Human serum albumin (HSA), a versatile protein carrier for drug delivery, is an ideal material to fabricate nanoparticles for drug delivery systems. These nanoparticles can accumulate in tumor interstitium due to the enhanced permeability and retention effect. The most important characteristics of HSA nanoparticles are particle size, shape, and zeta potential. A chemometric approach was applied for HSA nanoparticles’ size optimization in this study. The effects of three experimental parameters; pressure (P) or power, organic solvent volume (V), and time (T), were investigated under sonication and high-pressure homogenization, using multivariate analysis. The trials were performed based on the Box–Behnken experimental design. The criteria for the appraisal of the descriptive ability of a multinomial were R2 = 0.819, standard error = 20.420, and F-ratio = 19.550. The method was optimized with respect to the nanoparticles’ size as a response. The Box–Behnken experimental design was applied to optimize and trial the robustness of the HSA nanoparticle preparation method.
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