Abstract INTRODUCTION: In the present study, we determined the chemical composition of Clinopodium macrostemum essential oil obtained by microwave-assisted hydrodistillation and its effect on the growth and development of Culex quinquefasciatus mosquito larvae. METHODS: The essential oil compounds were identified by gas chromatography-mass spectrometry analysis, and bioassays were conducted to evaluate the influence on the developmental stages of early second instar larvae of Cx. quinquefasciatus until the emergence of adults, using essential oil concentrations of 50, 100, 200, 400, and 800mg/L. RESULTS: The gas chromatography-mass spectrometry analyses revealed that the leaf essential oil of C. macrostemum contained 32 compounds and the major chemical compounds identified were linalool (55.4%), nerol (6.4%), caryophyllene (6.25%), menthone (5.8%), geraniol acetate (4.1%), terpineol (3.7%), and pulegone (2.8%). The essential oil yield obtained by microwave-assisted hydrodistillation was 0.8% at 20 min. The treatments showed lethal concentrations (LC50 and LC90) of 22.49 and 833.35mg/L, respectively, after the final measurement of the total number of dead larvae (second, third, and fourth instars), and LC50 and LC90 of 6.62 and 693.35mg/L, respectively, at the end of the experimental period. The essential oil inhibited the growth and development of the mosquito larvae by 32% (relative growth index = 0.68) at 50mg/L, and up to 47% (relative growth index = 0.53) at 800mg/L. CONCLUSION: The present study demonstrated the larvicidal effect of C. macrostemum essential oil on Cx. quinquefasciatus, which can be attributed to the oxygenate compounds obtained by the extraction method.