Development and validation of RP HPLC method for simultaneous determination of picroside I, picroside II, phyllanthin and boeravinone B in a polyherbal hepatoprotective tablet formulation

• Published in: Tropical journal of pharmaceutical research 2022-03, Vol.21 (3), p.619-627
• Main Authors: Shanbhag, Sunil V, Bachute, Madhusudan T
• Format: Article
• Language: English
• ISSN: 1596-5996; 1596-9827
• DOI: 10.4314/tjpr.v21i3.23

Purpose: To develop and validate RP-HPLC method for simultaneous determination of four active ingredients: picroside I (PSI), picroside II (PSII), phyllanthin (PHY) and boeravinone-B (BVB) in a polyherbal hepatoprotective tablet formulation.Methods: The study was carried out using Waters X-Bridge, C18 (250 mm x 4.6 mm, 5 μm) column with mobile phase consisting of 5 mM ammonium acetate in 10 % methanol and acetonitrile, with gradient programme at dual wavelengths of 220 nm and 274 nm and flow speed of 1 mL.min-1. The procedure was validated with respect to specificity, linearity, precision, accuracy, system suitability, limit of detection (LOD) and of quantification (LOQ), and robustness in line with ICH specifications.Results: The method was linear within the concentration range of 25 to 200 %, and the values of correlation coefficients (R2) were > 0.999. Intra-day and inter-day RSDs of PAs and RTs were < 5.0 %, with recovery in the range of 100.0 - 106.0 %.Conclusion: The four active ingredients have with good resolution with regard to the method used. The method is rapid, simple, highly selective, sensitive and cost effective, which make it an efficient method for quality assurance.