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Commutable whole blood reference materials for hemoglobin A1c validated on multiple clinical analyzers

  • Hong Liu , Lingkai Wong , Sharon Yong , Qinde Liu ORCID logo EMAIL logo , Tang Lin Teo , Tong Kooi Lee , Tze Ping Loh , Sunil Kumar Sethi , Moh Sim Wong , Shelia Marie Delos Santos Cosio , Clement K.M. Ho , Johnson Weng Sung Setoh , Sok Fong Maria Lim , Grace Lay Lay Lee , Hafash Khalid , Sharine Lim , Choong-Weng Lam , Mee-Yin Lee , Chin Pin Yeo , Pallavi Ravikiran Chincholkar , Robert Hawkins and Bih-Yann Ng
Published/Copyright: December 13, 2018
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Clinical Chemistry and Laboratory Medicine (CCLM)
From the journal Clinical Chemistry and Laboratory Medicine (CCLM) Volume 57 Issue 5

Abstract

Background

The measurement of hemoglobin A1c (HbA1c) is important for diagnosing diabetes mellitus as well as assessing glycemic control in diabetic patients. Commutable whole blood certified reference materials (CRMs) are needed in the measurement of HbA1c for method validation and/or as quality controls.

Methods

We developed three levels of hemolyzed whole blood CRMs for HbA1c. The certified values were determined using liquid chromatography-isotope dilution tandem mass spectrometry method (LC-IDMS/MS) where two “signature” hexapeptides of HbA1c and hemoglobin A0 (HbA0) were used as the calibration standards. The concentrations of the hexapeptide solutions were determined by amino acid analysis by the LC-IDMS/MS method using amino acid CRMs as the calibration standards. The commutability study was conducted by measuring 25 patient specimens and the whole blood CRMs by both LC-IDMS/MS method and various routine methods using six different clinical analyzers.

Results

The certified values were determined to be 35.1±2.0, 50.3±1.9 and 65.8±2.6 mmol/mol, respectively. These CRMs showed good commutability on five of the six clinical analyzers but showed poor commutability on one of the clinical analyzers that used similar method as two other analyzers where good commutability was observed.

Conclusions

With certified target values based on metrological traceability and good commutability on most of the clinical analyzers, the developed whole blood CRMs can be used for method validation or as quality control materials in the measurement of HbA1c. The commutability study results also underscored the need of commutability testing of clinical CRMs using various clinical analyzers.

Keywords: certified reference material; commutability; diabetes; hemoglobin A1c; isotope dilution mass spectrometry

Acknowledgments

The authors are grateful to the Health Sciences Authority, Singapore, for the support of this project. The authors would also like to thank Ms Teng Kah Lee of Mount Alvernia Hospital for participating in the commutability study and Ms Pui Sze Cheow of HSA for valuable discussion.

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

  2. Research funding: None declared.

  3. Employment or leadership: None declared.

  4. Honorarium: None declared.

  5. Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.

References

1. World Health Organization. Abbreviated report of a WHO consultation: use of glycated hemoglobin (HbA1c) in the diagnosis of diabetes mellitus. http://www.who.int/diabetes/publications/report-hba1c_2011.pdf. Accessed: 24 May 2018.Search in Google Scholar

2. Weykamp C. HbA1c: a review of analytical and clinical aspects. Ann Lab Med 2013;33:393–400.10.3343/alm.2013.33.6.393Search in Google Scholar PubMed PubMed Central

3. Bi J, Wu L, Yang B, Yang Y. Development of hemoglobin A1c certified reference material by liquid chromatography isotope dilution mass spectrometry. Anal Bioanal Chem 2012; 403:549–54.10.1007/s00216-012-5834-9Search in Google Scholar PubMed

4. Reference Material Institute for Clinical Chemistry Standards (ReCCS) Certificate of Analysis Certified Reference Material for Measurement of HbA1c JDS HbA1c Lot 2. http://www.reccs.or.jp/images/hba1clot2.pdf. Accessed: 24 May 2018.Search in Google Scholar

