Perennial grasses are hosts to an extremely diverse assemblage of endophytic fungi, but their significance is still underexplored. In the present study, an endophytic fungus was isolated from the aerial regions of Digitaria bicornis (Lam.) Roem. & Schult. and was characterized by morphological and molecular methods (ITS rDNA region), as Penicillium citrinum Thom. The crude extracts of endophytic fungus and host were recovered and evaluated for their antioxidant potential by spectrophotometric and electrochemical methods. The present study was also an attempt to compare the anti-radical power of extracts by spectrophotometric (2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid (ABTS), hydrogen peroxide and nitric oxide radical scavenging assay) and electrochemical (cyclic voltammetry) methods and suggested that cyclic voltammetry could be used routinely instead of assaying by more number of spectrophotometric methods. The results indicated that the ethyl acetate extract of P. citrinum and methanolic extract of D. bicornis has potential compounds with antioxidant and other pharmaceutical activities. Nine and 17 antioxidant biomolecules, respectively, in P. citrinum and D. bicornis extracts were detected by OHR-LC-MS and the presence of function group of the bioactive compounds was confirmed by FTIR spectroscopy. Finally, the study also reported that the extracts of P. citrinum and D. bicornis have several bioactive compounds with application in commercial pharmaceutical industries.