Greenness Assessment and Quantitative Profiling of Glycyrrhizic Acid, Piperine and Ursolic Acid Content in Polyherbal Cough Syrup Using High-Performance Liquid Chromatographic Analysis

Abstract The increasing demand for safe, effective and sustainable alternatives to traditional cough suppressants has brought polyherbal formulations in respiratory ailment management. This report outlines the development and greenness evaluation of a high-performance liquid chromatography method for the simultaneous quantification of glycyrrhizic acid, piperine and ursolic acid from crude plant extracts and commercial polyherbal cough syrups. Chromatographic separation was performed using Agilent Eclipse C18 column (250 × 4.6 mm, 5 μm) under gradient elution using 0.1% formic acid in water and acetonitrile as mobile phases. The optimal absorbance wavelengths during quantification were 254 nm for glycyrrhizic acid, 342 nm for piperine, and 210 nm for ursolic acid at retention times of 21.3, 24.5 and 31.0 min, respectively. Results indicated a method that was sensitive, reproducible and specific to the three analytes quantified as well as validated according to ICH Q2(R1). Evaluating the greenness of the method (using the Analytical GREEnness metric, Green Analytical Procedure Index, and Eco-Scale Assessment confirmed that this method could be considered “green”. Upon completion of method validation, the method was used to quantify each marker compound from commercially available products in an effort to better understand production methodologies and directed our quality control and standardization efforts. The results of this systematic investigation point to the advantage of applying green analytical techniques for the assessments of polyherbal medicines, potentially establishing them as credible, environmentally friendly therapeutic options for respiratory ailments.