Quantification of Ellagic Acid, Gallic Acid and Picroside-I from Phalatrikadi kvatha Churna by HPTLC

Objective: Phytochemical evaluation of an ayurvedic formulation Phalatrikadi kvatha churna and establishment of multiple marker based methods for its standardization using marker compounds/biomarkers viz. picroside-I, gallic acid and ellagic acid. Materials and methods: Preliminary phytochemical analysis, development of fingerprint profiles and estimation of picroside - I, gallic acid and ellagic acid from the authentic sample and two market samples of the formulation. Results: Preliminary phytochemical screening showed the presence of phenols, tannins, flavonoids and sterols. The solvent systems developed for establishing fingerprint profiles and picroside-I, gallic acid, and ellagic acid gave good resolution of these compounds from the other components of the extract. The HPTLC methods developed for the estimation of marker compounds were validated in terms of instrumental precision, accuracy and repeatability. Picroside-I content of the authentic sample was found to be 0.16 %, whereas the two market samples contained very less picroside-I (~0.038%). The amount of gallic acid in the authentic sample was found to be 0.78%, while the other two market samples contained only about half the amount. The amount of ellagic acid in the authentic sample was found to be 0.12%, while the market samples contained 0.085% and 0.065% of free ellagic acid respectively. Conclusion: Qualitative and quantitative phytochemical evaluation of Phalatrikadi kvatha churna was carried out for multimarker based standardization using picroside-I, gallic acid, and ellagic acid by HPTLC. The TLC fingerprint profiles and TLC densitometric methods for the quantification of different markers can be used in the quality control of the formulation. The TLC densitometric methods developed can also be adopted for the quantification of these marker compounds in other preparations as well.