The influence of different application methods on UV protective properties of white and dyed cotton functionalized with ZnO nanoparticles (nano-ZnO) was investigated. The methods differ in application procedure, time of treatment and auxiliaries used in the treating bath. The ultraviolet protection factor (UPF) was determined for untreated and functionalized samples. The presence of nano-ZnO on fibres was investigated using scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR). The content of Zn was determined with energy-dispersive X-ray spectroscopy (EDS) and inductively coupled plasma mass spectrometry (ICP-MS). Dynamic light scattering (DLS) was used for particle size measurements in the prepared solutions. The results show that UV protection of cotton increases with a higher content and uniform distribution of nano-ZnO on the samples and that dyeing increases the loading capacity of cotton towards nano-ZnO. One of the methods (Method IV) gave remarkable results giving cotton an excellent UV protection whether it was dyed or not.