Preparation of silibinin NPs

Preparation of silibinin NPs through the APSP method was carried out using a previously reported method,12 with a slight modification. Briefly, a saturated solution of unprocessed silibinin was prepared in ethanol. A syringe was filled with the prepared drug solution and was quickly injected at a fixed flow rate (2 mL/min) into deionized water (antisolvent) of definite volume under mechanical stirring (3,000 rpm). Different volumes of deionized water were mixed with the same volume of saturated drug solution (1:10, 1:15, and 1:20, v/v), which was evaporated quickly in a vacuum using a rotary evaporator.

For the EPN method, a solution of the unprocessed silibinin was prepared in ethanol followed by quick addition of hexane, which served as an antisolvent, to the prepared saturated drug solution under mechanical stirring (3,000 rpm). Different solvent: antisolvent ratios (1:10, 1:15, and 1:20, v/v) were used. Nanosized drug particles from the nanosuspensions were obtained by quick evaporation of the solvent and antisolvent in a vacuum for both methods (APSP and EPN).