This study investigated capsaicin (CAP) and dihydrocapsaicin (DHC) contents in different Capsicum cultivars using high-performance liquid chromatography (HPLC). Fourteen Capsicum cultivars ('SUNGIL-C', '26B', '26C', 'DELB', 'DELC', 'MIGP-C', 'MI4H', 'MI2F', 'MI2H', 'MI8H', were dried, extracted with ethanol, and concentrated. Results from the HPLC analysis revealed notable differences in the capsaicinoid (CAP and DHC) levels. For instance, the cultivar 'SUNGIL-C' exhibited the highest CAP and DHC content of 1.92 mg/g extract and 0.88 mg/g extract, respectively. However, cultivars '26B', 'DELB', 'MIGP-C', and 'MI8H' showed trace amounts of capsaicinoids or were not detected at all. In addition to the capsaicinoid analysis, total polyphenol and flavonoid content analyses were performed by selecting extracts from only those cultivars in which CAP and DHC were detected, and the effects of CAP and DHC and phenolic compounds on antioxidant activity were discussed. This analysis provides insight into the potential pungency and physiological effects of different Capsicum cultivars and highlights the variability in capsaicinoid and DHC content. Understanding this variation can clarify the biological meaning and practical applications of capsaicinoids and warrants further research into their usefulness.