Monazzah Maryam, Herrmann Cedric, Morlock Gertrud E, Fuchs Jannika, Lachenmeier Dirk W
Chemisches und Veterinäruntersuchungsamt (CVUA) Karlsruhe, Weissenburger Strasse 3, 76187 Karlsruhe, Germany.
Department of Comparative Biomedicine and Food Science, University of Padua, Viale dell'Università 16, 35020 Legnaro, Italy.
Toxics. 2025 Aug 31;13(9):739. doi: 10.3390/toxics13090739.
The mutagenic potential of coffee by-products, including leaves, blossoms, cherries, and silverskin, was studied using thin-layer chromatography (TLC) coupled with the recent planar Ames bioassay via pH indicator endpoint. The 2LabsToGo-Eco allowed for the separation and detection of mutagens in complex samples. Hot water was the most effective extraction solvent in terms of yield and closely simulated the typical human consumption of coffee by-products. Separation was performed on TLC plates with a mixture of ethyl acetate, -propanol, and water, followed by bioassay detection. The positive control 4-nitroquinoline 1-oxide exhibited clear mutagenic responses, confirming the proper bioassay performance. In the Ames bioautogram, none of the tested coffee by-products showed mutagenic zones, suggesting the absence of strongly acting, acute mutagens under the applied test conditions; however, given the only 5 h short incubation and the use of TA98 strain only, a longer incubation time and testing with additional strains is recommended. The results provide new safety data for leaves and blossoms and are consistent with some previous studies demonstrating the safety of coffee by-products. However, further improvements in the sensitivity and selectivity of the planar Ames bioassay are demanded, and further in vivo and long-term safety studies are recommended. Considering natural variability, the different uses of pesticides and treatments, and the fluctuating supply chains, coffee by-products may differ highly. The planar bioassay technology using the affordable 2LabsToGo-Eco is a powerful toxicological screening option for the coffee industry, considering the increasing interest in utilizing coffee by-products.
利用薄层色谱法（TLC）结合最近的平面艾姆斯生物测定法（通过pH指示剂终点），研究了包括叶子、花朵、樱桃和银皮在内的咖啡副产品的致突变潜力。2LabsToGo-Eco可用于分离和检测复杂样品中的诱变剂。就产量而言，热水是最有效的提取溶剂，并且能紧密模拟人类对咖啡副产品的典型消费量。在涂有乙酸乙酯、正丙醇和水混合物的TLC板上进行分离，然后进行生物测定检测。阳性对照4-硝基喹啉1-氧化物表现出明显的诱变反应，证实了生物测定的正常性能。在艾姆斯生物自显影片中，所测试的咖啡副产品均未显示诱变区，这表明在所应用的测试条件下不存在强作用的急性诱变剂；然而，鉴于仅5小时的短孵育时间且仅使用TA98菌株，建议延长孵育时间并使用其他菌株进行测试。这些结果为叶子和花朵提供了新的安全数据，并且与之前一些证明咖啡副产品安全性的研究一致。然而，需要进一步提高平面艾姆斯生物测定法的灵敏度和选择性，并且建议进一步开展体内和长期安全性研究。考虑到自然变异性、农药的不同用途和处理方式以及供应链的波动，咖啡副产品可能存在很大差异。考虑到对利用咖啡副产品的兴趣日益增加，使用价格合理的2LabsToGo-Eco的平面生物测定技术是咖啡行业一种强大的毒理学筛选选择。
