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Potential of MALDI-TOF mass spectrometry to overcome the interference of hemoglobin variants on HbA1c measurement

  • Anping Xu ORCID logo EMAIL logo , Weijie Xie , Yajun Wang and Ling Ji
Published/Copyright: July 17, 2020
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Clinical Chemistry and Laboratory Medicine (CCLM)
From the journal Clinical Chemistry and Laboratory Medicine (CCLM) Volume 59 Issue 1

Abstract

Objectives

Hemoglobin (Hb) variants remain an important cause of erroneous HbA1c results. We present an approach to overcome the interference of Hb variants on HbA1c measurements using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS).

Methods

Samples containing or not containing Hb variants were analyzed for HbA1c using an MALDI-TOF MS system (QuanTOF) and a boronate affinity comparative method (Ultra2). For QuanTOF, two sets of HbA1c values were obtained through α- and β-chain glycation.

Results

A robust correlation between the glycation degrees of the α- and β-chains was found, and HbA1c values derived from α- and β-chain glycation correlated well with the Ultra2 results. Statistically significant differences (p<0.01) were found for all the Hb variants tested. When using the conventional β-chain glycation to determine HbA1c, clinically significant differences were only found among samples containing β-chain variants detected by QuanTOF (i.e., Hb J-Bangkok, Hb G-Coushatta, and Hb G-Taipei). In contrast, based on α-chain glycation, no clinically significant differences were found for these three variants.

Conclusions

In addition to conventional β-chain glycation, α-chain glycation can be used to calculate HbA1c values. The interference of Hb variants on HbA1c quantification can be overcome by employing the glycation of the globin chain without a genetic variant to estimate HbA1c values.

Keywords: α-chain glycation; glycation degree; HbA1c; hemoglobin variant; interference; matrix-assisted laser desorption/ionization time-of-flight mass spectrometry
Corresponding author: Anping Xu, Department of Laboratory Medicine, Peking University Shenzhen Hospital, Lian Hua Road No. 1120, Futian District, Shenzhen, Guangdong, 518036, PR China, E-mail:

Funding source: Research Foundation of Peking University Shenzhen Hospital

Award Identifier / Grant number: JCYJ2018005

  1. Research funding: This study was supported by Research Foundation of Peking University Shenzhen Hospital (JCYJ2018005).

  2. Author contributions: All authors have accepted responsibility for the entire content of this manuscript and approved its submission.

  3. Competing interests: Authors state no conflict of interest.

  4. Informed consent: Informed consent was obtained from all individuals included in this study.

  5. Ethical approval: The study was approved by the Ethics Committee of the Peking University Shenzhen Hospital.

References

1. The Diabetes Control and Complications Trial Research Group. The effect of intensive treatment of diabetes on the development and progression of long-term complications in insulin-dependent diabetes mellitus. N Engl J Med 1993;329:977–86.10.1056/NEJM199309303291401Search in Google Scholar PubMed

2. U.K. Prospective Diabetes Study (UKPDS) Group. Intensive blood-glucose control with sulphonyl ureas or insulin compared with conventional treatment and risk of complications in patients with type 2 diabetes (UKPDS 33). Lancet 1998;352:837–53.10.1016/S0140-6736(98)07019-6Search in Google Scholar

3. American Diabetes Association. Standards of medical care in diabetes–2011. Diabetes Care 2011;34:S11–61.10.2337/dc11-S011Search in Google Scholar PubMed PubMed Central

4. Little, RR, Rohlfing, C, Sacks, DB. The National Glycohemoglobin Standardization Program: over 20 years of improving hemoglobin A1c measurement. Clin Chem 2019;65:839–48. https://doi.org/10.1373/clinchem.2018.296962.Search in Google Scholar PubMed PubMed Central

5. Yun, YM, Ji, M, Ko, DH, Chun, S, Kwon, GC, Lee, K, et al. Hb variants in Korea: effect on HbA1c using five routine methods. Clin Chem Lab Med 2017;55:1234–42. https://doi.org/10.1515/cclm-2016-0865.Search in Google Scholar PubMed

