Solvent-based washing removes lipophilic contaminant interference with phospholipid fatty acid analysis of soil communities

Phospholipid fatty acid (PLFA) analysis is an informative method for characterising and quantifying changes in the phenotypic profile of the soil microbial community when soils are exposed to chemical toxicants and other xenobiotics. However, where such materials are lipophilic, a range of non-polar compounds can be co-extracted with PLFAs and can consequently mask PLFA chromatograms. We found this to be the case with the lipophilic anti-microbial compound triclosan, which can enter the soil via the addition of sewage sludge. A simple method of washing soil in solvent prior to extraction was developed in order to remove triclosan without altering the relative abundance of PLFAs. Three contrasting soils were spiked with 500 mg kg−1 of triclosan before being washed with methanol (MeOH), dichloromethane (DCM), hexane or aqueous solutions of these solvents. PLFAs were then extracted and analysed. All treatments were found to remove triclosan effectively, allowing all peaks to be identifiable. Whilst the polar solvents MeOH and DCM significantly altered the relative abundance of extracted fatty acids in most of the soils tested, soil washing with a small quantity of hexane was able to remove triclosan whilst best preserving the fidelity of the PLFA profiles.

► A method for removing pollutants from soil prior to PLFA analysis is described.
► Triclosan is used as an example pollutant, but the method is generally applicable.
► All treatments removed triclosan effectively but some also altered PLFA abundance.
► Washing with hexane removed triclosan and preserved the fidelity of PLFA profiles.