CM-Cellulose as an ion-exchange medium with Bis-tris as buffer and a gradient of sodium chloride provides a versatile system for the chromatography of hemoglobins. Changes in pH, Bis-tris concentration, and slope of the sodium chloride grandient provide means for markedly altering chromatographic behavior for special separations. Examples are given of the application of the method to normal samples and to those with hemoglobinopathies.