Chili peppers are globally cultivated for their rich bioactive compound profile. This study investigates the impact of two biostimulants, Humix® and Energen, on Capsicum chinense ‘Habanero Orange’ and Capsicum annuum ‘Kristian’, focusing on quantitative and qualitative parameters. Conducted over two years with three annual harvests, the research assesses the effects of biostimulant application on yield, fresh fruit number, fruit weight, drying ratio, capsaicin, dihydrocapsaicin, and ascorbic acid content (via HPLC-DAD analysis), as well as carotenoid levels (via spectrophotometric analysis). Biostimulant application significantly increased (p ≤ 0.05) total yields and capsaicin levels. Harvest timing also influenced dihydrocapsaicin and capsaicin levels, with the third harvest showing the highest values (p ≤ 0.001). The effects on ascorbic acid and carotenoids were variable and depended on genotype, harvest, and treatment. Thus, our study provides insights into the dynamic responses of Capsicum species to biostimulants under variable climatic conditions, contributing new knowledge to agricultural practices and the scientific understanding of biostimulant effects in Capsicum production.