Analysis of phospholipids in microalga Nitzschia closterium by UPLC-Q-TOF-MS |
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Authors: | Xiaojun Yan Haiying Li Jilin Xu Chengxu Zhou |
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Institution: | (1) Spence Biochemical Genetics Unit, Department of Child Health, Sir James Spence Institute of Child Health, University of Newcastle upon Tyne, Royal Victoria Infirmary, Newcastle upon Tyne, NE1 4LP, UK |
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Abstract: | Precise structural identification of phospholipids in the microalga Nitzschia closterium has been established using ultra performance liquid chromatography-electrospray ionization-quadrupole-time of flight-mass
spectrometry (UPLC-ESI-Q-TOF-MS) for direct analysis of total lipid extracts. Mass spectrometry was performed in reflective
time-of-flight using electron spraying ionization in negative mode. Phospholipid molecular species identification was based
on the characteristic product ions and neutral loss yielded by different phospholipids under ESI-MS/MS mode. The molecular
species were confirmed by the carboxylate anions produced by phospholipids in negative mode; the regiospecificity of the two
acyl chains was determined from the ratio of sn-1 to sn-2 carboxylate anion abundances. As a result, 18 lipid molecular species were identified for the first time in this microalga,
comprising seven phosphatidylcholines (PC), two phosphatidylethanolamines (PE), two phosphatidylinositols (PI), and seven
phosphatidylglycerols (PG). Lipid standards of PC, PE, PI, and PG were added to the total lipids as internal standards for
semiquantitative analysis, revealing concentrations of phospholipids in this species between 0.09 and 3.37 nmol/mg. This method
can produce a full structural profile of intact phospholipid molecular species and can be used for study of the physiological
and ecological functions of lipids by monitoring their individual changes over time. |
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