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Anal Chem. 2018 Apr 17;90(8):5358-5365. doi: 10.1021/acs.analchem.8b00503. Epub 2018 Mar 26.

Faster Protocol for Endogenous Fatty Acid Esters of Hydroxy Fatty Acid (FAHFA) Measurements.

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Clayton Foundation Laboratories for Peptide Biology , Salk Institute for Biological Studies , 10010 North Torrey Pines Road , La Jolla , California 92037 , United States.
Skaggs School of Pharmacy and Pharmaceutical Sciences , University of California-San Diego , 9500 Gilman Drive , La Jolla , California 92093 , United States.
Department of Clinical Sciences, Division of Experimental Vascular Research , Lund University , 221 00 Lund , Sweden.
Härröd Research , Frans Persons väg 6 , 40229 Gothenburg , Sweden.
Division of Endocrinology, Diabetes and Metabolism, Department of Medicine, Beth Israel Deaconess Medical Center , Harvard Medical School , Boston , Massachusetts 02215 , United States.


Fatty acid esters of hydroxy fatty acids (FAHFAs) are a recently discovered class of endogenous lipids with antidiabetic and anti-inflammatory activities. Interest in these lipids is due to their unique biological activites and the observation that insulin-resistant people have lower palmitic acid esters of hydroxystearic acid (PAHSA) levels, suggesting that a FAHFA deficiency may contribute to metabolic disease. Rigorous testing of this hypothesis will require the measurement of many clinical samples; however, current analytical workflows are too slow to enable samples to be analyzed quickly. Here we describe the development of a significantly faster workflow to measure FAHFAs that optimizes the fractionation and chromatography of these lipids. We can measure FAHFAs in 30 min with this new protocol versus 90 min using the older protocol with comparable performance in regioisomer detection and quantitation. We also discovered through this optimization that oleic acid esters of hydroxystearic acids (OAHSAs), another family of FAHFAs, have a much lower background signal than PAHSAs, which makes them easier to measure. Our faster workflow was able to quantify changes in PAHSAs and OAHSAs in mouse tissues and human plasma, highlighting the potential of this protocol for basic and clinical applications.

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