Quantitative Estimation of Andrographolide by Reverse Phase-High Liquid Chromatography Method from Andrographis Paniculata Nees.

Abstract

Reverse Phase High performance liquid chromatographic method with UV array detection was established for the determination of Andrographolide. The Andrographolide was separated using isocratic solvent system consisting of isopropyl alcohol, formic acid and water (70:10:20 v/v) at flow rate of 1.0 ml/min and the detection wavelength of 223 nm. The method was validated for linearity, precision, accuracy, limit of detection (LOD), and limit of quantitation (LOQ). The linearity of the proposed method was obtained in the range of 4.5-70 ?g/ml for andrographolide with regression coefficient of 0.9999. Intraday and interday precision studies showed the relative standard deviation less than 2.5%. The accuracy of the proposed method was determined by a recovery study conducted at 3 different levels. The average recovery was 97-99%. The LOD and LOQ were 0.03 and 0.05 ?g/ml for andrographolide, respectively. The contents of andrographolide obtained in the dried leaves powder was within the ranges of 0.98 1.15% w/w, respectively. The proposed method is simple, sensitive yet reproducible. It is therefore suitable for routine analysis of andrographolide in A. Paniculata Nees.