Optimization of Synergistic Anti–Helicobacter pylori Activity of Musa paradisiaca, Curcuma longa, Brassica oleracea, and Zingiber officinale Hydroethanolic Extracts Using Response Surface Methodology

Abstract

Helicobacter pylori infection remains a major global health challenge, with rising antibiotic resistance necessitating alternative therapeutic strategies. This study optimized the synergistic anti–H. pylori activity of hydroethanolic extracts of unripe plantain (Musa paradisiaca), turmeric (Curcuma longa), cabbage (Brassica oleracea), and ginger (Zingiber officinale) using Response Surface Methodology (RSM). Hydroethanolic (70% ethanol) extracts were prepared from authenticated plant materials. Phytochemical composition was determined qualitatively and quantitatively, while GC-MS identified bioactive constituents. Minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations (MBC) were determined against H. pylori isolate 35BH. A four-factor, five-level Central Composite Design (CCD) with 30 experimental runs was employed to optimize extract combinations. Response surface plots and ANOVA were used to model interactions. Model validation was performed using optimized concentrations. Ginger and turmeric exhibited the lowest MIC (125 mg/mL each), while unripe plantain and cabbage had MIC of 500 mg/mL. The quadratic model was highly significant (R² = 0.9536, adjusted R² = 0.9103, p < 0.0001), with turmeric (p < 0.0001) and ginger (p < 0.0001) demonstrating dominant linear effects. The lack of fit was non-significant (p = 0.2421). Optimal concentrations were: unripe plantain 375 mg/mL, cabbage 375 mg/mL, turmeric 187.5 mg/mL, and ginger 187.5 mg/mL, yielding a predicted inhibition zone of 21.87 mm. Experimental validation produced 21.87 ± 0.26 mm (1.17% deviation). The most pronounced synergy occurred between turmeric and ginger. RSM successfully optimized a synergistic polyherbal formulation with potent anti–H. pylori activity. This provides a rational framework for developing standardized phytotherapeutic interventions against H. pylori infection.