Evaluating the Surface Charge of C18 Stationary Phases

The surface charge of four C18 stationary phases was investigated by measuring the flow induced streaming potential, a well known electrokinetic property of charged surfaces. Three of the stationary phases (Symmetry, Gemini, and Xterra-MS) had significantly positive streaming potentials at both pH 3 and 4.5. The fourth (Zorbax-SB) appeared to be essentially neutral at pH 3 and became negative at pH 4.5. Apparent zeta potentials ranged from approximately +16 to -4 mV. The retention behavior was also investigated using chloride as model anion and glycinamide (in its protonated form) as model cation. When the retention factor (k) of glycinamide was subtracted from k of chloride anion, the resulting delta k values showed very similar trends as apparent zeta potential values, suggesting that the simple chromatographic method could be used to estimate zeta potential values, or that the zeta potential values could be useful for ranking columns according to ion exchange or exclusion behavior. The anion exchange capacity of the Symmetry and Gemini columns was also estimated, using a published chromatographic procedure, and the results suggest about 2 microEq. capacity per gram of packing.