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基于 SDS-PAGE 的生物样本中炭黑定量方法。

An SDS-PAGE based method for the quantification of carbon black in biological samples.

机构信息

Key Laboratory of Environmental Biotechnology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China.

出版信息

Analyst. 2020 May 7;145(9):3370-3375. doi: 10.1039/d0an00046a. Epub 2020 Apr 1.

DOI:10.1039/d0an00046a
PMID:32236243
Abstract

Carbon black (CB) has a wide range of industrial applications and recently has been used as the basic model for environmental health studies on airborne particulate matters (PM). Exposure characterization of CB is always the first and most important step towards a better understanding of its effects on human health. However, efforts were largely limited by the lack of valid methods capable of quantifying CB in biological samples. Here, we developed a new method based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) for CB quantification in bio-samples, with a detection limit of 4.3 ng. The method is so economical and convenient that it can be performed in most biology labs. The application of the method was successfully demonstrated in three different cell models (mouse macrophage cells (Raw264.7), human epithelial cells (A549) and mouse mesenchymal stem cells (MSC)) and the results showed that the uptake rates decrease in the order of MSC > Raw264.7 > A549. The surprisingly highest uptake rate of MSC deserves further investigation. The novel method provides a complementary quantitative tool to the use of conventional methods such as radioactive and fluorescent labeling and may facilitate related toxicological studies.

摘要

炭黑 (CB) 具有广泛的工业应用,最近已被用作环境健康研究中关于空气颗粒物 (PM) 的基础模型。对 CB 的暴露特征的描述始终是更好地了解其对人类健康影响的第一步,也是最重要的一步。然而,由于缺乏能够定量生物样本中 CB 的有效方法,因此这方面的努力受到了很大的限制。在这里,我们开发了一种基于十二烷基硫酸钠-聚丙烯酰胺凝胶电泳 (SDS-PAGE) 的新方法,用于生物样本中的 CB 定量,检测限为 4.3ng。该方法经济、方便,大多数生物学实验室都可以进行。该方法在三种不同的细胞模型(小鼠巨噬细胞 (Raw264.7)、人上皮细胞 (A549) 和小鼠间充质干细胞 (MSC))中的应用得到了成功验证,结果表明摄取率按 MSC>Raw264.7>A549 的顺序降低。间充质干细胞出人意料的高摄取率值得进一步研究。该新方法为放射性和荧光标记等传统方法的使用提供了一种补充的定量工具,并可能有助于相关的毒理学研究。

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