The water extraction of phenolic compounds from two varieties ("Mahan" and "Marameck") of pecan nutshells (Carya illinoinensis) without and with sonication, varying the solvent/solid ratio (S), the pH, and the refluxing time (t), was studied. Additionally, the in vitro cytotoxicity and the determination of the cell death mechanism of the extracts against the colon cancer cell line HT-29 were investigated. The content of total phenolic compounds (TPC) of "Marameck" nutshells resulted higher than for the "Mahan" variety, and the pH increase resulted in higher TPC contents for both cultivars. The optimized conditions for TPC extraction without and with sonication resulted: S = 33 ml/g, pH = 12, and t = 9.6 min, and yielded ≈ 70 and 90 mg/g of TPC for "Mahan" and "Marameck" nutshells, respectively. The optimized extracts of pecan nutshells without sonication from both cultivars presented similar cytotoxicity against HT-29 colon cancer cells (IC50 ≈ 50 µg/ml), higher than for sonicated extracts (IC50 ≈ 88 and 138 µg/ml for "Mahan" and "Marameck," respectively). Cell death through apoptosis was the main mechanism of cell death induced by the nutshell extracts. PRACTICAL APPLICATION: The extraction of phenolic compounds (TPC) from the residues of two varieties of pecan nutshells ("Mahan" and "Marameck") was studied. An optimal combination of variables within the pH range that minimizes the solvent-to-solid ratio (S) and the time of refluxing (t), saving at the same time, water and energy, was set up. The phenolic compound extracts obtained from the residues of the pecan nuts exhibit cytotoxic effects against colon cancer cells and could be of interest as an alternative treatment of different types of cancer. Additionally, these extracts may be of importance to the food industry as they can be used as antioxidant agents in food formulation. Also, the high levels of anthocyanidins obtained from the pecan nut extracts after proanthocyanidins' strong acid hydrolysis can be purified and employed as natural red dyes.
Keywords: HT-29 colon cancer cell line; cytotoxicity; pecan nutshells; phenolic compounds; ultrasound-assisted extraction.
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