Robust and high-throughput lipidomic quantitation of human blood samples using flow injection analysis with tandem mass spectrometry for clinical use
Robust and high-throughput lipidomic quantitation of human blood samples using flow injection analysis with tandem mass spectrometry for clinical use
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DOI：
10.1007/s00216-022-04490-w
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发表时间：
2023-01-04
影响因子：
4.3
通讯作者：
Holcapek, Michal
中科院分区：
文献类型：
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作者：
Idkowiak, Jakub;Jirasko, Robert;Holcapek, Michal
将脂质提取物直接注入质谱仪的离子源是一种成熟的脂质分析方法。在大多数情况下，纳米流体装置用于样品引入。然而，基于从色谱泵注入样品的流动注射分析（FIA）可以提供基于鸟枪的方法的简单替代方案。在这里，我们描述了基于FIA和串联质谱（MS/MS）的方法的重要修改。我们专注于最大限度地减少FIA/MS的污染，以使脂质组学平台更加稳健，并提高其在临床应用中所需的长序列测量的能力和适用性。对所开发方法的稳健性验证证实了其适用于人血浆分析中的脂质定量。标准人血浆参比物质（NIST SRM 1950）和从肾癌患者和健康志愿者中采集的一组血浆样品的测量在FIA-MS/MS和超高效超临界流体色谱（UHPSFC）/MS之间产生了高度相似的结果，从而证明所有修改对统计输出几乎没有影响。新修改的FIA-MS/MS允许在5.7 min内定量血浆中的141种脂质（11种主要脂质）。最后，我们在布拉格的综合大学医院的临床实验室中测试了该方法。在临床环境中，方法能力达到257个样品/天。我们还展示了基于在临床环境和Pardubice大学分析实验室中获得的结果训练的分类模型的类似性能。总之，这些研究结果表明，在临床实验室应用的高潜力的改进FIA-MS/MS测量血浆和血清脂质谱。
Direct infusion of lipid extracts into the ion source of a mass spectrometer is a well-established method for lipid analysis. In most cases, nanofluidic devices are used for sample introduction. However, flow injection analysis (FIA) based on sample infusion from a chromatographic pump can offer a simple alternative to shotgun-based approaches. Here, we describe important modification of a method based on FIA and tandem mass spectrometry (MS/MS). We focus on minimizing contamination of the FIA/MS both to render the lipidomic platform more robust and to increase its capacity and applicability for long-sequence measurements required in clinical applications. Robust validation of the developed method confirms its suitability for lipid quantitation in human plasma analysis. Measurements of standard human plasma reference material (NIST SRM 1950) and a set of plasma samples collected from kidney cancer patients and from healthy volunteers yielded highly similar results between FIA-MS/MS and ultra-high-performance supercritical fluid chromatography (UHPSFC)/MS, thereby demonstrating that all modifications have practically no effect on the statistical output. Newly modified FIA-MS/MS allows for the quantitation of 141 lipid species in plasma (11 major lipid classes) within 5.7 min. Finally, we tested the method in a clinical laboratory of the General University Hospital in Prague. In the clinical setting, the method capacity reached 257 samples/day. We also show similar performance of the classification models trained based on the results obtained in clinical settings and the analytical laboratory at the University of Pardubice. Together, these findings demonstrate the high potential of the modified FIA-MS/MS for application in clinical laboratories to measure plasma and serum lipid profiles.