Further Experiments in the Separation of Globin Chains by High Performance Liquid Chromatography

Abstract

The applicability of various column packings, solvents, and gradients for the separation of globin chains by high performance liquid chromatography has been examined. Although numerous combinations may be applied to advantage in special situations, a particularly effective system uses a Waters μBondepak C₁₈ packing with a slight gradient of perchlorate-phosphate-methanol-acetonitrile mixtures. A chromatogram with human β, α, ^Aγ^T, ^Gγ, and ^Aγ^I chains is finished in 80 min, and quantitation of the various chains is possible.