Firstly, an ethanol solution of RhoB was prepared (2.25 μmol L-1) and aliquots of this solution were diluted with ACN in volumetric flasks. Calibration curves were constructed within a range of 0.108 to 0.539 μmol L-1. A fixed concentration of the product 1 (0.152 mg mL-1) was maintained in all samples used to construct the calibration curve. The fluorescence intensity (I f) was measured using a rectangular cuvette (Hellma Quartz Suprasil®, 10 mm, Sigma-Aldrich) with the maximum excitation (λ max-ex) and λ em wavelengths observed for the product 1. The

I f was plotted as a function of the molar concentration of rhodamine B. The linear coefficient value for the linear regression corresponded to the amount of RhoB presented in purified product 1. The experiment was Dactolisib datasheet replicated three times. Preparation of the fluorescent nanocapsules The fluorescent-labeled polymeric nanocapsules Entospletinib solubility dmso were prepared by the solvent displacement method [8, 24]. The polymers Eudragit RS100 and Eudragit S100 were used to prepare the nanocapsule formulations NC-RS100 [25] and NC-S100 [26], respectively, and the polymer poly(ϵ-caprolactone) (PCL) was used to obtain the lipid-core nanocapsule formulation LNC-PCL [27]. To prepare

the nanocapsule formulations (NC-RS100 and NC-S100), an organic phase (27 mL of acetone), containing the polymer (100.0 mg), CCT/product 1 (9:1, w/w) (333 μL), and sorbitan monooleate (76.6 mg) (except for NC-RS100), was injected using moderate stirring into a polysorbate 80 aqueous phase (76.6 mg in 53 mL). The organic solvent was removed by evaporating the suspension under reduced pressure.

The suspension was evaporated until a final volume of 10 mL. The LNC-PCL formulation was obtained by the same procedure. Rho However, in this case, the organic phase was Adriamycin datasheet composed of the polymers, PCL116 (90.0 mg) and PCL14 (10.0 mg), CCT/product 1 (9:1, w/w) (160 μL), and sorbitan monostearate (40.0 mg) dissolved in acetone (27 mL). Three batches of each formulation were prepared. Characterization of the fluorescent-labeled nanocapsules The pH of the formulations was measured without dilution of the suspensions using a potentiometer, model B474 (Micronal, Brazil). Laser diffraction analysis was performed with a Malvern Mastersizer® 2000 instrument (Malvern Instruments, Worcestershire, UK) and used to determine the particle size distribution profile, volume-weighted mean diameter (D 4.3), and polydispersity (SPAN). Photon correlation spectroscopy (PCS) was used to characterize the nanometric population by determining the average diameter (z-average) and polydispersity index. Electrophoretic mobility (EM) analysis was performed to determine the zeta potential values.