Phytochemical analysis of Diospyros buxifolia (Blume) Hiern. bark extract via LC-MS/MS and evaluation of its antiproliferative activity against human colon carcinoma cells

Bioactive phytochemicals have shown a very significant role in pharmacological ailments in humans. The literature review indicates the extensive use of Diospyros extracts for the treatment of diseases related to the oral cavity and gastrointestinal disorders. Further, the bark, fruit, and root are the most commonly utilised components in traditional preparations. Due to a lack of awareness about Diospyros buxifolia (DB) and its underutilized status, relatively few studies have been conducted on its nutraceutical potential. Hence, the present study aimed to identify bioactive compounds in DB bark extracts using liquid chromatography mass spectroscopy (LC-MS/MS) and evaluate their antiproliferative activity against human colon carcinoma cells. The LC-MS/MS chromatogram showed the presence of various phytochemical compounds such as O-Phosphoryl ethanolamine, Dulcitol, 2-(2-Aminoethyl) pyridine, Alpha-Lipoic acid, 14-(Hydroxymethyl)-5,9-dimethyltetracyclo-hexadecan-5-ol, Geranyl acetone, Retinol, Retinoids, Pimelic acid, 3-Methyl-2-oxovaleric acid sodium, and Prolinamide, etc. An extensive literature survey revealed various pharmaceutical properties. The extract demonstrated dose-dependent antiproliferative effects against HT-29 and HCT-116 cells with IC50 values of 43.5 ± 4.2 and 48 ± 3.7 µg/ml, respectively, by MTT assay. Moreover, the extract enhanced the reactive oxygen species (ROS) level, altered cellular morphology, and induced apoptosis as evaluated by fluorescence microscopy. Current findings suggest DB produces significant levels of ROS (1.8-fold), which may cause apoptosis by disrupting the balance between the oxidant and antioxidant enzyme systems to alter the redox balance in the carcinoma cells tested. Interestingly, DB at the highest concentrations significantly enhances the release of lactate dehydrogenase (2.7-fold) in HT-29 and HCT-116 cells. This result suggests the involvement of necrosis in the cytotoxic effect of DB. In conclusion, DB includes strong anticancer agents with notable antiproliferative activity, which makes it suitable for application in medicine and the food industry.