High-speed countercurrent chromatography (HSCCC) is widely used for isolation of biologically active compounds of plant extracts and essential oils (EOs) in preparative scale. The objective of this work was the prefractionation of EOs from Lippia alba, Aloysia citriodora, Lantana armata and Xylopia aromatica by HSCCC and the evaluation of the antioxidant activity in vitro of their EOs and fractions. The antioxidant capacity was determined by two different in vitro assays: 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical and 2,2Ê¹-Azinobis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical cation. EOs and their fractions showed a low-moderate ability to trap the radical cation ABTS+ and DPPH radical.