4.6. Particle Size Distribution, PDI, and ζ-Potential

The two generations of nano conjugates were tested to characterize the physicochemical properties, which include particle size distribution, zeta potential, and PDI. The size of particles was assessed according to light scattering and the data were analyzed by an attached software, giving a measure of distribution of particle size. The particle size was measured at neutral pH (water). For the measurement, one mL of each formulation was diluted 100 times with water and subjected to particle size measurement. The ζ-potential was measured at PBS pH 5.4 and 7.4. For ζ-potential measurement, one mL of each formulation was diluted 100 times with each buffer and subjected for the ζ-potential measurement. ζ-potential determined the net charge presented on particles, which were moving in an electric field [42].