Development and Validation of a Novel HPLC Method for the Analysis of Andrographolide in Marketed Herbal Formulation

Abstract

A specific and straightforward HPLC method is developed and validated for the analysis of andrographolide in crude material. The method was performed utilizing Poroshell 120 EC C18 column (3.0 x 50mm i.d, 2.7µm molecule size), Column temperature was kept 25ºC, isocratic elution with mobile phase of methanol and water (53:47) observed at 223nm wavelength with infusion volume of 20µl. The purposed technique is particular to isolate the peak of andrographolide from different segments, at retention time of 6.6 min with r2 value of 0.9992 in the range of 230 to 1000ppm. The LOD and LOQ were found to be 0.013 ppm and 0.041 ppm respectively. Correlation coefficient was valued 0.9992 in the 230 to 1000 ppm concentration range, hence calibration plots showed good linear relationship. The recovery of the method was found to be between 109.06-116.23%. The purposed technique is appropriate for the quality control of the raw material of Andrographis paniculata.