Blood Res 2017; 52(2): 130-134
Comparison of the characteristics of two hemoglobin variants, Hb D-Iran and Hb E, eluting in the Hb A2 window
Jasmita Dass, Aastha Gupta, Suchi Mittal, Amrita Saraf, Sabina Langer, and Manorama Bhargava

Department of Hematology, Sir Ganga Ram Hospital, New Delhi, India.

Correspondence to: Aastha Gupta, M.D. Department of Hematology, Sir Ganga Ram Hospital, Rajinder Nagar, New Delhi 110060, India.
Received: February 18, 2017; Revised: April 17, 2017; Accepted: May 24, 2017; Published online: June 22, 2017.
© The Korean Journal of Hematology. All rights reserved.



Cation exchange-high performance liquid chromatography (CE-HPLC) is most commonly used to evaluate hemoglobin (Hb) variants, which elute in the Hb A2 window. This study aimed to assess prevalence of an uncommon Hb variant, Hb D-Iran, and compare its red cell parameters and peak characteristics with those of Hb E that commonly elutes in the Hb A2 window.


Generally, we assess abnormal Hb using CE-HPLC as the primary technique along with alkaline and acid electrophoresis. All cases with Hb A2 window >9%, as assessed by CE-HPLCs during 2009?2013, were selected.


Twenty-nine cases with Hb D-Iran variant were identified?25 heterozygous, 2 homozygous, 1 compound heterozygous Hb D-Iran/β-thalassemia, and 1 Hb D-Iran/Hb D-Punjab. Overall prevalence of Hb D-Iran was 0.23%. Compared to patients with Hb E, those with Hb D-Iran had significantly higher Hb (12.1 vs. 11.3 g/dL, P=0.03), MCV (82.4 vs. 76.4 fL, P=0.0044), MCH (27.9 vs. 25.45 pg, P =0.0006), and MCHC (33.9 vs. 33.3 g/dL, P=0.0005). Amount of abnormal Hb (40.7 vs. 26.4%, P=0.0001) was significantly higher while retention time (3.56 vs. 3.70 min, P=0.0001) was significantly lower in Hb D-Iran than in Hb E.


Hb D-Iran peak can be easily missed if area and retention time of the Hb A2 window are not carefully analyzed. To distinguish between variants, careful analysis of peak area and retention time is sufficient in most cases and may be further confirmed by the second technique?alkaline electrophoresis.

Keywords: CE-HPLC, Hb D-Iran, Hb E, Hb A2 window


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