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Identification of hemoglobin variants by HPLC.

We read with interest the recent article by Joutovsky et al. (1) in which the authors evaluated ion-exchange HPLC for identification of hemoglobin (Hb) variants. The authors concluded that “confirmatory testing by electrophoresis can be eliminated in the majority of cases by use of retention time, proportion of total hemoglobin, and peak characteristics of HPLC.” We disagree with their conclusion, which was based on the evaluation of only 34 Hb variants.

A recent search of the hemoglobin variant database (http://globin.cse.psu.edu) showed that there are currently 482 known β-chain variants and 297 variants of either the α1- or α2-globin chain. Our laboratory has extensive experience in the evaluation of Hb variants by HPLC (Table 1⇓ ). All 268 variants included in Table 1⇓ were evaluated by multiple methods and were confirmed by DNA sequencing …