To date, there is no information in the literature regarding the bioaccessibility of capsaicinoids from natural sources. The effect of ripening and heat-processing on the in vitro bioaccessibility of capsaicin and dihydrocapsaicin was studied in the absence and presence of two dietary fat types. The capsaicinoid bioaccessibility was also studied during the frozen storage of peppers for 6 months. Fresh green peppers showed the highest capsaicinoid bioaccessibility, as compared with that of other experimental groups. The bioaccessibility of capsaicinoids from green peppers decreased as the intensity of heat treatment increased. The dietary fat increased the bioaccessibility of capsaicin and dihydrocapsaicin in digestions with red peppers, especially that of dihydrocapsaicin. The bioaccessibility of capsaicinoids was altered by frozen storage. The Caco-2 cells incorporated capsaicin and dihydrocapsaicin (8.4% and 10.9%, respectively) but they were probably metabolized by cells. (C) 2015 Elsevier Ltd. All rights reserved.