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Developing an Enzyme-Assisted Derivatization Method for Analysis of C27 Bile Alcohols and Acids by Electrospray Ionization-Mass Spectrometry

Eylan Yutuc Orcid Logo, Jonas Abdel-Khalik, Peter J. Crick, Eylan Yutuc, Yuqin Wang, William Griffiths Orcid Logo

Molecules, Volume: 24, Issue: 3, Start page: 597

Swansea University Authors: Eylan Yutuc Orcid Logo, William Griffiths Orcid Logo

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DOI (Published version): 10.3390/molecules24030597

Abstract

Enzyme-assisted derivatization for sterol analysis (EADSA) is a technology designed to enhance sensitivity and specificity for sterol analysis using electrospray ionization–mass spectrometry. To date it has only been exploited on sterols with a 3β-hydroxy-5-ene or 3β-hydroxy-5α-hydrogen structure, u...

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Published in: Molecules
ISSN: 1420-3049
Published: 2019
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URI: https://cronfa.swan.ac.uk/Record/cronfa48910
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spelling 2019-05-17T18:27:51.9476006 v2 48910 2019-02-19 Developing an Enzyme-Assisted Derivatization Method for Analysis of C27 Bile Alcohols and Acids by Electrospray Ionization-Mass Spectrometry 99332f073ce913a9b7d8b6441b17516d 0000-0001-9971-1950 Eylan Yutuc Eylan Yutuc true false 3316b1d1b524be1831790933eed1c26e 0000-0002-4129-6616 William Griffiths William Griffiths true false 2019-02-19 BMS Enzyme-assisted derivatization for sterol analysis (EADSA) is a technology designed to enhance sensitivity and specificity for sterol analysis using electrospray ionization–mass spectrometry. To date it has only been exploited on sterols with a 3β-hydroxy-5-ene or 3β-hydroxy-5α-hydrogen structure, using bacterial cholesterol oxidase enzyme to convert the 3β-hydroxy group to a 3-oxo group for subsequent derivatization with the positively charged Girard hydrazine reagents, or on substrates with a native oxo group. Here we describe an extension of the technology by substituting 3α-hydroxysteroid dehydrogenase (3α-HSD) for cholesterol oxidase, making the method applicable to sterols with a 3α-hydroxy-5β-hydrogen structure. The 3α-HSD enzyme works efficiently on bile alcohols and bile acids with this stereochemistry. However, as found by others, derivatization of the resultant 3-oxo group with a hydrazine reagent does not go to completion in the absence of a conjugating double bond in the sterol structure. Nevertheless, Girard P derivatives of bile alcohols and C27 acids give an intense molecular ion ([M]+) upon electrospray ionization and informative fragmentation spectra. The method shows promise for analysis of bile alcohols and 3α-hydroxy-5β-C27-acids, enhancing the range of sterols that can be analyzed at high sensitivity in sterolomic studies. Journal Article Molecules 24 3 597 1420-3049 31 3 2019 2019-03-31 10.3390/molecules24030597 COLLEGE NANME Biomedical Sciences COLLEGE CODE BMS Swansea University UKRI, BB/I001735/1 2019-05-17T18:27:51.9476006 2019-02-19T15:44:29.5927529 Eylan Yutuc 0000-0001-9971-1950 1 Jonas Abdel-Khalik 2 Peter J. Crick 3 Eylan Yutuc 4 Yuqin Wang 5 William Griffiths 0000-0002-4129-6616 6 0048910-19022019154638.pdf 48910.pdf 2019-02-19T15:46:38.3470000 Output 5771119 application/pdf Version of Record true 2019-02-18T00:00:00.0000000 Released under the terms of a Creative Commons Attribution License (CC-BY). true eng 0048910-13032019142654.pdf 48910.s001.pdf 2019-03-13T14:26:54.6870000 Output 2255221 application/pdf Not Applicable (or Unknown) true 2019-03-13T00:00:00.0000000 Supplementary material true eng
title Developing an Enzyme-Assisted Derivatization Method for Analysis of C27 Bile Alcohols and Acids by Electrospray Ionization-Mass Spectrometry
spellingShingle Developing an Enzyme-Assisted Derivatization Method for Analysis of C27 Bile Alcohols and Acids by Electrospray Ionization-Mass Spectrometry
Eylan Yutuc
William Griffiths
title_short Developing an Enzyme-Assisted Derivatization Method for Analysis of C27 Bile Alcohols and Acids by Electrospray Ionization-Mass Spectrometry
title_full Developing an Enzyme-Assisted Derivatization Method for Analysis of C27 Bile Alcohols and Acids by Electrospray Ionization-Mass Spectrometry
title_fullStr Developing an Enzyme-Assisted Derivatization Method for Analysis of C27 Bile Alcohols and Acids by Electrospray Ionization-Mass Spectrometry
title_full_unstemmed Developing an Enzyme-Assisted Derivatization Method for Analysis of C27 Bile Alcohols and Acids by Electrospray Ionization-Mass Spectrometry
title_sort Developing an Enzyme-Assisted Derivatization Method for Analysis of C27 Bile Alcohols and Acids by Electrospray Ionization-Mass Spectrometry
author_id_str_mv 99332f073ce913a9b7d8b6441b17516d
3316b1d1b524be1831790933eed1c26e
author_id_fullname_str_mv 99332f073ce913a9b7d8b6441b17516d_***_Eylan Yutuc
3316b1d1b524be1831790933eed1c26e_***_William Griffiths
author Eylan Yutuc
William Griffiths
author2 Eylan Yutuc
Jonas Abdel-Khalik
Peter J. Crick
Eylan Yutuc
Yuqin Wang
William Griffiths
format Journal article
container_title Molecules
container_volume 24
container_issue 3
container_start_page 597
publishDate 2019
institution Swansea University
issn 1420-3049
doi_str_mv 10.3390/molecules24030597
document_store_str 1
active_str 0
description Enzyme-assisted derivatization for sterol analysis (EADSA) is a technology designed to enhance sensitivity and specificity for sterol analysis using electrospray ionization–mass spectrometry. To date it has only been exploited on sterols with a 3β-hydroxy-5-ene or 3β-hydroxy-5α-hydrogen structure, using bacterial cholesterol oxidase enzyme to convert the 3β-hydroxy group to a 3-oxo group for subsequent derivatization with the positively charged Girard hydrazine reagents, or on substrates with a native oxo group. Here we describe an extension of the technology by substituting 3α-hydroxysteroid dehydrogenase (3α-HSD) for cholesterol oxidase, making the method applicable to sterols with a 3α-hydroxy-5β-hydrogen structure. The 3α-HSD enzyme works efficiently on bile alcohols and bile acids with this stereochemistry. However, as found by others, derivatization of the resultant 3-oxo group with a hydrazine reagent does not go to completion in the absence of a conjugating double bond in the sterol structure. Nevertheless, Girard P derivatives of bile alcohols and C27 acids give an intense molecular ion ([M]+) upon electrospray ionization and informative fragmentation spectra. The method shows promise for analysis of bile alcohols and 3α-hydroxy-5β-C27-acids, enhancing the range of sterols that can be analyzed at high sensitivity in sterolomic studies.
published_date 2019-03-31T03:59:36Z
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score 11.013148

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