Recently, various plant species and different extracts obtained from them got more attention analyzing their biological activities. The in-vitro study wasdone for the first time to determine anticancer, anti-inflammatory, cytotoxicity, antifungal, antibacterial, leishmanicidal in Peucedanum ferulafolium whole plant (stem, root, flower and leaves).Our investigations revealed that, the antifungal activity in whole plant Peucedanum ferulaefolium aqueous fraction (WPFAF) anf butanol fraction (WPFBF) was significantly active against C. albicans with 89% of inhibition and C. glabarata with 89% of inhibition. For antibacterial activity, whole plant Peucedanum ferulaefolium methanol extract (WPFME) exhibited remarkable activity against S. aureus with 74% of inhibition and aqueous fraction (WPFAF) showed moderate activity with 60%of inhibition against S. aureus. After getting remarkable antifungal and antibacterial activity results from the aqueous fraction of Peucedanum ferulafolium we have done GC-MS of same fraction (WPFAF) and isolated 34 compounds, which were identified by GC-MS. Whole plant Peucedanum ferulaefoliumextract and fractions showed very low cytotoxic effect. Unfortunately whole plant Peucedanum ferulaefolium exhibited inactive against cancer cell line (HeLa and 3T3), antileishmanial and anti-inflammatory. In conclusion, Peucedanum ferulafolium aqueous fraction showed astounding amount of effective antifungal and antibacterial activity.