Objective: The present study was aimed to prepare and evaluate niosomal formulations for controlled delivery of an antiretroviral drug; tenofovir disoproxil fumarate (TDF). Methods: Niosomes were prepared by using various non-ionic surfactants (span-20, span-60 and span-80) and cholesterol in different ratios by ether injection technique to achieve increased permeation of drug at the particular site of action which significantly reduces dosage frequency hence increase patient compliance. The prepared vesicles were evaluated for entrapment efficiency, vesicle size, zeta potential, surface morphology by transmission electron microscopy (TEM) and in-vitro release. Results: TEM results confirmed that, the niosomes formed were white and spherical in shape and have a definite internal aqueous space with uniform particle size. A formulation F5 composed of span 60 and cholesterol (2:1) gave the most advantageous entrapment (92.46±1.62%) and slower release results after 8 hrs (Q8h=55.35±1.93%) as compared to other compositions. Conclusion: Thus, the niosomes may be considered as a promising carrier for the controlled delivery of tenofovir disoproxil fumarate (TDF).