Simultaneous spectrophotometric eco-friendly analysis of triple-drug H.pylori regimen (Vonoprazan, Amoxicillin, Metronidazole) for quality control and in vitro dissolution testing

The bacterium Helicobacter pylori is considered a major factor in the pathogenesis of gastric ulcers, gastrointestinal disorders, and gastric cancer, leading to the approval of multiple eradication strategies. Among these is the triple therapy regimen comprising vonoprazan, a potassium-competitive acid blocker, with the antimicrobials amoxicillin and metronidazole. This study introduces Dual-wavelength in ratio spectra, Double Divisor Ratio Spectra Derivative, and Mean Centering of Double Divisor Ratio Spectra methods as advanced spectrophotometric techniques that effectively resolved overlapping spectra of the studied mixture. It is the first instance of using these methodologies to determine the cited mixture. The concentration ranges analyzed were 2–60 µg/mL, 2–32 µg/mL, and 2–25 µg/mL for vonoprazan, amoxicillin, and metronidazole, respectively. Greener non-organic solvent was used to ensure environmental sustainability of analytical methods, significantly reducing ecological and health hazards. Greenness assessment was evaluated using GAPI and AGREE metrics, and blueness was assessed by BAGI. One-way ANOVA comparative assessment revealed no statistically significant differences from the reported method. Validation parameters were evaluated according to ICH guidelines, demonstrating sensitivity and selectivity; therefore, the methods are suitable for routine quality control analysis of the analytes in their ternary mixture and pharmaceutical formulation without prior interference from excipients.