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

Rankin, N. J. , Burgess, K., Weidt, S., Wannamethee, G., Sattar, N. and Welsh, P. (2019) High-throughput quantification of carboxymethyl lysine in serum and plasma using high-resolution accurate mass Orbitrap mass spectrometry. Annals of Clinical Biochemistry, (doi:10.1177/0004563219830432) (PMID:30832481) (Early Online Publication)

Rankin, N. J. , Burgess, K., Weidt, S., Wannamethee, G., Sattar, N. and Welsh, P. (2019) High-throughput quantification of carboxymethyl lysine in serum and plasma using high-resolution accurate mass Orbitrap mass spectrometry. Annals of Clinical Biochemistry, (doi:10.1177/0004563219830432) (PMID:30832481) (Early Online Publication)

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Abstract

Background: 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. Methods: 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). Results: 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. Conclusion: This high-throughput sample preparation method makes it possible to analyse large cohorts required to determine the potential of carboxymethyl lysine as a biomarker.

Item Type:Articles
Status:Early Online Publication
Refereed:Yes
Glasgow Author(s) Enlighten ID:Burgess, Dr Karl and Weidt, Dr Stefan and Welsh, Dr Paul and Sattar, Professor Naveed and Rankin, Dr Naomi
Authors: Rankin, N. J., Burgess, K., Weidt, S., Wannamethee, G., Sattar, N., and Welsh, P.
College/School:College of Medical Veterinary and Life Sciences > Institute of Cancer Sciences
College of Medical Veterinary and Life Sciences > Institute of Cardiovascular and Medical Sciences
College of Medical Veterinary and Life Sciences > Institute of Infection Immunity and Inflammation
Journal Name:Annals of Clinical Biochemistry
Publisher:SAGE Publications
ISSN:0004-5632
ISSN (Online):1758-1001
Published Online:04 March 2019
Copyright Holders:Copyright © 2019 The Authors
First Published:First published in Annals of Clinical Biochemistry 2019
Publisher Policy:Reproduced under a Creative Commons License

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
617341The application of a high-throughput NMR metabolomics system to the study of insulin resistance (ISSF Catalyst)David PreissWellcome Trust (WELLCOTR)097821/Z/11/ZRI CARDIOVASCULAR & MEDICAL SCIENCES
632234Funding SchemesAnna DominiczakWellcome Trust (WELLCOTR)105614/Z/14/ZRI CARDIOVASCULAR & MEDICAL SCIENCES
690421Glasgow Molecular Pathology (GMP) NodeKarin OienMedical Research Council (MRC)MR/N005813/1ICS - EXPERIMENTAL THERAPEUTICS