Electrophoretic measurement of water charge density and ion hydration

Water exchange between bulk water and water-ion complexes will be at equilibrium when the charge density of the complex surface equals the charge density of bulk water, producing a constant radius water-ion complex. This complex will migrate in an electric field at a velocity proportional to the complex radius. CE velocity is the sum of the complex charge-dependent velocity and the buffer electro-osmotic flow. Simultaneous use of both a base (1.07 mM imidazole) and an acid (1.5 mM MOPS) buffer negates EOF at pH 7.4. Electric fields below 300 V/cm (potassium, calcium) and 400 V/cm (magnesium) yield migration velocities with no dehydration of the water-ion complexes. The number of waters per complex increase with the ion charge density: K+ 1.90, Ca++ 5.90, Mg++ 6.59 waters/ion. The charge densities of the complexes are similar: K+ 1.24, Ca++ 1.43, Mg++ 1.21 e/nm(2), for an average bulk water charge density of 1.29 +/- 0.11 (SD) e/nm(2). The addition of 0.1% Triton increases the number of waters for Mg++ to 25.33 and lowers the charge density to 0.497 e/nm(2). High electric field dehydration shows that calcium will be fully dehydrated at 638.3 V/cm and magnesium fully dehydrated at 925.5 V/cm, which occur at 6.15 and 5.78 nm from the membrane. Dehydrated magnesium will then bind to calcium channels leading to decreased smooth muscle activation.