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HAEMOGLOBINOPATHY CASES on HPLC Dr. Archana Vazifdar Head Pathologist Hindlabs, HLL Lifecare Ltd.

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Presentation on theme: "HAEMOGLOBINOPATHY CASES on HPLC Dr. Archana Vazifdar Head Pathologist Hindlabs, HLL Lifecare Ltd."— Presentation transcript:

1 HAEMOGLOBINOPATHY CASES on HPLC Dr. Archana Vazifdar Head Pathologist Hindlabs, HLL Lifecare Ltd.

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3 HPLC Automated system precalibrated column and gradient Direction of flow Detector

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

5 CHROMATOGRAMSOutputTime Peak RT RT (min) Area Total Area Count: 1-3 million HbF: 1-2%RT: HbA2: % (2-3.6%)RT:

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

7 8 mnth/M, pallor, failure to thrive

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9 Homozygous beta thalassemia

10 22/Female

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12 Heterozygous Beta thalassemia Mother of 8 mnth old child

13 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

14 HbA2% HbF% ≤ 2 Hb N MCV N Symptoms - FINAL NORMAL 4-10≤ 2/ mBorderline Asymptoma tic β THAL TRAIT NN Asymptoma tic Hb LEPORE TRAIT 25-35≤ 1NN Asymptoma tic HbE TRAIT 40-48≤ 2NN Asymptoma tic HbD IRAN Heterozygo us 50-60≥ 10Severe HbE/Hbβ double heterozygou s ≥ mAsymptoma tic HbE homozygou s

15 Hb Lepore Trait

16 34/M, Kolkata

17 Heterozygous E thalassemia

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

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

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

21 Recently transfused c/o homozygous beta thalassemia

22 Case 2: 28/F, Severe anemia

23 Borderline HbA2 levels: Silent carrier of thalassemia β thal with superadded IDA Macrocytosis αβ thalassemia

24 Borderline elevated HbA2 due to macrocytosis

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

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

27 Double heterozygous for HbE and β thalassaemia

28 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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