Aim:The aim of the study was to evaluate the local anesthetic activity of root extracts of the Anacyclus pyrethrum, and to check its effect of interaction at the site of injection in guinea pigs.Materials and Methods:The study sample included thirty guinea pigs each weighing 450–500 g, maintained under standard conditions. The root extracts were prepared using three solvents, in 1% and 2% concentration and injected in guinea pigs. The animals were divided into five groups, six in each group based on the type of extract used along with a control and a standard drug. All the animals received 0.5 ml of intradermal injection of the prepared extract, with 1% concentration in the left and 2% in the right dorsal flank of the animal and were checked for local anesthetic activity by a pinprick test. After 72 h, biopsy was done from the injected site to check for drug interaction.Results:The number of negative response obtained from 2% ethanol extract is more effective when compared to other extracts. Histological samples showed inflammatory changes in 1% aqueous extract in a single animal.Conclusion:Among the test compounds, 2% ethanol showed more significant effect; hence, it is suggested to synthesize more compounds in this series and evaluate their pharmacological actions.