The current study was designed to evaluate the properties of various fractions of methanolic extract prepared from aerial parts of Swertia chirata. Five fractions, RA-78, RA-91, RA-112, RA-14 and RA-15 showed significantly high DPPH scavenging capacity. In finding out antimicrobial activities, different fractions have shown mild activities against Bordetella bronchiseptica and Micrococcus luteus, while all fractions showed significant results against Aspergillus fumigatus followed by Mucor species and five fractions were active against Aspergillus niger. In measurement of brine shrimp cytotoxic potential of different fractions, three fractions RA-12, RA-34 and RA-78 had shown the best cytotoxic results with zero LD50. In determining antileishmanial properties of different fractions, the best results were exhibited by fraction RA-78 and RA-112 with zero percent survival of Leishmania parasite. Various fractions of methanolic extract prepared from aerial parts of Swertia chirata promisingly possess antioxidant, antimicrobial, brine shrimp cytotoxic, and antileishmanial activities.