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Title: Evaluation of novel derivatisation reagents for the analysis of oxysterols

Graphical abstract: - Highlights: • New derivatisation reagents for LC–MS analysis of oxysterols. • New reagents based on Girard P give high ion-currents and informative LC–MS{sup n} spectra. • Permanent charge is vital for efficient MS{sup n} fragmentation. • New reagents offer greater scope for incorporation of isotope labels. - Abstract: Oxysterols are oxidised forms of cholesterol that are intermediates in the synthesis of bile acids and steroid hormones. They are also ligands to nuclear and G protein-coupled receptors. Analysis of oxysterols in biological systems is challenging due to their low abundance coupled with their lack of a strong chromophore and poor ionisation characteristics in mass spectrometry (MS). We have previously used enzyme-assisted derivatisation for sterol analysis (EADSA) to identify and quantitate oxysterols in biological samples. This technique relies on tagging sterols with the Girard P reagent to introduce a charged quaternary ammonium group. Here, we have compared several modified Girard-like reagents and show that the permanent charge is vital for efficient MS{sup n} fragmentation. However, we find that the reagent can be extended to include sites for potential stable isotope labels without a loss of performance.
Authors:
 [1] ; ;  [1] ;  [2] ;  [1] ;  [1]
  1. Institute of Mass Spectrometry, College of Medicine, Swansea University, Singleton Park, Swansea SA2 8PP (United Kingdom)
  2. College of Engineering, Swansea University, Singleton Park, Swansea SA2 8PP (United Kingdom)
Publication Date:
OSTI Identifier:
22416370
Resource Type:
Journal Article
Resource Relation:
Journal Name: Biochemical and Biophysical Research Communications; Journal Volume: 446; Journal Issue: 3; Other Information: Copyright (c) 2014 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved.; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; BILE ACIDS; CHOLESTEROL; COMPARATIVE EVALUATIONS; ESTROGENS; GAS CHROMATOGRAPHY; GENES; GTP-ASES; LIQUID COLUMN CHROMATOGRAPHY; LIVER; MASS SPECTRA; MASS SPECTROSCOPY; REAGENTS; RECEPTORS; RETINOIC ACID; SYNTHESIS; VIRUSES