The development of resistance by mosquitoes to current synthetic compounds has resulted in reduced effectiveness of prevention and control methods worldwide. An alternative nonchemical based control tools are needed to be evaluated particularly plant-derived essential oils. Several components of vetiver oil have been documented as insect repellents. However, detailed knowledge of those components action against insect remains unknown. In this study, behavioral response of Anopheles minimus to four constituents of vetiver oil (valencene, terpinen-4-ol, isolongifolene, vetiverol) was evaluated by using the high-throughput screening assay system. Vetiverol and isolongifolene exhibited strong contact irritancy action at 1.0% (80.2% escaping) and 5.0% (81.7% escaping) concentration, respectively, while moderate action was found in both valencene and terpinen-4-ol at 5.0% (57.6% escaping). Only at 1.0% (0.7 spatial activity index [SAI]) and 5.0% (1.0 SAI) of valencene and 0.5% (0.7 SAI) of isolongifolene showed spatial repellency activity. High mortality (58.9–98.2%) was recorded in all concentration of vetiverol and isolongifolene. Meanwhile, valencene exhibited high mortality only at 5.0%, terpinen-4-ol showed very low toxic action (0–4.3%) in all concentration. These proved that valencene in vetiver oil is the promising constituent that can be developed as an alternative mosquito control mean in efforts to prevent disease transmission.