Identification of Abnormal Hemoglobins Ⅵ．Separation of Peptides by Reversed Phase High Performance Liquid Chromatography． Application to a New Case of Hb M Iwate

Reversed phase high performance liquid chromarography on a C18 column (чBondapak,Waters) was applied to the identification of Hb M Iwate found in a newborn girl. Theabnormal αchain was retained in the column even longer than the　βchain, and well separated peaks for the normal αβabnormal α,and chains were eluted in sequence by the A/B solvent system of Huisman et al. It was co-eluted with the chain by C/D solvent system of Shelton et al. The normal and abnormal αT9s were easily purified by a single peptide mapping of the tryptic hydrolysate of the total α chain mixture. Each of them was hydrolyzed with chymotrypsin, and the products were purifiedby peptide mapping on the same column. The father and grandfather of the baby carried the same abnormal hemoglobin. The abnormal α chain comprised about 24%of the total α chain in the three carries.

High performance liquid chromatography

Abnormal hemoglobin