5. Tran TT, Lim J, Kim J, Kwon H-J, Kwon GC, Jeong JS. Fully international system of units-traceable glycated hemoglobin quantification using two stages of isotope-dilution high-performance liquid chromatography-tandem mass spectrometry. J Chromatogr A 2017;1513:183–93.10.1016/j.chroma.2017.07.056Search in Google Scholar PubMed

6. Miller WG, Schimmel H, Rej R, Greenberg N, Ceriotti F, Burns C, et al. IFCC Working Group recommendations for assessing commutability. Part 1: general experimental design. Clin Chem 2018;64:447–54.10.1373/clinchem.2017.277525Search in Google Scholar PubMed PubMed Central

7. Nilsson G, Budd JR, Greenberg N, Delatour V, Rej R, Panteghini M, et al. IFCC Working Group recommendations for assessing commutability. Part 2: using the difference in bias between a reference material and clinical samples. Clin Chem 2018; 64:455–64.10.1373/clinchem.2017.277541Search in Google Scholar PubMed PubMed Central

8. Budd JR, Weykamp C, Rej R, MacKenzie F, Ceriotti F, Greenberg N, et al. IFCC Working Group recommendations for assessing commutability. Part 3: using the calibration effectiveness of a reference material. Clin Chem 2018;64:465–74.10.1373/clinchem.2017.277558Search in Google Scholar PubMed

9. ISO REMCO. Information on commutability of reference materials (2014). http://isotc.iso.org/livelink/livelink/fetch/-8854933/8854951/8854960/279217/Commutability_document_final.pdf?nodeid=16787892&vernum=-2. Accessed: 24 May 2018.Search in Google Scholar

10. Vesper HW, Miller WG, Myers GL. Reference materials and commutability. Clin Biochem Rev 2007;28:139–47.Search in Google Scholar

11. Korzun WJ, Nilsson G, Bachmann LM, Myers GL, Sakurabayashi I, Nakajima K, et al. Difference in bias approach for commutability assessment: application to frozen pools of human serum measured by 8 direct methods for HDL and LDL cholesterol. Clin Chem 2015;61:1107–13.10.1373/clinchem.2015.240861Search in Google Scholar PubMed

12. Jeppsson J-O, Kobold U, Barr J, Finke A, Hoelzel W, Hoshino T, et al. Approved IFCC reference method for the measurement of HbA1c in human blood. Clin Chem Lab Med 2002;40:78–89.10.1515/CCLM.2002.016Search in Google Scholar PubMed

13. Kaiser P, Akerboom T, Molnar P, Reinauer H. Modified HPLC-electrospray ionization/mass spectrometry method for HbA1c based on IFCC reference measurement procedure. Clin Chem 2008;54:1018–22.10.1373/clinchem.2007.100875Search in Google Scholar PubMed

14. Kaiser P, Akerboom T, Dux L, Reinauer H. Modification of the IFCC reference measurement procedure for determination of HbA1c by HPLC-ESI-MS. Ger Med Sci 2006;4:Doc06.Search in Google Scholar

15. IFCC Standardization of HbA1c. http://www.ngsp.org/ifccngsp.asp. Accessed: 24 May 2018.Search in Google Scholar

16. Milton MJ, Quinn TJ. Primary methods for the measurement of amount of substance. Metrologia 2001;38:289–96.10.1088/0026-1394/38/4/1Search in Google Scholar

17. De Bièvre P, Peiser HS. Basic equations and uncertainties in isotope-dilution mass spectrometry for traceability to SI of values obtained by this primary method. Fresenius J Anal Chem 1997;359:523–5.10.1007/s002160050625Search in Google Scholar

18. Alonso JI, Moldovan M, González PR. Isotope dilution mass spectrometry. Cambridge: Royal Society of Chemistry, 2013.10.1039/9781839168925Search in Google Scholar

