The identification of conjugated linoleic acid (CLA) in Datura species represents a novel and promising avenue for biomedical exploration. CLA comprises a group of positional and geometric isomers of linoleic acid, renowned for their wide-ranging health benefits, including anti-obesity, anti-diabetogenic, and anti-carcinogenic properties. While CLA is conventionally obtained from ruminant-derived dairy and meat products, the discovery of its potential occurrence in Datura offers a compelling plant-based alternative source. Preclinical studies have established CLA’s ability to inhibit tumorigenesis via multiple mechanisms, including the induction of apoptosis, suppression of cell proliferation, modulation of immune responses, and inhibition of angiogenesis. However, clinical trials in human subjects have yielded inconsistent outcomes, highlighting the need for continued investigation into its efficacy and therapeutic scope. Building upon these insights, the present study aims to extract, purify, and characterize CLA from Datura seeds, followed by evaluation of its anticancer potential. This will involve in vitro cytotoxicity assays against human cancer cell lines and mechanistic studies to elucidate its molecular mode of action. The overarching goal is to develop a CLA-based natural formulation with potential application as an anticancer therapeutic agent