Development and validation of hplc method for determination of andrographolide in raw material and tablet of ethyl acetate fractions of Andrographis paniculata Ihsan Bachtiar Rifai Pratita1,4, Hafid Achmad Fuad2,3, Primaharinastiti Riesta1, Widyawaruyanti Aty2, Yuwono Mochammad1,* 1Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Airlangga University, Indonesia 2Department of Pharmacognosy and Phytochemistry, Faculty of Pharmacy, Airlangga University, Indonesia 3Natural Product Medicines Research and Development, Institute of Tropical Disease, Airlangga University, Indonesia 4Department of Pharmaceutical Chemistry Pharmacy, Faculty of Medicines, Brawijaya University, Malang, Indonesia *Corresponding Author E-mail: mochammad.yuwono@gmail.com
Online published on 16 September, 2020. 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. Top Keywords High Performance Liquid Chromatography, Method validation, Andrographis paniculata, Andrographolide, Ethyl Acetate Fraction. Top |