Dual herbal combination of Withania somnifera and five Rasayana herbs: A phytochemical, antioxidant, and chemometric profiling

BACKGROUND: Traditional medicine adequately emphasis plant resources for addressing a wide variety of human ailments by utilizing the naturally occurring phytoconstituents; in particular medicinal plants or parts of plants in combination have prodigious antioxidant potentials. OBJECTIVE: The present study aims to analyze methanolic extract of W. somnifera (W) individually, and in dual combination with five Rasayana herbs P. emblica - (W:P), B. monnieri - (W:B), T. sinensis - (W:T), O. basilicum - (W:O), A. racemosus - (W:A) in three dual ratios [4:1, 1:1, and 1:4]. The efficacy of the combinations is assessed based on their chemometric profiling. MATERIAL AND METHODS: A total of 15 dual combinatorial methanolic extracts together with W. somnifera were evaluated for their preliminary phytochemical profiles, antioxidant potentials using DPPH and FRAP assays. Five dual samples were selected and analyzed for High-Performance Thin-Layer Chromatography (HPTLC) image-based chemometric profiling followed by Principal Component Analysis (PCA) and Hierarchical Cluster Analysis (HCA)-Heatmaps. RESULTS: Qualitative phytochemical analysis of combinatorial extracts exhibited a richness for a variety of phytoconstituents. The antioxidant activity was significantly higher for DPPH IC(50) (μg/mL): W = 11.56 ± 3.69; W:P/1:4 = 7.89 ± 1.52; W:O/1:4 = 8.995 ± 2.64 and FRAP (μM FeE/g): W = 4.56 ± 0.54; W:P/1:4 = 138.34 ± 9.25; W:O/1:4 = 15.32 ± 1.64. Chemometric data acquisition displayed improved secondary metabolite close cluster combination with W:O/1:4 and W:P/1:4 than W. somnifera (W) alone. CONCLUSION: The dual herbal combinatorial study revealed that the methanolic combinatorial extracts phytoconstituents correlated with an increase in the antioxidant potential and would serve as a promising source for phytomedicine.