Blood plasma has gained protagonism in lipidomics studies due to its availability, uncomplicated collection and preparation, and informative readout of physiological status. At the same time, it is also technically challenging to analyze due to its complex lipid composition affected by many factors, which can hamper the throughput and/or lipidomics coverage. To tackle these issues, we developed a comprehensive, high throughput, and quantitative mass spectrometry-based shotgun lipidomics platform for blood plasma lipid analyses. The main hallmarks of this technology are (i) it is comprehensive, covering 22 quantifiable different lipid classes encompassing more than 200 lipid species; (ii) it is amenable to high-throughput, with less than 5 min acquisition time allowing the complete analysis of 200 plasma samples per day; (iii) it achieves absolute quantification, by inclusion of internal standards for every lipid class measured; (iv) it is highly reproducible, achieving an average coefficient of variation of <10% (intra-day), approx. 10% (inter-day), and approx. 15% (inter-site) for most lipid species; (v) it is easily transferable allowing the direct comparison of data acquired in different sites. Moreover, we thoroughly assessed the influence of blood stabilization with different anticoagulants and freeze-thaw cycles to exclude artifacts generated by sample preparation. Practical applications: This shotgun lipidomics platform can be implemented in different laboratories without compromising reproducibility, allowing multi-site studies and inter-laboratory comparisons. This possibility combined with the high-throughput, broad lipidomic coverage and absolute quantification are important aspects for clinical applications and biomarker research.

中文翻译：

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用于血浆完整脂质的高通量、全面和定量分析的自动化鸟枪脂质组学平台

血浆因其可用性、简单的收集和制备以及生理状态的信息读出而在脂质组学研究中占据主导地位。同时，由于其复杂的脂质成分受多种因素影响，分析在技术上也具有挑战性，这可能会阻碍通量和/或脂质组学的覆盖。为了解决这些问题，我们开发了一个全面的、高通量的、基于定量质谱的鸟枪脂质组学平台，用于血浆脂质分析。该技术的主要特点是 (i) 它是全面的，涵盖 22 种可量化的不同脂质类别，包括 200 多种脂质；(ii) 适合高通量，采集时间少于 5 分钟，允许每天完整分析 200 个血浆样本；(iii) 它实现了绝对量化，通过为每个测量的脂质类别包含内部标准；(iv) 它是高度可重复的，平均变异系数 <10%（日内），大约 10%（日间），大约 大多数脂质种类为 15%（位点间）；(v) 它易于转移，允许直接比较在不同地点获得的数据。此外，我们彻底评估了不同抗凝剂和冻融循环对血液稳定的影响，以排除样品制备产生的伪影。实际应用：这种鸟枪式脂质组学平台可以在不同的实验室中实施，而不会影响可重复性，允许多站点研究和实验室间比较。这种可能性与高通量相结合，