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The volatile oil of leaves and barks of Calycopteris floribunda was examined by gas chromatography-mass spectrometry (GC-MS). 52 volatile chemical components in leaves were identified. The antimicrobial assay of oils in the leaves and barks was carried out by disk diffusion method in vitro. The major components (mass fraction) in leaves are caryophyllcne oxide (13.79%), n-hexadecanoic acid (11.91%) and β-caryophyllene (10.45%). Ten constituents are identified accounting for about 99.98% of the total volatile oil in the bark. Among these components, n-hexadecanoic acid (59.18%), linolic acid (12.70%) and butyl octyl phthalate (8.21%) are the major constituents. The oils exhibit strong antimicrobial activity and display more potent against bacteria than fungi.