Freeze-drying has been considered as an advanced and reliable technique to improve the long-term stability of colloidal nanoparticles. Stavudine loaded solid lipid nanoparticles (SLN) suitable for parenteral administration were converted into dry product. The lipid matrix Trimyristin (2.0 w/w %) was stabilized using combination surfactant system. The protective effect of various types and concentrations of cryoprotectants (e.g. carbohydrates, polymers etc) was studied on stability of the particle size. The sugar trehalose proved to be most effective in preventing particle growth during freeze-drying process. Changes in particle size distribution during lyophilization could be minimized by optimizing the right concentration of cryoprotectants. Reconstituted lyophilized stavudine lipid nanoparticles were suitable for i.v. injection with regard to the size distribution. The mean particle size of freeze-dried lipid nanoparticles after reconstitution was significantly increased in comparison to that of the preparations before freeze-drying.