Callus cultures derived from leaves of Vitex negundo L. were assayed for the production of secondary metabolites. Earlier studies revealed that the compounds produced by calli maintained in Murashige and Skoog medium with 2,4-D and BA did not correspond to those isolated from the intact plant. These differences further extend to the volatile oil and hexane fractions of leaves and calli as evidenced by GC-MS analysis. In this paper, comparisons of the gas chromatographic profiles of leaf and callus extracts and the identities of some of the major metabolites were reported.