Affiliations 

  • 1 Specialist Hematologist, Department of Hematology, MMCH (Tertiary Care Hospital and Thalassemia Referral Centre), Madinah Munawarah, KSA
  • 2 Prof. and Head Department of Anatomy LUC Malaysia, MMCH (Tertiary Care Hospital and Thalassemia Referral Centre), Madinah Munawarah, KSA
  • 3 Head Cytogenetics, MMCH (Tertiary Care Hospital and Thalassemia Referral Centre), Madinah Munawarah, KSA
  • 4 Prof. Cytogenetic Department, MMCH (Tertiary Care Hospital and Thalassemia Referral Centre), Madinah Munawarah, KSA
  • 5 Laboratory Head and Director, MMCH (Tertiary Care Hospital and Thalassemia Referral Centre), Madinah Munawarah, KSA
  • 6 Deputy Lab. Director, MMCH (Tertiary Care Hospital and Thalassemia Referral Centre), Madinah Munawarah, KSA
  • 7 Supervisor Laboratory, MMCH (Tertiary Care Hospital and Thalassemia Referral Centre), Madinah Munawarah, KSA
  • 8 Senior Staff, Department of Special Chemistry, MMCH (Tertiary Care Hospital and Thalassemia Referral Centre), Madinah Munawarah, KSA
Indian J Pathol Microbiol, 2021 8 4;64(3):518-523.
PMID: 34341263 DOI: 10.4103/IJPM.IJPM_709_20

Abstract

Background: : HPLC is one of the most important tools for accurate diagnosis of hemoglobinopathies and thalassemias. The advantage of the HPLC system is the excellent resolution, reproducibility &quantification of several normal and abnormal hemoglobin.

Results: BIO RAD Variant II analyzer was used. Sickle cell syndromes including double heterozygous states accounted for 56.13% of total cases. HbSS, HbS/β0-th, HbS/β+-th β-thal trait comprises 29%, 6.5%, 5.1%& 10% of total cases respectively with mean MCV (fl) = 84, 68,71,64 respectively. The Mean HbA2 for β-thal trait, HbE trait &HbE-β thal showed 5.1 ± 1.1, 19 ± 9 & 24 ± 8 respectively. HbF is increased in 8.6% case (excluding SC syndromes & β-thal disorders), of these 5.5% were infants & 12 cases of Aplastic Anemias. Peak P2 >7% (2.4% cases) was seen in uncontrolled diabetes mellitus which on quantification showed HbA1C = 8 ± 2.1 mmol/L.

Discussion: : HPLC in correlation with CBC parameters & family studies can aid in the diagnosis of majority of Hemoglobinopathies and thalassemic syndrome. The CBC & HPLC parameters of the present study are in good correlation with the research conducted by Tejinder Sing, RiouJ & Alla Joutovsky. Present study showed HPLC comprehensively characterizing HbS, A, A2, F, S, C, D from each other & was also applicable for the quantification of HbA1c for the monitoring of Diabetes Mellitus.

Conclusion: : The merits of HPLC are small quantity of sample required, economical, less TAT, accurate categorization of HbS, HbA2 & F. But one has to be aware of the limitations and problems associated with this method due to variant hemoglobin within the same retention windows. The present findings show HPLC as an excellent & powerful diagnostic tool for the direct identification of hemoglobin variants with a high degree of precision in the quantification of normal and abnormal hemoglobin fractions.

* Title and MeSH Headings from MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.