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

doi:10.3724/SP.J.1123.2019.12009

Abstract

Abstract:

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.

Key words: lipidomics, extraction, mass spectrometry (MS), fragmentation pathway, hyperthermophiles, archaea

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.

Figures/Tables 13

References

1	Ebaid R , Wang H , Sha C , et al. J Clean Prod, 2019, 238: 117925
2	Michoud G , Jebbar M . Sci Rep, 2016, 6(1): 27289
3	Mathai J C , Sprott G D , Zeidel M L . J Biol Chem, 2001, 276(29): 27266
4	Schouten S , Hopmans E C , Damste J S S . Org Geochem, 2013, 54: 19
5	Han X L . Nat Rev Endocrinol, 2016, 12(11): 668
6	Bruegger B . Annu Rev Biochem, 2014, 83: 79
7	Paglia G , Angel P , Williams J P , et al. Anal Chem, 2015, 87(2): 1137
8	Zehethofer N , Pinto D M . Anal Chim Acta, 2008, 627(1): 62
9	Mubarak M , Shaija A , Suchithra T V , et al. Algal Res, 2015, 7: 117
10	Patel A , Mikes F , Matsakas L . Molecules, 2018, 23(7): 1562
11	Liu H W , Bai Y . Chinese Journal of Chromatography, 2017, 35(1): 86
11	刘虎威, 白玉色谱, 2017, 35(1): 86
12	Bligh E G , Dyer W J . Can J Biochem Physiol, 1959, 37(8): 911
13	Kim R , Bleijerveld O B , Gestel R A , et al. Rapid Commun Mass Sp, 2010, 22(12): 1853
14	Vitali M , Gerhard L , Kurzchalia T V , et al. J Lipid Res, 2008, 49(5): 1137
15	Leinonen A , Kuuranne T , Kotiaho T , et al. Steroids, 2004, 69(2): 101
16	Zhukov A V , Vereschagin A G . Adv Lipid Res, 1981, 18: 247
17	Ostermann A I , Muller M , Willenberg I , et al. Prostag Leukotr Ess, 2014, 91(6): 235
18	Satomi Y , Hirayama M , Kobayashi H . J Chromatogr B, 2017, 1063: 93
19	Teo C C , Chong W P K , Tan E , et al. TRAC-Trends Anal Chem, 2015, 66: 1
20	Pan X H , Yin S , Liu Y L , et al. Chinese Journal of Chromatography, 2018, 36(4): 356
20	潘小红, 殷帅, 刘玉玲, et al. 色谱, 2018, 36(4): 356
21	Afshar F H , Asgharian P , Khodaie L , et al. J Nat Prod, 2017, 12(2): e36903
22	Khan K M , Nahar L , Mannan A , et al. Nat Prod Res, 2018, 32(3): 346
23	Tulipani S , Llorach R , Urpi-Sarda M , et al. Anal Chem 85(1): 341
24	Song S Y , Bai Y , Liu H W . Chinese Journal of Chromatography, 2020, 38(1): 66
24	宋诗瑶, 白玉, 刘虎威色谱, 2020, 38(1): 66
25	Wang H Y , Liu W G , Zhang C L , et al. Journal of Earth Environment, 2017, 8(2): 176
25	王欢业, 刘卫国, 张传伦, et al. 地球环境学报, 2017, 8(2): 176
26	Zerrouk Z , Alexandre S , Lafontaine C , et al. Colloid Surface B, 2008, 63(2): 306
27	Angles E , Jaouen P , Pruvost J , et al. Algal Res, 2017, 21: 27
28	Zheng S N , Zhang Y G , Lü J , et al. Chinese Journal of Chromatography, 2017, 35(2): 169
28	郑姝宁, 张延国, 吕军, et al. 色谱, 2017, 35(2): 169
29	Reid C W , Twine S M , Reid A N . Bacterial Glycomics: Current Research. 1st ed. Wymondham: Caister Academic Press, 2012: 179
30	Birrien J L , Zeng X , Jebbar M , et al. Int J Syst Evol Microbiol, 2011, 61: 2827
31	Dunn W B , Broadhurst D , Begley P , et al. Nat Protoc 6(7): 1060
32	Marteinsson V T , Moulin P , Birrien J L , et al. Appl Environ Microbiol, 1997, 63(4): 1230
33	Lesser M P . Annu Rev Physiol, 2006, 68: 253

Extraction	Sample	Extracted lipid	Reference
GDGT: glycerol dialkyl glycerol tetraethers; MTBE: methyl tert-butyl ether; SPE: solid phase extraction.
Bligh-Dyer method	archaea, bacteria	GDGT	[25]
Folch method	Escherichia coli	Membrane lipid	[26]
MTBE method	microalgae	Fatty acid	[27, 28]
SPE method	bacteria	lipid-linked	[29]
		oligosaccharide

Extraction	Sample	Extracted lipid	Reference
GDGT: glycerol dialkyl glycerol tetraethers; MTBE: methyl tert-butyl ether; SPE: solid phase extraction.
Bligh-Dyer method	archaea, bacteria	GDGT	[25]
Folch method	Escherichia coli	Membrane lipid	[26]
MTBE method	microalgae	Fatty acid	[27, 28]
SPE method	bacteria	lipid-linked	[29]
		oligosaccharide

Parameter	Setting value
Rt: retention time; ppm: 10^-6.
Search type	Product QEX
Precursor tol	5.0 ppm
Product tol	5.0 ppm
Rt range	±0.5 min
Adducts	+H, +NH₄, +Na, +2H

Parameter	Setting value
Rt: retention time; ppm: 10^-6.
Search type	Product QEX
Precursor tol	5.0 ppm
Product tol	5.0 ppm
Rt range	±0.5 min
Adducts	+H, +NH₄, +Na, +2H

Compound name	Formula	Adduct	m/z	Mass error/ppm	Fragments (m/z)
TG(15:0/14:0/16:0)	C₄₈H₉₂O₆	[M+NH₄]⁺	782.7220	-0.92	509.4564, 523.4719, 537.4870
		[M+Na]⁺	787.6772	-1.761
TG(15:0/16:1/17:1)	C₅₁H₉₄O₆	[M+NH₄]⁺	820.7387	-1.136	561.4870, 549.4878
TG(16:1/14:0/16:1)	C₄₇H₈₈O₆	[M+NH₄]⁺	792.70681	-0.953	547.4711, 521.4559
		[M+Na]⁺		-0.877
DG(18:0/16:0)	C₃₇H₇₂O₅	[M+Na]⁺	619.5254	-2.745	341.3045, 313.2734
DG(18:0/18:0)	C₃₉H₇₆O₅	[M+NH₄]⁺	642.6023	-1.143	607.5656, 341.3050