title: Profile and quantification of human stratum corneum ceramides by normal-phase liquid chromatography coupled with dynamic multiple reaction monitoring of mass spectrometry: development of targeted lipidomic method and application to human stratum corneum of different age groups.
abstract: Skin, the largest organ of the human body, serves as the primary barrier to the external environment. Ceramides are one of the main constituents of stratum corneum (SC), playing an important role in skin barrier function. Therefore, comprehensive profiling and quantification of SC ceramide is important. Herein, a new targeted lipidomic method for human SC ceramide profiling and quantification is presented and tested. Normal-phase high-performance liquid chromatography coupled with dynamic multiple reaction monitoring mass spectrometry (NP-HPLC-dMRM-MS) was used to separate ceramides into subclasses and then characterize different ceramides within each subclass on the basis of their characteristics. In total, 483 ceramides were quantified in a single run within 20 min, covering 12 subclasses as well as some glycosylated ceramides not previously reported. Each subclass had typical standard substances (if available) that served to establish representative standard curves and were used for related substances with no standards. Linearity range, limit of quantification (LOQ), limit of detection (LOD), precision, accuracy, stability, and matrix effects were validated. dMRM increased sensitivity and accuracy greatly compared with common MRM (cMRM). This method was successfully applied to the study of human SC from different age groups. A total of 193 potential biomarkers were found to indicate age differences between children and adults. This method is an innovative approach for high-throughput quantification of SC ceramide. Graphical Abstract Method establishment (MRM spectra by the established method) and method application (score scatter plots of authentic samples).
authors.affiliation: State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, No. 2 Nanwei Road, Xicheng District, Beijing, 100050, China.
authors.forename: Zhi-Xin
authors.initials: ZX
authors.lastname: Jia
authors.affiliation: State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences & Peking Union Medical College, No. 2 Nanwei Road, Xicheng District, Beijing, 100050, China. zhjl@imm.ac.cn.
authors.forename: Jin-Lan
authors.initials: JL
authors.lastname: Zhang
authors.affiliation: Beijing Children's Hospital, Capital Medical University, 56 South Lishi Road, Xicheng District, Beijing, 100045, China.
authors.forename: Chun-Ping
authors.initials: CP
authors.lastname: Shen
authors.affiliation: Beijing Children's Hospital, Capital Medical University, 56 South Lishi Road, Xicheng District, Beijing, 100045, China.
authors.forename: Lin
authors.initials: L
authors.lastname: Ma
mesh_terms: D000328:Adult; D002518:Ceramides / Q000032:analysis*; D002648:Child; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004817:Epidermis / Q000737:chemistry*; D006801:Humans; D057230:Limit of Detection; D013058:Mass Spectrometry / Q000379:methods*
chemical_list: D002518:Ceramides
keywords: Different age groups; Dynamic multiple reaction monitoring (dMRM); Lipidomics; Normal-phase high-performance liquid chromatography (NPLC); Stratum corneum (SC)