Background: In order to produce an effective callus in Echinacea purpurea L.; determination of the explant type and growth regulators that best respond to callus induction and the optimization of the culture conditions to increase the amount of caffeic acid derivatives (CADs) in the obtained callus. CADs contents of callus cultures of E. purpurea were evaluated by establishing an effective callus induction system in vitro. Results: Various medium containing different growth regulators were tested using leaf, petiole, cotyledon and root as the explants. The best callus development was achieved in MS medium with 1.0 mg l−1 2,4-D + 2.0 mg l−1 BAP in leaf, 1.0 mg l−1 NAA + 0.5 mg l−1 TDZ in petiole, 2.0 mg l−1 NAA + 1.0 mg l−1 TDZ in cotyledon and 0.5 mg l−1 NAA + 0.5 mg l−1 BAP in roots. Upon optimisation of callus growth, each type of explant was cultured for 4, 6, 8 and 10 weeks in medium for the analyses of caftaric acid, chlorogenic acid, caffeic acid and chicoric acid contents. The highest amounts of caftaric acid (4.11 mg/g) and chicoric acid (57.89 mg/g) were found from petiole explants and chlorogenic acid (8.83 mg/g) from root explants at the end of the 10-week culture time. Conclusions: As a result of the present study, the production of caffeic acid derivatives was performed by providing the optimization of E. purpurea L. callus cultures. Effective and repeatable protocols established in this study may offer help for further studies investigating the production of caffeic acid derivatives in vitro. How to cite: Tanur Erkoyuncu M, Yorgancilar M. Optimization of callus cultures at Echinacea purpurea L. for the amount of caffeic acid derivatives. Electron J Biotechnol 2021;51. https://doi.org/10.1016/j.ejbt.2021.02.003.