Cochlospermum regium is a shrub plant species from the Cerrado domain used in traditional medicine. This study aimed to evaluate the chemical profile and antioxidant, antibacterial and cytotoxic activities on Artemia salina from the essential oil of fresh leaves and xylopodium. Fresch leaves and xylopodium of C. regium were collected in an area of Cerrado domain in Goiás State, Brazil, 2021. The essential oil was obtained by hydrodistillation, the yield was quantified and the chemical profile determined by gas chromatography with mass spectrometry (GC-MS). Physicochemical analyzes were carried out for organoleptic analysis (color and appearance), solubility, relative density (g mL-1), refractive index, optical rotation (αD), antioxidant activity in DPPH radical reduction (IC50 µL mL-1), antibacterial activity on Escherichia coli, Staphylococcus aureus, Enterococcus faecalis, Salmonella serovar Enteritidis and Salmonella serovar Thyphymurium by the disc method (mm), and cytotoxicity bioassay on Artemia salina (LC50 µg mL-1). The major compounds for the essential oil of fresh leaves were viridiflorol 10.21%, Copaen-4-α-ol <β>, longiborneol 9.07 and β-bisabolene 11.48%, and for the essential oil of xylopodium β-selinene 26.17%, aromadendrene 8.66 % and thujopsene 8.09%. The yield was 0.58 and 0.33%, color slightly yellow and yellow for fresh leaves and xylopodium, respectively. Positive solubility, refractive index of 1.3468 and 1.3347, optical rotation +48.8 and +21.5, relative density 0.932 and 0.936 g mL-1, antioxidant activity IC50 = 47.65 and 111.16 µL mL-1 for fresh leaves essential oil and xylopodium, respectively. The essential oil from fresh leaves showed high antibacterial potential for all strains, as well as for cytotoxic activity on A. salina with LC50 = 90.17 and 625.08 µg mL-1, respectively.