Optimisation of high-speed lipidome analysis by nanoflow ultrahigh-performance liquid chromatography-tandem mass spectrometry: Application to identify candidate biomarkers for four different cancers
Optimisation of high-speed lipidome analysis by nanoflow ultrahigh-performance liquid chromatography-tandem mass spectrometry: Application to identify candidate biomarkers for four different cancers
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DOI：
10.1016/j.jchromb.2021.122739
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发表时间：
2021-05-13
影响因子：
3
通讯作者：
Moon, Myeong Hee
中科院分区：
文献类型：
--
作者：
Lee, Gwang Bin;Kim, Young Beom;Moon, Myeong Hee
脂质分析是一种强大的工具，可以阐明脂质在代谢疾病中的致病作用，并促进潜在生物标志物的开发。大规模脂质组学的脂质分析需要高速、高通量的分析平台。在本研究中，通过研究柱流速、泵流速、停留时间、初始二元流动相组成和梯度持续时间对标准脂质混合物分离效率的影响，优化了使用纳流超高效液相色谱-电喷雾电离-串联质谱（nUHPLC-ESI-MS/MS）进行脂质分析的高速分析方法。高速脂质分离的最小梯度时间是通过检查基于时间的分离效率和同量异位脂质种类的光谱重叠来确定的，其中鞘磷脂和磷脂酰胆碱的第二同位素的分子量仅相差 1 Da，基于反应监测的定量。最后，应用优化的 nUHPLC-ESI-MS/MS 方法分析来自肝癌、胃癌、肺癌和结直肠癌患者的 200 份血浆样本，通过测量先前确定的候选脂质生物标志物来评估其性能。大约 73% 的报告候选标志物（肝脏 7 个中的 6 个、胃 5/9、肺癌 4/6 和结直肠癌 6/7）可以使用优化方法进行分配，支持其用于高通量脂质分析。
Lipid analysis is a powerful tool that can elucidate the pathogenic roles of lipids in metabolic diseases, and facilitate the development of potential biomarkers. Lipid analysis by large-scale lipidomics requires a high-speed and high-throughput analytical platform. In the present study, a high-speed analytical method for lipid analysis using nanoflow ultrahigh-performance liquid chromatography-electrospray ionisation-tandem mass spectrometry (nUHPLC-ESI-MS/MS) was optimised by investigating the effects of column flow rate, pump flow rate, dwell time, initial binary mobile phase composition, and gradient duration on the separation efficiency of standard lipid mixtures. The minimum gradient time for high-speed lipid separation was determined by examining the time-based separation efficiency and spectral overlap of isobaric lipid species during selected reaction monitoring-based quantification of sphingomyelin and a second isotope of phosphatidylcholine, which differ in molecular weight by only 1 Da. Finally, the optimised nUHPLC-ESI-MS/MS method was applied to analyse 200 plasma samples from patients with liver, gastric, lung, and colorectal cancer to evaluate its performance by measuring previously identified candidate lipid biomarkers. About 73% of the reported marker candidates (6 out of 7 in liver, 5/9 in gastric, 4/6 in lung, and 6/7 in colorectal cancer) could be assigned using the optimised method, supporting its use for high-throughput lipid analysis.