人体脸部皮肤角质层中神经酰胺提取及含量测定  

Extraction and Determination of Ceramide in Human Facial Cuticle

作　　者：粟雨桑 何海鸥 明新林 周立武 Su Yusang;He Haiou;Ming Xinlin;Zhou Liwu(Zhuhai Easycare Technology Co.,Ltd.,Zhuhai 519000,China)

机构地区：[1]伊斯佳科技股份有限公司,广东珠海519000

出　　处：《广东化工》2020年第9期58-60,共3页Guangdong Chemical Industry

摘　　要：建立人体脸部角质层中神经酰胺的提取及含量测定方法。本研究采用透明聚酯胶带对脸部皮肤角质层进行采样,采样面积为4.5cm^2;在角质层神经酰胺的提取上,选择甲醇作为溶解剂,角质细胞采用超声波细胞破碎仪进行破碎,使神经酰胺充分释放,确定提取时间为20min。本研究采用了高效液相色谱(HPLC)测定脸部皮肤角质层神经酰胺,色谱条件为ZORBAX系列SB-C18柱(4.6 mm×250 mm,5μm),流动相为甲醇/异丙醇(70/30),流速为1.0 mL/min,柱温为30℃,检测波长为205 nm,进样量10μL。结果标准品神经酰胺mix浓度在5~200 mg/L范围内与峰面积呈良好线性关系,相关系数R^2为0.99968(n=5),检出限为2.5 mg/L。该法对皮肤角质层神经酰胺测定的加标回收率为100.99%,相对标准偏差(RSD)分别为1.55%。此法可快速高效的对脸部皮肤角质层神经酰胺定量分析,得出测试者神经酰胺的含量为3.601μg/cm^2。结论本文将神经酰胺的高效液相色谱检测方法,应用到人体脸部皮肤角质层神经酰胺的检测中。从皮肤角质层的采样、角质层神经酰胺的提取及高效液相色谱检测的条件上等作出了优化,从而达到取更少的样本量亦能简便高效的检测。为皮肤无创检测特定物质提供了科学方法。To establish a method for the extraction and determination of ceramide in human facial cuticle.In this study,transparent polyester tape was used to sample the cuticle of facial skin.The sampling area is 4.5 cm^2.Extraction of ceramide from stratum corneum,selection of Methanol as solvent.The keratinocytes were broken by ultrasonic cell breaker,determine the extraction time for 20 minutes to full release of ceramide.A high performance liquid chromatography(HPLC)method was developed for the determination of ceramide in human skin keratinocytes.The chromatographic conditions are ZORBAX SB-C18 column(4.6 mm×250 mm,5μm),the mobile phase was methanol and isopropanol solution,the flow rate was 1.0 mL/min,the column temperature was 30,the detection wavelength was 205 nm and the injection volume was 10μL.Result Ceramide mix had a good linear relationship in the range of 0.005~0.200 g/L with the correlation coefficient of 0.99968(n=5).The limit of detection was 2.5 mg/L.The recovery rates of ceramide in human shin keratinocytes was 100.99%.The relative standard deviation(RSD)was 1.55%,The method is rapid and accurate for quantitative analysis of ceramide in the sample,the content of ceramide was 3.601μg/cm^2.Conclusion In this study,the determination of ceramide by high performance liquid chromatography was applied to the detection of ceramide in human facial cuticle.The conditions of the sampling of cuticle,the extraction of ceramide and the detection of HPLC were optimized.Therefore,it is easy and efficient to take fewer samples.It provides a scientific method for non-invasive detection of specific substances in skin.

关 键 词：脸部皮肤角质层 高效液相色谱法 神经酰胺 

分 类 号：TQ[化学工程]