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High-throughput quantification of carboxymethyl lysine in serum and plasma using high-resolution accurate mass Orbitrap mass spectrometry.

Authors
  • Rankin, Naomi J1, 2
  • Burgess, Karl3
  • Weidt, Stefan2
  • Wannamethee, Goya4
  • Sattar, Naveed1
  • Welsh, Paul1
  • 1 1 Institute of Cardiovascular and Medical Sciences (ICAMS), BHF Glasgow Cardiovascular Research Centre, University of Glasgow, Glasgow, UK.
  • 2 2 Glasgow Polyomics, University of Glasgow, Glasgow, UK.
  • 3 3 Institute of Quantitive Biology, Biochemistry and Biotechnology, University of Edinburgh, Edinburgh, UK.
  • 4 4 Primary Care and Population Health, Royal Free Campus, University College London Medical School, London, UK.
Type
Published Article
Journal
Annals of Clinical Biochemistry International Journal of Laboratory Medicine
Publisher
SAGE Publications
Publication Date
May 01, 2019
Volume
56
Issue
3
Pages
397–407
Identifiers
DOI: 10.1177/0004563219830432
PMID: 30832481
Source
Medline
Keywords
Language
English
License
Unknown

Abstract

Carboxymethyl lysine is an advanced glycation end product of interest as a potential biomarker of cardiovascular and other diseases. Available methods involve ELISA, with potential interference, or isotope dilution mass spectrometry (IDMS), with low-throughput sample preparation. A high-throughput sample preparation method based on 96-well plates was developed. Protein-bound carboxymethyl lysine and lysine were quantified by IDMS using reversed phase chromatography coupled to a high-resolution accurate mass Orbitrap Exactive mass spectrometer. The carboxymethyl lysine concentration (normalized to lysine concentration) was measured in 1714 plasma samples from the British Regional Heart Study (BRHS). For carboxymethyl lysine, the lower limit of quantification (LLOQ) was estimated at 0.16 μM and the assay was linear between 0.25 and 10 μM. For lysine, the LLOQ was estimated at 3.79 mM, and the assay was linear between 2.5 and 100 mM. The intra-assay coefficient of variation was 17.2% for carboxymethyl lysine, 9.3% for lysine and 10.5% for normalized carboxymethyl lysine. The inter-assay coefficient of variation was 18.1% for carboxymethyl lysine, 14.8 for lysine and 16.2% for normalized carboxymethyl lysine. The median and inter-quartile range of all study samples in each batch were monitored. A mean carboxymethyl lysine concentration of 2.7 μM (IQR 2.0-3.2 μM, range 0.2-17.4 μM) and a mean normalized carboxymethyl lysine concentration of 69 μM/M lysine (IQR 54-76 μM/M, range 19-453 μM/M) were measured in the BRHS. This high-throughput sample preparation method makes it possible to analyse large cohorts required to determine the potential of carboxymethyl lysine as a biomarker.

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