CytogeneticsRapid Resolution of Medically Relevant Polyunsaturated, Very Long-, Odd-, and Branched-Chain Fatty Acid Methyl Esters

Abstract

Abstract Fatty acids play an important role in human health and disease. Nearly 100 years ago, the Burrs demonstrated that polyunsaturated fatty acids (PUFAs) were essential for development; subsequent work uncovered the role of PUFAs as precursors to important signaling molecules such as prostaglandins, leukotrienes, and thromboxanes. Very long chain fatty acids (VLCFAs) originally sparked interest over 40 years ago for being elevated in humans with peroxisomal disorders. More recent studies have shown that VLCFAs are elevated in healthy human controls, compared to their diabetic counterparts. In a similar vein, odd chain fatty acids (OCFAs) and branched-chain fatty acids (BCFAs) of the iso and anteiso variety are higher in humans with insulin sensitivity than in insulin resistant individuals. These families of fatty acids are most frequently analyzed as their fatty acid methyl esters (FAMEs) by gas chromatography (GC). Regrettably, even recently published methods resolving BCFAs from their linear correlates have run-times in excess of an hour and, in some cases, over 100 minutes. Objective: The goal of this study was to decrease the GC run-time for the analysis of FAMEs relevant to human health and disease. Methods: Baseline resolution of FAMEs required careful combination of the right gas chromatograph, column dimensions, stationary phase, carrier gas, and temperature programming. Results: We cleanly resolved the FAMEs of all common polyunsaturated, very long-, odd-, and branched-chain fatty acids with baseline separation in only ~15 minutes – a 4-fold improvement over many existing methods. Conclusion: This method will significantly improve analysis of fatty acids in health and disease, especially in large-scale, clinical studies, by quartering the run time necessary to resolve PUFAs, VLCFAs, OCFAs, and BCFAs.