19. Nakanishi T, Iguchi K, Shimizu A. Method for haemoglobin A1c measurement based on peptide analysis by electrospray ionization mass spectrometry with deuterium-labeled synthetic peptides as internal standards. Clin Chem 2003;49:829–31.10.1373/49.5.829Search in Google Scholar PubMed

20. Iguchi K, Nakanishi T, Miyazaki A, Shimizu A, Ota A. Development of an isotope dilution mass spectrometry assay for HbA1c based on enzyme-cleaved peptide analysis. J Chromatogr B 2004;806:25–31.10.1016/j.jchromb.2004.01.038Search in Google Scholar PubMed

21. Kaiser P, Akerboom T, Ohlendorf R, Reinauer H. Liquid chromatography-isotope dilution-mass spectrometry as a new basis for the reference measurement procedure for hemoglobin A1c determination. Clin Chem 2010;56:750–4.10.1373/clinchem.2009.139477Search in Google Scholar PubMed

22. Wong L, Liu H, Yong S, Liu Q, Lee TK. Improved reference measurement method for hemoglobin A1c by use of liquid chromatography-isotope dilution-tandem mass spectrometry. Clin Chem 2015;62:435–6.10.1373/clinchem.2014.231340Search in Google Scholar PubMed

23. Liu H, Wong L, Yong S, Liu Q, Lee TK. Achieving comparability with IFCC reference method for the measurement of hemoglobin A1c by use of an improved isotope-dilution mass spectrometry method. Anal Bioanal Chem 2015;407:7579–87.10.1007/s00216-015-8961-2Search in Google Scholar PubMed

24. Westwood S, Josephs RD, Choteau T, Daireaux A, Wielgosz R, Davies SR, et al. Final report on key comparison CCQM-K55.c (L-(+)-Valine): characterization of organic substances for chemical purity. Metrologia 2014;51:08010.10.1088/0026-1394/51/1A/08010Search in Google Scholar

25. ISO 13485:2016 Medical devices – quality management systems – requirements for regulatory purposes. Geneva, Switzerland: International Organization for Standardization.Search in Google Scholar

26. Chen Y, Liu Q, Yong S, Lee TK. High accuracy analysis of glucose in human serum by isotope dilution liquid chromatography-tandem mass spectrometry. Clin Chim Acta 2012;413:808–13.10.1016/j.cca.2012.01.025Search in Google Scholar PubMed

27. Josephs RD, Li M, Song D, Westwood S, Stoppacher N, Daireaux A, et al. Key comparison study on peptide purity — synthetic human C-peptide. Metrologia 2017;54:08007.10.1088/0026-1394/54/1A/08007Search in Google Scholar

28. ISO Guide 35:2006 Reference materials — general and statistical principles for certification. Geneva, Switzerland: International Organization for Standardization.Search in Google Scholar

29. ISO 17034:2016 General requirements for the competence of reference material producers. Geneva, Switzerland: International Organization for Standardization.Search in Google Scholar

30. Clinical and Laboratory Standards Institute. EP30-A characterization and qualification of commutable reference materials for laboratory medicine; approved guideline. May 2010.Search in Google Scholar

31. Delatour V, Liu Q, Vesper HW. Commutability assessment of external quality assessment materials with the difference in bias approach: are acceptance criteria based on medical requirements too strict? Clin Chem 2016;62:1670–1.10.1373/clinchem.2016.261008Search in Google Scholar PubMed PubMed Central

32. College of American Pathologists (CAP) GH5 Survey Data. http://www.ngsp.org/CAP/CAP18b.pdf. Accessed: 5 October 2018.Search in Google Scholar

33. Allowable Limits of Performance — RCPAQAP. http://www.rcpaqap.com.au/docs/2014/chempath/ALP.pdf. Accessed: 5 Oct 2018.Search in Google Scholar

