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

Eylan Yutuc Orcid Logo, Alison Dickson, Manuela Pacciarini, Lauren Griffiths Orcid Logo, Paul R.S. Baker, Lisa Connell, Anders Öhman, Lars Forsgren, Miles Trupp, Sílvia Vilarinho, Youssef Khalil, Peter T. Clayton, Sinan Sari, Buket Dalgic, Philip Höflinger, Ludger Schöls, William Griffiths Orcid Logo, Yuqin Wang Orcid Logo

Analytica Chimica Acta, Volume: 1154, Start page: 338259

Swansea University Authors: Eylan Yutuc Orcid Logo, Alison Dickson, Manuela Pacciarini, Lauren Griffiths Orcid Logo, William Griffiths Orcid Logo, Yuqin Wang Orcid Logo

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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...

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Published in: Analytica Chimica Acta
ISSN: 0003-2670
Published: Elsevier BV 2021
Online Access: Check full text

URI: https://cronfa.swan.ac.uk/Record/cronfa56163
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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.
Keywords: Cholesterol; Hydroxycholesterol; Cholestenoic acid; Bile acid; LC-MS; Derivatisation; Isotope-labelled standard
College: Faculty of Medicine, Health and Life Sciences
Funders: UKRI, BB/S019588/1
Start Page: 338259