The aim of this work was to study the influence of essential oil fractionation on acaricidal activity against the cattle tick Rhipicephalus (Boophilus) microplus. The citronella (Cymbopogon winterianus J.) and pepper tree (Schinus molle L.) essential oils were fractionated by vacuum distillation yielding fractions that were analyzed by the GC/MS. Laboratory tests were carried out to determine the effect of the total essential oil and fractions on larvae of the cattle tick R. (B.) microplus. The fractions 04 and 05 of the C. winterianus essential oil were the most active showing LC50 values of 1.20 and 1.34 μL/mL, respectively. The LC50 of the total oil was 3.30 μL/mL while the effect of the fractions 01, 02 and 03 was less pronounced, with LC50 values of 4.37, 4.24 and 3.49 μL/mL, respectively. The fraction 03 of the S. molle essential oil was the most active showing LC50 value of 8.80 μL/mL while the fractions 01 and 02 did not show toxic effects on the larvae.