints for the different species. Multiple chemometric methods, including hierarchical cluster analysis (HCA), principal component analysis (PCA), and partial least-squares discriminant analysis (PLS-DA), were employed to analyze the chemical differences between the two species and to identify chemical markers. Finally, the differences in the content of the three chemical markers in the two plantago samples were determined and analyzed. RESULTS: The analysis revealed a distinct classification of PAL and PDW into two groups. Significant differences were observed in the content of key components such as plantamajoside, acteoside, and isoacteoside between the two species. CONCLUSION: This study provides a reliable scientific basis for the rational application and quality evaluation of plantago. The findings contribute to ensuring the safe and effective use of plantago in various applications. HIGHLIGHTS: HPLC fingerprinting combined with chemometrics identified three key markers distinguishing PAL and PDW, supporting quality control and medicinal development.