In this study, the effects of immobilization and high hydrostatic pressure (HHP) on capsaicin production in cell suspension culture of pepper seeds at different application times were investigated. Callus cultures were obtained from in vitro germinated hypocotyl explants of pepper seedlings and cell suspensions were prepared from calluses. Immobilized cell suspension cultures with calcium alginate and free cell suspension cultures were obtained by using these cell suspensions. 50, 100, 200, 300 and 400 MPa were applied to both free and immobilized cell suspensions as an elicitor. When all the results were compared, the highest amount of capsaicin was achieved in immobilized cell suspensions on the tenth day as 293.187 mu g/g f.w at 400 MPa pressure. As a result, it can be concluded that the combined application of immobilization and elicitor (HHP) caused significant increases (p < .05) in the amount of capsaicin.