The content of protoanemonin, a known biologically active constituent of Clematis spp., was determined by GC-MS in the leaf, stem and root extracts of one Chinese species and four Australian Clematis taxa. The results showed that protoanemonin concentrations varied between different plants and that leaves contained higher concentrations than stems and roots. To our knowledge this is the first study to determine protoanemonin content variation in leaf, stem and root of Clematis spp.