METHOD DEVELOPMENT AND VALIDATION OF RP-HPLC METHOD FOR THE SIMULTANEOUS ESTIMATION OF ANDROGRAPHOLIDE IN HERBAL FORMULATIONS AND HUMAN PLASMA

Nandhini Priya V.*, Kamalakannan D., Manivannan R., Ragul M., Kannagi A., Mahavishnu B., Nivash E.

Andrographolide, a bioactive diterpenoid lactone isolated from Andrographis paniculata, exhibits significant pharmacological activities such as anti-inflammatory, antioxidant, and antidiabetic effects. Accurate estimation of andrographolide in herbal formulations and biological matrices is essential for quality control and pharmacokinetic evaluation. The present study aimed to develop and validate a simple, sensitive, and reliable reverse-phase high-performance liquid chromatography (RP-HPLC) method for the simultaneous estimation of andrographolide in herbal formulations and human plasma using gallic acid as an internal standard. Chromatographic separation was achieved under optimized conditions using a suitable mobile phase, controlled flow rate, and UV detection, resulting in well-resolved and symmetrical peaks. The developed method was validated according to ICH guidelines for parameters including specificity, linearity, accuracy, precision, limit of detection, limit of quantification, robustness, and stability. The method demonstrated good linearity over the selected concentration range, with acceptable accuracy and precision values. Stability studies confirmed that andrographolide remained stable under various conditions. The validated RP-HPLC method was found to be reliable, reproducible, and suitable for routine analysis of andrographolide in pharmaceutical formulations and biological samples.