Phytochemical Screening of Diospyros malabarica Bark Extracts via GC-MS/MS and Assessment of In Vitro Antiproliferative Activity Against Human Colon Carcinoma Cells

Abstract

Diospyros malabarica bark extract is used in Ayurvedic medicine, particularly to reduce gastrointestinal motility and diarrhoea. This study aims to identify the bioactive compounds of D. malabarica using GC-MS/MS and to assess antiproliferative activity against human colon carcinoma cells (HT-29 and HCT-116). The GC-MS/MS chromatogram indicated the presence of 16 phytocompounds. A comprehensive review indicated that 8 bioactives (Hydroquinone(1,4-dihydroxybenzene), 3-Methoxycatechol, 2-Methoxyresorcinol, 2,6-Dimethoxyphenol (Syringol), 1,2,3-Benzenetriol (Pyrogallol), 2,4,6-Trihydroxybenzoic acid, 3,4,5-Trimethoxyphenyl acetate, and Glycerol 1-palmitate) possess anticancer properties. The extract demonstrated dose-dependent in vitro antiproliferative effects against HT-29 and HCT-116, with IC₅₀ values of 53 ± 4.2 and 58 ± 5.3 μg/ml, as determined by the MTT assay. The extract increased ROS levels (2.6-fold, p<0.01), altered cellular morphology, and triggered apoptosis, as revealed by fluorescence microscopy. Results indicate that D. malabarica generates substantial ROS to promote apoptosis by disrupting the balance between oxidant and antioxidant enzyme systems, thereby shifting the cellular redox balance. Notably, at higher concentrations, D. malabarica significantly elevated lactate dehydrogenase (3.2-fold), suggesting late apoptosis and consequent cytotoxicity. Overall, our findings suggest that D. malabarica contains strong anticancer agents that exhibit marked antiproliferative and apoptotic activity against human colon carcinoma cells; hence, extracts may offer benefits for gastrointestinal disorders.