6. Xu, A, Chen, W, Xia, Y, Zhou, Y, Ji, L. Effects of common hemoglobin variants on HbA1c measurements in China: results for α- and β-globin variants measured by six methods. Clin Chem Lab Med 2018;56:1353–61. https://doi.org/10.1515/cclm-2017-1211.Search in Google Scholar PubMed

7. Little, RR, La’ulu, SL, Hanson, SE, Rohlfing, CL, Schmidt, RL. Effects of 49 different rare Hb variants on HbA1c measurement in eight methods. J Diabetes Sci Technol 2015;9:849–56. https://doi.org/10.1177/1932296815572367.Search in Google Scholar PubMed PubMed Central

8. Jaisson, S, Leroy, N, Meurice, J, Guillard, E, Gillery, P. First evaluation of Capillarys 2 Flex Piercing (Sebia) as a new analyzer for HbA1c assay by capillary electrophoresis. Clin Chem Lab Med 2012;50:1769–75. https://doi.org/10.1515/cclm-2012-0017.Search in Google Scholar PubMed

9. Jaisson, S, Leroy, N, Guillard, E, Desmons, A, Gillery, P. Analytical performances of the D-100TM hemoglobin testing system (Bio-Rad) for HbA1c assay. Clin Chem Lab Med 2015;53:1473–9. https://doi.org/10.1515/cclm-2015-0288.Search in Google Scholar PubMed

10. John, WG, Little, R, Sacks, DB, Weykamp, C, Lenters-Westra, E, Hornsby, T, et al. Multicentre evaluation of the premier Hb9210 HbA1c analyser. Clin Chem Lab Med 2015;53:319–27. https://doi.org/10.1515/cclm-2014-0589.Search in Google Scholar PubMed PubMed Central

11. Lin, M, Wang, Q, Zheng, L, Huang, Y, Lin, F, Lin, CP, et al. Prevalence and molecular characterization of abnormal hemoglobin in eastern Guangdong of southern China. Clin Genet 2012;81:165–71. https://doi.org/10.1111/j.1399-0004.2011.01627.x.Search in Google Scholar PubMed

12. Zhang, J, Li, P, Yang, Y, Yan, Y, Zeng, X, Li, D, et al. Molecular epidemiology, pathogenicity, and structural analysis of haemoglobin variants in the Yunnan province population of Southwestern China. Sci Rep 2019;9:8264. https://doi.org/10.1038/s41598-019-44793-0.Search in Google Scholar PubMed PubMed Central

13. American Diabetes Association. 2. Classification and diagnosis of diabetes: standards of medical care in diabetes-2019. Diabetes Care 2019;42:S13–8. https://doi.org/10.2337/dc19-s002.Search in Google Scholar

14. Roberts, NB, Amara, AB, Morris, M, Green, BN. Long-term evaluation of electrospray ionization mass spectrometric analysis of glycated hemoglobin. Clin Chem 2001;47:316–21. https://doi.org/10.1093/clinchem/47.2.316.Search in Google Scholar

15. Biroccio, A, Urbani, A, Massoud, R, di Ilio, C, Sacchetta, P, Bernardini, S, et al. A quantitative method for the analysis of glycated and glutathionylated hemoglobin by matrix-assisted laser desorption ionization-time of flight mass spectrometry. Anal Biochem 2005;336:279–88. https://doi.org/10.1016/j.ab.2004.10.002.Search in Google Scholar PubMed

16. Hattan, SJ, Parker, KC, Vestal, ML, Yang, JY, Herold, DA, Duncan, MW. Analysis and quantitation of glycated hemoglobin by matrix assisted laser desorption/ionization time of flight mass spectrometry. J Am Soc Mass Spectrom 2016;27:532–41. https://doi.org/10.1007/s13361-015-1316-6.Search in Google Scholar PubMed

17. Xu, A, Wang, Y, Li, J, Liu, G, Li, X, Chen, W, et al. Evaluation of MALDI-TOF MS for the measurement of glycated hemoglobin. Clin Chim Acta 2019;498:154–60. https://doi.org/10.1016/j.cca.2019.08.025.Search in Google Scholar PubMed

