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Analysis of Polar Lipids in Chlamydomonas reinhardtii Using Nanoelectrospray Direct Infusion Method and Gas Chromatography and Mass Spectrometric Detection
Yang Dawei1,2,3; Fan Xiaolei1; Kind, Tobias3; Friehn, Oliver3; Guo Rongbo1
2013-04-15
发表期刊ACTA CHIMICA SINICA
卷号71期号:4页码:663-669
摘要Recently, microalgae as feedstock for lipid-based biofuel research has evoked great concern around the world. Previous lipid analyses focused on fatty acid esters, triglyceride and total lipid content in algae, but ignored the polar lipids analysis. Here, we present a comprehensive method to cover polar lipids and free fatty acids in Chlamydomonas reinhardtii. Firstly, lipid extraction was performed by methyl-tert-butyl ether which is well suited for shotgun lipidomics method. Afterwards, we developed a fast detection method in polar lipids analysis of C. reinhardtii by using a chip-based nanoelectrospray direct infusion with ion trap mass spectrometry (Nanomate-LTQ). A data-dependent MS/MS method acquired tandem mass spectra in positive and negative mode with a range of 10 minutes infusion time. MS/MS data obtained from Nanomate-LTQ were searched against automated MS/MS database with a precursor window of 0.4 Da and a product ion search window of 0.8 Da. Hit score represents the level of confidence from the Lipidblast library search which can be achieved by free availably NIST MS Search GUI. Therefore, thirty-five lipids were annotated by means of mass precursor search and subsequent in-silico MS/MS mass spectral database (Lipidblast) search, including most polar lipid species which are most common lipid species reported for C. reinhardtii. Among them, twenty classes of diacylglyceryltrimethylhomoserine that are very special lipid species in C. reinhardtii belonging to betaine lipids were annotated with in-house Lipidblast database. In order to validate this Lipidblast match, we performed a manual spectral interpretation of every experimental MS/MS spectrum to ensure authentic ester annotations. Besides, we also applied a gas chromatography tandem mass spectrometric method to detect free fatty acids in C. reinhardtii and found that linolenic acid, linoleic acid and palmitic acid are most abundant free fatty acids in C. reinhardtii. This study provides a potential comprehensive lipidomic method for further lipid analysis in algal research.
文章类型Article
关键词Polar Lipids Chlamydomonas Reinhardtii Direct Infusion Method Gas Chromatography-mass Spectrometry Free Fatty Acids
WOS标题词Science & Technology ; Physical Sciences
DOI10.6023/A13010152
关键词[WOS]FATTY-ACID
收录类别SCI
语种英语
WOS研究方向Chemistry
WOS类目Chemistry, Multidisciplinary
WOS记录号WOS:000318258800028
引用统计
文献类型期刊论文
条目标识符http://ir.qibebt.ac.cn/handle/337004/5952
专题工业生物燃气研究组
作者单位1.Chinese Acad Sci, Key Lab Biofuel, Qingdao Inst Bioenergy & Bioproc Technol, Qingdao 266101, Peoples R China
2.Chinese Acad Sci, Grad Univ, Beijing 100039, Peoples R China
3.UC Davis Genome Ctr Metabol, Davis, CA 95616 USA
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Yang Dawei,Fan Xiaolei,Kind, Tobias,et al. Analysis of Polar Lipids in Chlamydomonas reinhardtii Using Nanoelectrospray Direct Infusion Method and Gas Chromatography and Mass Spectrometric Detection[J]. ACTA CHIMICA SINICA,2013,71(4):663-669.
APA Yang Dawei,Fan Xiaolei,Kind, Tobias,Friehn, Oliver,&Guo Rongbo.(2013).Analysis of Polar Lipids in Chlamydomonas reinhardtii Using Nanoelectrospray Direct Infusion Method and Gas Chromatography and Mass Spectrometric Detection.ACTA CHIMICA SINICA,71(4),663-669.
MLA Yang Dawei,et al."Analysis of Polar Lipids in Chlamydomonas reinhardtii Using Nanoelectrospray Direct Infusion Method and Gas Chromatography and Mass Spectrometric Detection".ACTA CHIMICA SINICA 71.4(2013):663-669.
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