Profiling analysis of phospholipid fatty acids in serum as a complement to the comprehensive fatty acids method

Abstract

Fatty acids (FAs) are mostly found in blood as triglycerides, phospholipids (PLs) and cholesteryl esters. Determination of FAs is typically carried out in serum or plasma by a comprehensive method (known as the classical FAMEs method since FAs are determined as Fatty Acids Methyl Esters), which is based on liquid-liquid extraction, derivatization by transesterification, and determination by gas chromatography (GC) coupled to a suited detection technique. However, this method does not favor the determination of FAs that are chemically conjugated in PLs due to kinetics impediment. For this reason, we have developed a selective method to determine the FAs profile of PLs in serum based on solid-phase extraction (SPE) for isolation of PLs and determination of the FAME derivatives by GC-mass spectrometry (GC-MS). The method was applied to serum samples collected from twenty-five individuals to compare the FAs profile versus that provided by the non-selective protocol based on liquid-liquid extraction of lipid families. Statistical analysis revealed compositional changes in the FAs profile with special emphasis on the content of saturated (SFAs) and monounsaturated FAs (MUFAs). Thus, SFAs passed from 34.0% with the classical method to 49.3% in PLs while MUFAs went from 24.4% to 11.4%. This study proves that the proposed method provides complementary results to the comprehensive method and, therefore, both methods can be combined to evaluate the effect of intervention diets and their connection to metabolic diseases.

Keywords: FAMEs, solid-phase extraction; Fatty acids; Gas chromatography; Mass spectrometry; Phospholipids.