Limitations of the ultracentrifugal approach for measuring the effective net charge of a macroion.

Limitations have been detected in a recently published method for macroion valence determination by an ultracentrifugal procedure for quantifying the Donnan distribution of small ions in macroion solutions dialyzed against buffer supplemented with chromate as an indicator ion. The limitation reflects an implicit assumption that sedimentation velocity affords an unequivocal means of separating effects of chromate binding from those reflecting the Donnan redistribution of small ions. Although the assumed absence of significant Donnan redistribution of small ions across the sedimenting macroion boundary seemingly holds for some systems, this approximation is demonstrably invalid for others. Despite preliminary signs of promise, the ultracentrifugal procedure does not afford a simple, readily applied solution to the problem of unequivocal macroion valence determination.