Parthenolide, a biologically active sesquiterpene lactone found in feverfew [Tanacetum parthenium (L.) Schultz. Bip.], has been indirectly linked to the antimigraine action of feverfew preparations. Commercial products of feverfew leaves vary widely in parthenolide content (0-1.0%/g dwt). No comprehensive studies have quantified parthenolide variation among feverfew populations or cultivars, and whether morphological traits are correlated with this natural product. In this study, 30 feverfew accessions were examined for parthenolide content, morphological traits, and seed origin. Statistically significant differences in parthenolide levels were found among the populations studied. Parthenolide content ranged from (0.012% ± 0.017 to 2.0% ± 0.97 /g dwt) as determined by HPLC-UV-MS. Higher parthenolide levels tended to be in wild material (0.41% ± 0.27) as opposed to cultivated material (0.19% ± 0.09). Parthenolide levels correlated with flower morphology: disc flower (0.49% = B1 0.36), semi-double (0.38% ± 0.13), double (0.29% ± 0.16), and pompon-like flower (0.22 ± 0.14). Leaf color also appeared to be indicative of parthenolide levels, with the light-green/golden leafed accessions showing significantly higher parthenolide content than darker-leafed varieties, but whether this was due to inadvertent original selection of a high parthenolide-containing golden leaf selection is not yet known. This study does show that further selection for improved horticultural attributes and natural product content is promising to improve feverfew lines for the botanical/ medicinal plant industry.