Ethnopharmacological relevanceThe safety assessment of herbal products is critical for their appropriate pharmacological applications. Garcinia cowa Roxb., commonly known as Cha-muang in Thai, has ethnopharmacological relevance in inflammation, infectious diseases, and diabetes. The leaf extracts of G. cowa have been extensively reported for their anticancer, anti-inflammatory, antimicrobial and antioxidative effects. Notably, chamuangone is their major active constituent that contributes to various pharmacological properties. Aim of the studyThe current study aims to establish a standardized chamuangone enriched extract (CEE) and assess its acute and sub-acute toxicities in animal models. MethodologyCEE was established from G. cowa leaves using microwave-assisted extraction (MAE), followed by fractionation and enrichment through silica gel vacuum and column chromatography. The concentration of chamuangone in the leaves was quantified using validated quantitative high-performance liquid chromatography (HPLC) method. The safety profile of CEE was thoroughly evaluated in rodents according to the Organization for Economic Cooperation and Development (OECD) guidelines, specifically 425 and 407. The effect on oxidative stress markers such as superoxide dismutase (SOD), reduced glutathione (GSH), catalase (CAT), and malondialdehyde (MDA) levels were also evaluated in various organs. ResultsBase on the quantitative HPLC analysis the CEE containing 73.0 ± 2% w/w of chamuangone in the extract. In the acute toxicity study, following up and down procedure the female rats were dosed with CEE at 1750 and 550 mg/kg body weight (b.w.), with CEE 1750 mg/kg b.w. identified as toxic, causing mortality, while CEE 550 mg/kg b.w. was deemed safe. The LD50 value was calculated according to the standard protocols, resulting in 970 mg/kg b.w. In histopathological examination, CEE 550 mg/kg b.w. were safe in all the selected organs while the CEE 1750 mg/kg b.w. treated rats exhibited toxic effects in histological tissues sections in the form of necrosis in brain, cardiac muscle hypertrophy, liver inflammation, mild untoward in the spleen, fibrosis in lungs, pancreatitis, pyelonephritis, and ovaries cyst. Administration of CEE at doses of 550 mg/kg b.w. (single dose) in acute and 100 mg/kg b.w. (regularly 28-days) in sub-acute toxicities studies significantly (p < 0.05) decreased levels of uric acid, triglycerides, and cholesterol. Importantly, the CEE 550 and 100 mg/kg b.w. also significantly increased the levels of antioxidant enzymes (SOD, GSH and CAT) and decreased malondialdehyde levels. Normal histopathology was observed in the sub-acute toxicity study in all treated groups. ConclusionThe study successfully concludes that CEE at a dose of 100 mg/kg b.w. is safe for therapeutic application or use as a chemo preventive functional food utilizing green extraction methods. However, chronic toxicity studies are recommended to validate safety concerns over an extended period.