18. Rohlfing, C, Hanson, S, Weykamp, C, Siebelder, C, Higgins, T, Molinaro, R, et al. Effects of hemoglobin C, D, E and S traits on measurements of hemoglobin A1c by twelve methods. Clin Chim Acta 2016;455:80–3. https://doi.org/10.1016/j.cca.2016.01.031.Search in Google Scholar PubMed PubMed Central

19. Bisse, E, Huaman-Guillen, P, Wieland, H. Chromatographic evaluation of minor hemoglobins: clinical significance of hemoglobin A1d, comparison with hemoglobin A1c, and possible interferences. Clin Chem 1995;41:658–63. https://doi.org/10.1093/clinchem/41.5.658.Search in Google Scholar

20. Shapiro, R, McManus, MJ, Zalut, C, Bunn, HF. Sites of nonenzymatic glycosylation of human hemoglobin A. J Biol Chem 1980;255:3120–7.10.1016/S0021-9258(19)85860-XSearch in Google Scholar

21. Castagnola, M, Caradonna, P, Bertollini, A, Cassiano, L, Rossetti, DV, Salvi, ML. Determination of the non-enzymatic glycation of hemoglobin by isoelectrofocusing of its globin chains. Clin Biochem 1985;18:327–31. https://doi.org/10.1016/s0009-9120(85)80069-2.Search in Google Scholar PubMed

22. Weykamp, C, Kemma, E, Leppink, S, Siebelder, C. Glycation rate of haemoglobins S, C, D, E, J and G, and analytical interference on the measurement of HbA1c with affinity chromatography and capillary electrophoresis. Clin Chem Lab Med 2015;53:e207–10. https://doi.org/10.1515/cclm-2014-1134.Search in Google Scholar PubMed

23. Kleinert, P, Schmid, M, Zurbriggen, K, Speer, O, Schmugge, M, Roschitzki, B, et al. Mass spectrometry: a tool for enhanced detection of hemoglobin variants. Clin Chem 2008;54:69–76. https://doi.org/10.1373/clinchem.2007.089961.Search in Google Scholar PubMed

Supplementary Material

The online version of this article offers supplementary material (https://doi.org/10.1515/cclm-2020-0724).

Received: 2020-05-14
Accepted: 2020-06-27
Published Online: 2020-07-17
Published in Print: 2021-01-26

© 2020 Walter de Gruyter GmbH, Berlin/Boston

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https://doi.org/10.1515/cclm-2020-0724
Keywords for this article
α-chain glycation; glycation degree; HbA1c; hemoglobin variant; interference; matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

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  2. Mario Plebani turns 70
  3. Editorial
  4. Measurement uncertainty and the importance of correlation
  5. Reviews
  6. Untangling the association between prostate-specific antigen and diabetes: a systematic review and meta-analysis
  7. The emerging role of cell senescence in atherosclerosis
  8. Mini Review
  9. Minimum retesting intervals in practice: 10 years experience
  10. Opinion Paper
  11. Cardiotoxic effects and myocardial injury: the search for a more precise definition of drug cardiotoxicity
  12. EFLM Papers
  13. The CRESS checklist for reporting stability studies: on behalf of the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM) Working Group for the Preanalytical Phase (WG-PRE)
  14. Thoughts and expectations of young professionals about the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM)
  15. Guidelines and Recommendations
  16. Evidence on clinical relevance of cardiovascular risk evaluation in the general population using cardio-specific biomarkers
  17. Genetics and Molecular Diagnostics
  18. A systematic evaluation of stool DNA preparation protocols for colorectal cancer screening via analysis of DNA methylation biomarkers
  19. General Clinical Chemistry and Laboratory Medicine
  20. The new IVD Regulation 2017/746: a case study at a large university hospital laboratory in Belgium demonstrates the need for clarification on the degrees of freedom laboratories have to use lab-developed tests to improve patient care
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  22. Feasibility for aggregation of commutable external quality assessment results to evaluate metrological traceability and agreement among results
  23. Sample size and rejection limits for detecting reagent lot variability: analysis of the applicability of the Clinical and Laboratory Standards Institute (CLSI) EP26-A protocol to real-world clinical chemistry data
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