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KMID : 0043320090320121795
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Archives of Pharmacal Research
2009 Volume.32 No. 12 p.1795 ~ p.1801
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Simultaneous HPLC Analysis of Ceramide and Dihydroceramide in Human Hairs
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Yoo Hwan-Soo
Lee Youn-Sun
Choi Kyeong-Mi
Choi Mi-Hwa
Ji So-Young
Yoo Jae-Myung
Lee Yong-Moon
Hong Jin-Tae
Yun Yeo-Pyo
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Abstract
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Ceramide, a major class of hair lipid, can help determine the physicochemical properties of human hairs such as the chemical diffusion barrier and water retention. In this study, we developed a quantitation method for ceramide and dihydroceramide, a saturated form of ceramide, in human hairs. Lipids were extracted with ethanol from human hairs spiked with N-oleoyl-Derythro- C17 sphingosine, an internal standard. Ceramide and dihydroceramide were resolved by TLC and deacylated by sphingolipid-ceramide deacylase to release sphingosine and dihydrosphingosine, respectively. The hair content of ceramide was measured by HPLC following derivatization with o-phthalaldehyde. The limits of detection and quantification for ceramide extracted from hair fibers were 0.1 and 1 pmol, respectively. The linear range of hair weight for determining ceramide and dihydroceramide contents was 1 to 50 mg, with R2 values of 0.9695 and 0.9898, respectively. The recoveries of ceramide and dihydroceramide from intra-day and interday assays were 99.55% to 98.53%, respectively. Concentrations of dihydroceramide from the hair roots to distal tip ends ranged from 10.42 ± 2.19 to 1.20 ± 0.11 nmol/g hair, while those of ceramide ranged from 2.27 ± 0.22 to 1.47 ± 0.15 nmol/g hair. The present analytical method provides a simultaneous and reproducible quantification of ceramide and dihydroceramide, and may be used as a potential biomarker for lipid abnormality-related diseases.
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KEYWORD
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Ceramide, Dihydroceramide, Sphingolipid, Hairs, HPLC
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