Monoclonal antibodies (MAbs) against lactucin and lactucopicrin (two of the bitter principles of chicory;Cichorium intybusL.) have been prepared. In competitive ELISA, the sera from mice immunized with keyhole limpet haemocyanin (KLH)‐lactucin and KLH‐ lactucopicrin have been screened for cross‐reactivity to various sesquiterpene lactones with a guaiane skeleton that are present in chicory, such as lactucin, 8‐deoxylactucin, and their 11β,13‐dihydro derivatives, and lactucopicrin. Mice showing low cross‐reactivity were used to produce hybridomas that were screened for cross‐reactivity in the same way. The screening resulted in a lactucopicrin‐specific hybridoma (no. 4H10) showing a low cross‐reactivity (≪1%)for lactucin, 8‐deoxylactucin and their 11β,13‐dihydro derivatives. From KLH‐lactucin immunized mice, hybridoma no. 9F12 showed the best characteristics, i.e. 25% cross‐reactivity to 8‐deoxylactucin and its 11β,13‐dihydro derivative, and a 10% cross‐reactivity to lactucopicrin. The affinity constants of MAbs nos 9F12 and 4H10 were 2.2X104and 2.8X105M‐1respectively. Chicory extracts spiked with free sesquiterpene lactones showed an inhibition level equal to the sum of the inhibition level of the extract and the level of sesquiterpene lactones added.