haemoglobinopathy cases on hplc dr. archana vazifdar head pathologist hindlabs, hll lifecare ltd
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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
