HAEMOGLOBINOPATHY CASESon HPLC

Dr. Archana VazifdarHead Pathologist

Hindlabs, HLL Lifecare Ltd.

HPLC Automated system precalibrated column and gradient

Direction of flow Detector

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

CHROMATOGRAMS

Output

Time

Peak

RT (min)

Area

Total Area Count: 1-3 million

HbF: 1-2% RT: 1.03-1.13HbA2: 1.75-3.25% (2-3.6%) RT: 3.63-3.64

P2- Glycemic status, upto 6% acceptableP3- 6% acceptable, 6-15% deterioration

15-25% HbJ

8 mnth/M, pallor, failure to thrive

Homozygous beta thalassemia

22/Female

Heterozygous Beta thalassemia

Mother of 8 mnth old child

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

HbA2%

2-3.6

HbF%

≤ 2

Hb

N

MCV

N

Symptoms

-

FINAL

NORMAL

4-10 ≤ 2/ m Borderline

Asymptomatic

β THAL TRAIT

10-18 2- 10 N N Asymptomatic

Hb LEPORE TRAIT

25-35 ≤ 1 N N Asymptomatic

HbE TRAIT

40-48 ≤ 2 N N Asymptomatic

HbD IRAN

Heterozygous

50-60 ≥ 10 SevereHbE/Hbβ double

heterozygous

≥ 60 2-10 m Asymptomatic

HbE homozygo

us

Hb Lepore Trait

34/M, Kolkata

Heterozygous E thalassemia

• All criteria fit provided there is NO history of recent blood transfusion

• ALWAYS CORRELATE with clinical history with CBC & peripheral blood picture

• Degenerated sample

Hb 7MCV 84 MCH 28MCHC 32.4RDW 20.2

Case 1: 2 yr/M, anemia, hepatosplenomegaly

Elevated HbF:

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

Recently transfused c/o homozygous beta thalassemia

Case 2:28/F, Severe anemia

Borderline HbA2 levels:

•Silent carrier of thalassemia•β thal with superadded IDA•Macrocytosis•αβ thalassemia

Borderline elevated HbA2 due to macrocytosis

P3: 6% acceptable, 6-12% deterioration15-25% HbJ

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

Double heterozygous for HbE and β thalassaemia

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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