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Black carrot (Daucus carotaL. ssp. sativusvar. atrorubensAlef.), an indigenous vegetable of India, has been widely used for the preparation of traditional fermented product Kanji, and traditional medicine for treating various kinds of diseases. Although previous research has reported its biochemical composition and some in vitro functional properties, very scanty information is available to date on the in vivo effect of black carrot. Given this gap of knowledge, this study was aimed to investigate the effect of black carrot on antioxidant defense, glucose metabolism and renal, as well as hepatic toxicity in normal Charles Foster albino rats. In addition, secondary metabolites were identified by high-resolution LCMS and in vitro antioxidant activity was also assessed. LC/MS/MS analysis revealed the presence of nine anthocyanins and two flavonols. Among these, four new compounds were putatively identified for the first time. Under in vivo condition, feeding of black carrot juice for 30 days resulted in the reduction in the superoxide dismutase activity, both in dose- and time-dependent manner. It was also observed that feeding of rats with black carrot juice for 30 days reduced their blood triglyceride concentration, both in dose- and time-dependent manners. No significant effect was observed on plasma glucose concentration, and also, no toxicity was noted with regard to the functioning of the kidney and liver.