No Cover Image

Journal article 136 views 24 downloads

Deep mining of oxysterols and cholestenoic acids in human plasma and cerebrospinal fluid: Quantification using isotope dilution mass spectrometry / Eylan Yutuc, Alison Dickson, Manuela Pacciarini, Lauren Griffiths, 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, Yuqin Wang

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

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

  • 56163.pdf

    PDF | Version of Record

    © 2021 The Author(s). This is an open access article under the CC BY license

    Download (5.89MB)

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...

Full description

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
Tags: Add Tag
No Tags, Be the first to tag this record!
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: Swansea University Medical School
Funders: UKRI, BB/S019588/1
Start Page: 338259