†Center for Proteomics and Metabolomics, Leiden University Medical Center, Albinusdreef 2, 2300RC Leiden, The Netherlands.
‡Department of Rheumatology, Leiden University Medical Center, Albinusdreef 2, 2300RC Leiden, The Netherlands.
Anal Chem. 2015;87(10):5036-40. doi: 10.1021/acs.analchem.5b00786. Epub 2015 Apr 30.
Differential mobility spectrometry (DMS) is capable of separating stereoisomeric molecular ions based on their mobility in an oscillating electrical field with an asymmetric waveform. Thus, it is an "orthogonal" technique to chromatography and (tandem) mass spectrometry. Bioactive lipids, particularly of the eicosanoid and docosanoid class feature numerous stereoisomers, which exhibit a highly specific structure-activity relationship. Moreover, the geometry of these compounds also reflects their biochemical origin. Therefore, the unambiguous characterization of related isomers of the eicosanoid and docosanoid classes is of fundamental importance to the understanding of their origin and function in many biological processes. Here we show, that SelexION DMS technology coupled to μLC-MS/MS is capable of differentiating at least five closely related leukotrienes partially coeluting and (almost) unresolvable using LC-MS/MS only. We applied the developed method to the separation of LTB4 and its coeluting isomer 5S,12S-diHETE in murine peritoneal exudate cells, showing that LTB4 is present only after zymosan A injection while its isomer 5S,12S-diHETE is produced after saline (PBS) administration. Additionally, we show that the SelexION technology can also be applied to the separation of PD1 and PDX (10S,17S-diHDHA), two isomeric protectins.
差分式迁移谱（DMS）能够根据分子离子在具有不对称波形的振荡电场中的迁移率来分离对映异构体。因此，它是一种与色谱和（串联）质谱“正交”的技术。生物活性脂质，特别是类二十烷酸和类二十二烷酸，具有许多对映异构体，这些对映异构体表现出高度特异性的结构-活性关系。此外，这些化合物的几何形状也反映了它们的生化起源。因此，明确表征类二十烷酸和类二十二烷酸相关的异构体对于理解它们在许多生物过程中的起源和功能至关重要。在这里，我们展示了 SelexION DMS 技术与 μLC-MS/MS 相结合，能够区分至少五种密切相关的白三烯，这些白三烯部分共洗脱，并且仅使用 LC-MS/MS 几乎无法分辨。我们将开发的方法应用于分离鼠腹膜渗出液细胞中的 LTB4 及其共洗脱异构体 5S,12S-二 HETE，结果表明只有在注射酵母聚糖 A 后才存在 LTB4，而其异构体 5S,12S-二 HETE 是在生理盐水（PBS）给药后产生的。此外，我们还表明，SelexION 技术还可应用于分离 PD1 和 PDX（10S,17S-二 HDHA），这两种异构体保护素。
