Development of validated HPTLC method for quantitation of phyllanthin, hypophyllanthin and picroside-II in marketed herbal formulation (Valiliv capsules)

Abstract

A rapid and sensitive high-performance thin-layer chromatographic (HPTLC) method was developed and validated for quantitative estimation of Picroside-II, Phyllanthin and Hypophyllanthin simultaneously in marketed formulation (Valliliv capsule) containing Picrorhiza kurroa and Phyllanthus niruri. The sample (Valliliv capsule) were chromatographed on silica gel 60F₂₅₄- TLC plates, using solvent system (Hexane: Ethyl acetate, 6:3 v/v) and scanned at 278nm. The linearity range for Picroside was 200-1600ng/spot with average recovery of 99.90%, Phyllanthin was 200-1600ng/spot with average recovery 100.41% and Hypophyllanthin 200-1600ng/spot with average recovery of 99.99% in formulation. The limit of detection and limit of quantification for Picroside-II, Phyllanthin and Hypophyllanthin were found to be 40ng/spot- 120ng/spot, 90-270ng/spot and 100ng/spot-300ng/spot. The developed method was successfully applied for the assay of market formulation containing Picrorhiza kurroa and Phyllanthus niruri.