Research Unit Analytical BioGeoChemistry, Helmholtz Zentrum München, Neuherberg, Germany.
Chair of Analytical Food Chemistry, TUM School of Life Sciences, Technical University of Munich, Freising-Weihenstephan, Germany.
Metabolomics. 2023 Jun 23;19(7):61. doi: 10.1007/s11306-023-02025-7.
Polar metabolites in Caenorhabditis elegans (C. elegans) have predominantly been analyzed using hydrophilic interaction liquid chromatography coupled to mass spectrometry (HILIC-MS). Capillary electrophoresis coupled to mass spectrometry (CE-MS) represents another complementary analytical platform suitable for polar and charged analytes.
We compared CE-MS and HILIC-MS for the analysis of a set of 60 reference standards relevant for C. elegans and specifically investigated the strengths of CE separation. Furthermore, we employed CE-MS as a complementary analytical approach to study polar metabolites in C. elegans samples, particularly in the context of longevity, in order to address a different part of its metabolome.
We analyzed 60 reference standards as well as metabolite extracts from C. elegans daf-2 loss-of-function mutants and wild-type (WT) samples using HILIC-MS and CE-MS employing a Q-ToF-MS instrument.
CE separations showed narrower peak widths and a better linearity of the estimated response function across different concentrations which is linked to less saturation of the MS signals. Additionally, CE exhibited a distinct selectivity in the separation of compounds compared to HILIC-MS, providing complementary information for the analysis of the target compounds. Analysis of C. elegans metabolites of daf-2 mutants and WT samples revealed significant alterations in shared metabolites identified through HILIC-MS, as well as the presence of distinct metabolites.
CE-MS was successfully applied in C. elegans metabolomics, being able to recover known as well as identify novel putative biomarkers of longevity.
秀丽隐杆线虫（C. elegans）中的极性代谢物主要使用亲水相互作用液相色谱与质谱联用（HILIC-MS）进行分析。毛细管电泳与质谱联用（CE-MS）是另一种适用于极性和带电分析物的互补分析平台。
我们比较了 CE-MS 和 HILIC-MS 对 60 种参考标准品的分析，这些标准品与 C. elegans 相关，特别是研究了 CE 分离的优势。此外，我们将 CE-MS 作为一种互补的分析方法，用于研究 C. elegans 样品中的极性代谢物，特别是在寿命方面，以解决其代谢组的不同部分。
我们使用 Q-ToF-MS 仪器，通过 HILIC-MS 和 CE-MS 分析了 60 种参考标准品以及 daf-2 功能丧失突变体和野生型（WT）C. elegans 样品的代谢物提取物。
CE 分离显示出较窄的峰宽和估计响应函数在不同浓度下更好的线性，这与 MS 信号的饱和程度较低有关。此外，CE 在化合物的分离中表现出与 HILIC-MS 不同的选择性，为目标化合物的分析提供了互补信息。分析 daf-2 突变体和 WT 样品的 C. elegans 代谢物，揭示了通过 HILIC-MS 鉴定的共享代谢物以及存在的独特代谢物的显著变化。
CE-MS 成功应用于 C. elegans 代谢组学，可以恢复已知的以及鉴定与长寿相关的新的潜在生物标志物。
