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HAEMOGLOBINOPATHY CASES on HPLC

HAEMOGLOBINOPATHY CASES on HPLC. Dr. Archana Vazifdar Head Pathologist Hindlabs , HLL Lifecare Ltd. Detector. Direction of flow. HPLC Automated system precalibrated column and gradient. Hemoglobin is eluted in a stepped manner by Buffers of Increasing Ionic strength .

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HAEMOGLOBINOPATHY CASES on HPLC

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  1. HAEMOGLOBINOPATHY CASESon HPLC Dr. ArchanaVazifdar Head Pathologist Hindlabs, HLL Lifecare Ltd.

  2. Detector Direction of flow HPLC Automated system precalibrated column and gradient

  3. Hemoglobin is eluted in a stepped manner by Buffers of Increasing Ionic strength

  4. Output Time CHROMATOGRAMS Peak Area RT (min) Total Area Count: 1-3 million HbF: 1-2% RT: 1.03-1.13 HbA2: 1.75-3.25% (2-3.6%) RT: 3.63-3.64

  5. P2- Glycemic status, upto 6% acceptable P3- 6% acceptable, 6-15% deterioration 15-25% HbJ

  6. 8 mnth/M, pallor, failure to thrive

  7. Homozygous beta thalassemia

  8. 22/Female

  9. Mother of 8 mnth old child Heterozygous Beta thalassemia

  10. HbA2- Normal • RBC indices- Normal • Silent β thalassaemia • RBC indices s/o thal • Co existing IDA • Co inheritance of α thalassaemia • δβ thalassaemia HbA2 LOW in α thalassaemia

  11. HbLepore Trait

  12. 34/M, Kolkata

  13. Heterozygous E thalassemia

  14. All criteria fit provided there is NO history of recent blood transfusion • ALWAYS CORRELATE with clinical history with CBC & peripheral blood picture • Degenerated sample

  15. Case 1: 2 yr/M, anemia, hepatosplenomegaly Hb 7 MCV 84 MCH 28 MCHC 32.4 RDW 20.2

  16. Elevated HbF: • HPFH (HbF 5-30%, CBC N, asymp) • Heterozygous δβthal • (HbF 3-20%, asymp) • Homozygous beta thal • Pregnancy

  17. Recently transfused c/o homozygous beta thalassemia

  18. Case 2: 28/F, Severe anemia

  19. Borderline HbA2 levels: • Silent carrier of thalassemia • βthal with superadded IDA • Macrocytosis • αβthalassemia

  20. Borderline elevated HbA2 due to macrocytosis

  21. P3: 6% acceptable, 6-12% deterioration 15-25% HbJ

  22. Hb: 6.5 MCV: 63 MCH: 23 MCHC: 26 RDW: 19.6

  23. Double heterozygous for HbE and β thalassaemia

  24. Tests may not be accurate if… • Patient had a blood transfusion within the past four months. • Patient has polycythemia (increased red blood cell production) or underlying anemia • If the patient is on certain medications • Aged/ degenerated sample

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