Stütz Lena, Weiss Stefan C, Schulz Wolfgang, Schwack Wolfgang, Winzenbacher Rudi
Laboratory for Operation Control and Research, Zweckverband Landeswasserversorgung, Am spitzigen Berg 1, 89129 Langenau, Germany; Institute of Food Chemistry, University of Hohenheim, Garbenstraße 28, 70599 Stuttgart, Germany.
Laboratory for Operation Control and Research, Zweckverband Landeswasserversorgung, Am spitzigen Berg 1, 89129 Langenau, Germany.
J Chromatogr A. 2017 Nov 17;1524:273-282. doi: 10.1016/j.chroma.2017.10.009. Epub 2017 Oct 6.
There are thousands of organic trace substances in the environment that are not fully characterized, and evaluation of their relevance to the ecosystem is difficult. Effect-directed analysis (EDA) is a suitable tool to assess the effects of a substance via in-vitro bioassays, which can provide information about the relevance of the substance. High-performance thin-layer chromatography (HPTLC) has been shown to be a good method for fractionation. Environmental samples, however, often have high complexity, which is why the peak capacity of HPTLC is not sufficient. Therefore, this study focused on the development of selective two-dimensional (2D) HPTLC-EDA to increase the peak capacity and facilitate the identification of effective compounds. Thus, only effective zones were selected in the first dimension in terms of heart-cutting and were transferred to the second dimension through elution head-based extraction. Three 2D approaches were developed and validated. The best results in terms of peak capacity and orthogonality were achieved when the retardation factors of the first dimension were used to adjust the mobile phase (MP) for the second dimension. Applying the acetylcholinesterase (AChE) inhibition assay as an example EDA, analysis of spiked surface water by 2D HPTLC-EDA allowed zones with neurotoxic effects to responsible substances to be assigned. The 2D separation reduced the complexity of effective zones and thus facilitated the subsequent identification of effective compounds. Knowledge about a substancés effects enabled assessment of its relevance to the environment.
环境中存在数千种未被充分表征的有机痕量物质，评估它们与生态系统的相关性具有难度。效应导向分析（EDA）是一种通过体外生物测定来评估物质效应的合适工具，它能够提供有关该物质相关性的信息。高效薄层色谱法（HPTLC）已被证明是一种良好的分离方法。然而，环境样品通常具有高度复杂性，这就是HPTLC的峰容量不足的原因。因此，本研究聚焦于开发选择性二维（2D）HPTLC-EDA，以提高峰容量并便于鉴定有效化合物。因此，在第一维中仅根据中心切割选择有效区域，并通过基于洗脱头的萃取转移到第二维。开发并验证了三种二维方法。当使用第一维的保留因子来调整第二维的流动相（MP）时，在峰容量和正交性方面取得了最佳结果。以乙酰胆碱酯酶（AChE）抑制测定为例进行EDA，通过二维HPTLC-EDA对加标地表水进行分析，可将具有神经毒性效应的区域归因于相关物质。二维分离降低了有效区域的复杂性，从而便于后续鉴定有效化合物。关于物质效应的知识有助于评估其与环境的相关性。
