Stability Indicating RP- HPLC Method for Determination of Andrographolide

Author(s)

Damle, M., Gujar, N.

Author's Affiliation

Abstract

Andrographis Paniculata (Kalmegh) is used since ancient time for treatment of various diseases viz cancer, malaria, diabetes etc. Andrographolide is one of the major phytoconstituent responsible for the activity, which is a diterpene lactone. An approach for the stress degradation was successfully applied for the development of stability indicating HPLC method for the determination of Andrographolide. The system comprised of HiQ Sil C₁₈ (150*4.6 mm particle size 5μ) column as stationary phase and 65:35 v/v mixture of methanol:water as mobile phase(flow rate 1ml/min) and a UV detector at wavelength of 226nm. The retention time of Andrographolide was found to be 3.85 ± 0.2min. The marker was subjected to acid, base, neutral hydrolysis, oxidation, thermal degradation and photolysis. Stress testing of Andrographolide was carried out according to the International Conference of Harmonization (ICH) guideline Q1A (R2). The method was successfully validated according to ICH guidelines Q2 (R1). The calibration curve was linear over the concentration range of 10 -50 μg/ml (r²=0.993). The accuracy of the method was established based on the recovery studies. The LOD and LOQ of Andrographolide was found to be 0.60 µg/ml and 1.82 µg/ml respectively. Among various stressed conditions, Andrographolide showed considerable degradation under alkali and acid catalyzed hydrolysis, oxidative and photolytic condition.

Keywords

Andrographis Paniculata, Andrographolide High Performance Liquid Chromatography, Stability Indicating Method

Cite This Article

Damle, M., Gujar, N. (2016). Stability Indicating RP- HPLC Method for Determination of Andrographolide, International Journal for Pharmaceutical Research Scholars (IJPRS), 5(3), 47-53.