Activity-guided Fractionation and in Vitro Anticancer Evaluation of Bioactive Phytoconstituents From Clitoria Ternatea L. Flowers Against HT-29 Human Colorectal Cancer
Abstract
Abstract Natural products continue to play a significant role in the discovery of anticancer agents because of their diverse bioactive constituents. Clitoria ternatea L. (Butterfly pea) is a medicinal plant rich in flavonoids, anthocyanins, phenolic compounds, and other phytochemicals with reported therapeutic potential, although the specific constituents responsible for its anticancer activity remain inadequately explored. In this study, the ethanolic flower extract was fractionated by liquid–liquid partitioning using n-hexane, chloroform, and ethyl acetate, followed by phytochemical screening and thin-layer chromatography (TLC) fingerprinting. The cytotoxic potential of the fractions was assessed against HT-29 human colorectal adenocarcinoma cells using the MTT assay (1–1000 µg/mL), and IC₅₀ values were calculated with GraphPad Prism. Among the fractions, the ethyl acetate fraction showed the highest yield and was enriched with flavonoids, anthocyanins, phenolics, glycosides, and saponins. TLC analysis revealed five distinct bands, including one with an Rf value comparable to quercetin. This fraction also demonstrated the strongest cytotoxic effect (IC₅₀ = 79.29 µg/mL), followed by the hexane (99.40 µg/mL) and chloroform (169.0 µg/mL) fractions, with concentration-dependent morphological changes in HT-29 cells. These findings indicate that activity-guided fractionation effectively enriched bioactive constituents, highlighting the ethyl acetate fraction as a promising…
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