The first CE method enabling the enantiomeric separation of the synthetic pyrethroid cis-bifenthrin (BF) was developed in this work. CD-MEKC was the CE mode employed for this purpose. The influence of several experimental parameters such as temperature, voltage, type and concentration of surfactant (chiral and achiral) and CD was investigated. The use of the bile salt sodium cholate at a concentration of 100 mM in the presence of 20 mM heptakis (2,3,6-tri-O-methyl)-beta-CD enabled the separation of cis-BF enantiomers in less than 10 min and with a resolution of 2.8. The analytical characteristics of the developed methodology were evaluated allowing its application to the quantitation of cis-BF in a polyvalent commercial insecticide formulation.