Excited state proton transfer (ESPT) of pyranine (8-hydroxypyranine-1,3,6-trisulfonate, HPTS) in a niosome is studied by fluorescence correlation spectroscopy (FCS) and femtosecond up-conversion. The niosome consists of a neutral surfactant ant triton X-100 (TX-100) and cholesterol. FCS studies suggest that in the presence of niosome almost all of the HPTS is transferred to the niosome and the amount of free HPTS present in bulk water is negligible. The time constant of initial proton transfer (tau(PT)) in niosome (40 ps) is similar to 8 times slower than that (5 ps) in bulk water, while the time constants of recombination (tau(rec)) and dissociation (tau(diss)) are similar to 4 times and similar to 1.5 times slower in niosome, respectively. On addition of NaCl, the rate of ESPT is markedly retarded both in free water and in niosome. In the niosome, tau(PT) slows down to 80 ps in 1 M NaCl and 225 ps in 4 M NaCl.