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Nano-liquid chromatography–tandem mass spectrometry analysis of oxysterols in brain: monitoring of cholesterol autoxidation

Kersti Karu, John Turton, Yuqin Wang Orcid Logo, William Griffiths Orcid Logo

Chemistry and Physics of Lipids, Volume: 164, Issue: 6, Pages: 411 - 424

Swansea University Authors: Yuqin Wang Orcid Logo, William Griffiths Orcid Logo

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DOI (Published version): 10.1016/j.chemphyslip.2011.04.011

Abstract

Oxysterols are present in mammalian brain at ng/g–μg/g levels while cholesterol is present at the mg/g level. This makes oxysterol analysis of brain challenging. In an effort to meet this challenge we have developed, and validated, an isolation method based on solid phase extraction and an analytica...

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Published in: Chemistry and Physics of Lipids
ISSN: 0009-3084
Published: 2011
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URI: https://cronfa.swan.ac.uk/Record/cronfa10760
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spelling 2011-10-01T00:00:00.0000000 v2 10760 2012-05-21 Nano-liquid chromatography–tandem mass spectrometry analysis of oxysterols in brain: monitoring of cholesterol autoxidation c92729b58622f9fdf6a0e7d8f4ce5081 0000-0002-3063-3066 Yuqin Wang Yuqin Wang true false 3316b1d1b524be1831790933eed1c26e 0000-0002-4129-6616 William Griffiths William Griffiths true false 2012-05-21 BMS Oxysterols are present in mammalian brain at ng/g–μg/g levels while cholesterol is present at the mg/g level. This makes oxysterol analysis of brain challenging. In an effort to meet this challenge we have developed, and validated, an isolation method based on solid phase extraction and an analytical protocol involving oxidation/derivatisation (i.e., charge-tagging) followed by nano-flow liquid chromatography (nano-LC) combined with tandem mass spectrometry utilising multi-stage fragmentation (MSn). The oxidation/derivatisation method employed improves detection limits by two orders of magnitude, while nano-LC–MSn provides separation of isomers and allows oxysterol quantification. Using this method 13 different oxysterols have been identified in rat brain including 24S-hydroxycholesterol, 24S,25-epoxycholesterol and 7α,26-dihydroxycholest-4-en-3-one. The level of 24S-hydroxycholesterol in rat brain was determined to be 20.3 ± 3.4 μg/g and quantitative estimates were made for the other oxysterols identified. The presence of a large excess of cholesterol over oxysterol in brain raises the problem of autoxidation during sterol isolation and sample preparation. Thus, in parallel to identification studies, the degree of cholesterol autoxidation occurring during sterol isolation and analysis has been evaluated with the aid of [2H7]-labelled cholesterol and cholesterol autoxidation products identified. Journal Article Chemistry and Physics of Lipids 164 6 411 424 0009-3084 31 12 2011 2011-12-31 10.1016/j.chemphyslip.2011.04.011 COLLEGE NANME Biomedical Sciences COLLEGE CODE BMS Swansea University 2011-10-01T00:00:00.0000000 2012-05-21T15:43:18.5604145 Faculty of Medicine, Health and Life Sciences Swansea University Medical School - Medicine Kersti Karu 1 John Turton 2 Yuqin Wang 0000-0002-3063-3066 3 William Griffiths 0000-0002-4129-6616 4
title Nano-liquid chromatography–tandem mass spectrometry analysis of oxysterols in brain: monitoring of cholesterol autoxidation
spellingShingle Nano-liquid chromatography–tandem mass spectrometry analysis of oxysterols in brain: monitoring of cholesterol autoxidation
Yuqin Wang
William Griffiths
title_short Nano-liquid chromatography–tandem mass spectrometry analysis of oxysterols in brain: monitoring of cholesterol autoxidation
title_full Nano-liquid chromatography–tandem mass spectrometry analysis of oxysterols in brain: monitoring of cholesterol autoxidation
title_fullStr Nano-liquid chromatography–tandem mass spectrometry analysis of oxysterols in brain: monitoring of cholesterol autoxidation
title_full_unstemmed Nano-liquid chromatography–tandem mass spectrometry analysis of oxysterols in brain: monitoring of cholesterol autoxidation
title_sort Nano-liquid chromatography–tandem mass spectrometry analysis of oxysterols in brain: monitoring of cholesterol autoxidation
author_id_str_mv c92729b58622f9fdf6a0e7d8f4ce5081
3316b1d1b524be1831790933eed1c26e
author_id_fullname_str_mv c92729b58622f9fdf6a0e7d8f4ce5081_***_Yuqin Wang
3316b1d1b524be1831790933eed1c26e_***_William Griffiths
author Yuqin Wang
William Griffiths
author2 Kersti Karu
John Turton
Yuqin Wang
William Griffiths
format Journal article
container_title Chemistry and Physics of Lipids
container_volume 164
container_issue 6
container_start_page 411
publishDate 2011
institution Swansea University
issn 0009-3084
doi_str_mv 10.1016/j.chemphyslip.2011.04.011
college_str Faculty of Medicine, Health and Life Sciences
hierarchytype
hierarchy_top_id facultyofmedicinehealthandlifesciences
hierarchy_top_title Faculty of Medicine, Health and Life Sciences
hierarchy_parent_id facultyofmedicinehealthandlifesciences
hierarchy_parent_title Faculty of Medicine, Health and Life Sciences
department_str Swansea University Medical School - Medicine{{{_:::_}}}Faculty of Medicine, Health and Life Sciences{{{_:::_}}}Swansea University Medical School - Medicine
document_store_str 0
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description Oxysterols are present in mammalian brain at ng/g–μg/g levels while cholesterol is present at the mg/g level. This makes oxysterol analysis of brain challenging. In an effort to meet this challenge we have developed, and validated, an isolation method based on solid phase extraction and an analytical protocol involving oxidation/derivatisation (i.e., charge-tagging) followed by nano-flow liquid chromatography (nano-LC) combined with tandem mass spectrometry utilising multi-stage fragmentation (MSn). The oxidation/derivatisation method employed improves detection limits by two orders of magnitude, while nano-LC–MSn provides separation of isomers and allows oxysterol quantification. Using this method 13 different oxysterols have been identified in rat brain including 24S-hydroxycholesterol, 24S,25-epoxycholesterol and 7α,26-dihydroxycholest-4-en-3-one. The level of 24S-hydroxycholesterol in rat brain was determined to be 20.3 ± 3.4 μg/g and quantitative estimates were made for the other oxysterols identified. The presence of a large excess of cholesterol over oxysterol in brain raises the problem of autoxidation during sterol isolation and sample preparation. Thus, in parallel to identification studies, the degree of cholesterol autoxidation occurring during sterol isolation and analysis has been evaluated with the aid of [2H7]-labelled cholesterol and cholesterol autoxidation products identified.
published_date 2011-12-31T03:12:14Z
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score 11.013148

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