Simultaneous HPLC analysis of ceramide and dihydroceramide in human hairs.
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-D-erythro-C(17) 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 R(2) 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.
Lee
Youn-Sun
YS
College of Pharmacy and CBITRC, Chungbuk National University, Cheongju, 361-763, Korea.
Choi
Kyeong-Mi
KM
Choi
Mi-Hwa
MH
Ji
So-Young
SY
Yoo
Jae-Myung
JM
Lee
Yong-Moon
YM
Hong
Jin-Tae
JT
Yun
Yeo-Pyo
YP
Yoo
Hwan-Soo
HS
D000375:Aging / Q000502:physiology; D015415:Biomarkers / Q000032:analysis; D002518:Ceramides / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D003640:Dealkylation; D006197:Hair / Q000737:chemistry*; D006801:Humans; D008055:Lipids / Q000737:chemistry / Q000302:isolation & purification; D012015:Reference Standards; D015203:Reproducibility of Results; D012727:Sex Characteristics; D013050:Spectrometry, Fluorescence; D013107:Sphingolipids / Q000737:chemistry
D015415:Biomarkers; D002518:Ceramides; D008055:Lipids; D013107:Sphingolipids; C109343:dihydroceramide