Comparison of high performance liquid chromatography and turbidimetric inhibition immunoassay methods for measurement of hemoglobin A1c

Year 2022, Volume: 5 Issue: 5, 1281 - 1285, 25.09.2022

Murat Cihan , Abdullah Üner , Muhammed Fevzi Kılınçkaya , Tevfik Noyan

https://doi.org/10.32322/jhsm.1139162

Abstract

Aim: Hemoglobin A1c is a valuable parameter for the diagnosis and follow-up of its diabetes mellitus since its biological variation is low, does not require preparation before the test, is not affected by acute stress, and has high preanalytical stability. HbA1c measurement by HPLC has been determined as the reference method by National Glycohemoglobin Standardization Program (NGSP) in USA; after that The International Federation of Clinical Chemistry (IFCC) defined another reference method which could be related with NGSP. In our study, we aim to compare the two NGSP-certified methods of HbA1c, which are high-performance liquid chromatography (HPLC) and turbidimetric inhibition immunoassay (TINIA).
Material and Method: HbA1c levels of the patients were measured using two HPLC and one TINIA method in three different hospitals (Lab A, Lab B (Both are HPLC), and Lab C (TINIA), in which Lab A was served as a reference). Because of the lower precision values of LabB, we firstly conducted a method comparison study of 40 volunteers (Group 1). After that, corrective and preventive activities carried out and the precision values in LabB reached the desired range. Following this, another method comparison study consisting of 60 new volunteers (Group 2) was conducted. The statistical flow of this study complied with Clinical Laboratory Standards Institute (CLSI) EP09-A3; Precision studies, Blant-Altman and Passing Bablok regression analysis were performed.
Results: The percentage of the mean difference between the two HPLC methods (LabA and LabB) was 3.1%. After corrective and preventive actions had been taken, the mean difference between the two HPLC methods decreased to 2.0%. A decrease in systematic bias was found in our study. Two HPLC methods can be used interchangeably in both Group 1 and Group 2. In Group 1; 95% CI of intercept and slope were found as (-1.41 to -0.30) and (1.03 to 1.22), respectively. In Group 2; 95% CI of intercept and slope were found as (-1.33 to -0.31) and (1.01 to 1.17), respectively. HPLC and TINIA methods could not be used interchangeable without affecting patient results and outcome in both Group 1 and Group 2.
Conclusion: Our study concluded that TINIA and HPLC methods could not be used interchangeably without affecting patient results and outcome. Because of the methodology that clinical laboratories are used to, clinicians and clinical biochemists should collaborate on managing diabetes mellitus regarding diagnosis, treatment, and follow-up.

Keywords

Diabetes mellitus, HPLC, immunoturbidimetry, HbA1c

Supporting Institution

None

Project Number

None

Thanks

None

References

• Bunn HF, Haney DN, Kamin S, Gabbay KH, Gallop PM. The biosynthesis of human hemoglobin A1c. Slow glycosylation of hemoglobin in vivo. J Clin Invest 1976; 57: 1652-9.
• Kaiser P, Reinauer H. Diabetes mellitus: The long way of standardization of HbA1c to the level of highest metrological order. GMS German Med Sci 2011; 9: 1-4.
• EuBIVAS. The EFLM Biological Variation Database: Aarsand AK, Fernandez-Calle P, Webster C, et al.; 2022 [cited 2022 29.06]. Available from: https: //biologicalvariation.eu.
• International Expert C. International Expert Committee report on the role of the A1C assay in the diagnosis of diabetes. Diabetes Care 2009; 32: 1327-34.
• Jeppsson JO, Kobold U, Barr J, et al. Approved IFCC reference method for the measurement of HbA1c in human blood. Clin Chem Lab Med 2002; 40: 78-89.
• Karami A, Baradaran A. Comparative evaluation of three different methods for HbA1c measurement with High-performance liquid chromatography in diabetic patients. Adv Biomed Res 2014; 3: 6-10.
• Penttila I, Penttila K, Holm P, et al. Methods, units and quality requirements for the analysis of haemoglobin A1c in diabetes mellitus. World J Methodol 2016; 6: 133-42.
• Halwachs-Baumann G, Katzensteiner S, Schnedl W, Purstner P, Pieber T, Wilders-Truschnig M. Comparative evaluation of three assay systems for automated determination of hemoglobin A1c. Clin Chem 1997; 43: 511-7.
• Consensus C. Consensus statement on the worldwide standardization of the hemoglobin A1C measurement: the American Diabetes Association, European Association for the Study of Diabetes, International Federation of Clinical Chemistry and Laboratory Medicine, and the International Diabetes Federation. Diabetes Care 2007; 30: 2399-400.
• Hoelzel W, Weykamp C, Jeppsson JO, et al. IFCC reference system for measurement of hemoglobin 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.
• Urrechaga E. Analytical evaluation of the ADAMS () A1c HA8180T analyzer for the measurement of HbA1c. J Clin Lab Anal 2018; 32: 1-6.
• Wu X, Chao Y, Wan Z, et al. A comparative evaluation of the analytical performances of Capillarys 2 Flex Piercing, Tosoh HLC-723 G8, Premier Hb9210, and Roche Cobas c501 Tina-quant Gen 2 analyzers for HbA1c determination. Biochem Med (Zagreb) 2016; 26: 353-64.
• Syed IA, Khan WA. Glycated haemoglobin--a marker and predictor of cardiovascular disease. J Pak Med Assoc 2011; 61: 690-5.
• Genc S, Omer B, Aycan-Ustyol E, Ince N, Bal F, Gurdol F. Evaluation of turbidimetric inhibition immunoassay (TINIA) and HPLC methods for glycated haemoglobin determination. J Clin Lab Anal 2012; 26: 481-5.
• Kın Tekçe B, Tekçe H, Aktaş G, M. T. HbA1c Ölçümünde Architect C 8000 ile MQ-2000PT Sonuçlarının Karşılaştırılması. J Acad Res Med 2015: 52-5.
• Davari Edalat Panah S, Karimian Tousi N, Rahimi L, Sabouri G, Mirsalehi A, Zahedi Avval F. Comparison of Two Methods for Measurement of HbA1c in Two University Hospitals of Mashhad. Journal of Patient Safety & Quality Improvement 2015; 3: 262-5.
• CLSI. EP09-A3: Measurement Procedure Comparison and Bias Estimation Using Patient Samples. USA: CLSI; 2018. p. 15-9.
• Pum J. A practical guide to validation and verification of analytical methods in the clinical laboratory. Adv Clin Chem 2019; 90: 215-81.
• Ozcelik F, Yiginer O, Serdar M, et al. Comparison of three methods for measurement of HbA1c. Turk J Biochem 2010; 35: 344-9.
• Hamwi A, Schweiger CR, Veitl M, Schmid R. Quantitative measurement of HbA1c by an immunoturbidimetric assay compared to a standard HPLC method. Am J Clin Pathol 1995; 104: 89-95.
• Groche D, Hoeno W, Hoss G, Vogt B, Herrmann Z, Witzigmann A. Standardization of two immunological HbA1c routine assays according to the new IFCC reference method. Clin Lab 2003; 49: 657-61.
• Sağlık Bakanlığı. Sağlık Uygulama Tebliği 2022 [cited 2022 15.07]. Available from: https://www.mevzuat.gov.tr/File/Pdf?mevzuatNo=17229&mevzuatTur=Teblig&mevzuatTertip=5.