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J Lipid Res. 2014 Aug;55(8):1668-77. doi: 10.1194/jlr.M046995. Epub 2014 Jun 17.

Characterization of acyl chain position in unsaturated phosphatidylcholines using differential mobility-mass spectrometry.

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School of Chemistry University of Wollongong, New South Wales 2522, Australia.
School of Medicine, University of Wollongong, New South Wales 2522, Australia.
Central Analytical Research Facility, Queensland University of Technology, Queensland 4000, Australia.
AB SCIEX, Concord, Ontario L4K 4V8, Canada.


Glycerophospholipids (GPs) that differ in the relative position of the two fatty acyl chains on the glycerol backbone (i.e., sn-positional isomers) can have distinct physicochemical properties. The unambiguous assignment of acyl chain position to an individual GP represents a significant analytical challenge. Here we describe a workflow where phosphatidylcholines (PCs) are subjected to ESI for characterization by a combination of differential mobility spectrometry and MS (DMS-MS). When infused as a mixture, ions formed from silver adduction of each phospholipid isomer {e.g., [PC (16:0/18:1) + Ag](+) and [PC (18:1/16:0) + Ag](+)} are transmitted through the DMS device at discrete compensation voltages. Varying their relative amounts allows facile and unambiguous assignment of the sn-positions of the fatty acyl chains for each isomer. Integration of the well-resolved ion populations provides a rapid method (< 3 min) for relative quantification of these lipid isomers. The DMS-MS results show excellent agreement with established, but time-consuming, enzymatic approaches and also provide superior accuracy to methods that rely on MS alone. The advantages of this DMS-MS method in identification and quantification of GP isomer populations is demonstrated by direct analysis of complex biological extracts without any prior fractionation.


differential mobility spectrometry; lipid isomers; mass spectrometry; sn-positional isomers

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