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Friday, February 26, 2016

A great way to do phospholipid analysis

 In a recent article published in the Journal of Chromatography A, Cífková et al. have investigated the chromatographic behavior of various phospholipids such as (lyso)phosphatidic acids and (lyso)phosphatidylserines. If you have ever done these kinds of compounds then you know they present a challenge to say the least; they contain polar functional groups which make them less suitable for chromatographic methods used to retain more hydrophobic lipids.
Furthermore, identification of discreet lipid classes is greatly facilitated by the high specificity of LC-MS, but previous methods reported in the literature tend to use ion pair reagents, which are not amenable to MS.  This can be painful.
The authors present a comparison of columns for the LC-MS separation of these compounds in real world samples (porcine brain and kidney extracts). In terms of peak symmetry, the Cogent Diamond Hydride™ produced tailing factors in the range 1.0–1.6 for all the studied analytes. Two analytes in particular tailed so severely on HILIC columns that a tailing factor could not be calculated; in contrast, the same two compounds produced only moderate tailing (1.4 and 1.6) on the Diamond Hydride™.
If you struggle with phospholipid analysis, this paper is well worthwhile reading as it is full of new insights.
REFERENCE:
E. Cífková, R. Hájek, M. Lísa, M. Holčapek, Hydrophilic interaction liquid chromatography–mass spectrometry of (lyso)phosphatidic acids, (lyso)phosphatidylserines and other lipid classes, J. Chromatogr. A (2016), in press.

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