Published: 2019-10-24

Pattern distribution of abnormal hemoglobin variants by cation exchange High Performance Liquid Chromatography: a study of 9,116 subjects

Sejal Kalyanbhai Gamit, Arpita Nishal, Pinal Shah, Archana Patel


Background: The present study was conducted to identify pattern distribution of abnormal haemoglobin variants by using HPLC method in a tertiary care hospital, Surat, Gujrat, India.

Methods: A cross sectional study of one-year duration was conducted including 9,116 patients screened for the presence of abnormal haemoglobin variants. Blood samples were initially tested for solubility test and run on automated haemoglobin analyzer for complete haemogram. All the suspected and family study cases were processed for HPLC (Bio-Rad Variant II) for conclusive diagnosis. Patients with a history of recent blood transfusion of less than 3 months duration were excluded from the study.

Results: A total of 9,116 cases (1390 males, 7726 females) were included in the present study. The age group of patients ranged from 1 month to 95 years. Solubility test and complete haemogram were performed in all the cases. Out of the 9,116 cases, 8409(92.24%)cases had normal HPLC pattern. 492(5.40%)cases were diagnosed as sickle cell trait, 176(1.93%) cases as sickle cell disease, 29(0.32%) cases as β thalassaemia trait, 1(0.01%) case as β thalassaemia major, 2(0.02%)cases as Hb E heterozygous and 03 (0.07%) cases as Hb D Punjab heterozygous. One case of double heterozygous for Hb E-β thalassaemia was also found.

Conclusions: HPLC is a rapid, accurate and useful method for diagnosing haemoglobinopathies. It serves as an reliable tool in diagnosing the presence of abnormal haemoglobin variants in suspected cases on routine haematology in developing countries like India, where the resources for detection of haemoglobinopathies are limited. Early diagnosis may help in proper management of patients.


Blood transfusion, Haemoglobinopathies, Haemogram, High performance liquid chromatography

Full Text:



Vaz FE, Thakur CB, Banerjee MK, Gangal SG. Distribution of beta-thalassaemia mutations in the indian population referred to a diagnostic center. Haemoglobin. 2000;24(3):181-94.

Balgir RS. The burden of haemoglobinopathies in india and the challenges ahead. Current Sci Association. 2000;79(11):1536-47.

Balgir RS. The genetic burden of haemoglobinopathies with special references to community health in India and the challanges ahead. Indian J Hemat Blood Transfuse. 2002;20:2-7.

Baruah MK, Saikia M, Baruah A. Pattern of haemoglobinopathies and thalassaemia’s in upper Assam region of North Eastern India: high performance liquid chromatography studies in 9000 patients. Ind J Pathol Microbiol. 2014;57(2):236-43.

Wild BJ, Stephens AD. The use of automated HPLC to detect and quantitate haemoglobins. Clin Laborat Haematol. 1997;19(3):171-6.

Ou CN, Rognerud CL. Diagnosis of hemoglobinopathies: electrophoresis vs. HPLC. Clinica Chimica Acta. 2001;313(1-2):187-94.

Bhatia HM, Shanbagh S.R, Baxi AJ, Bapat J.P, Sharma R.S. Genetic studies among the endogamous groups of Lohanas of North and West India. Hum Hered.1976;26(4):298-305.

Mukherjee MB, Gangakhedkar RR, Sathe MS. Abnormal hemoglobin, G6PD deficiency and their pattern of interaction in the tribal population of Valsad district (Gujarat). Indian J Hematol Blood Transf. 1993;11:227-31.

Vyas GN, Bhatia HM, Sukumaran PK, Balkrishnan V, Sanghvi LD. Study of blood groups, abnormal hemoglobins and other genetical characters in some tribes of Gujarat. Am J Phys Anthropol. 1962;20(3):255-65.

Joutovsky A, Hadzi-Nesic J, Nardi MA. HPLC retention time as a diagnostic tool for haemoglobin variants and haemoglobinopathies: a study of 60000 sample in a clinical diagnostic laboratory clinical chemistry. 2004;50(10):1736-47.

Verma IC, Choudhary VP, Jain PK. Prevention of thalassemia: a necessity in India. Indian J Pediatr. 1992;5(6):649-54.

Manglani M, Lokeshwar MR, Vani VG, Bhatia N, Mhaskar V. NESTROFT-an effective screening test for β thalassemia trait. Indian Pediatr. 1997;34(8):702-7.

Sharma RS, Parekh JG, Shah KM. Hemoglobin opathies in western India. J Assoc Physic India. 1963;11:969-973.

Misra RC, Ram B, Mohapatra BC, Das SN, Misra SC. High prevalence and heterogenicity of thalassemias in Orissa. Indian J Med Res. 1991;94:391-394.

Dash S. Beta thalassemia trait in the Punjab (North India) Br J Haematol. 1985;61(1):185-6.