Aims: The present study aimed to investigate the local anesthetic activity of Chrysanthemum cinerarifolium extract as well as to formulate a topical local anesthetic gel containing the extract to be used for human. Methods: The local anesthetic activity was assessed using the intradermal wheal method in eighteen guinea pigs divided into three groups, six in each group. Pain response was elicited by gentle pinprick. Two concentrations (1.0 and 0.3mg/ml) of lidocaine were injected intradermally into the left and right dorsal flank of the animal respectively to form wheals, which were encircled with a marker. Similarly, two concentrations (100 and 50mg/ml) of the plant extract were injected into the left and right dorsal flank respectively of another group. Normal saline was injected into the control group. The encircled areas were each pricked with a pin 6 times at 5 minutes interval for 30 minutes starting with zero-time before the injection. To confirm the animal study, prepared gels of the plant extract (1%) and lidocaine (1%) were applied on two groups each containing 6 human volunteers, and the local anesthetic activity was assessed. Results: The plant extracts exhibited a local anesthetic activity on guinea pigs. Comparable results were also observed of the local anesthetic activities of both topical gels prepared from the plant extract or lidocaine in human. No adverse effects were observed either in animals or in human. Conclusions: Chrysanthemum cinerarifolium extract possesses a local anesthetic activity in both animals and human