Development and Application of Mass Spectroscopy Assays for Nε-(1-Carboxymethyl)-L-Lysine and Pentosidine in Renal Failure and Diabetes
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Background: Advanced glycation end products (AGEs) are formed via the nonenzymatic glycation of sugars with amino acids. Two AGEs, Nε-(1-carboxymethyl)-L-Lysine (CML) and pentosidine, have been observed to be elevated in subjects suffering from a multitude of chronic disease states, and accumulation of these compounds may be related to the pathophysiology of disease progression and aging. Methods: We describe here the development and validation of a specific and reproducible LC-MS/MS method to quantify CML and pentosidine in human serum with lower limits of quantitation of 75 ng/mL and 5 ng/mL, respectively. The analyte calibration curve exhibited excellent linearity at a range of 0–10 900 ng/mL for CML and 0–800 ng/mL for pentosidine. High-low linearity of 5 serum pairs was assessed, with a mean recovery of 103% (range 94—116%) for CML, and 104% (range 97—116%) for pentosidine. Results: Serum concentrations of CML and pentosidine were quantified in 30 control and 30 subjects with chronic renal insufficiency. A significant increase in both analytes was observed in renal failure compared to control subjects (2.1-fold and 8.4-fold, respectively; P < 0.001 for both). In a separate cohort of 49 control versus 95 subjects with type 2 diabetes mellitus (T2DM), serum CML but not serum pentosidine, was significantly elevated in the T2DM patients, and CML was also correlated with glycemic control, as assessed by hemoglobin A1c (r = 0.34, P < 0.001). Conclusions: These mass spectroscopy-based assays for serum CML and pentosidine should be useful in accurately evaluating circulating levels of these key AGEs in various disease states.
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O'Grady et al. found higher CML levels in plasma of T2D patients compared to healthy subjects but similar pentosidine concentrations.
“O'Grady et al. found higher CML levels in plasma of T2D patients compared to healthy subjects but similar pentosidine concentrations [165].”
From Protein oxidation - Formation mechanisms, detection and relevance as biomarkers in human diseases (2021) · cites this paper as [165]
The “Control versus T2DM patients” results (page unavailable) report significantly higher CML in T2DM patients and no significant difference in pentosidine. However, the paper identifies the measured samples as serum, not plasma, so it does not establish the claim specifically for plasma. (LLM, conf=0.88)
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Cites
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“Advanced glycation endproducts (AGEs) accumulate in various tissues with aging and may contribute to the pathology of diabetes mellitus, atherosclerosis, and induction of oxidative stress and inflammation (1, 2).”
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Protein oxidation - Formation mechanisms, detection and relevance as biomarkers in human diseases
Kehm, R., Baldensperger, T., Raupbach, J., & Höhn, A. (2021) · Redox Biology · doi:10.1016/j.redox.2021.101901 · Cited by 348 · see citing works
Accelerated osteocyte senescence and skeletal fragility in mice with type 2 diabetes
Eckhardt, B. A., Rowsey, J. L., Thicke, B. S., Fraser, D. G., O’Grady, K. L., Bondar, O. P., Hines, J. M., Singh, R. J., Thoreson, A. R., Rakshit, K., et al. (2020) · JCI Insight · doi:10.1172/jci.insight.135236 · Cited by 130 · see citing works
Self-citation
- Authors
- O’Grady, K. L., Khosla, S., Farr, J. N., Bondar, O. P., Atkinson, E. J., Achenbach, S. J., Eckhardt, B. A., Thicke, B. S., Tweed, A. J., Volkman, T. L., Drake, M. T., Hines, J. M., & Singh, R. J.
- Affiliations
- Mayo Clinic (US)
- Corresponding author
- Khosla, Sundeep
- Singh, Ravinder J
- ORCID
- Venue
- The Journal of Applied Laboratory Medicine
- Publisher
- Oxford University Press (OUP)
- Published
- 2020-02-08
- Volume
- 5
- Issue
- 3
- Pages
- 558-568
- ISSN
- 2475-7241 (ISSN-L)
- 2576-9456
- PMC
- PMC7192546
- OpenAlex
- W3014718637
- Cited by
- 19
- References
- 2
- Citations by year
- 2026: 3 · 2025: 4 · 2024: 2 · 2023: 1 · 2022: 2 · 2021: 6 · 2020: 1
- FWCI
- 1.66
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model (vor)
- Locations
- The Journal of Applied Laboratory Medicine (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- NIH (AR027065, UL1TR002377)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Clinical Biochemistry › Advanced Glycation End Products research
- Health Sciences › Medicine › Nephrology › Chronic Kidney Disease and Diabetes
- Health Sciences › Medicine › Nephrology › Dialysis and Renal Disease Management
- Keywords
- pentosidine, advanced glycation end products, glycation, chronic renal insufficiency, type 2 diabetes, serum biomarkers, LC-MS/MS method development, glycemic control, HbA1c
- MeSH terms
- Arginine, Diabetes Mellitus, Type 2, Humans, Lysine, Mass Spectrometry, Reproducibility of Results, Glycation End Products, Advanced, Renal Insufficiency
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