Development and validation of hplc method for determination of andrographolide in raw material and tablet of ethyl acetate fractions of Andrographis paniculata

Abstract

A simple and selective HPLC method for the determination of andrographolide in raw material and tablet formulation of Andrographis paniculata was developed and validated. The Method was performed using a Poroshell 120 EC-C18 column (3.0 x 50 mm i.d., 2.7 μm particle size), column temperature was maintained at 30°C, isocratic elution with mobile phase of methanol: water (pH 3.05 with phosphoric acid) (50:50 v/v) with flow rate of 0.3 ml/min., 0.5 μl injection volume and detected at 228 nm. The proposed method is selective to separate the peak of andrographolide from other components, generating symmetry peak and good resolution with the retention time of andrographolide of 2.6 min. The calibration plots showed a good linear relationship with r2 = 0.9996 in the concentration range from 50 to 1000 ppm. The LOD and LOQ were 4.89 ppm and 16.19 ppm, respectively. The recovery of the method was found between 93.76 and 101.72% and the relative standard deviations of method was found between 1.60% and 2.39%. The proposed method is suitable for quality control of raw material and tablet formulation of Andrographis paniculata.