Citation: WANG Xiaoxue, HE Zhi'an, LI Xin, SONG Qinghao, ZOU Xinwei, SONG Xueyao, FENG Lei. Comparison of pretreatment methods in lipid analysis and ultra-performance liquid chromatography-mass spectrometry analysis of archaea[J]. Chinese Journal of Chromatography, ;2020, 38(8): 914-922. doi: 10.3724/SP.J.1123.2019.12009 shu

Comparison of pretreatment methods in lipid analysis and ultra-performance liquid chromatography-mass spectrometry analysis of archaea

WANG Xiaoxue ^1,2 ,
HE Zhi'an ^1,2 ,
LI Xin ² ,
SONG Qinghao ³ ,
ZOU Xinwei ^2,4 ,
SONG Xueyao ^2,4 ,
FENG Lei ^1,2,,

1.
School of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, China;
2.
Instrumental Analysis Center, Shanghai Jiao Tong University, Shanghai 200240, China;
3.
State Key Laboratory of Microbial Metabolism, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China;
4.
School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China

Corresponding author: FENG Lei, fiona.fenglei@sjtu.edu.cn
Received Date: 4 December 2019

Fund Project: Youth Program of National Natural Science Foundation of China (No. 21106082)

Archaea are single-cell microorganisms, structurally and biochemically similar to bacteria and fungi. Most of them live in extreme environments, such as high salt, extremely acidic, extremely hot, and anaerobicenvironments. The membrane structure and related metabolic pathways of archaea are different from those of other microorganisms. Therefore, studying the lipid metabolism of archaea is of great significance for exploring the life activities in extreme environments. As the first step in lipidomic analysis, lipid extraction and pretreatment methods play an important role, as they influence the accuracy and reliability of the final results. We harnessed ultra-performance liquid chromatography coupled with high-resolution mass spectrometry (UPLC-HRMS) to detect the total normal lipids. The hyperthermophilic archaeon Pyrococcus yayanosii was selected as the model. The Bligh-Dyer acidic method, Folch method, methyl tert-butyl ether (MTBE) method, and solid-phase extraction (SPE) method were compared by multi-component analysis in terms of extraction efficiency, reproducibility, and extraction discrimination. Comprehensive analysis revealed that the SPE and MTBE methods showed the best extraction repeatability and extraction efficiency, and were suitable for high-throughput microbial lipid extraction. Finally, normal lipid components of P. yayanosii were comprehensively analyzed by SPE coupled with UPLC-HRMS. A total of 1402 lipid components were identified. This article aims to provide a reference for non-targeted lipidomic analysis of archaea and other microorganisms towards understanding their lipid metabolism.
- lipidomics,
- extraction,
- mass spectrometry (MS),
- fragmentation pathway,
- hyperthermophiles,
- archaea
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