A HPLC procedure was developed to separate vanillylamine, L-phenylalanine, caffeic acid, coumaric acid, ferulic acid, cinnamic acid, capsaicin, and dihydrocapsaicin. Using the HPLC method, concentrations of intermediates were determined in placenta, pericarp, cell cultures, immobilized cells, and immobilized placenta. The profile of intermediates showed higher capability of biotransformation in immobilized placenta over immobilized cells. Vanillylamine was not a limiting intermediate for capsaicin production. Three-week-old placenta separated from fruit had a higher quantity of capsaicin than did pericarp.