During the past decade, it has been reported that the consumption of certain foods and spices such as pepper may have a positive effect on health. The present study evaluates the influence of fruit ripening on total phenols, flavonoids, carotenoids and capsaicinoids content and antioxidant, hypoglycaemic and anticholinesterase activities of Capsicum chinense Jacq. cv Habanero. The chemical investigation showed a different composition between the two stages of ripening (immature and mature). Generally, the concentration of carotenoids and capsaicinoids increased as the peppers reached maturity, whereas the concentration of phenols declined. The immature fruits showed the highest radical scavenging activity (IC 50 of 97.14 μg/ml). On the contrary, the antioxidant activity evaluated by the β-carotene bleaching test showed a significant activity for mature peppers (IC 50 value of 4.57 μg/ml after 30 min of incubation). Mature peppers inhibited α-amylase with an IC 50 of 130.67 μg/ml. The lipophilic fractions of both mature and immature peppers exhibited an interesting and selective inhibitory activity against α-amylase with IC 50 values of 29.58 and 9.88 μg/ml, respectively. Both total extracts of mature and immature peppers inhibited butyrylcholinesterase selectively. The obtained results underline the potential health benefits as a result of consuming C. chinense Habanero and suggest that it could be used as new valuable flavour with functional properties for food or nutriceutical products on the basis of the high content of phytochemicals and found biological properties.