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Keywords = planar chromatography coupled with bioassay

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12 pages, 1627 KB  
Article
Coffee By-Products Studied by the Planar Ames Bioassay with pH Indicator Endpoint Using the 2LabsToGo-Eco
by Maryam Monazzah, Cedric Herrmann, Gertrud E. Morlock, Jannika Fuchs and Dirk W. Lachenmeier
Toxics 2025, 13(9), 739; https://doi.org/10.3390/toxics13090739 - 31 Aug 2025
Viewed by 681
Abstract
The mutagenic potential of coffee by-products, including Coffea 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. [...] Read more.
The mutagenic potential of coffee by-products, including Coffea 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, n-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 Salmonella strains is recommended. The results provide new safety data for Coffea 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. Full article
(This article belongs to the Special Issue Health Risk Evaluation of Hazardous Substances in Food)
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15 pages, 3919 KB  
Article
Incorporation of Metabolic Activation in the HPTLC-SOS-Umu-C Bioassay to Detect Low Levels of Genotoxic Chemicals in Food Contact Materials
by Emma Debon, Paul Rogeboz, Hélia Latado, Gertrud E. Morlock, Daniel Meyer, Claudine Cottet-Fontannaz, Gabriele Scholz, Benoît Schilter and Maricel Marin-Kuan
Toxics 2022, 10(9), 501; https://doi.org/10.3390/toxics10090501 - 27 Aug 2022
Cited by 21 | Viewed by 3416
Abstract
The safety evaluation of food contact materials requires excluding mutagenicity and genotoxicity in migrates. Testing the migrates using in vitro bioassays has been proposed to address this challenge. To be fit for that purpose, bioassays must be capable of detecting very low, safety [...] Read more.
The safety evaluation of food contact materials requires excluding mutagenicity and genotoxicity in migrates. Testing the migrates using in vitro bioassays has been proposed to address this challenge. To be fit for that purpose, bioassays must be capable of detecting very low, safety relevant concentrations of DNA-damaging substances. There is currently no bioassay compatible with such qualifications. High-performance thin-layer chromatography (HPTLC), coupled with the planar SOS Umu-C (p-Umu-C) bioassay, was suggested as a promising rapid test (~6 h) to detect the presence of low levels of mutagens/genotoxins in complex mixtures. The current study aimed at incorporating metabolic activation in this assay and testing it with a set of standard mutagens (4-nitroquinoline-N-oxide, aflatoxin B1, mitomycin C, benzo(a)pyrene, N-ethyl nitrourea, 2-nitrofluorene, 7,12-dimethylbenzanthracene, 2-aminoanthracene and methyl methanesulfonate). An effective bioactivation protocol was developed. All tested mutagens could be detected at low concentrations (0.016 to 230 ng/band, according to substances). The calculated limits of biological detection were found to be up to 1400-fold lower than those obtained with the Ames assay. These limits are lower than the values calculated to ensure a negligeable carcinogenic risk of 10−5. They are all compatible with the threshold of toxicological concern for chemicals with alerts for mutagenicity (150 ng/person). They cannot be achieved by any other currently available test procedures. The p-Umu-C bioassay may become instrumental in the genotoxicity testing of complex mixtures such as food packaging, foods, and environmental samples. Full article
(This article belongs to the Special Issue Risk Assessment of Food Contact Materials/Articles)
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