文献类型：期刊文献

英文题名：Improved Sphingolipidomic Approach Based on Ultra-High Performance Liquid Chromatography and Multiple Mass Spectrometries with Application to Cellular Neurotoxicity

作者：Wang, Jing-Rong[1,2];Zhang, Hongyang[1,3];Yau, Lee Fong[1];Mi, Jia-Ning[1];Lee, Stephanie[4];Lee, Kim Chung[4];Hu, Ping[3];Liu, Liang[1,2];Jiang, Zhi-Hong[1,2]

机构：[1]Macau Univ Sci & Technol, Macau Inst Appl Res Med & Hlth, State Key Lab Qual Res Chinese Med, Taipa, Macau, Peoples R China;[2]Hong Kong Baptist Univ, Sch Chinese Med, Kowloon Tong, Hong Kong, Peoples R China;[3]E China Univ Sci & Technol, Sch Chem & Mol Engn, Shanghai 200237, Peoples R China;[4]Agilent Technol Hong Kong Ltd, North Point, Hong Kong, Peoples R China

年份：2014

卷号：86

期号：12

起止页码：5688

外文期刊名：ANALYTICAL CHEMISTRY

收录：;EI(收录号：20142617872383);WOS:【SCI-EXPANDED(收录号:WOS:000337643500018)】；

基金：This work was financially supported by the Macao Science and Technology Development Fund, Macau Special Administrative Region (039/2011/A2 to Z.-H.J.).

语种：英文

外文关键词：Isotopes - Cell culture - High performance liquid chromatography - Neurodegenerative diseases - Cells

摘要：The emerging field of sphingolipidomics calls for accurate quantitative analyses of sphingolipidome. Existing analytical methods for sphingolipid (SPL) profiling often suffer from isotopic/isomeric interference, leading to the low-abundance, but biologically important SPLs being undetected. In the current study, we have developed an improved sphingolipidomic approach for reliable and sensitive quantification of up to 10 subclasses of cellular SPLs. By integratively utilizing high efficiency chromatographic separation, quadrupole time-of-flight (Q-TOO and triple quadrupole (QQQ) mass spectrometry (MS), our approach facilitated unambiguous identification of several groups of potentially important but low-abundance SPLs that are usually masked by isotopic/isomeric species and hence largely overlooked in many published methods. The methodology, which featured a modified sample preparation and optimized MS parameters, permitted the measurement of 86 individual SPLs in PC12 cells in a single run, demonstrating great potential for high throughput analysis. The improved characterization, along with increased sensitivity for low-abundance SPL species, resulted in the highest number of SPLs being quantified in a single run in PC12 cells. The improved method was fully validated and applied to a lipidomic study of PC12 cell samples with or without amyloid beta peptide (A beta) treatment, which presents a most precise and genuine sphingolipidomic profile of the PC12 cell line. The adoption of the metabolomics protocol, as described in this study, could avoid misidentification and bias in the measurement of the analytically challenging low-abundance endogenous SPLs, hence achieving informative and reliable sphingolipidomics data relevant to discovery of potential SPL biomarkers for A beta-induced neurotoxicity and neurodegenerative disease.