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

Objective: The present study was proposed to quantitatively estimate the amount of three marker compounds; curcuminoids, piperine, and gallic acid in a multicomponent ayurvedic formulation using high-performance thin layer chromatographic (HPTLC) method for routine analytical work. Methods: TLC separation was performed on silica gel 60 F plates using toluene:ethyl acetate:formic acid:methanol (5.6:2.2:1.2:1.0 v/v/v/v) as mobile phase. Plate was developed by to a distance of 90 mm at ambient room temperature with 20 minutes saturation time. Densitometric analysis was performed at 327 nm. Method was validated as per International Conference on Harmonization Q2 (R1) guideline also. 254 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 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 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, 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% w/w, and 0.8% w/w, respectively, when analyzed quantitatively by developed validated HPTLC method. Conclusion: The method can be used as a tool for quality control of herbal formulation. Keywords: Curcuminoids, Piperine, Gallic acid, High-performance thin layer chromatographic.