Antioxidant Activity, HPTLC Fingerprint, and Discriminant Analysis of Plantago major Leaves from Diverse Origins in Indonesia

Introduction: Plantago major L. (Plantaginaceae) is a perennial herb having contribution to the folk medicine all around the world, including Indonesia with wide geographical distribution. Plant materials origin is one factor that significantly influences the quality of herbal medicines. Materials and Methods: In this paper, High-Performance Thin Layer Chromatography (HPTLC) method using pattern-oriented approach has been employed to evaluate the quality of Plantago major leaves collected from seven origins in Indonesia. To differentiate the antioxidant capacities of those plant materials, the crude extracts were tested using 1,1-diphenyl-2-picrylhydrazyl (DPPH), total phenolics, and total flavonoids assay methods. Results: The results showed that radical scavenging activity, total phenolics, and total flavonoids of plant material from seven origins were significantly different. Moreover, HPTLC fingerprints analyzed with chemometrics showed an ability to discriminate the leaves samples from various origins as well as detect chemicals responsible for discrimination. Two models using principal component analysis (PCA) and partial least squares (PLS-DA) were built in chemometrics test. The PCA model was able to describe the studied samples by using four principal components with a value of explained variance of 95%, whereas PLS-DA model accurately classified the leaves samples with prediction ability of 100%. In the PCA, loading plot of the first PC showed that peaks number 10 and 12 are the most important peaks for clustering of the samples. Conclusions: Plantago major collected from different origins revealed different radical scavenging activity and concentration of total phenolics as well as total flavonoids. HPTLC fingerprints coupled with chemometrics analysis can be used as an alternative to marker-oriented method for the quality control of Plantago major. Key words: Chemometrics, Flavonoids, Herbal medicines, Pattern-oriented, Phenolics, PLSDA