Phenolic Profiling and OECD-Guided Acute and Subacute Oral Toxicity Assessment of Cannabis sativa L. Seed Hydroethanolic Extract

Cannabis sativa L. seeds have been traditionally used in Moroccan folk medicine and are increasingly recognized as a valuable source of nutrients and bioactive phytochemicals with promising therapeutic properties. Despite their widespread traditional and nutritional use, scientific evidence supporting their toxicological safety remains limited. This study aimed to characterize the phenolic profile of a hydroethanolic extract of Cannabis sativa L. seeds and to evaluate its acute and 28-day repeated-dose oral toxicity in mice following the Organization for Economic Cooperation and Development (OECD) guidelines. The phenolic composition of the extract was analyzed using high-performance liquid chromatography coupled with diode-array detection (HPLC-DAD). Acute oral toxicity was assessed according to OECD Guideline 423 after a single oral administration of 5000 mg/kg body weight. Repeated-dose oral toxicity was evaluated following OECD Guideline 407 by administering the extract orally at doses of 500 and 1000 mg/kg/day for 28 consecutive days. Clinical observations, body weight, food and water intake, relative organ weights, hematological and biochemical parameters, and histopathological examinations of the liver and kidneys were performed. HPLC-DAD analysis identified seven major phenolic compounds, namely catechin, chlorogenic acid, caffeic acid, vanillin, p-coumaric acid, quercetin, and kaempferol. No mortality, behavioral abnormalities, or treatment-related clinical signs were observed during either the acute or repeated-dose studies. The extract produced no significant alterations in body weight gain, food and water consumption, relative organ weights, liver and kidney histology, or most hematological and biochemical parameters. A significant reduction in serum total cholesterol and triglyceride levels was observed in treated animals, while platelet counts were significantly reduced only in female mice receiving 1000 mg/kg/day. Under the experimental conditions employed, the hydroethanolic extract of Cannabis sativa L. seeds exhibited a favorable oral safety profile, with an estimated LD50 greater than 5000 mg/kg. These findings support the traditional use and potential nutraceutical applications of Cannabis sativa seeds and provide a scientific basis for further pharmacological investigations and long-term safety studies.