Submitted on October 31, 2007
Revised on December 18, 2007
Accepted on December 18, 2007
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. Reily
Bioanalytical and Discovery Analytical Science, Bristol-Myers Squibb, Princeton, NJ 08543
Corresponding Author: michael.reily{at}bms.com
A nuclear magnetic resonance (NMR) spectroscopic method is described that enables 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 and triglycerides 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-ELSD 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 wax esters 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 wax esters, cholesterol esters, triglycerides 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 where fatty acid-independent quantification of lipid classes is desired.
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