Optimal conditions for the reversed-phase chromatography of proteins

Summary Optimum stationary phases for reversed phase chromatography should be based on a silica with a large average pore diameter, preferably above 30nm. To exclude anormalous elution behaviour of proteins, octadecyl groups should be bonded to the surface and the residual silanol groups should be reacted with a highly active silanization agent like bistrimethylsilyl acetamide. The total amount of bonded carbon should not exceed 5% w/w. Because of the low diffusion coefficients of proteins, the particle diameter should be as small as possible. Protein retention can hardly be influenced by changes of the organic modifier or by temperature. Because of solubility and viscosity, however, acetonitrile is to be prefered. Presented at the 15th International Symposium on Chromatography, Nürnberg, October 1984