Extraction parameters significantly influence the quantity and the profile of PLFAs extracted from soils

Phospholipid fatty acid (PLFA) analysis is a robust method for characterizing soil microbial communities. We determined the effects of extraction solvent (chloroform vs dichloromethane) and buffer (phosphate vs citrate) on the yield and the profile of PLFAs extracted from an acidic (pH 5.5) and an alkaline (pH 8.6) soil following two 2-h sequential extractions. The yield and the profile of the PLFAs obtained separately by the first and the second 2-h extraction were compared to identify the impact of sequential extractions on the PLFA extraction efficiency. Chloroform-citrate and a 2 × 2-h extraction maximized PLFA yields in both soils. Multivariate analysis of the data showed that the choice of the extraction mixture did not significantly influence the profile of the PLFAs obtained by the first 2-h extraction, whereas it had a profound effect on the profile of the PLFAs obtained by the second 2-h extraction. Most PLFAs were extracted during the first extraction except 18:2ω6,9 and 22:0 which were almost equally extracted by the two sequential extractions. The choice of organic solvent significantly influenced the profile of the PLFAs extracted; their yield increased with chloroform with the exception of 18:2ω6,9 and 22:0 which were favored by dichloromethane. Overall, a 2 × 2-h extraction with chloroform/methanol/citrate is expected to provide maximum PLFA yields.