Abstract：
High-performance thin-layer chromatography hyphenated with planar multiplex bioassays and high-resolution tandem mass spectrometry contributes to the non-target detection or even identification of active compounds in complex mixtures such as food, feed, cosmetics, commodities, and environmental samples. It can be used to discover previously unknown harmful or active substances in complex samples and to tentatively assign molecular formulas. This method is already faster than the commonly used in vitro assays along with liquid chromatographic separations, but overnight cell cultivation still prevents a planar bioassay from being performed within one day. There is also still potential for optimization in terms of sustainability. To achieve this, the planar bioassay protocols for the detection of androgen-like and estrogen-like compounds were harmonized. The successful minimization of the cell culture volume enabled accelerated cell cultivation, which allowed the bioassay to be performed within one day. This was considered a milestone achieved, as up to 23 samples per plate can now be analyzed from the start of cultivation to the biological endpoint on the same day. Doubling the substrate amount and increasing the pH of the silica gel layer led to a more sensitive and selective bioassay due to the enhanced fluorescence of the formed end-product. The faster and more sustainable bioassay protocol was applied to complex samples such as sunscreen and red wine to detect estrogen-like compounds. The developed method was validated by comparison with a standard method.
摘要：
高效薄层色谱与平面多重生物测定和高分辨率串联质谱联用有助于非目标检测甚至鉴定复杂混合物如食品中的活性化合物，饲料，化妆品,商品,和环境样本。它可用于在复杂样品中发现先前未知的有害或活性物质，并初步指定分子式。这种方法已经比常用的体外测定和液相色谱分离更快，但是过夜细胞培养仍然阻止平面生物测定在一天内进行。在可持续性方面仍有优化的潜力。为了实现这一点，统一了检测雄激素样和雌激素样化合物的平面生物测定方案.细胞培养体积的成功最小化使得能够加速细胞培养。这使得生物测定可以在一天内进行。这被认为是一个里程碑，现在，从培养开始到同一天的生物学终点，每个板可以分析多达23个样品。由于所形成的最终产物的荧光增强，将底物量加倍并增加硅胶层的pH会导致更灵敏和选择性的生物测定。更快，更可持续的生物测定方案被应用于复杂的样品，如防晒霜和红酒，以检测雌激素样化合物。通过与标准方法的比较验证了所开发的方法。
