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Deep mining of oxysterols and cholestenoic acids in human plasma and cerebrospinal fluid : quantification using isotope dilution mass spectrometry

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Date
15/04/2021
Author
Yutuc, Eylan
Dickson, Alison L
Pacciarini, Manuela
Griffiths, Lauren
Baker, Paul R S
Connell, Lisa
Öhman, Anders
Forsgren, Lars
Trupp, Miles
Vilarinho, Sílvia
Khalil, Youssef
Clayton, Peter T
Sari, Sinan
Dalgic, Buket
Höflinger, Philip
Schöls, Ludger
Griffiths, William J
Wang, Yuqin
Keywords
Cholesterol
Derivatisation
Isotope-labelled standard
Cholestenoic acid
Bile acid
LC-MS
Hydroxycholesterol
QD Chemistry
QH301 Biology
DAS
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Abstract
Both plasma and cerebrospinal fluid (CSF) are rich in cholesterol and its metabolites. Here we describe in detail a methodology for the identification and quantification of multiple sterols including oxysterols and sterol-acids found in these fluids. The method is translatable to any laboratory with access to liquid chromatography - tandem mass spectrometry. The method exploits isotope-dilution mass spectrometry for absolute quantification of target metabolites. The method is applicable for semi-quantification of other sterols for which isotope labelled surrogates are not available and approximate quantification of partially identified sterols. Values are reported for non-esterified sterols in the absence of saponification and total sterols following saponification. In this way absolute quantification data is reported for 17 sterols in the NIST SRM 1950 plasma along with semi-quantitative data for 8 additional sterols and approximate quantification for one further sterol. In a pooled (CSF) sample used for internal quality control, absolute quantification was performed on 10 sterols, semi-quantification on 9 sterols and approximate quantification on a further three partially identified sterols. The value of the method is illustrated by confirming the sterol phenotype of a patient suffering from ACOX2 deficiency, a rare disorder of bile acid biosynthesis, and in a plasma sample from a patient suffering from cerebrotendinous xanthomatosis, where cholesterol 27-hydroxylase is deficient.
Citation
Yutuc , E , Dickson , A L , Pacciarini , M , Griffiths , L , Baker , P R S , Connell , L , Öhman , A , Forsgren , L , Trupp , M , Vilarinho , S , Khalil , Y , Clayton , P T , Sari , S , Dalgic , B , Höflinger , P , Schöls , L , Griffiths , W J & Wang , Y 2021 , ' Deep mining of oxysterols and cholestenoic acids in human plasma and cerebrospinal fluid : quantification using isotope dilution mass spectrometry ' , Analytica Chimica Acta , vol. 1154 , 338259 . https://doi.org/10.1016/j.aca.2021.338259
Publication
Analytica Chimica Acta
Status
Peer reviewed
DOI
https://doi.org/10.1016/j.aca.2021.338259
ISSN
0003-2670
Type
Journal article
Rights
Copyright © 2021 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
Description
This work was supported by the UKRI via the Biotechnology and Biological Sciences Research Council (BBSRC, grant numbers BB/I001735/1, BB/N015932/1 and BB/S019588/1 to WJG, BB/L001942/1 to YW), the European Union through European Structural Funds (ESF), as part of the Welsh Government funded Academic Expertise for Business project (to WJG and YW) and the Michael J. Fox Foundation for Parkinson’s Research (Grant ID: 16231 to WJG). ALD was supported via a KESS2 award with Markes International from the Welsh Government and European Social Fund. MP and LG are supported by PhD fellowships from Swansea University.
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  • University of St Andrews Research
URI
http://hdl.handle.net/10023/26355

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