Cardiovascular disease is one of the major causes of deaths worldwide. Increased arginase activity is associated with cardiovascular disease. The literature shows that plants are a good source of arginase inhibitors. Hence in the present work arginase inhibitor activity is studied from Ficus religiosa leaves. A fine powder of F. religiosa leaves was serially extracted in various solvents, viz. hexane, chloroform, ethyl acetate and methanol. Out of those four solvent extracts, the one showing highest arginase inhibitor activity was loaded onto the column for further fractionation. Among the collected fractions, the one showing the highest activity was subjected to identification of metabolites by using LC–HRMS. Total compounds including acipimox, edoxudine, levulinic acid, hydroxyhydroquinone, ramiprilglucuronide, berberine, antimycin A, swietenine and some short peptides were identified from the fraction showing the highest arginase inhibitory activity. Identification of these metabolites from F. religiosa and their biological importance may help to promote its use as medicinal plant. Further purification and characterization of therapeutically novel molecules will be the subject of future work.