A high-throughput method for profiling fatty acids in plant seeds based on one-step acid-catalyzed methylation followed by gas chromatography-mass spectrometry

Abstract

Fatty acids (FAs) in oilseeds are important for human health. Gas chromatography-mass spectrometry (GC-MS) is a popular analytical technology for FAs measurement. FAs need to be derivatized into fatty acid methyl esters (FAMEs), which is essential for GC-MS analysis. The previous methods for methylation of FAs usually include reaction at high temperature and need specific vessels, for which samples with a large scale cannot be analyzed in a short time, leading to low efficiency. Here we report a high-throughput method for profiling fatty acids in plant seeds based on one-step acid-catalyzed methylation followed by GC-MS. This method for FA methylation via derivatization by sulfuric acid with methanol (5%, v/v) and incubation at 64 °C for 4 h in a more convenient 2 mL tube, is efficient. Satisfactory methodological performance was achieved by optimization of the parameters. Only 5 mg seeds were needed as a microscale sample usage by this method. The method is a successful approach to increase the efficiency of sample preparation and analysis for large-scale experiments.

Keywords

• fatty acids
• acid-catalyzed methylation
• gas chromatography-mass spectrometry
• plant seeds