Development of a high-throughput method for soil fatty acid derivatization and analysis

Abstract

The Bligh and Dyer method of extraction and separation of phospholipid fatty acids (PLFAs) and neutral lipid fatty acids (NLFAs) has been extensively used in soil microbial community studies. A high throughput method of soil fatty acid analysis was developed which involves the extraction of fatty acids, separation of the fatty acids to PLFAs and NLFAs using solid-phase extraction technique, and subjecting the fatty acids to mild alkaline methanolysis prior to analysis using a GC-FID. Where possible stages were modified to be carried on a 96 well plate multi-tier unit, increasing the sample throughput and decreasing the overall consumption of solvents, cost of analysis, and time. The novel high throughput method showed comparable biomass and community characteristics to that of the standard method and thus, could be proposed as a novel method of fatty acid analysis.