COMPARATIVE EXTRACTION, MCF-7 CYTOTOXICITY SCREENING, AND BIOASSAY-GUIDED FRACTIONATION OF TURRAEA VILLOSA ROOT EXTRACTS

Abstract

Background:Turraea villosa,an underexplored member of the Meliaceae family, may contain cytotoxic secondary metabolites. Extraction methods can markedly affect yield, phytochemical enrichment, and biological activity.

Objective: This study compared seven root extract preparations of Turraea villosa and used MCF-7 breast cancer cell viability reduction to guide selection and fractionation of the superior extract.

Methods: Authenticated Turraeavillosaplant material collected from Gaganbavada forest, Kolhapur, Maharashtra, India, was extracted by Soxhlet extraction, ultrasonication-assisted extraction, accelerated solvent extraction (ASE), and supercritical fluid extraction (SFE). Methanol and water were used for solvent-based methods, while SFE was performed using supercritical CO2 at 350 bar and 55°C for 90 min with a CO2 flow rate of 30 g/min and ethanol as a co-solvent. Extract yields were calculated as % w/w. MCF-7 cells were treated with extracts and SFE fractions, and cell viability was determined using the MTT assay.

Results: SFE produced the highest yield (12.15% w/w) and showed the greatest extract-level reduction in MCF-7 cell viability, with 54.04% viability at 10 µg/mL and the lowest estimated IC₅₀ (11.02 µg/mL). ASE-water and Soxhlet-water also showed notable activity, with estimated IC₅₀ values of 14.18 and 15.60 µg/mL, respectively. SFE was fractionated into ten fractions. F2 and F3 produced the greatest reduction in MCF-7 cell viability, with viability values of 32.39% and 31.88%, respectively, at the screening concentration. F3 showed the lowest viability among the tested fractions and was prioritised for further phytochemical profiling.

Conclusion: SFE was the most suitable extraction method for enriching constituents that reduced MCF-7 cell viability under the tested conditions. These findings support further LC-MS-guided identification and mechanistic validation of active SFE fractions. However, because untreated cells served as the assay control and no reference anticancer drug or assessment of normal-cell selectivity was included, the results should be interpreted as preliminary cytotoxicity screening evidence rather than as confirmed anticancer efficacy.