Liquid chromatography-mass spectrometric analysis of lipids present in human meibomian gland secretions

Igor Butovich, Eduardo Uchiyama, Mario A. Di Pascuale, James P McCulley

Research output: Contribution to journalArticlepeer-review

73 Scopus citations

Abstract

The purpose of the study was to qualitatively characterize the major lipid species present in human meibomian gland secretions (MGS) by means of high-performance liquid chromatography with atmospheric pressure ionization mass spectrometric detection of the analytes (NP HPLC-MS). Two different NP HPLC-MS methods have been developed to analyze lipid species that were expected to be present in MGS. The first method was optimized for the analysis of relatively nonpolar lipids [wax esters (WE), di- and triacyl glycerols (DAG and TAG), cholesterol (Chl) and its esters (Chl-E), and ceramides (Cer)], while the second method was designed to separate and detect phospholipids. The major lipid species in MGS were found to be WE, Chl-E, and TAG. A minor amount of free Chi (less then 0.5% of the Chl-E fraction) was detected in MGS. No appreciable amounts of DAG and Cer were found in MGS. The second NP HPLC-MS method, capable of analyzing model mixtures of authentic phospholipids (e.g. phosphatidylglycerol, phosphatidylethanolamine, phosphatidic acid, phosphatidylinositol, phosphatidylserine, phosphatidylcholine, and sphingomyelin) in submicrogram/mL concentrations, showed little or no presence of these species in the MGS samples. These observations suggest that MGS are a major source of the nonpolar lipids of the WE and Chl-E families for the tear film lipid layer (TFLL), but not of the previously reported phospholipid components of the TFLL.

Original languageEnglish (US)
Pages (from-to)765-776
Number of pages12
JournalLipids
Volume42
Issue number8
DOIs
StatePublished - Aug 2007

Keywords

  • HPLC
  • Human meibomian gland
  • Lipids
  • Mass spectrometry
  • Tear film lipid layer

ASJC Scopus subject areas

  • Biochemistry
  • Organic Chemistry
  • Cell Biology

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