Separation of core and intact polar archaeal tetraether lipids using silica columns: Insights into living and fossil biomass contributions

Abstract

Intact polar lipids (IPLs) are frequently used as biomarkers for living microbial cells and can be separated from core lipids (i.e. lipids without polar headgroups), which are mainly derived from fossil (i.e. dead) cell material, using column chromatography. We have compared the effect of various silica column conditions on the separation and recovery of archaeal glycerol dialkyl glycerol tetraether (GDGT) core lipids, glycolipids and phosphoglycolipids using authentic standards and direct analysis with various high performance liquid chromatography-mass spectrometry (HPLC-MS) techniques. The commonly used procedure to separate these compound classes using dichloromethane, acetone and methanol as eluents, respectively, did not separate core GDGTs from glyco- and phosphoglyco-GDGTs. In contrast, a recently described procedure using hexane:ethyl acetate (3:1, v:v), ethyl acetate and methanol achieved both high recovery and successful separation of core GDGTs from the other IPLs. Application of the method to a geothermally heated soil and suspended particulate matter from the North Sea showed that it separates most of the core GDGTs from the other IPLS and that considerable qualitative and quantitative differences can occur between core and IPL-GDGTs. We conclude that the method is therefore appropriate for the separation of intact archaeal IPLs and their fossil analogues.