High-performance liquid chromatography/electrospray ionization ion-trap tandem mass spectrometric analysis and quantification of phosphatidylcholine molecular species in the serum of cystic fibrosis subjects supplemented with docosahexaenoic acid

Deborah Pacetti, Marco Malavolta, Fabio Bocci, Emanuele Boselli, Natale G. Frega

Research output: Contribution to journalArticle

Abstract

Since phosphatidylcholine (PC) is the most abundant phospholipid (PL) class in human serum, its concentration represents an important marker for the evaluation of lipid absorption and metabolism. High-performance liquid chromatography coupled on-line with electrospray ionization ion-trap tandem mass spectrometry (HPLC/ESI-MS/MS) was successfully applied to the quantitative analysis of PC molecular species from serum of cystic fibrosis (CF) subjects before and after supplementation with docosahexaenoic acid (DHA). Seven molecular species of PC (containing C 16:0/C 20:4, C 16:0/C 22:6, C 18:0/C 20:4, C 18:0/C 22:6, C 16:0/C 18:1, C 16:0/C 18:2 and C 18:0/C 18:2, respectively) were quantified using MS in the negative scan mode with 1,2-diundecanoyl-sn-glycero-phosphocholine as the internal standard. The molecular species containing DHA, C 16:0/C 22:6 and C 18:0/C 22:6, increased from 41.3 ± 31.7 and 33.1 ± 18.2 to 85.4 ± 20.4 and 52.1 ± 20.7 μg/mL serum, respectively, after a 3-month supplementation. Interestingly, the species containing arachidonic acid (C 18:0/C 20:4 and C 16:0/C 20:4) decreased from 115 ± 55 and 139 ± 57 to 58.1 ± 22.5 and 70.5 ± 28.1, respectively. HPLC/ ESI-MS/MS allowed the direct analysis of the lipid extract without previous purification of PLs, thus it is a useful analytical support in CF research in order to understand the extent of lipid dysfunctions typical of CF or other diseases. The present method might also be used for quantitative analysis of each serum phospholipid class molecular species. However, the instrument response was found to be very dependent on the phospholipid class considered, and thus the use of appropriate standards for each class of PLs is recommended.

Original languageEnglish
Pages (from-to)2395-2400
Number of pages6
JournalRapid Communications in Mass Spectrometry
Volume18
Issue number20
DOIs
Publication statusPublished - 2004

ASJC Scopus subject areas

  • Analytical Chemistry
  • Spectroscopy

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