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

Eylan Yutuc, Alison L 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 J Griffiths*, Yuqin Wang

*Corresponding author for this work

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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.

Original languageEnglish
Article number338259
Number of pages16
JournalAnalytica Chimica Acta
Volume1154
Early online date18 Feb 2021
DOIs
Publication statusPublished - 15 Apr 2021

Keywords

  • Cholesterol
  • Derivatisation
  • Isotope-labelled standard
  • Cholestenoic acid
  • Bile acid
  • LC-MS
  • Hydroxycholesterol

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