Which method is used as a reference method for HbA1c analysis in clinical labs?

Which method is used as a reference method for HbA1c analysis in clinical labs?

• A. Affinity chromatography
• B. HPLC
• C. Immunoassay
• D. Gel filtration

Answer: (B)

High-performance liquid chromatography is used as the reference method because it physically separates the glycated form of hemoglobin from the non-glycated form and reports HbA1c as a percentage of total hemoglobin. This separation provides a direct and highly reproducible measurement that labs can standardize against, which is essential for comparing results across different platforms and over time. Immunoassay methods, while convenient, rely on antibodies and can be affected by hemoglobin variants or other interferences, making them less robust as a universal reference. Chromatography-based approaches, particularly ion-exchange HPLC, have long been the benchmark for accuracy and traceability, forming the basis for standardized HbA1c reporting.

High-performance liquid chromatography is used as the reference method because it physically separates the glycated form of hemoglobin from the non-glycated form and reports HbA1c as a percentage of total hemoglobin. This separation provides a direct and highly reproducible measurement that labs can standardize against, which is essential for comparing results across different platforms and over time. Immunoassay methods, while convenient, rely on antibodies and can be affected by hemoglobin variants or other interferences, making them less robust as a universal reference. Chromatography-based approaches, particularly ion-exchange HPLC, have long been the benchmark for accuracy and traceability, forming the basis for standardized HbA1c reporting.