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Quantitative Charge-Tags for Sterol and Oxysterol Analysis

P. J. Crick, T. William Bentley, J. Abdel-Khalik, I. Matthews, P. T. Clayton, A. A. Morris, B. W. Bigger, C. Zerbinati, L. Tritapepe, L. Iuliano, Y. Wang, W. J. Griffiths, William Griffiths Orcid Logo, Yuqin Wang Orcid Logo

Clinical Chemistry

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

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DOI (Published version): 10.1373/clinchem.2014.231332

Abstract

Background. Global sterol analysis is challenging owing to the extreme diversity of sterol natural products, the tendency of cholesterol to dominate in abundance over all other sterols, and the structural lack of a strong chromophore or readily ionized functional group. We developed a method to over...

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Published in: Clinical Chemistry
Published: 2014
URI: https://cronfa.swan.ac.uk/Record/cronfa19854
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fullrecord <?xml version="1.0"?><rfc1807><datestamp>2017-02-15T10:53:08.4067393</datestamp><bib-version>v2</bib-version><id>19854</id><entry>2015-01-05</entry><title>Quantitative Charge-Tags for Sterol and Oxysterol Analysis</title><swanseaauthors><author><sid>3316b1d1b524be1831790933eed1c26e</sid><ORCID>0000-0002-4129-6616</ORCID><firstname>William</firstname><surname>Griffiths</surname><name>William Griffiths</name><active>true</active><ethesisStudent>false</ethesisStudent></author><author><sid>c92729b58622f9fdf6a0e7d8f4ce5081</sid><ORCID>0000-0002-3063-3066</ORCID><firstname>Yuqin</firstname><surname>Wang</surname><name>Yuqin Wang</name><active>true</active><ethesisStudent>false</ethesisStudent></author></swanseaauthors><date>2015-01-05</date><deptcode>BMS</deptcode><abstract>Background. Global sterol analysis is challenging owing to the extreme diversity of sterol natural products, the tendency of cholesterol to dominate in abundance over all other sterols, and the structural lack of a strong chromophore or readily ionized functional group. We developed a method to overcome these challenges by using different isotope-labeled versions of the Girard P reagent (GP) as quantitative charge-tags for the LC-MS analysis of sterols including oxysterols. Methods. Sterols/oxysterols in plasma were extracted in ethanol containing deuterated internal standards, separated by C18 solid-phase extraction, and derivatized with GP, with or without prior oxidation of 3&#x3B2;-hydroxy to 3-oxo groups. Results. By use of different isotope-labeled GPs, it was possible to analyze in a single LC-MS analysis both sterols/oxysterols that naturally possess a 3-oxo group and those with a 3&#x3B2;-hydroxy group. Intra- and interassay CVs were &amp;#60;15%, and recoveries for representative oxysterols and cholestenoic acids were 85%&#x2013;108%. By adopting a multiplex approach to isotope labeling, we analyzed up to 4 different samples in a single run. Using plasma samples, we could demonstrate the diagnosis of inborn errors of metabolism and also the export of oxysterols from brain via the jugular vein. Conclusions. This method allows the profiling of the widest range of sterols/oxysterols in a single analytical run and can be used to identify inborn errors of cholesterol synthesis and metabolism</abstract><type>Journal Article</type><journal>Clinical Chemistry</journal><publisher/><keywords/><publishedDay>31</publishedDay><publishedMonth>12</publishedMonth><publishedYear>2014</publishedYear><publishedDate>2014-12-31</publishedDate><doi>10.1373/clinchem.2014.231332</doi><url/><notes/><college>COLLEGE NANME</college><department>Biomedical Sciences</department><CollegeCode>COLLEGE CODE</CollegeCode><DepartmentCode>BMS</DepartmentCode><institution>Swansea University</institution><apcterm/><lastEdited>2017-02-15T10:53:08.4067393</lastEdited><Created>2015-01-05T11:04:34.6264563</Created><path><level id="1">Swansea University Medical School</level><level id="2">Medicine</level></path><authors><author><firstname>P. J.</firstname><surname>Crick</surname><order>1</order></author><author><firstname>T.</firstname><surname>William Bentley</surname><order>2</order></author><author><firstname>J.</firstname><surname>Abdel-Khalik</surname><order>3</order></author><author><firstname>I.</firstname><surname>Matthews</surname><order>4</order></author><author><firstname>P. T.</firstname><surname>Clayton</surname><order>5</order></author><author><firstname>A. A.</firstname><surname>Morris</surname><order>6</order></author><author><firstname>B. W.</firstname><surname>Bigger</surname><order>7</order></author><author><firstname>C.</firstname><surname>Zerbinati</surname><order>8</order></author><author><firstname>L.</firstname><surname>Tritapepe</surname><order>9</order></author><author><firstname>L.</firstname><surname>Iuliano</surname><order>10</order></author><author><firstname>Y.</firstname><surname>Wang</surname><order>11</order></author><author><firstname>W. J.</firstname><surname>Griffiths</surname><order>12</order></author><author><firstname>William</firstname><surname>Griffiths</surname><orcid>0000-0002-4129-6616</orcid><order>13</order></author><author><firstname>Yuqin</firstname><surname>Wang</surname><orcid>0000-0002-3063-3066</orcid><order>14</order></author></authors><documents/><OutputDurs/></rfc1807>
