Department of Metabolism, Digestion and Reproduction, Imperial College London, South Kensington, London SW7 2AZ, U.K.
AnalytiCon Discovery GmbH, Hermannswerder Haus 17, 14473 Potsdam, Germany.
J Proteome Res. 2022 Jun 3;21(6):1428-1437. doi: 10.1021/acs.jproteome.2c00030. Epub 2022 May 10.
Liquid chromatography coupled to mass spectrometry is a key metabolomics/metabonomics technology. Reversed-phase liquid chromatography (RPLC) is very widely used as a separation step, but typically has poor retention of highly polar metabolites. Here, we evaluated the combination of two alternative methods for improving retention of polar metabolites based on 6-aminoquinoloyl--hydroxysuccinidimyl carbamate derivatization for amine groups, and ion-pairing chromatography (IPC) using tributylamine as an ion-pairing agent to retain acids. We compared both of these methods to RPLC and also to each other, for targeted analysis using a triple-quadrupole mass spectrometer, applied to a library of ca. 500 polar metabolites. IPC and derivatization were complementary in terms of their coverage: combined, they improved the proportion of metabolites with good retention to 91%, compared to just 39% for RPLC alone. The combined method was assessed by analyzing a set of liver extracts from aged male and female mice that had been treated with the polyphenol compound ampelopsin. Not only were a number of significantly changed metabolites detected, but also it could be shown that there was a clear interaction between ampelopsin treatment and sex, in that the direction of metabolite change was opposite for males and females.
液相色谱-质谱联用是代谢组学/代谢组学的关键技术。反相液相色谱（RPLC）作为分离步骤得到了广泛的应用，但通常对高极性代谢物的保留性较差。在这里，我们评估了两种替代方法的结合，以改善基于 6-氨基喹啉酰基 - 羟基琥珀酰亚胺基碳酸酯衍生化的胺基和使用三丁胺作为离子对试剂的离子对色谱（IPC）对极性代谢物的保留，用于使用三重四极杆质谱仪进行靶向分析，应用于约 500 种极性代谢物的文库。就其覆盖范围而言，IPC 和衍生化是互补的：组合使用时，与单独使用 RPLC 相比，将具有良好保留性的代谢物的比例提高到 91%。通过分析一组用多酚化合物安石榴苷处理的老年雄性和雌性小鼠的肝提取物来评估组合方法。不仅检测到了一些明显变化的代谢物，还可以表明安石榴苷处理和性别之间存在明显的相互作用，即雄性和雌性的代谢物变化方向相反。
