Capsaicin purification from Samandag (Hatay) pepper with affinity chromatography

Within the scope of this thesis, it was aimed to purify capsaicin, which has an important place in the health sector, from Samandağ pepper, a local product. The capsaicin content of Samandağ pepper, which is an endemic species, was examined in dry and wet form and compared with the contents of green, stuffed and capia peppers. Capsaicin determination was started with Soxhlet extraction and three different solvents, methanol, ethanol and acetonitrile, were selected for this purpose. In order to optimize the extraction, three different temperature and three different time parameters were studied in each selected solvent. It was determined that capsaicin yield as a result of Soxhlet extraction was obtained at 1780.9 µg/g at 80oC and by using methanol solvent for 3 hours. The extracts obtained as a result of the extraction were first analyzed by one-dimensional HPLC for the determination and purification of capsaicin content, and it was observed that only capsaicin and dihydrocapsaicin could be detected among capsaicinoids. As a result of the analysis of dried red pepper, it was calculated that 1780.9 µg/g capsaicin and 897.8 µg/g dihydrocapsaicin could be obtained, and considering the total amount of capsaicinoids, the pungency value was calculated as 40185.5 SU (Scoville Units). In fresh red peppers, 105.4 µg/g capsaicin and 66.48 µg/g dihydrocapsaicin could be obtained. As a result of one-dimensional HPLC analysis, capsaicinoid was not detected in stuffed and capia peppers. In order to obtain and purify capsaicinoids with a higher yield, two-dimensional HPLC analysis was performed and the p(HEMA-MATrp) monolithic column synthesized and characterized during the analysis was used as the first dimensional column. Thus, both preconcentration was made and the amount of capsaicinoid held in the column was increased. Commercial C-18 column was used as the second dimensional column and it was observed that four different capsaicinoids, namely capsaicin, dihydrocapsaicin, nordihydrocapsaicin, and homocapsaicin, could be detected at the end of the analysis. As a result of the two-dimensional HPLC analysis, it was revealed that the total amount of capsaicinoid increased to 3853.7 µg/g. As a conclusion of the thesis study, it has been proven that capsaicin, an important bioactive molecule, can be obtained in pure form from Samandağ pepper which is a local product.