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J. Lipid Res.
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Originally published In Press as doi:10.1194/jlr.D700035-JLR200 on December 19, 2007

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Journal of Lipid Research, Vol. 49, 686-692, March 2008
Copyright © 2008 by American Society for Biochemistry and Molecular Biology

Methods

Quantitative evaluation of sebum lipid components with nuclear magnetic resonance

Lora C. Robosky, Kimberly Wade, Dayna Woolson, John David Baker, Matthew L. Manning, Douglas A. Gage and Michael D. Reily1,2

Pfizer Global Research and Development, Michigan Laboratories, Ann Arbor, MI 48105

Published, JLR Papers in Press, December 19, 2007.

1 Present address of M. D. Reily: Bristol-Myers Squibb, MS L14-05, Route 206 and Province Line Road, Princeton, NJ 08543-4000.

2 To whom correspondence should be addressed. e-mail: michael.reily{at}bms.com

A NMR spectroscopic method is described that enables the quantitation of specific lipid classes and components, independent of fatty acid composition. We demonstrate this method for measuring cholesterol, squalene, and pools of sterol esters, wax esters (WEs), and triglyceride (TG) components in sebum and meibum. When 600 MHz NMR equipment is used in conjunction with highly sensitive cryogenically cooled probes, this method has adequate sensitivity, and for some applications, advantages over commonly used HPLC-evaporative light-scattering detection and mass spectrometry-based approaches. This method is shown to be useful for preclinical and clinical monitoring of the efficacy of sebum-reducing agents in animals and humans. In Syrian hamsters, 3% topical flutamide and 20 mg/kg oral isotretinoin reduced sterol esters by 18.7% and 30.0%, respectively, and reduced WEs by 32.9% and 31.8%, respectively, as measured in a punch biopsy of the ear. In a 72 patient clinical methodology study, the assay delivered reproducible and noninvasive measurements of WEs, cholesteryl esters, TGs, and squalene from Sebutape® skin blots. The quantitative results of sebum analysis obtained by the NMR method correlate well with those obtained with HPLC-based approaches. This approach may be broadly applicable to cases in which fatty acid-independent quantification of lipid classes is desired.

Supplementary key words skin lipid • meibum lipid • high-performance liquid chromatography-evaporative light-scattering detection • human sebum • hamster sebum • lipid class measurement

Abbreviations: AA, arachidonic alcohol; CE, cholesteryl ester; ELSD, evaporative light-scattering detection; HSE, hamster sterol ester; TG, triglyceride; WE, wax ester


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