The quantity of chitosan adsorbed to the surface of NPs was determined by the fluorescamine assay. Chitosan coated NPs were ultracentrifuged at 80,000 × g and the supernatant was obtained. A 50 µL aliquot of fluorescamine in DMSO (2% w/v) was added to 20 µL of the supernatant sample in wells of a 96-well plate. The reaction continued for 3 h in the dark and the fluorescence in the wells were measured at 390 nm excitation and 515 nm emission, using a microplate reader (CLARIOstar® BMG Labtech, Germany). A standard curve was prepared for each type of chitosan by spiking blank supernatant with predefined quantities of the chitosans. The adsorption was calculated by the difference between the initial amount of chitosan and the amount detected in the supernatant

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