Abstract
Few high-performance liquid chromatography–tandem mass spectrometry (LC-MS/MS) methods have been developed for the full quantitation of fatty acids from human plasma without derivatization. Therefore, we propose a method that requires fewer sample preparation steps, which can be used for the quantitation of several polyunsaturated fatty acids in human plasma. The method offers rapid, accurate, sensitive, and simultaneous quantification of omega 3 (α-linolenic, eicosapentaenoic, and docosahexaenoic acids) and omega 6 fatty acids (arachidonic and linoleic acids) using high-performance LC-MS/MS. The selected fatty acids were analysed in lipid extracts from both free and total forms. Chromatographic separation was achieved using a reversed phase C18 column with isocratic flow using ammonium acetate for improving negative electrospray ionization (ESI) response. Mass detection was performed in multiple reaction monitoring (MRM) mode, and deuterated internal standards were used for each target compound. The limits of quantification were situated in the low nanomolar range, excepting linoleic acid, for which the limit was in the high nanomolar range. The method was validated according to the U.S. Department of Health and Human Services guidelines, and offers a fast, sensitive, and reliable quantification of selected omega 3 and 6 fatty acids in human plasma.
Highlights
Polyunsaturated fatty acids (PUFAs) play essential roles in human physiology, and are both obtained from foods and synthesized endogenously
By comparing it to the Bligh–Dyer method, we found that the two methods gave similar results for free FAs extraction
A previous study by Reis et al [20] showed that the hexane/isopropanol method is very efficient for extracting triacylglycerols, ceramides, and free fatty acids, and fairly efficient for extracting phosphatidylcholine
Summary
Polyunsaturated fatty acids (PUFAs) play essential roles in human physiology, and are both obtained from foods and synthesized endogenously. Dietary habits have changed in the last decades, especially in the western world where the intake ratio between polyunsaturated omega 3 (n-3) and omega 6 (n-6) fatty acids has shifted from 1:1 to 20:1 in favour of n-3. Studies revealed that this modified ratio has several health implications, influencing health outcomes through several mechanisms such as systemic inflammation, adipogenesis, browning of adipose tissue, lipid homeostasis, and the brain–gut–adipose-tissue communication [1]. The main dietary sources of DHA and ALA are marine algae and fish oil
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