Analytical and Bioanalytical Chemistry ( IF 3.8 ) Pub Date : 2023-02-25 , DOI: 10.1007/s00216-023-04597-8 Aaron Boughbina-Portolés 1 , Lorenzo Sanjuan-Navarro 1 , Lusine Hakobyan 2 , Marta Gómez-Ferrer 2 , Yolanda Moliner-Martínez 1 , Pilar Sepúlveda 2 , Pilar Campíns-Falcó 1
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Carbon black nanomaterial (CB-NM), as an industrial product with a large number of applications, poses a high risk of exposure, and its impact on health needs to be assessed. The most common testing platform for engineered (E)NMs is in vitro toxicity assessment, which requires prior ENM dispersion, stabilization, and characterization in cell culture media. Here, asymmetric flow field-flow fractionation (AF4) coupled to UV–Vis and dynamic light scattering (DLS) detectors in series was used for the study of CB dispersions in cell culture media, optimizing instrumental variables and working conditions. It was possible to disperse CB in a non-ionic surfactant aqueous solution due to the steric effect provided by surfactant molecules attached on the CB surface which prevented agglomeration. The protection provided by the surfactant or by culture media alone was insufficient to ensure good dispersion stability needed for carrying out in vitro toxicity studies. On the other hand, cell culture media in combination with the surfactant improved dispersion stability considerably, enabling the generation of shorter particles and a more favourable zeta potential magnitude, leading to greater stability due to electrostatic repulsion. It was demonstrated that the presence of amino acids in the culture media improved the monodisperse nature and stability of the CB dispersions, and resulted in a turn towards more negative zeta potential values when the pH was above the amino acid isoelectric point (IEP). Culture media used in real cell culture scenarios were also tested, and in vitro toxicity assays were developed optimizing the compatible amount of surfactant.
Graphical abstract
中文翻译：
通过不对称流场-流动分馏可靠地评估各种细胞培养基的炭黑纳米材料，用于体外毒性测定
炭黑纳米材料（CB-NM）作为一种具有大量应用的工业产品，具有很高的暴露风险，需要评估其对健康的影响。工程化 (E)NM 最常见的测试平台是体外毒性评估，这需要预先在细胞培养基中进行 ENM 分散、稳定和表征。在这里，不对称流场流分馏 (AF4) 耦合到串联的紫外-可见光和动态光散射 (DLS) 检测器，用于研究细胞培养基中的 CB 分散体，优化仪器变量和工作条件。由于附着在 CB 表面的表面活性剂分子提供的空间效应防止团聚，因此可以将 CB 分散在非离子表面活性剂水溶液中。表面活性剂或培养基单独提供的保护不足以确保进行体外毒性研究所需的良好分散稳定性。另一方面，细胞培养基与表面活性剂的结合显着提高了分散稳定性，能够生成更短的颗粒和更有利的 zeta 电位大小，从而由于静电排斥而导致更高的稳定性。结果表明，培养基中氨基酸的存在改善了 CB 分散体的单分散性和稳定性，并导致当 pH 值高于氨基酸等电点 (IEP) 时，zeta 电位值转向更负的 zeta 电位值。还测试了真实细胞培养场景中使用的培养基，
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