Electropheresis can be used to separate different hemoglobins based on their electrical charge, which can be useful in the diagnosis of the different hemoglobinopathies.
(1) electropheresis at alkaline pH (cellulose acetate at pH 8.6)
(2) electropheresis at acid pH (citrate agar at 6.2)
Key bands on electropheresis at alkaline pH
point of application
C, C-Harlem, E, O
S, D, G
Key bands on electropheresis at acid pH
E, D, G, Lepore, Barts
(1) There is some variability between the sources for the band patterns, especially for electropheresis at acid pH.
(2) The precise location of S and O hemoglobins relative to the application point at acid pH appears to vary with conditions, with some showing the point of application between O and A1, while others between S and O.
(3) Hemoglobin E may be masked by A2 in both acid and alkaline conditions.
(1) solubility tests to confirm hemoglobin S
(2) hemoglobin A2 by DEAE cellulose column
(3) alkali denaturation of hemoglobin F
(4) testing for iron deficiency
If iron deficient, repeat testing after iron replacement complete
• Iron deficiency can mask beta-thalassemia.
0-3% in adults
< 2% F in adults
> 60% F during 1st week; no abnormal bands
normal or increased
microcytic despite normal iron stores
34-42% S; positive solubility test
AS + alpha thalassemia
< 34% S; positive solubility test; microcytic
AS + beta thalassemia
> 34% S; positive solubility test; microcytic
none unless transfused
predominantly S; positive solubility test
AC + alpha thalassemia
< 30% C; microcytic
AC + beta thalassemia
> 30% C; microcytic
AE + alpha thalassemia
< 30% E; microcytic
AE + beta thalassemia
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Specialty: Hematology Oncology, Clinical Laboratory