Simultaneous Quantification of Three Bioactive Lignans, viz., Phyllanthin, Hypophyllanthin, and Niranthin from Phyllanthus amarus Using High-Performance Thin-Layer Chromatography

Summary

Phyllanthus amarus, a well-known drug of the Indian Systems of Medicine, possesses a wide variety of pharmacological activities including anticancer, anti-inflammatory, hepatoprotective, immunomodulatory, nephroprotective, and radioprotective activity. The pharmacological activity of P. amarus is due to the presence of various lignans and flavonols, the major ones being phyllanthin, hypophyllanthin, and niranthin. A simple high-performance thin-layer chromatographic method has been developed for the simultaneous quantification of these three lignans from the whole plant of P. amarus. The optimized mobile phase n-hexane-ethyl acetate-glacial acetic acid (9:3:0.3, v/v) gave good separation of phyllanthin, hypophyllanthin, and niranthin on thin-layer chro-matography (TLC) silica gel 60 F₂₅₄ layers when quantified at 230 nm. The developed method was validated according to the International Conference on Harmonization (ICH) guidelines for precision, repeatability, accuracy, and linearity. Peak identity and purity were determined. This is the first report of simultaneous quantification of phyllanthin, hypophyllanthin, and niranthin from P. amarus using normal phase TLC silica gel 60 F₂₅₄ plates. The developed method is simple, precise, accurate, cost-effective, and less time-consuming.