Dr Ben Jenkins
- LC-MS methods
Typical lipidomics methods incorporate a liquid–liquid extraction with LC–MS quantitation;
however, the classic sample extraction methods are not high-throughput and do not perform well
at extracting the full range of lipids especially, the relatively polar species (e.g., acyl-carnitines and
glycosphingolipids). In this manuscript, wepresent a novel sample extraction protocol, which produces
a single phase supernatant suitable for high-throughput applications that oers greater performance
in extracting lipids across the full spectrum of species. We applied this lipidomics pipeline to a
ruminant fat dose–response study to initially compare and validate the dierent extraction protocols
but also to investigate complex lipid biomarkers of ruminant fat intake (adjoining onto simple odd
chain fatty acid correlations). We have found 100 lipids species with a strong correlation with
ruminant fat intake. This novel sample extraction along with the LC–MS pipeline have shown to
be sensitive, robust and hugely informative (>450 lipids species semi-quantified): with a sample
preparation throughput of over 100 tissue samples per day and an estimated ~1000 biological fluid
samples per day. Thus, this work facilitating both the epidemiological involvement of ruminant
fat, research into odd chain lipids and also streamlining the field of lipidomics (both by sample
preparation methods and data presentation).