Journal
JOURNAL OF CHROMATOGRAPHY A
Volume 1351, Issue -, Pages 37-45Publisher
ELSEVIER
DOI: 10.1016/j.chroma.2014.04.069
Keywords
Phosphatidylcholine isomers; Two dimensional liquid chromatography; Mass spectrometry; Ozonolysis; Double bond location
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In this study, two dimensional (2D) liquid chromatography (LC) was coupled in-line with ozonolysis-mass spectrometry (O-3-MS) for the elucidation of phosphatidylcholine (PC) structures in phospholipid extracts. In O-3-MS, PC molecules carried by mobile phase pass through a length of gas-permeable tubing surrounded by ozone vapor resulting in oxidative cleavage of double bonds. The characteristic aldehyde products are then directly analyzed by eletrospray ionization (ESI)-MS, allowing assignment of the double bond positions within the PC fatty acyl chains. The on-line 2D-LC method employs hydrophilic interaction chromatography as the first dimension to separate phospholipid classes and reversed phase chromatography in the second dimension to further separate PC molecular species. Thus, by combing the separation power of 2D-LC with in-line O-3-MS (2D-LC/O-3-MS), PC species in complex mixtures can be identified including the detailed structure of their two fatty acid chains. When the 2D-LC/O-3-MS method was applied to a rat liver phospholipid extract, 19 PC molecular species were identified with fatty acid isomers unambiguously assigned. (C) 2014 Elsevier B.V. All rights reserved.
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