A high performance liquid chromatographic (HPLC) method was established to determine coenzyme Q10 in queen bee larvae. Samples were ground and extracted with absolute ethanol under the assistance of ultrasonic and the resulting extract was then reextracted with n-hexane. The n-hexane layer was finally condensed by rotation evaporation and rediluted with absolute ethanol. The chromatographic separation was achieved on an XBridge shield RP18 column using methanol as a mobile phase prior to UV detection at 275 nm wavelength. Results showed that a good separation of coenzyme Q10 was observed within 12 min. The developed standard curve exhibited good linearity. Average recoveries for coenzyme Q10 in a real sample spiked at 3 levels, each of which was performed in 6 replicates, were within the range of 95.0% to 101.8%, with a RSD of less than 8%. The limit of detection for coenzyme Q10 was 0.23 mg/L. The analytical method proved to be simple, accurate, reliable and sensitive so as to be most suitable for the determination of coenzyme Q10 in queen bee larvae. Satisfying results from the analysis of coenzyme Q10 in some real samples by the method were obtained.