Development and validation of simultaneous equation method for the estimation of andrographolide and apocynin in hepatoprotective polyherbal formulation using UV–visible spectrophotometry

Abstract

The aim of this study was to develop a simple, accurate, and precise UV–Vis spectrophotometric method for the simultaneous determination of Andrographolide and Apocynin, bioactive compounds derived from Andrographis paniculata and Picrorhiza kurroa, respectively. These compounds are known for their hepatoprotective and antioxidant properties, and accurate quantification is essential for quality control of herbal formulations. A simultaneous equation method was developed based on the additive absorbances of Andrographolide and Apocynin at their respective λmax values (224 nm for Andrographolide and 274 nm for Apocynin). Linearity was established over a concentration range of 0.5 to 2.5 µg/mL for both compounds. Validation was conducted for parameters such as limits of detection (LOD), limits of quantification (LOQ), precision, and recovery. The method was applied to the analysis of three marketed herbal formulations. The method exhibited excellent linearity with correlation coefficients of 0.999 for Andrographolide and 0.998 for Apocynin. LODs were 0.012 µg/mL for Andrographolide and 0.066 µg/mL for Apocynin, with LOQs of 0.039 µg/mL and 0.202 µg/mL, respectively. Precision, confirmed by relative standard deviation (RSD) values below 2%, and recovery studies (98.66 %–102 % for Andrographolide, 98 %–101.60 % for Apocynin) demonstrated the method’s accuracy. The method successfully quantified the compounds in marketed herbal formulations, with results consistent with the label claims. The UV–Vis spectrophotometric method offers a reliable, precise, and cost-effective approach for the simultaneous quantification of Andrographolide and Apocynin in herbal formulations. Its simplicity and efficiency make it suitable for routine quality control in the pharmaceutical industry.