Development
of a comprehensive LC method for signaling lipid analysis.
(A) Extracted ion chromatograms (XICs) of free phosphate group-containing
analytes LPA, S1P, and CerP. Three different columns only differing
in the type of column hardware were tested using 0.1% HAc as an eluent
additive. “SS” (stainless steel) corresponds to YMC-Triart,
“PEEK” to YMC-Triart metal-free, and “coated”
to YMC-Accura Triart. (B) Relative signal intensities for the three
tested columns (n = 4). (C) Carryover of free phosphate
group-containing analytes in dependency of the column coating. Carryover
was calculated as the ratio between the peak area of the blank injection
after the standard mix injection and the peak area of the standard
mix injection itself and is expressed as a percentage. (D) Signal
intensities for different salt and acid concentrations using the YMC-Accura
Triart column (n = 4). (E) Carryover of LPA, S1P,
and CerP using the YMC-Accura Triart column and different salt and
acid concentrations. (F) Example of improved peak shape of LPA 17:1
after final method optimization.