Chemical markers in marine food web: A simple workflow based on methyl tert-butyl ether extraction for fatty acids and stable isotopes assessment in plankton samples

Fatty acids (FAs) are used, often in combination with stable isotopes (SIs), as chemical biomarkers to assess the contribution of different prey to the diet of consumers and define food web structure and dynamics. Extraction of lipids is traditionally carried out using methanol (MeOH) combined with chloroform or dichloromethane, these latter being well-known environmental pollutant and potential carcinogenic agents. Recently, extraction protocols based on methyl tert-butyl ether (MTBE) and MeOH have been proposed as an alternative to halogenated solvents in lipidomic studies. However, no specific investigation has been performed to assess MTBE suitability in marine ecological studies including FA analysis together with SI measurements. We used an analytical workflow for qualitative and quantitative analysis of FAs and SIs in field samples of phytoplankton, zooplankton and the scyphomedusa Pelagia noctiluca, applying MTBE in comparison with chloroform- and dichloromethane-based protocols for total lipid extraction. Our analysis suggested that MTBE is a reliable substitute for lipid extraction in trophic ecology studies in marine planktonic organisms.