A Simple Method for Detection of Chain Anomaly of Abnormal Hemoglobin in Hemolysate without Preliminary Purification

A new simple and rapid method for determining chain anomaly of abnormal hemoglobin was invented. Hemolysate, without preliminary separation and purification of the abnormal hemoglobin, is treated with p-chloromercuribenzoic acid (PCMB) in acidic solution, and subjected to starch gel electophoresis (pH 8.56). As control materials the hemolysate (of the same hemoglobinopathy) without PCMB treatment and the PCMB-treated normal hemolysate are applied to the same starch gel. Three main groups of electrophoretic stripes appear in the PCMB-treated normal hemolysate: (1) the ”anodal” representing β chain, (2) the ”intermediate” referring to undissociated Hb A, and (3) the ”cathodal” referring to α chain. In the PCMB-treated hemolysate of hemoglobinopathy another group of stripes, namely the ”new”, makes its appearance in addition to the three groups. The chain anomaly of the abnormal hemoglobin is presumed by examining the electrophoretic migration of the ”new” in comparison with those of the ”anodal” and the ”cathodal”. The criteria of the presumption is summarized as follows. If the relevant abnormal hemoglobin is fast-moving electrophoretically, the ”new” possessing the migration intermediate of the ”anodal” (β chain) and the ”cathodal” (α chain) indicates α chain anomaly, and the ”new” migrating more closely to the anode than the ”anodal” (β chain) referrs to β chain anomaly. If the abnormal hemoglobin is slow-moving, the ”new” situating more closely to the cathode than the ”cathodal” (α chain) is indicative of α chain anomaly, and the ”new” migrating between the ”anodal” (β chain) and the ”cathodal” (α chain) is suggestive of β chain anomaly.