To prepare niosomes containing rifampicin were prepared using various nonionic surfactants of sorbitan ester class and cholesterol in 50:60 (1:1.2) percent mol fraction ratios for sustain release. To improve the dissolution rate of noisome are prepare handshaking method using Surfactants and cholesterol (150 μmol) in 50:60 (1:1.2) percent mol fraction ratio. The percent of drug estimated to be entrapped was noted to decrease progressively for various sorbitan esters used in the order of Span-85>Span-80>Span-60>Span-40>Span-20. In vitro release rate studies revealed that the cumulative percent rifampicin released was maximum for Span-20- based niosomes and minimum for Span-85-based niosomes. The handshaking method is a simple and efficient technique for designing functional niosomes for hydrophobic or amphiphilic drugs.