Comparative analysis of cellulose acetate hemoglobin electrophoresis and high performance liquid chromatography for quantitative determination of hemoglobin A2

Abstract

BackgroundThe present study is designed to evaluate the reliability and cost effectiveness of cellulose acetate Hb electrophoresis and high performance liquid chromatography (HPLC) in the determination of HbA2 levels.MethodsThe test population comprised 160 individuals divided into four groups: normal individuals, β-thalassemia trait (BTT) patients, iron deficiency anemia (IDA) patients, and co-morbid patients (BTT with IDA). HbA2 levels determined using cellulose acetate Hb electrophoresis and HPLC were compared.ResultsHbA2 levels were found to be diagnostic for classical BTT using either method. In co-morbid cases, both techniques failed to diagnose all cases of BTT. The sensitivity, specificity, and Youden's index for detection of the co-morbid condition was 69% and 66% for HPLC and cellulose acetate Hb electrophoresis, respectively.ConclusionThis study revealed that semi-automated cellulose acetate Hb electrophoresis is more suitable for use in β-thalassemia prevention programs in low-income countries like Pakistan. This technique is easily available, simple and cost effective.

Detection of Haemoglobinopathies Using Haemoglobin Electrophoresis in Microcytic Hypochromic Anaemia in Paediatric Population of Southern Odisha

“…But, to quantitate the different Hb variants, HPLC was done and the results of haemoglobin electrophoresis is comparable with that of HPLC as was the observations in the studies of Shafi Mohammad Khosa, Muhammad Usman, Moinuddin Moinuddin, Hassan Osman Mehmood and Khansa Qamar in 2015. 13 With the help of electrophoresis, all cases of β-thalassaemia major were picked up correctly, but some thalassaemia traits and heterozygous cases could not be diagnosed for which quantification of Hb variants is a must. In 2002, ICSH 14 also recommends that β-thalassaemia minor should be diagnosed by determining HbA2 levels using techniques such as cellulose acetate Hb electrophoresis or HPLC.…”

Tunisian Newborn's Cord Blood: Reference Values of Complete Blood Count and Hemoglobin Fractions

“…18 The instruments used for the analysis differ among the studies while they are also important, not only for CBC but also for Hb profiles. 32,33 For this analysis, for example, the Omani used high-performance liquid chromatography, the Sudani used alkaline agarose gel electrophoresis, and we used capillary electrophoresis which is well known to be more accurate for Hb quantification. Discordance between parameters' values depends also on inclusion/exclusion criteria.…”

Hb A2 Episkopi – a novel δ-globin chain variant [HBD:c.428C>T] in a family of mixed Cypriot–Lebanese descent

“…access to HPLC equipment, where complementary electrophoresis methods might be used instead. For instance, the combination of methods detecting changes in pI or in charge at a given pH, such as isoelectric focussing (IEF) or CAE [18,19], could be combined with methods detecting changes in molecular weight, such as acid-urea-Triton X-100 polyacrylamide gel electrophoresis (AUT-PAGE) [19], to minimize false-negative variant detection and thus the risk of misdiagnosis of haemoglobinpathies. The known Hb A 2 tetramer structure (PDB 1si4 [9], left panel) has been energy-optimized using FoldX to accommodate the A143V amino-acid change (right panel), which is located five amino acids from the carboxy-terminus of δ-globin.…”