DEVELOPMENT AND VALIDATION OF HPTLC METHOD FOR ESTIMATION OF GALLIC ACID AND ANDROGRAPHOLIDE IN POLYHERBAL FORMULATION.

Shingala, Dhara; Gaudani, Rashmi; Nayak, Bhavesh; Ghodasara, Trupti

A Gallic acid and Andrographolide possesses a wide range of biological activities and is used for the treatment of liver diseases. A simple HPTLC method has been developed for the quantification of Gallic acid and Andrographolide. Analysis of Gallic acid and Andrographolide were performed on TLC aluminium plates pre-coated with silica gel 60F-254 as stationary phase. Linear ascending development were carried out in twin trough glass chamber saturated with mobile phase consisting of Toluene: Acetone: Formic acid [9: 7: 1 v/v/v] for Gallic acid and Andrographolide at room temperature [25 ± 2°C]. Spectrodensitometric scanning was performed by TLC scanner III (CAMAG) in absorbance mode at the wavelength of 254nm for Gallic acid and Andrographolide. The system was found to give compact spots for Gallic acid (Rf value of 0.45) and Andrographolide (Rf value of 0.51). The data for calibration plots showed good linear relationship with R² = 0.996 for Gallic acid and R² = 0.996 for Andrographolide in the concentration range of 200-1200 ng/spot and 100-600 ng/spot with respect to peak area. Gallic acid and Andrographolide were found to be 0.65% and 0.47% respectively in Polyherbal (Capsule) Formulation. The method was validated for accuracy, precision and recovery. The limits of detection and limits of quantification were determined. Statistical analysis of the data showed that the method is reproducible and selective for estimation of Gallic acid and Andrographolide.

GALLIC acid; DITERPENES; THIN layer chromatography; ALUMINUM plates; SILICA gel; ABSORPTION spectra; DRUG formularies; DETECTION limit

Pharma Science Monitor, 2012, Vol 3, Issue 4, p2673

0976-9242

Academic Journal