Hemoglobin A2 quantification by capillary zone electrophoresis

Area of Science:

• Biochemistry
• Analytical Chemistry
• Hematology

Background:

• Hemoglobin A2 (HbA2) is a minor hemoglobin component, typically ~2.2% of total erythrocytes.
• Accurate quantification of HbA2 is crucial for diagnosing hemoglobinopathies, including sickle cell trait.

Purpose of the Study:

• To describe and validate a capillary electrophoresis (CE) method for HbA2 separation and quantification.
• To investigate factors influencing the accuracy and precision of HbA2 measurement by CE.
• To compare the performance of CE with microcolumn chromatography for HbA2 quantification.

Main Methods:

• Capillary electrophoresis (CE) utilizing an arginine Tris buffer system.
• Investigation of variables impacting quantification accuracy and precision.
• Comparative analysis against a microcolumn chromatography method.

Main Results:

• A robust CE method was developed for HbA2 separation and quantification.
• Key variables affecting CE accuracy and precision were identified.
• CE demonstrated superior suitability for HbA2 measurement in sickle cell trait compared to microcolumn chromatography.

Conclusions:

• Capillary electrophoresis provides an accurate and precise method for HbA2 quantification.
• The developed CE method is advantageous for diagnosing sickle cell trait.
• CE offers a more suitable alternative to microcolumn chromatography for measuring HbA2.