Characteristic Properties of Hemoglobin Variants by Hemoglobin Separation and Quantitation Analysis

Abstract

Hemoglobin variants are caused by the synthesis of abnormal globins structure or abnormal type of amino acids. This research was conducted to study the retention time of hemoglobin variants by automated HPLC and LPLC. All samples were confirmed by DNA sequencing. In this study, 23 hemoglobin variants were identified. Nine α-globin variants are Hb I, Hb J-Norfolk, Hb Q-Thailand, Hb Grey Lynn, Hb O-Indonesia, Hb Westmead, Hb Quong Sze, Hb Pakse and Hb CS. Fourteen β-globin variants are Hb C, Hb G-Makassar, Hb S, Hb Malay, Hb E, Hb J-Bangkok, Hb J-Kaohsiung, Hb Korle Bu, Hb Pyrgos, Hb D Los Angeles, Hb Tende, Hb Dhonburi, Hb Hope and Hb Tak. The results from Hb typing by automated HPLC and LPLC shows hemoglobin elution peak with retention time 1.41-5.14 minutes and 71-367 seconds, respectively. In the separation of hemoglobin variants by HPLC and LPLC, three identical patterns of elution peaks were observed. The first group had retention times in the S-window such as Hb S, Hb Q-Thailand and Hb G Makassar, the second group was in the D-window such as Hb D Los Angeles, Hb Tak and Hb Korle Bu, and the third group was in the A window such as Hb Dhonburi, Hb Malay, Hb Quong Sze and Hb Westmead. Therefore, it is recommended that routine clinical laboratory reports should not specify the type of rare hemoglobin variants and simply report as abnormal hemoglobin. Identification of the type of hemoglobins can only be made through DNA sequencing or amino acid analysis.