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Hemoglobin Electrophoresis

Hemoglobin (Hb) electrophoresis is probably the most useful laboratory method for separating and measuring normal and some abnormal Hb. Through electrophoresis, different types of Hb are separated to form a series of distinctly pigmented bands in a medium (cellulose acetate or starch gel). Results are then compared with those of a normal sample.

Hb A, A2, S, C, and F are routinely checked, but the laboratory may change the medium or its pH to expand the range of this test.

Purpose

• To measure the amount of Hb A and to detect abnormal Hb
• To aid diagnosis of thalassemia
• To aid diagnosis of sickle cell disease or trait.

Patient preparation

• Explain to the patient that this test is used to evaluate Hb.
• Tell him that a blood sample will be taken Explain who will perform the venipuncture and when.
• Reassure him that drawing a blood sample will take less than 3 minutes.
• Explain that he may feel slight discomfort from the tourniquet pressure and the needle puncture.
• If the patient is an infant or child, explain to the parents that a small amount of blood will be taken from the finger or earlobe.
• Inform the patient that food or fluids need not be restricted before the test

Procedure and posttest care

• Perform a venipuncture, and collect the sample in a 7-ml lavender-top tube.
• For very young children, collect capillary blood in a microcollection device.
• If a hematoma develops at the venipnncture site, apply warm soaks.

Precautions

• Completely fill the collection tube, and Invert it gently several times to mix the sample and the anticoagulant. Do not shake the tube vigorously.
• Handle the sample gently to prevent hrmolysis.

Reference values

In adults, Hb A accounts for 95% to 98% of all Hb; HbA2 1.5% to 3%; and HF, less than 2%. In neonates, Hb F normally accounts for half the total. Hb S and Hb C are normally absent.

Interfering factors

• Failure to fill the tube completely, to use the proper anticoagulant, or to adequately mix the sample and the anticoagulant.
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• Hemolysis due to rough handling of the sample.
• Blood transfusion within the past 4 months.

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