Leibniz-Institut für Analytische Wissenschaften-ISAS e.V , Otto-Hahn-Str. 6b , 44227 Dortmund , Germany.
Anal Chem. 2018 Jul 3;90(13):8093-8101. doi: 10.1021/acs.analchem.8b01275. Epub 2018 Jun 11.
Nano-liquid chromatography (nLC)-nanoelectrospray (NSI) is one of the cornerstones of mass-spectrometry-based bioanalytics. Nevertheless, the application of nLC is not yet prevalent in lipid analyses. In this study, we established a reproducible nLC separation for global lipidomics and describe the merits of using such a miniaturized system for lipid analyses. In order to enable comprehensive lipid analyses that is not restricted to specific lipid classes, we particularly optimized sample preparation conditions and reversed-phase separation parameters. We further benchmarked the developed nLC system to a commonly used high flow HPLC/ESI MS system in terms of lipidome coverage and sensitivity. The comparison revealed an intensity gain between 2 and 3 orders of magnitude for individual lipid classes and an increase in the linear dynamic range of up to 2 orders of magnitude. Furthermore, the analysis of the yeast lipidome using nLC/NSI resulted in more than a 3-fold gain in lipid identifications. All in all, we identified 447 lipids from the core phospholipid lipid classes (PA, PE, PC, PS, PG, and PI) in Saccharomyces cerevisiae.
纳升级液相色谱（nLC）-纳升级电喷雾（NSI）是基于质谱的生物分析的基石之一。然而，nLC 的应用在脂质分析中尚未普及。在这项研究中，我们建立了一种可重现的用于全局脂质组学的 nLC 分离方法，并描述了使用这种微型系统进行脂质分析的优点。为了能够进行不受特定脂质类别限制的综合脂质分析，我们特别优化了样品制备条件和反相分离参数。我们还根据脂质组覆盖范围和灵敏度，将开发的 nLC 系统与常用的高通量 HPLC/ESI MS 系统进行了基准测试。比较结果显示，个别脂质类别的强度增益在 2 到 3 个数量级之间，线性动态范围增加了 2 个数量级。此外，使用 nLC/NSI 分析酵母脂质组可使脂质鉴定数量增加 3 倍以上。总的来说，我们从酿酒酵母的核心磷脂脂质类（PA、PE、PC、PS、PG 和 PI）中鉴定出 447 种脂质。
