Abstract: Background and Objective: Egypt produced 236,314 t of artichoke in 2016, which produce a huge amount of useless by-product, which can be used as cheaper source for many active compounds can be applied for some medical application. The objective of this study was to assess the toxicity of the artichoke by-product extract through its effect on rats’ kidney, brain and liver biomarkers. Materials and Methods: Chemical composition of artichoke by-product (crude protein, crude fiber, crude fat and minerals) was determined. Conventional extraction (CE), microwave-assisted extraction (MAE) and ultrasonic-assisted extraction (UAE) extraction methods were used for artichoke by-product and comparison between them were performed according to antioxidant activity using DPPH and the phenolic profile identity using HPLC technique. Chronic oral gavage of thirty adult male albino rats for 4 weeks in the concentrations of (0.1, 0.5, 1 and 5 g kg–1) artichoke by-product extract was used for evaluation of its toxicity. Results: MAE with ethanol more suitable for extraction of the polyphenols (193.63±4.9 μg gallic acid equivalents (GAE) mg–1) and showed IC50 = 159.7 mg mL–1. Three major active phenolic compounds were identified benzoic acid, ellagic acid and caffeine. Rats administrated 5 g kg–1 artichoke extract have no changes in brain, liver and kidney parameters (p<0.05). Histology of brain and liver exhibited normal architecture. Conclusion: The results showed that the artichoke by-product extract had no any toxic effect on rats and considered be safe for human use even at a high level of doses (up to 5 g kg–1).