A Process for Eco-Sustainable Extraction of Cytotoxic Flavonoids from Muntingia calabura L. using Sonication Method Optimized by Quality by Design Approach through Central Composite Model

Background

The present invention relates to an eco-sustainable, Quality by Design (QbD)- and Design of Experiments (DoE)-guided sonication-assisted extraction process for isolating cytotoxic flavonoid compounds from the leaves of Muntingia calabura L.

Materials and Methods

The invention strategically integrates green chemistry principles with statistical optimization to enhance extraction efficiency, reproducibility, and environmental compatibility. The process employs an optimized methanol-water solvent system (50-100% v/v) for selective solubilization of flavonoids, assisted by ultrasonic cavitation at controlled amplitude and temperature to facilitate rapid solvent diffusion and efficient mass transfer. The Central Composite Design (CCD) under the QbD framework was used to identify and optimize critical parameters influencing extraction yield, while statistical validation through Response Surface Methodology (RSM) and Analysis of Variance (ANOVA) confirmed model significance (R²=0.956, p<0.05).

Results

The optimized conditions-75% methanol, 60 min of sonication, and 50±2ºC-yielded the maximum flavonoid recovery (23.7±0.3%) with high purity. The ethyl acetate fraction exhibited enhanced flavonoid concentration and potent cytotoxicity against colon cancer (HCT-116) cell lines, with an IC₅₀ value of 185.6 µg/mL compared to 218.2 µg/mL for the crude extract.

Conclusion

This invention provides a scalable, reproducible, and environmentally sustainable process capable of generating bioactive flavonoid-rich fractions with significant anticancer potential, thereby advancing green extraction methodologies in phytopharmaceutical and nutraceutical development.