Ultrasonic bath 5.7l

Iva-BSA-NPs were produced by the antisolvent method with some modifications^{20 , 30 (link), 31} . Initially, BSA and Iva with mass ratios of 20:2, 25:2 or 30:2 were dissolved in ultra-pure water and dimethyl sulfoxide (DMSO), respectively. The volume ratio of aqueous phase to organic phase was set at 3:1. The nanoparticles were obtained by a continuous dropwise addition of Iva solution into BSA solution at a constant rate of 1.0 mL/min under an ultrasonic dispersion (Ultrasonic Bath 5.7L, Thermo Fisher scientific Inc.). After the antisolvent process, glutaraldehyde solution (8.0%, v/v) was added into the nano-suspension at a volume ratio of 1:100 with magnetic stirring (400 rpm) for 24 hours at ambient temperature to fully crosslink the amino groups of BSA. Finally, the resulting Iva-BSA-NPs were obtained by centrifugation (16,000 g for 20 min), and were washed three times with ultra-pure water to eliminate free BSA, Iva and excess glutaraldehyde.

Iva-Col-NPs were produced by modifying the antisolvent precipitation-ultrasonication method (Abdelbary et al., 2015 (link)). Briefly, 0.75 mL of 67 mg/mL ivacaftor DMF solution was slowly added to 10 mL aqueous solution at a constant rate of 1.0 mL/min under an ice bath and ultrasonic dispersion at 930 W (Ultrasonic Bath 5.7L, Thermo Fisher Scientific Inc.). The concentrations of ivacaftor, stabilizers and colistin are listed in the Table 1. Fig. 2 shows the manufacturing process of Iva-Col-NPs.