1 research outputs found
Separation of Cis–Trans Phospholipid Isomers Using Reversed Phase LC with High Resolution MS Detection
The increased presence of synthetic trans fatty acids
into western
diets has been shown to have deleterious effects on physiology and
raising an individual’s risk of developing metabolic disease,
cardiovascular disease, and stroke. The importance of these fatty
acids for health and the diversity of their (patho) physiological
effects suggest that not only should the free trans fatty acids be
studied but also monitoring the presence of these fats into the side
chains of biological lipids, such as glycerophospholipids, is also
essential. We developed a high resolution LC-MS method that quantitatively
monitors the major lipid classes found in biospecimens in an efficient,
sensitive, and robust manner while also characterizing individual
lipid side chains through the use of high energy collisional dissociation
(HCD) fragmentation and chromatographic alignment. We herein show
how this previously described reversed phase method can baseline separate
the cis–trans isomers of phosphatidylglycerol and phosphatidylcholine
(PC) with two 18:1 side chains, in both positive and negative mode,
as neat solutions and when spiked into a biological matrix. Endogenous
PC (18:1/18:1)-cis and PC (18:1/18:1)-trans isomers were examined
in mitochondrial and serum profiling studies, where rats were fed
diets enriched in either trans 18:1 fatty acids or cis 18:1 fatty
acids. In this study, we determined the cis:trans isomer ratios of
PC (18:1/18:1) and related this ratio to dietary composition. This
generalized LC-MS method enables the monitoring of trans fats in biological
lipids in the context of a nontargeted method, allowing for relative
quantitation and enhanced identification of unknown lipids in complex
matrixes