Analysis of Polar Lipids in Chlamydomonas reinhardtii Using Nanoelectrospray Direct Infusion Method and Gas Chromatography and Mass Spectrometric Detection

doi:10.6023/A13010152

QIBEBT-IR > 工业生物燃气研究组

	Analysis of Polar Lipids in Chlamydomonas reinhardtii Using Nanoelectrospray Direct Infusion Method and Gas Chromatography and Mass Spectrometric Detection
	Yang Dawei 1,2,3; Fan Xiaolei1 ; Kind, Tobias 3; Friehn, Oliver 3; 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
DOI	10.6023/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
推荐引用方式 GB/T 7714	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.