SIMULTANEOUS ESTIMATION OF CURCUMINOIDS, PIPERINE, AND GALLIC ACID IN AN AYURVEDIC FORMULATION BY VALIDATED HIGH-PERFORMANCE THIN LAYER CHROMATOGRAPHIC METHOD

Abstract

<p>ABSTRACT<br />Objective: The present study was proposed to quantitatively estimate the amount of three marker compounds; curcuminoids, piperine, and gallic<br />acid in a multicomponent ayurvedic formulation using high-performance thin layer chromatographic (HPTLC) method for routine analytical work.<br />Methods: TLC separation was performed on silica gel 60 F<br /> plates using toluene:ethyl acetate:formic acid:methanol (5.6:2.2:1.2:1.0 v/v/v/v) as<br />mobile phase. Plate was developed by to a distance of 90 mm at ambient room temperature with 20 minutes saturation time. Densitometric analysis<br />was performed at 327 nm. Method was validated as per International Conference on Harmonization Q2 (R1) guideline also.<br />254<br />Results: Piperine, curcuminoids, and gallic acid were separated on TLC at retention factor values of 0.71, 0.61, and 0.29, respectively. The described<br />method was linear over the range of 300-700 ng/spot, 100-300 ng/spot, and 250-550 ng/spot, respectively, for curcuminoids, piperine, and gallic<br />acid. The accuracy of the method was assessed by recovery studies and was found to be 101.71%, 99.67%, and 99.59% for curcuminoids, piperine,<br />and gallic acid, respectively. The amount of curcuminoids, piperine, and gallic acid in the ayurvedic formulation was found to be 3.99% w/w, 1.9%<br />w/w, and 0.8% w/w, respectively, when analyzed quantitatively by developed validated HPTLC method.<br />Conclusion: The method can be used as a tool for quality control of herbal formulation.<br />Keywords: Curcuminoids, Piperine, Gallic acid, High-performance thin layer chromatographic.</p>