College of Pharmacy and CBITRC, Chungbuk National University, Cheongju, 361-763, Korea.
Arch Pharm Res. 2009 Dec;32(12):1795-801. doi: 10.1007/s12272-009-2219-5. Epub 2010 Feb 17.
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.
神经酰胺是头发脂质的主要类别之一，它可以帮助确定人类头发的理化性质，如化学扩散屏障和保水能力。在这项研究中，我们开发了一种定量测定人类头发中神经酰胺和二氢神经酰胺（神经酰胺的饱和形式）的方法。将含有 N-油酰-D-赤式-C（17）神经鞘氨醇（内标）的人类头发用乙醇提取脂质。用薄层色谱法（TLC）分离神经酰胺和二氢神经酰胺，并用神经鞘脂酶-神经酰胺脱酰酶使它们分别释放神经醇和二氢神经醇。用邻苯二醛衍生化后，通过 HPLC 测定神经酰胺的含量。从头发纤维中提取的神经酰胺的检测限和定量限分别为 0.1 和 1 pmol。用于测定神经酰胺和二氢神经酰胺含量的头发重量线性范围为 1 至 50mg，相关系数（R(2)）分别为 0.9695 和 0.9898。日内和日间测定中神经酰胺和二氢神经酰胺的回收率分别为 99.55%至 98.53%。从发根到发梢的二氢神经酰胺浓度范围为 10.42±2.19 至 1.20±0.11 nmol/g 头发，而神经酰胺的浓度范围为 2.27±0.22 至 1.47±0.15 nmol/g 头发。本分析方法可同时且重现性地定量测定神经酰胺和二氢神经酰胺，可作为脂质异常相关疾病的潜在生物标志物。
