Hines Kelly M, Herron Josi, Xu Libin
Department of Medicinal Chemistry University of Washington, Seattle, WA 98195.
Department of Medicinal Chemistry University of Washington, Seattle, WA 98195; Department of Environmental and Occupational Health Sciences, University of Washington, Seattle, WA 98195.
J Lipid Res. 2017 Apr;58(4):809-819. doi: 10.1194/jlr.D074724. Epub 2017 Feb 6.
Ion mobility-mass spectrometry (IM-MS) has proven to be a highly informative technique for the characterization of lipids from cells and tissues. We report the combination of hydrophilic-interaction liquid chromatography (HILIC) with traveling-wave IM-MS (TWIM-MS) for comprehensive lipidomics analysis. Main lipid categories such as glycerolipids, sphingolipids, and glycerophospholipids are separated on the basis of their lipid backbones in the IM dimension, whereas subclasses of each category are mostly separated on the basis of their headgroups in the HILIC dimension, demonstrating the orthogonality of HILIC and IM separations. Using our previously established lipid calibrants for collision cross-section (CCS) measurements in TWIM, we measured over 250 CCS values covering 12 lipid classes in positive and negative modes. The coverage of the HILIC-IM-MS method is demonstrated in the analysis of Neuro2a neuroblastoma cells exposed to benzalkonium chlorides (BACs) with C10 or C16 alkyl chains, which we have previously shown to affect gene expression related to cholesterol and lipid homeostasis. We found that BAC exposure resulted in significant changes to several lipid classes, including glycerides, sphingomyelins, phosphatidylcholines, and phosphatidylethanolamines. Our results indicate that BAC exposure modifies lipid homeostasis in a manner that is dependent upon the length of the BAC alkyl chain.
离子淌度-质谱联用（IM-MS）已被证明是一种用于表征细胞和组织中脂质的高信息量技术。我们报道了亲水相互作用液相色谱（HILIC）与行波离子淌度质谱（TWIM-MS）联用进行全面脂质组学分析的方法。主要的脂质类别，如甘油脂、鞘脂和甘油磷脂，在离子淌度维度上根据其脂质骨架进行分离，而每类脂质的亚类大多在亲水相互作用液相色谱维度上根据其头部基团进行分离，这证明了亲水相互作用液相色谱和离子淌度分离的正交性。使用我们先前建立的用于在行波离子淌度质谱中测量碰撞截面（CCS）的脂质校准物，我们在正离子和负离子模式下测量了超过250个覆盖12类脂质的CCS值。在分析暴露于具有C10或C16烷基链的氯化苯甲烃铵（BACs）的Neuro2a神经母细胞瘤细胞时，展示了亲水相互作用液相色谱-离子淌度质谱联用方法的覆盖范围，我们之前已表明这些物质会影响与胆固醇和脂质稳态相关的基因表达。我们发现，BAC暴露导致几种脂质类别发生显著变化，包括甘油酯、鞘磷脂、磷脂酰胆碱和磷脂酰乙醇胺。我们的结果表明，BAC暴露以一种依赖于BAC烷基链长度的方式改变脂质稳态。
