A comprehensive analysis of hemoglobin variants by high-performance liquid chromatography (HPLC).
BACKGROUND
High-performance liquid chromatography (HPLC) is a method commonly used for the detection of hemoglobin (Hb) variants. In addition to providing precise quantitation of Hb A2 and Hb F, the reported retention time and peak shape of a high number of hemoglobin (Hb) variants are very helpful for presumptive identification. However, there is a scarcity of summarized data in the literature of the mobility of Hb variants on this method.


METHODS
A total of 383 Hb variants were studied on the Bio-Rad Variant (™) Classic HPLC instrument. Hb variant identification used a number of methods, including confirmation by DNA sequencing in at least one case for all alpha and beta chain Hb variants.


RESULTS
Retention time data and the number of occurrences of each Hb variant were obtained. This showed that rare Hb variants can have similar retention times to the five most common alpha or beta chain Hb variants.


CONCLUSIONS
HPLC is a very powerful tool in the evaluation of Hb variants, particularly when combined with other methods. However, it should not be used as a stand-alone method for definitive identification of Hb variants.
Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, MN, USA.
J
J
Szuberski
Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, MN, USA.
J L
JL
Oliveira
Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, MN, USA.
J D
JD
Hoyer
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D014644:Genetic Variation; D006455:Hemoglobins, Abnormal / Q000032:analysis* / Q000235:genetics; D006801:Humans; D015203:Reproducibility of Results; D017422:Sequence Analysis, DNA; D055542:alpha-Globins / Q000032:analysis* / Q000235:genetics; D055544:beta-Globins / Q000032:analysis* / Q000235:genetics
D006455:Hemoglobins, Abnormal; D055542:alpha-Globins; D055544:beta-Globins