Application of molecular docking approach in a novel eco-friendly impurity profiling HPLC-UV method for the simultaneous estimation of ternary hypoglycemic pharmaceutical mixture

This study presents a novel, eco-friendly high-performance liquid chromatography-ultraviolet (HPLC-UV) method for the simultaneous quantification of a ternary mixture of hypoglycemic drugs—metformin hydrochloride (MTF), pioglitazone hydrochloride (PGT), and glibenclamide (GBC)—along with two toxic impurities, cyanoguanidine (CYG) and melamine (MEL). The method utilizes a VDSpher Pur 100 C18-E column (250 mm × 4.6 mm, 5 µm) with a gradient elution system comprising 0.1 M heptane sulfonic acid (pH 2.2) and acetonitrile as the mobile phase, at a flow rate of 1.5 mL/min. Detection was performed at 225 nm using a photodiode array (PDA) detector. The retention times for CYG, MEL, MTF, PGT, and GBC were 1.749, 2.950, 3.640, 5.062, and 7.788 min, respectively, demonstrating effective separation within a reasonable timeframe. The method was rigorously validated according to ICH Q2(R1) guidelines, confirming its specificity, precision, accuracy, limits of detection (LOD), and limits of quantification (LOQ). Furthermore, the study incorporated molecular docking simulations to assess the toxicity of melamine by evaluating its binding to albumin's arachidonic acid sites. The greenness of the analytical method was evaluated using the Green Analytical Procedure Index (GAPI) and an analytical greenness calculator, indicating its environmental acceptability. This approach offers a significant advancement by simultaneously analyzing these components, addressing a gap in existing literature for such complex mixtures and their impurities, while also emphasizing environmental sustainability in analytical practices.

This study presents a novel, eco-friendly high-performance liquid chromatography-ultraviolet (HPLC-UV) method for the simultaneous quantification of a ternary mixture of hypoglycemic drugs—metformin hydrochloride (MTF), pioglitazone hydrochloride (PGT), and glibenclamide (GBC)—along with two toxic impurities, cyanoguanidine (CYG) and melamine (MEL). The method utilizes a VDSpher Pur 100 C18-E column (250 mm × 4.6 mm, 5 µm) with a gradient elution system comprising 0.1 M heptane sulfonic acid (pH 2.2) and acetonitrile as the mobile phase, at a flow rate of 1.5 mL/min. Detection was performed at 225 nm using a photodiode array (PDA) detector. The retention times for CYG, MEL, MTF, PGT, and GBC were 1.749, 2.950, 3.640, 5.062, and 7.788 min, respectively, demonstrating effective separation within a reasonable timeframe. The method was rigorously validated according to ICH Q2(R1) guidelines, confirming its specificity, precision, accuracy, limits of detection (LOD), and limits of quantification (LOQ). Furthermore, the study incorporated molecular docking simulations to assess the toxicity of melamine by evaluating its binding to albumin's arachidonic acid sites. The greenness of the analytical method was evaluated using the Green Analytical Procedure Index (GAPI) and an analytical greenness calculator, indicating its environmental acceptability. This approach offers a significant advancement by simultaneously analyzing these components, addressing a gap in existing literature for such complex mixtures and their impurities, while also emphasizing environmental sustainability in analytical practices. Research Background and Significance The pharmaceutical industry demands rigorous analytical methods for the accurate quantification of active pharmaceutical ingredients (APIs) a