{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,19]],"date-time":"2026-06-19T11:45:17Z","timestamp":1781869517374,"version":"3.54.5"},"reference-count":61,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2025,9,3]],"date-time":"2025-09-03T00:00:00Z","timestamp":1756857600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"European Union Next-Generation EU","award":["CN00000022"],"award-info":[{"award-number":["CN00000022"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Toxins"],"abstract":"<jats:p>The European Circular Economy Action Plan outlines a forward-looking strategy that emphasizes waste reduction and the acquisition of high-quality secondary resources. Previous research has shown that cocoa processing by-products contain compounds of interest for various industrial areas, making them an attractive matrix for reuse. However, a gap remains in our understanding of the safety of these by-products intended for feed. In this study, theobromine and caffeine were quantified by High-Performance Liquid Chromatography (HPLC-UV) in cocoa hulls for safety considerations, evaluating theobromine compliance with toxicological and safety levels, and considering their potential application as an ingredient in animal feed. In addition, the identification of phenolic components and associated antioxidant activity was conducted through High-Performance Thin-Layer Chromatography (HPTLC). This preliminary study indicates that theobromine content is a limiting factor for the inclusion of cocoa hulls in animal diets, as it restricts inclusion levels to remain within current regulatory limits. Examples of general estimates of dietary theobromine exposure at inclusion levels based on regulatory limits for dairy cows and veal calves confirmed a low risk for animal health. Furthermore, the detection of antioxidant activity linked to the presence of polyphenols highlights the potential of cocoa hulls as a sustainable food by-product for feed formulation.<\/jats:p>","DOI":"10.3390\/toxins17090441","type":"journal-article","created":{"date-parts":[[2025,9,3]],"date-time":"2025-09-03T12:07:32Z","timestamp":1756901252000},"page":"441","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Preliminary Assessment of Alkaloid Content in Cocoa (Theobroma cacao L.) Hulls for Safe Consumption as a Feed Ingredient"],"prefix":"10.3390","volume":"17","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-1785-0437","authenticated-orcid":false,"given":"Francesca","family":"Mercogliano","sequence":"first","affiliation":[{"name":"Department of Pharmacological and Biomolecular Sciences, Universit\u00e0 Degli Studi di Milano, Via Balzaretti 9, 20133 Milano, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9736-8746","authenticated-orcid":false,"given":"Corinne","family":"Bani","sequence":"additional","affiliation":[{"name":"Department of Pharmacological and Biomolecular Sciences, Universit\u00e0 Degli Studi di Milano, Via Balzaretti 9, 20133 Milano, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3317-4384","authenticated-orcid":false,"given":"Marco","family":"Tretola","sequence":"additional","affiliation":[{"name":"Institute for Livestock Sciences, Agroscope, La Tioleyre 4, 1725 Posieux, Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carla","family":"Landolfi","sequence":"additional","affiliation":[{"name":"ToxHub Srl, Via Ariberto 20, 20123 Milan, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0123-3888","authenticated-orcid":false,"given":"Matteo","family":"Ottoboni","sequence":"additional","affiliation":[{"name":"Department of Veterinary Medicine and Animal Sciences, Universit\u00e0 Degli Studi di Milano, Via Dell\u2019Universit\u00e0 6, 26900 Lodi, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2682-8685","authenticated-orcid":false,"given":"Federica","family":"Cheli","sequence":"additional","affiliation":[{"name":"Department of Veterinary Medicine and Animal Sciences, Universit\u00e0 Degli Studi di Milano, Via Dell\u2019Universit\u00e0 6, 26900 Lodi, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3008-5987","authenticated-orcid":false,"given":"Patrizia","family":"Restani","sequence":"additional","affiliation":[{"name":"Faculty of Pharmacy, Universit\u00e0 Degli Studi di Milano, 20133 Milano, Italy"},{"name":"Coordinating Research Center (CRC) \u201cInnovation for Well-Being and Environment\u201d, Universit\u00e0 Degli Studi di Milano, 20133 Milano, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0337-9426","authenticated-orcid":false,"given":"Luciano","family":"Pinotti","sequence":"additional","affiliation":[{"name":"Department of Veterinary Medicine and Animal Sciences, Universit\u00e0 Degli Studi di Milano, Via Dell\u2019Universit\u00e0 6, 26900 Lodi, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2798-8720","authenticated-orcid":false,"given":"Chiara","family":"Di Lorenzo","sequence":"additional","affiliation":[{"name":"Department of Pharmacological and Biomolecular Sciences, Universit\u00e0 Degli Studi di Milano, Via Balzaretti 9, 20133 Milano, Italy"},{"name":"Coordinating Research Center (CRC) \u201cInnovation for Well-Being and Environment\u201d, Universit\u00e0 Degli Studi di Milano, 20133 Milano, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,9,3]]},"reference":[{"key":"ref_1","unstructured":"European Commission Directorate-General for Communication (2020). 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