Test Detail
Overview
Interpreting Results
FAQ's

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Written by
Dr. Betina Chandolia
BDS, MDS - Oral Pathology and Microbiology
Reviewed by
Dr. Lalit Mohan Gupta
MBBS, MD - Microbiology
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Hemoglobin (Hb) Electrophoresis by HPLC

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Hemoglobin (Hb) Electrophoresis by HPLC
You need to provide
Blood
This test is for
Male, Female
Test Preparation
  1. No special preparation required

Overview


What is Hemoglobin (Hb) Electrophoresis by HPLC?

Hemoglobin (Hb) electrophoresis is used to identify the different types of hemoglobin present in the blood. This includes hemoglobin A1 (HbA1), hemoglobin A2 (HbA2), hemoglobin F (HbF; fetal hemoglobin), hemoglobin C (HbC), and hemoglobin S (HbS). It also detects abnormal hemoglobins which may abrupt the oxygen flow.  This test is used as the first step in the screening test.

Why is Hemoglobin (Hb) Electrophoresis by HPLC done?


  • As a part of a routine checkup

  • To diagnose blood disorders

  • To monitor the treatment in patients having abnormal hemoglobin

  • As a screening test for genetic conditions



What does Hemoglobin (Hb) Electrophoresis by HPLC Measure?

Hemoglobin (Hb) Electrophoresis by HPLC test measures and identifies different types of hemoglobin in the blood. A human body contains various types of hemoglobin in their blood. With age, the percentage of hemoglobin present in the body changes. Hemoglobin is the protein which is present inside the red blood cells. Its function is to transport oxygen to the tissues and organs of the body.

When there is a change in DNA sequence commonly called a mutation, the production of hemoglobin gets affected. These type of changes cause abnormality in hemoglobin and the normal working of healthy hemoglobin gets defected. This will result in a decreased amount of required oxygen transported to tissues and organs. 

The different types of hemoglobin include:

  • Hemoglobin F - Fetal hemoglobin

  • Hemoglobin A - Adult hemoglobin 

  • Hemoglobin C

  • Hemoglobin D

  • Hemoglobin E

  • Hemoglobin M

  • Hemoglobin S

Hemoglobin F is found in growing fetuses and newborns. It is soon replaced with hemoglobin A after birth. Hemoglobin A is found in children and adults. It is the most common type of hemoglobin. Hemoglobin C, D, E, M, and S are rare types of abnormal hemoglobin which occur due to genetic mutations.



Interpreting Hemoglobin (Hb) Electrophoresis by HPLC results


Interpretations

In children 

Age

Hemoglobin F

Newborn

60 to 80%

1+ Year

1 to 2%

 

In adults

Normal levels

 

Type of Hemoglobin

Percentage

Hemoglobin A

95% - 98%

Hemoglobin A2

2% - 3%

Hemoglobin F

1% - 2%

Hemoglobin S

0%

Hemoglobin C

0%

 All the results should be carefully analyzed and interpreted in correlation with the patient's signs and symptoms and other laboratory findings. 

  • Presence of high level of sickle cell hemoglobin (HbS) on HPLC indicates either sickle cell trait or sickle cell anemia 

  • Increased levels of fetal hemoglobin (HbF) on HPLC is associated with beta-thalassemia major. Sometimes, it can also be seen in sickle cell anemia 

  • High HbA2 levels are mostly associated with beta thalassemia minor or trait. Recent studies say that HbA2 level between 4% and 9% indicates beta thalassemia trait 

  • Other less common hemoglobins which can be detected are hemoglobin C, D, E, M, etc.



Answers to Patient Concerns & Frequently Asked Questions (FAQs) about Hemoglobin (Hb) Electrophoresis by HPLC


Frequently Asked Questions about Hemoglobin (Hb) Electrophoresis by HPLC

Q. Is there any preparation required before the test?
Inform the doctor about the medications you may be taking. No other specific preparations are usually required before this test.
Q. How is the blood sample taken?
The healthcare provider takes a blood sample from the arm. The site from where the blood is to be withdrawn is cleaned with a swab of rubbing alcohol. This is then followed by inserting a small needle which has a tube attached to it for collecting blood. Once the sufficient blood for analysis is withdrawn, the needle is removed. The site is then covered with a gauze pad.
Q. Is there any risk associated with the withdrawal of blood sample procedure?
As such there is no risk but in few cases, bruising, bleeding, and infection at the puncture site can be seen. In very few cases, there can be swelling of the vein after the blood is withdrawn.
Q. How is Hemoglobin (Hb) electrophoresis by HPLC performed?
This test is performed by a process called electrophoresis. In this, an electric current is passed through the hemoglobin in the collected blood sample. This will separate the charge particles of hemoglobin into different bands. The sample is then compared with healthy samples to detect the presence of any kind of abnormal hemoglobin.
Q. In which conditions abnormal hemoglobin levels can be seen?
The abnormal hemoglobin levels can be seen in the following conditions: -> Hemoglobin C disease - It is a genetic disorder which leads to severe anemia. -> Rare hemoglobinopathy - They are a group of genetic disorders which cause the abnormal production or structure of red blood cells. -> Thalassemia - It is a condition in which there is less production of hemoglobin than what is required by the body to function properly. -> Sickle cell anemia - In this, Red Blood Cells take irregular shape which results in blocking of passage for the blood to flow.
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