High-performance liquid chromatography as a method to identify haemoglobin abnormalities.
This short review summarizes data obtained with two different high-performance liquid chromatography (HPLC) methods, useful in the study of numerous haemoglobinopathies. The first concerns the use of a cation exchange HPLC column which, when developed with specific gradient elution systems, will allow the separation of many normal and abnormal haemoglobins in blood from adults and newborn babies. The method has great diagnostic value. The second procedure uses a reverse-phase HPLC system and acetonitrile-H2O-trifluoroacetic acid developers at low pH for the separation of globin chains (alpha, zeta, beta, delta, G gamma, A gamma, epsilon). The method can be applied as an analytical as well as a preparative tool; it is ideally suited to separate several normal and abnormal chains with neutral amino acid substitutions, including the three gamma chains (G gamma = gamma 75Ile; 136Gly, A gamma = gamma 75Ile; 136Ala, A gamma T = gamma 75Thr; 136Ala).
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D006455:Hemoglobins, Abnormal / Q000032:analysis*; D006801:Humans; D007525:Isoelectric Focusing
D006455:Hemoglobins, Abnormal