Investigations on the resolving power of hydroxyapatite columns

An investigation on the resolving power of hydroxyapatite columns is reported. The macromolecules chosen for this investigation have been five proteins endowed with a rigid structure and of different sizes: cytochrome c, lysozyme, β‐lactoglobulin A, collagen, and T2 phage. The following points have been investigated in detail: (1) the dependence of the elution molarity upon column length and slope of the gradient; (2) the dependence of the elution molarity and the width of the protein peak upon the load and the presence of other chromatographic components; (3) the optimal conditions for the resolution of macromolecules having the same size or different sizes.