34. Hoelzel W, Weykamp C, Jeppsson J-O, Miedema K, Barr JR, Goodall I, et al. IFCC reference system for measurement of haemoglobin A1c in human blood and the national standardization schemes in the United States, Japan, and Sweden: a method comparison study. Clin Chem 2004;50:166–74.10.1373/clinchem.2003.024802Search in Google Scholar PubMed

35. ISO 13528:2015 Statistical methods for use in proficiency testing by interlaboratory comparisons. Geneva, Switzerland: International Organization for Standardization.Search in Google Scholar

36. ISO Guide 98-3:2008 Uncertainty of measurement — part 3: guide to the expression of uncertainty in measurement (GUM:1995). Geneva, Switzerland: International Organization for Standardization.Search in Google Scholar

Supplementary Material

The online version of this article offers supplementary material (https://doi.org/10.1515/cclm-2018-0861).

Received: 2018-08-10
Accepted: 2018-11-05
Published Online: 2018-12-13
Published in Print: 2019-04-24

©2019 Walter de Gruyter GmbH, Berlin/Boston

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https://doi.org/10.1515/cclm-2018-0861
Keywords for this article
certified reference material; commutability; diabetes; hemoglobin A1c; isotope dilution mass spectrometry

Articles in the same Issue

  1. Frontmatter
  2. Editorial
  3. Cardiac biomarkers – 2019
  4. Reviews
  5. Current understanding and future directions in the application of TIMP-2 and IGFBP7 in AKI clinical practice
  6. Serum cytokines, adipokines and ferritin for non-invasive assessment of liver fibrosis in chronic liver disease: a systematic review
  7. Opinion Papers
  8. Detection capability of quantitative faecal immunochemical tests for haemoglobin (FIT) and reporting of low faecal haemoglobin concentrations
  9. Should phosphatidylethanol be currently analysed using whole blood, dried blood spots or both?
  10. IFCC Papers
  11. High sensitivity, contemporary and point-of-care cardiac troponin assays: educational aids developed by the IFCC Committee on Clinical Application of Cardiac Bio-Markers
  12. Cardiac troponin and natriuretic peptide analytical interferences from hemolysis and biotin: educational aids from the IFCC Committee on Cardiac Biomarkers (IFCC C-CB)
  13. Genetics and Molecular Diagnostics
  14. Droplet digital PCR for the simultaneous analysis of minimal residual disease and hematopoietic chimerism after allogeneic cell transplantation
  15. General Clinical Chemistry and Laboratory Medicine
  16. Commutable whole blood reference materials for hemoglobin A1c validated on multiple clinical analyzers
  17. When results matter: reliable creatinine concentrations in hyperbilirubinemia patients
  18. Mass spectrometry based analytical quality assessment of serum and plasma specimens with patterns of endo- and exogenous peptides
  19. Association of serum sphingomyelin profile with clinical outcomes in patients with lower respiratory tract infections: results of an observational, prospective 6-year follow-up study
  20. Effect of an activated charcoal product (DOAC Stop™) intended for extracting DOACs on various other APTT-prolonging anticoagulants
  21. Hematology and Coagulation
  22. Commutability assessment of reference materials for the enumeration of lymphocyte subsets
  23. Circulating platelet-neutrophil aggregates as risk factor for deep venous thrombosis
  24. Reference Values and Biological Variations
  25. A comparison of complete blood count reference intervals in healthy elderly vs. younger Korean adults: a large population study
  26. Indirect determination of hematology reference intervals in adult patients on Beckman Coulter UniCell DxH 800 and Abbott CELL-DYN Sapphire devices
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  28. Large platelet size is associated with poor outcome in patients with metastatic pancreatic cancer
  29. Cardiovascular Diseases
  30. Sample matrix and high-sensitivity cardiac troponin I assays
  31. Preoperative proteinuria and clinical outcomes in type B aortic dissection after thoracic endovascular aortic repair
  32. Infectious Diseases
  33. The rational specimen for the quantitative detection of Epstein-Barr virus DNA load
  34. Letters to the Editor
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