spelling 2017-02-15T10:53:08.4067393 v2 19854 2015-01-05 Quantitative Charge-Tags for Sterol and Oxysterol Analysis 3316b1d1b524be1831790933eed1c26e 0000-0002-4129-6616 William Griffiths William Griffiths true false c92729b58622f9fdf6a0e7d8f4ce5081 0000-0002-3063-3066 Yuqin Wang Yuqin Wang true false 2015-01-05 BMS Background. Global sterol analysis is challenging owing to the extreme diversity of sterol natural products, the tendency of cholesterol to dominate in abundance over all other sterols, and the structural lack of a strong chromophore or readily ionized functional group. We developed a method to overcome these challenges by using different isotope-labeled versions of the Girard P reagent (GP) as quantitative charge-tags for the LC-MS analysis of sterols including oxysterols. Methods. Sterols/oxysterols in plasma were extracted in ethanol containing deuterated internal standards, separated by C18 solid-phase extraction, and derivatized with GP, with or without prior oxidation of 3β-hydroxy to 3-oxo groups. Results. By use of different isotope-labeled GPs, it was possible to analyze in a single LC-MS analysis both sterols/oxysterols that naturally possess a 3-oxo group and those with a 3β-hydroxy group. Intra- and interassay CVs were &#60;15%, and recoveries for representative oxysterols and cholestenoic acids were 85%–108%. By adopting a multiplex approach to isotope labeling, we analyzed up to 4 different samples in a single run. Using plasma samples, we could demonstrate the diagnosis of inborn errors of metabolism and also the export of oxysterols from brain via the jugular vein. Conclusions. This method allows the profiling of the widest range of sterols/oxysterols in a single analytical run and can be used to identify inborn errors of cholesterol synthesis and metabolism Journal Article Clinical Chemistry 31 12 2014 2014-12-31 10.1373/clinchem.2014.231332 COLLEGE NANME Biomedical Sciences COLLEGE CODE BMS Swansea University 2017-02-15T10:53:08.4067393 2015-01-05T11:04:34.6264563 Swansea University Medical School Medicine P. J. Crick 1 T. William Bentley 2 J. Abdel-Khalik 3 I. Matthews 4 P. T. Clayton 5 A. A. Morris 6 B. W. Bigger 7 C. Zerbinati 8 L. Tritapepe 9 L. Iuliano 10 Y. Wang 11 W. J. Griffiths 12 William Griffiths 0000-0002-4129-6616 13 Yuqin Wang 0000-0002-3063-3066 14
title Quantitative Charge-Tags for Sterol and Oxysterol Analysis
spellingShingle Quantitative Charge-Tags for Sterol and Oxysterol Analysis
William Griffiths
Yuqin Wang
title_short Quantitative Charge-Tags for Sterol and Oxysterol Analysis
title_full Quantitative Charge-Tags for Sterol and Oxysterol Analysis
title_fullStr Quantitative Charge-Tags for Sterol and Oxysterol Analysis
title_full_unstemmed Quantitative Charge-Tags for Sterol and Oxysterol Analysis
title_sort Quantitative Charge-Tags for Sterol and Oxysterol Analysis
author_id_str_mv 3316b1d1b524be1831790933eed1c26e
c92729b58622f9fdf6a0e7d8f4ce5081
author_id_fullname_str_mv 3316b1d1b524be1831790933eed1c26e_***_William Griffiths
c92729b58622f9fdf6a0e7d8f4ce5081_***_Yuqin Wang
author William Griffiths
Yuqin Wang
author2 P. J. Crick
T. William Bentley
J. Abdel-Khalik
I. Matthews
P. T. Clayton
A. A. Morris
B. W. Bigger
C. Zerbinati
L. Tritapepe
L. Iuliano
Y. Wang
W. J. Griffiths
William Griffiths
Yuqin Wang
format Journal article
container_title Clinical Chemistry
publishDate 2014
institution Swansea University
doi_str_mv 10.1373/clinchem.2014.231332
college_str Swansea University Medical School
hierarchytype
hierarchy_top_id swanseauniversitymedicalschool
hierarchy_top_title Swansea University Medical School
hierarchy_parent_id swanseauniversitymedicalschool
hierarchy_parent_title Swansea University Medical School
department_str Medicine{{{_:::_}}}Swansea University Medical School{{{_:::_}}}Medicine
document_store_str 0
active_str 0
description Background. Global sterol analysis is challenging owing to the extreme diversity of sterol natural products, the tendency of cholesterol to dominate in abundance over all other sterols, and the structural lack of a strong chromophore or readily ionized functional group. We developed a method to overcome these challenges by using different isotope-labeled versions of the Girard P reagent (GP) as quantitative charge-tags for the LC-MS analysis of sterols including oxysterols. Methods. Sterols/oxysterols in plasma were extracted in ethanol containing deuterated internal standards, separated by C18 solid-phase extraction, and derivatized with GP, with or without prior oxidation of 3β-hydroxy to 3-oxo groups. Results. By use of different isotope-labeled GPs, it was possible to analyze in a single LC-MS analysis both sterols/oxysterols that naturally possess a 3-oxo group and those with a 3β-hydroxy group. Intra- and interassay CVs were &#60;15%, and recoveries for representative oxysterols and cholestenoic acids were 85%–108%. By adopting a multiplex approach to isotope labeling, we analyzed up to 4 different samples in a single run. Using plasma samples, we could demonstrate the diagnosis of inborn errors of metabolism and also the export of oxysterols from brain via the jugular vein. Conclusions. This method allows the profiling of the widest range of sterols/oxysterols in a single analytical run and can be used to identify inborn errors of cholesterol synthesis and metabolism
published_date 2014-12-31T03:30:04Z
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