Determination of Phyllanthin and Gallic Acid In Herbal Hepatoprotective Formulation By TLC-Densitometry Analysis

Introduction: Phyllanthus niruri, Andrographis paniculata and Embilica officinalis are most important and high value medicinal plant known for its hepatoprotective activity. Several analytical methods including HPTLC and HPLC are reported for respective phytoconstituents. However, there is no simultaneous estimation of phyllanthin and gallic acid reported. The Objective of present work is to develop and validate HPtlc method for simultaneous determination of phyllanthin and gallic acid in polyherbal hepatoprotective formulation. Method: The method is employed tlc alumina plates precoated with silica gel 60F-254 as a stationary phase and toluene: ethyl acetate: formic acid (5:3.5:0.5 v/v/v) as a mobile phase. The standard markers namely phyllanthin and galic acid were identified in polyherbal formulation containing P.niruri and E.officinalis by Rf value. Densitometric analysis of phyllanthin and gallic acid was carried out at 254nm. Results: The precision of method was confirmed by relative standard deviation (RSD) which was lower than the 2%. This method was found to give proper separation of phyllanthin and gallic acid. The linear regression data for calibration plot shown good relationship with r2=0.998 (phyllanthin) and 0.9991(gallic acid) in the concentration range 5-9 µg/band. The method was validated for precision, accuracy, sensitivity, and specificity. Conclusion: statistical analysis proves that method is accurate and reproducible. The method is economical and can be employed for routine analysis of marketed polyherbal hepatoprotective formulation containing P.niruri and E.officinalis.