Cordia globosa is used to treat gastrointestinal diseases in San Rafael Coxcatlan, Puebla, Mexico; however, its medicinal properties have not been investigated. This study reports the chemical composition and the antimicrobial effect of C. globosa essential oil. The essential oil of aerial parts was obtained by steam distillation and 25 compounds were identified by gas chromatography-mass spectrometry (GC-MS). The major constituent was α-pinene (38.4%). Antimicrobial activities were tested on 6 bacterial and 9 fungi strains. The most sensitive strain was Vibrio cholera (minimum inhibitory concentration [MIC], 0.060 mg/ml). These results show the chemical composition and biological properties of essential oil of C. globosa. The results validate the medicinal use of C. globosa.