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Journal of Lipid Research, Vol. 50, 501-513, March 2009
Copyright © 2009 by American Society for Biochemistry and Molecular Biology

Cholesteryl esters as a depot for very long chain fatty acids in human meibum^*

Igor A. Butovich¹

Department of Ophthalmology and Graduate School of Biomedical Sciences, University of Texas Southwestern Medical Center, Dallas, TX 75390

^* The author acknowledges support from Research to Prevent Blindness (New York, NY) and from the National Institutes of Health in the form of unrestricted core grant EY-016664.

Published, JLR Papers in Press, October 4, 2008.

¹ To whom correspondence should be addressed. e-mail: igor.butovich{at}utsouthwestern.edu

Human meibomian gland secretions (also known as meibum) were analyzed for the presence of cholesteryl esters (Chl-E) using HPLC in combination with atmospheric pressure chemical ionization mass spectrometry. A special procedure based on detection of the in-source generated ion m/z 369 was developed to monitor all Chl-E simultaneously. The structures of the detected compounds were studied using in-source and postsource fragmentation of the precursor (M+H)⁺ ions. In concordance with previous studies, Chl-E were found in all of the tested samples and comprised 31% of the entire lipid pool (w/w, dry weight). There were at least 20 different saturated and unsaturated Chl-E species observed, whose fatty acid residues ranged from C18 to C34. Monounsaturated fatty acids were the most visible components of the Chl-E pool. The eleven most prominent compounds were: C20:0-, C22:1-, C22:0-, C24:1-, C24:0, C25:0-, C26:1-, C26:0-, C28:1-, C28:0-, and C30:1-Chl-E. Other Chl-containing compounds were detected but not identified at the time. Therefore, Chl-E are a depot for very long chain saturated and monounsaturated fatty acids in human meibum.

Supplementary key words lipids • meibomian glands • HPLC • mass spectrometry

Abbreviations: APCI, atmospheric pressure chemical ionization; Chl-E, cholesteryl ester; CID, collision-induced dissociation; LCFA, long-chain fatty acid; MS, mass spectrometry; NP, normal phase; RP, reversed-phase; RT, retention time; SIM, single ion monitoring; SRM, single reaction monitoring; TFLL, tear film lipid layer; TIC, total ion chromatogram; VLCFA, very long chain fatty acid; WE, wax ester

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