Objective To analyze beauvericin and enniatins concentrations in the culture complex after Isaria cicadae Miquel was inoculated on different media for certain time. Methods One strain of Isaria cicadae Miquel was inoculated on 4 kinds of liquid culture media and 4 kinds of solid culture media. After incubation for 1-7 weeks under 25 ℃, high performance liquid chromatography tandem mass spectrometry was employed to detect beauvericin and enniatins. Results The detection rates of beauvericin in Isaria cicadae Miquel culture on 4 kinds of liquid media ranged between 42.9%-100.0%, and the beauvericin concentrations varied from 1.0 to 94.6 μg/L. The positive rates of beauvericin in Isaria cicadae Miquel culture on 4 kinds of solid media were 100.0%, and the beauvericin concentrations ranged from 60.9 to 44 677.5 μg/kg. Enniatins were negative for Isaria cicadae Miquel on 8 kinds of culture media. Conclusion The strain of Isaria cicadae Miquel used in our study could produce beauvericin in 8 kinds of culture media after incubation for certain time, and the beauvericin concentrations in solid media were much higher than in liquid media. The strain of Isaria cicadae Miquel didn't produce enniatins in 8 kinds of culture media after incubation for certain time.