Identification of unusual oxysterols and bile acids with 7-oxo or 3β,5α,6β-trihydroxy functions in human plasma by charge-tagging mass spectrometry with multistage fragmentation
7-Oxocholesterol (7-OC), 5,6-epoxycholesterol (5,6-EC), and its hydrolysis product cholestane-3β,5α,6β-triol (3β,5α,6β-triol) are normally minor oxysterols in human samples; however, in disease, their levels may be greatly elevated. This is the case in plasma from patients suffering from some lysoso...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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The American Society for Biochemistry and Molecular Biology
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5983402/ https://www.ncbi.nlm.nih.gov/pubmed/29626102 http://dx.doi.org/10.1194/jlr.D083246 |
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author | Griffiths, William J. Gilmore, Ian Yutuc, Eylan Abdel-Khalik, Jonas Crick, Peter J. Hearn, Thomas Dickson, Alison Bigger, Brian W. Wu, Teresa Hoi-Yee Goenka, Anu Ghosh, Arunabha Jones, Simon A. Wang, Yuqin |
author_facet | Griffiths, William J. Gilmore, Ian Yutuc, Eylan Abdel-Khalik, Jonas Crick, Peter J. Hearn, Thomas Dickson, Alison Bigger, Brian W. Wu, Teresa Hoi-Yee Goenka, Anu Ghosh, Arunabha Jones, Simon A. Wang, Yuqin |
author_sort | Griffiths, William J. |
collection | PubMed |
description | 7-Oxocholesterol (7-OC), 5,6-epoxycholesterol (5,6-EC), and its hydrolysis product cholestane-3β,5α,6β-triol (3β,5α,6β-triol) are normally minor oxysterols in human samples; however, in disease, their levels may be greatly elevated. This is the case in plasma from patients suffering from some lysosomal storage disorders, e.g., Niemann-Pick disease type C, or the inborn errors of sterol metabolism, e.g., Smith-Lemli-Opitz syndrome and cerebrotendinous xanthomatosis. A complication in the analysis of 7-OC and 5,6-EC is that they can also be formed ex vivo from cholesterol during sample handling in air, causing confusion with molecules formed in vivo. When formed endogenously, 7-OC, 5,6-EC, and 3β,5α,6β-triol can be converted to bile acids. Here, we describe methodology based on chemical derivatization and LC/MS with multistage fragmentation (MS(n)) to identify the necessary intermediates in the conversion of 7-OC to 3β-hydroxy-7-oxochol-5-enoic acid and 5,6-EC and 3β,5α,6β-triol to 3β,5α,6β-trihydroxycholanoic acid. Identification of intermediate metabolites is facilitated by their unusual MS(n) fragmentation patterns. Semiquantitative measurements are possible, but absolute values await the synthesis of isotope-labeled standards. |
format | Online Article Text |
id | pubmed-5983402 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | The American Society for Biochemistry and Molecular Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-59834022018-06-04 Identification of unusual oxysterols and bile acids with 7-oxo or 3β,5α,6β-trihydroxy functions in human plasma by charge-tagging mass spectrometry with multistage fragmentation Griffiths, William J. Gilmore, Ian Yutuc, Eylan Abdel-Khalik, Jonas Crick, Peter J. Hearn, Thomas Dickson, Alison Bigger, Brian W. Wu, Teresa Hoi-Yee Goenka, Anu Ghosh, Arunabha Jones, Simon A. Wang, Yuqin J Lipid Res Methods 7-Oxocholesterol (7-OC), 5,6-epoxycholesterol (5,6-EC), and its hydrolysis product cholestane-3β,5α,6β-triol (3β,5α,6β-triol) are normally minor oxysterols in human samples; however, in disease, their levels may be greatly elevated. This is the case in plasma from patients suffering from some lysosomal storage disorders, e.g., Niemann-Pick disease type C, or the inborn errors of sterol metabolism, e.g., Smith-Lemli-Opitz syndrome and cerebrotendinous xanthomatosis. A complication in the analysis of 7-OC and 5,6-EC is that they can also be formed ex vivo from cholesterol during sample handling in air, causing confusion with molecules formed in vivo. When formed endogenously, 7-OC, 5,6-EC, and 3β,5α,6β-triol can be converted to bile acids. Here, we describe methodology based on chemical derivatization and LC/MS with multistage fragmentation (MS(n)) to identify the necessary intermediates in the conversion of 7-OC to 3β-hydroxy-7-oxochol-5-enoic acid and 5,6-EC and 3β,5α,6β-triol to 3β,5α,6β-trihydroxycholanoic acid. Identification of intermediate metabolites is facilitated by their unusual MS(n) fragmentation patterns. Semiquantitative measurements are possible, but absolute values await the synthesis of isotope-labeled standards. The American Society for Biochemistry and Molecular Biology 2018-06 2018-04-06 /pmc/articles/PMC5983402/ /pubmed/29626102 http://dx.doi.org/10.1194/jlr.D083246 Text en Copyright © 2018 Griffiths et al. Published under exclusive license by The American Society for Biochemistry and Molecular Biology, Inc. http://creativecommons.org/licenses/by/4.0/ Author’s Choice—Final version open access under the terms of the Creative Commons CC-BY license. |
spellingShingle | Methods Griffiths, William J. Gilmore, Ian Yutuc, Eylan Abdel-Khalik, Jonas Crick, Peter J. Hearn, Thomas Dickson, Alison Bigger, Brian W. Wu, Teresa Hoi-Yee Goenka, Anu Ghosh, Arunabha Jones, Simon A. Wang, Yuqin Identification of unusual oxysterols and bile acids with 7-oxo or 3β,5α,6β-trihydroxy functions in human plasma by charge-tagging mass spectrometry with multistage fragmentation |
title | Identification of unusual oxysterols and bile acids with 7-oxo or 3β,5α,6β-trihydroxy functions in human plasma by charge-tagging mass spectrometry with multistage fragmentation |
title_full | Identification of unusual oxysterols and bile acids with 7-oxo or 3β,5α,6β-trihydroxy functions in human plasma by charge-tagging mass spectrometry with multistage fragmentation |
title_fullStr | Identification of unusual oxysterols and bile acids with 7-oxo or 3β,5α,6β-trihydroxy functions in human plasma by charge-tagging mass spectrometry with multistage fragmentation |
title_full_unstemmed | Identification of unusual oxysterols and bile acids with 7-oxo or 3β,5α,6β-trihydroxy functions in human plasma by charge-tagging mass spectrometry with multistage fragmentation |
title_short | Identification of unusual oxysterols and bile acids with 7-oxo or 3β,5α,6β-trihydroxy functions in human plasma by charge-tagging mass spectrometry with multistage fragmentation |
title_sort | identification of unusual oxysterols and bile acids with 7-oxo or 3β,5α,6β-trihydroxy functions in human plasma by charge-tagging mass spectrometry with multistage fragmentation |
topic | Methods |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5983402/ https://www.ncbi.nlm.nih.gov/pubmed/29626102 http://dx.doi.org/10.1194/jlr.D083246 |
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