Identification and Quantitation of Changes in the Platelet Activating Factor Family of Glycerophospholipids over the Course of Neuronal Differentiation by High-Performance Liquid Chromatography Electr

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• 作者：Shawn N. Whitehead ; Weimin Hou ; Martin Ethier ; Jeffrey C. Smith ; André ; Bourgeois ; Rachelle Denis ; Steffany A. L. Bennett ; Daniel Figeys
• 刊名：Analytical Chemistry
• 出版年：2007
• 出版时间：November 15, 2007
• 年：2007
• 卷：79
• 期：22
• 页码：8539 - 8548
• 全文大小：538K
• 年卷期：v.79,no.22(November 15, 2007)
• ISSN：1520-6882

文摘

Glycerophospholipids are important structural lipids inmembranes with changes associated with progressiveneurodegenerative disorders such as Alzheimer disease.Synthesis of the platelet activating factor (PAF) glycerophospholipid subclass is implicated in the control ofneuronal differentiation and death. In this article, wecombine nanoflow HPLC and mass spectrometry toscreen, identify, and quantitate changes in glycerophospholipid subspecies, specifically PAF family members,over the course of neuronal differentiation. Furthermore,precursor ion scans for fragments characteristic of PAFphosphocholine family members and the standard additions of PAF subspecies were combined to performabsolute quantitation of PAF lipids in undifferentiated anddifferentiated PC12 cells. Surprisingly, a marked asymmetry was detected in the two predominant PAF species(C16:0, C18:0) over the course of differentiation. Theseresults describe a new technique for the sensitive analysisof lipids combining nanoflow HPLC, ESI-MS, and precursor ion scan. Limits of detection of as little as 2 pg of PAFand LPC were obtained, and analysis of the lipidome ofas little as 70 000 cells was performed on this system.Furthermore, application to the PC12 model identified aquantifiable difference between PAF molecular speciesproduced over the course of neuronal differentiation.