jf5049595_si_001.pdf (354.86 kB)
Characterization of Phospholipids by Two-Dimensional Liquid Chromatography Coupled to In-line Ozonolysis–Mass Spectrometry
journal contribution
posted on 2015-02-11, 00:00 authored by Chenxing Sun, Yuan-Yuan Zhao, Jonathan M. CurtisIn this study, the characterization
of phospholipids (PL) was achieved
by using a combination of LC/MS/MS and two-dimensional LC/MS. A HILIC
LC column was used for PL class separation, while the further molecular
species separation of each PL class was achieved by using online HILIC
× C18 LC. The double bond positions along the fatty acyl chains
of these PL molecular species were also obtained by using the combination
of 2D-LC and in-line ozonolysis–MS analysis. The ozonolysis
device is composed of a gas-permeable, liquid-impermeable Teflon tube
passing through a glass chamber filled with ozone gas, which is then
placed in-line between the 2D-LC and the mass spectrometer. The eluting
PL molecules in the LC mobile phase passed through the device where
they rapidly reacted with the ozone that penetrated through the tubing
wall. The ozonolysis products were then detected by MS in real-time,
which allowed the localization of the double bonds along the fatty
acyl chains in these PL molecular species. This comprehensive method
was successfully applied to an egg yolk PL extract, which revealed
the detailed structures of the PL molecules.