Cajka Tomas, Fiehn Oliver
UC Davis Genome Center - Metabolomics, University of California, Davis, 451 Health Sciences Drive, Davis, 95616, CA, USA.
Biochemistry Department, Faculty of Science, King Abdulaziz University, Jeddah, 21589, Saudi Arabia.
Methods Mol Biol. 2017;1609:149-170. doi: 10.1007/978-1-4939-6996-8_14.
This protocol describes the analysis, specifically the identification, of blood plasma lipids. Plasma lipids are extracted using methyl tert-butyl ether (MTBE), methanol, and water followed by separation and data acquisition of isolated lipids using reversed-phase liquid chromatography coupled to quadrupole/time-of-flight mass spectrometry (RPLC-QTOFMS) operated in MS/MS mode. For lipid identification, acquired MS/MS spectra are converted to the mascot generic format (MGF) followed by library search using the in-silico MS/MS library LipidBlast. Using this approach, lipid classes, carbon-chain lengths, and degree of unsaturation of fatty-acid components are annotated.
本方案描述了血浆脂质的分析方法，特别是其鉴定方法。使用甲基叔丁基醚（MTBE）、甲醇和水提取血浆脂质，然后使用在MS/MS模式下运行的反相液相色谱-四极杆/飞行时间质谱（RPLC-QTOFMS）对分离出的脂质进行分离和数据采集。对于脂质鉴定，将采集到的MS/MS谱图转换为吉祥物通用格式（MGF），然后使用计算机模拟MS/MS库LipidBlast进行库检索。使用这种方法，可以注释脂质类别、碳链长度和脂肪酸成分的不饱和度。
