Development and optimization of a gas-chromatographic separation method of fatty acids in human serum

Abstract

In this paper, we present the development and optimization of an efficient GC-MS method for the separation of fatty acids in human serum collected from a healthy volunteer and from an adrenoleukodystrophic (ALD) patient. The method was optimized in every step, meaning derivatization, extraction of obtained fatty acid methyl-esters (FAME) and GC separation. For the separation, we used a non-polar stationary phase (DB-5ms column) and helium as carrier gas at 1 ml/min, under a temperature gradient (100-170-280°C). The method was applied for the separation and identification of 16 fatty acids and other 3 lipidic compounds. Very long chain fatty acids, such as C26:0, were identified in single ion monitoring (SIM) mode of the mass spectrometric detector.