Department of Environment and Health, Vrije Universiteit, De Boelelaan 1085, 1081 HV, Amsterdam, The Netherlands.
Anal Bioanal Chem. 2020 Jul;412(18):4313-4325. doi: 10.1007/s00216-020-02661-1. Epub 2020 Apr 29.
In this study, both conventional one-dimensional liquid chromatography (1DLC) and comprehensive two-dimensional liquid chromatography (2DLC) coupled to a high-resolution time-of-flight mass spectrometer (HR-TOF MS) were used for full-scale lipid characterization of lipid extracts from zebrafish embryos. We investigated the influence on annotated lipids and different separation mechanisms (HILIC, C18, and PFP), and their different orders arranged in the first and the second dimensions. As a result, the number of lipid species annotated by conventional one-dimensional LC-MS was between 212 and 448. In contrast, the number of individual lipids species annotated by C18×HILIC, HILIC×C18, and HILIC×PFP were 1784, 1059, and 1123, respectively. Therefore, it was evident that the performance of comprehensive 2DLC, especially the C18×HILIC method, considerably exceeded 1DLC. Interestingly, a comparison of the HILIC×C18 and C18×HILIC approaches showed, under the optimized conditions, similar orthogonality, but the effective separation power of the C18×HILIC was much higher. A comparison of the HILIC×C18 and the HILIC×PFP methods demonstrated that the HILIC×PFP separation had superior orthogonality with a small increase on its effective peak capacity, indicating that the HILIC×PFP combination maybe a promising platform for untargeted lipidomics in complex samples. Finally, from the comprehensive lipid profiling respective, the C18×HILIC was selected for further studies.
在这项研究中，我们使用传统的一维液相色谱（1DLC）和全面的二维液相色谱（2DLC）与高分辨率飞行时间质谱（HR-TOF MS）联用，对斑马鱼胚胎脂质提取物进行全谱脂质特征分析。我们研究了不同分离机制（亲水作用色谱、C18 和 PFP）以及在第一维和第二维中不同顺序排列对注释脂质的影响。结果表明，传统一维 LC-MS 注释的脂质种类数在 212 到 448 之间。相比之下，C18×HILIC、HILIC×C18 和 HILIC×PFP 注释的单个脂质种类数分别为 1784、1059 和 1123。因此，全面二维 LC 的性能，特别是 C18×HILIC 方法，明显优于一维 LC。有趣的是，对 HILIC×C18 和 C18×HILIC 方法的比较表明，在优化条件下，两者具有相似的正交性，但 C18×HILIC 的有效分离能力要高得多。对 HILIC×C18 和 HILIC×PFP 方法的比较表明，HILIC×PFP 分离具有更好的正交性，其有效峰容量略有增加，表明 HILIC×PFP 组合可能是复杂样品中靶向脂质组学的有前途的平台。最后，从全面的脂质分析来看，选择 C18×HILIC 进行进一步研究。
