[Use of HPLC in different stages of characterization of abnormal hemoglobin. Application to a case of hemoglobin Grady (alpha-118 [+Glu-Phe-Thr])].

In this work the structural abnormality of Hb Grady was determined using mainly the high performance liquid chromatography (HPLC) technology. The methods described here are now easily available in an increasing number of laboratories. A small amount of the abnormal Hb (ca. 10 mg) was isolated by isoelectric focusing and its subunits fractionated by reversed phase HPLC. After tryptic or chymotryptic digestion the peptides were isolated by HPLC. Their amino-acid compositions were also determined by HPLC procedures following a precolumn derivatization.