Laserpitium carduchorum is frequently used as a spice, and in Bane folk medicine, the aerial parts of this are used to treat urinary infections. Variation in the quantity and quality of the essential oil of Iranian L. carduchorum at different developmental growth stages including vegetative, flowering, and seed ripening is reported. In total, 33 compounds were identified and quantified in the oils of vegetative, flowering, and seed ripening stages, representing 97.8%, 98.8%, and 98.7% of the oils, respectively. α-Pinene (45.1, 61.4, and 46.4%), sabinene (16.5, 10.3, and 17.5%), and limonene (6.4, 8.5, and 20.4%) were the main compounds in all samples. The antioxidant activities of different extracts of L. carduchorum at different developmental growth stages were examined by employing various established in vitro experiments including DPPH, FRAP, and TEAC assays. The amounts of total phenolics were also determined spectrophotometerically. Antimicrobial activities of different extracts and essential oils of L. carduchorum at different developmental growth stages were examined against five Gram-positive and four Gram-negative bacteria, as well as two fungi. The results showed that maximum antioxidant and antimicrobial activity of extracts were at the flowering stage of the plant. Maximum antimicrobial activity of essential oils was at seed ripening stage.