HPTLC Method Development and Validation for Simultaneous Estimation of Berberine, Ellagic Acid and Ferulic Acid in Amrtadi Churna

The objective of the current study was to develop a new simple and precise High Performance Thin Layer Chromatography (HPTLC) method for standardization of three biomarkers i.e., berberine, ellagic acid and ferulic acid in Amrtadi churna. Amrtadi churna is mainly used for hyperacidity and contains dried plant parts of Gokshur, Amla and Guduchi. The method was developed and validated using precoated silica gel at 60 F254 as the stationary phase and toluene:ethyl acetate:formic acid:methanol (6:6:1.6:0.4, v/v/v/v) as the mobile phase. The detection and quantification were performed at 319 nm and the Rf value obtained was 0.35±2 for berberine, 0.5±2 for ellagic acid and 0.74±2 for ferulic acid. The method was validated as per ICH guidelines in terms of linearity, precision, specificity, accuracy and robustness.