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学科主题: 物理化学
题名: Large-scaled human serum sphingolipid profiling by using reversed-phase liquid chromatography coupled with dynamic multiple reaction monitoring of mass spectrometry: Method development and application in hepatocellular carcinoma
作者: Li, Jia1;  Hu, Chunxiu1;  Zhao, Xinjie1;  Dai, Weidong1;  Chen, Shili1;  Lu, Xin1;  Xu, Guowang1
通讯作者: 许国旺
关键词: Sphingolipid profiling ;  Lipidomics ;  Liquid chromatography-mass spectrometry ;  Multiple reaction monitoring ;  Tert-butyl methyl ether ;  Serum
刊名: JOURNAL OF CHROMATOGRAPHY A
发表日期: 2013-12-13
DOI: 10.1016/j.chroma.2013.10.064
卷: 1320, 页:103-110
收录类别: SCI
合作性质: 
文章类型: Article
部门归属: 18
项目归属: 1808
产权排名: 待补充
WOS标题词: Science & Technology ;  Life Sciences & Biomedicine ;  Physical Sciences
类目[WOS]: Biochemical Research Methods ;  Chemistry, Analytical
资助者: 1,1
研究领域[WOS]: Biochemistry & Molecular Biology ;  Chemistry
英文摘要: Sphingolipids are a family of bioactive molecules with high structural diversity and complexity. They not only serve as integral components of cellular membrane, but also play pivotal roles in signaling and other cellular events. It is desirable for the development of sensitive, robust and structural-specific analytical approaches enabling rapid determination of as many sphingolipid species as possible. Herein we present an analytical method for large-scaled profiling of sphigolipids in human serum, which consisted of an improved extraction protocol using tert-butyl methyl ether combined with mild alkaline hydrolysis, and an ultra high performance reversed-phase liquid chromatography-dynamic multiple reaction monitoring-mass spectrometric (RPLC-dynamic MRM-MS) method. In total 84 endogenous sphingolipid species covering six subcategories (i.e. free sphingoid base, dihydroceramide, ceramide, hexosylceramide, lactosylceramide, and sphingomyelin), were separated and quantified in a single run within 10 min. A broad linear range over 2.5-4 orders of magnitude (r(2)>0.99), a limit of detection of 0.01-0.17 pmol/mL, and a limit of quantitation of 0.02-0.42 pmol/mL were obtained for each subcategory. Average recovery of each subcategory was within 85.6-95.6%. Median values of coefficient of variation (CV) of all detected 84 sphingolipids were 3.9% and 6.8% for intraday and interday precision, respectively. This method was exemplarily applied in a study regarding dysregulated sphingolipid homeostasis in hepatocellular carcinoma. The establishment of this method provides a useful tool for serum-based high throughput screening of sphingolipid biomarkers and mechanism investigation of sphingolipid metabolic regulation in human disease. (C) 2013 Elsevier B.V. All rights reserved.
关键词[WOS]: SIMULTANEOUS QUANTITATIVE-ANALYSIS ;  TERT-BUTYL ETHER ;  CELLULAR-EXTRACTS ;  LIPID EXTRACTION ;  HIGH-THROUGHPUT ;  HUMAN PLASMA ;  CERAMIDE ;  METABOLISM ;  CANCER ;  QUANTIFICATION
语种: 英语
原文出处: 查看原文
WOS记录号: WOS:000327922100012
Citation statistics: 
内容类型: 期刊论文
URI标识: http://cas-ir.dicp.ac.cn/handle/321008/119653
Appears in Collections:中国科学院大连化学物理研究所_期刊论文

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作者单位: 1.Chinese Acad Sci, Dalian Inst Chem Phys, CAS Key Lab Separat Sci Analyt Chem, Dalian 116023, Peoples R China

Recommended Citation:
Li, Jia,Hu, Chunxiu,Zhao, Xinjie,et al. Large-scaled human serum sphingolipid profiling by using reversed-phase liquid chromatography coupled with dynamic multiple reaction monitoring of mass spectrometry: Method development and application in hepatocellular carcinoma[J]. JOURNAL OF CHROMATOGRAPHY A,2013,1320:103-110.
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