Didanosine was mixed, incubated for 1 h at room temperature. Ethanol was added dropwise at a rate of 1 ml/min into the BSA solutions as a desolvating agent until the solutions became just turbid. Thereafter 30 min of the desolvation process, 100 μl of an 8% v/v aqueous solution of glutaraldehyde was added to induce particle cross linking. This process was performed during stirring over a time period of 3 h at room temperature. The nanosuspensions were purified by two cycle centrifugation at 20,000 rpm for 30 min and then subjected to freeze drying after adding 2% (w/v) mannitol as a cryoprotectant for 8 h to obtain fine powder of nanoparticles. The dried nanoparticles obtained were then transferred

to vials and were stored at 4 °C. Coating was done for D1 immediately after cross linking by adding 1% polysorbate 80 and was http://www.selleckchem.com/products/wnt-c59-c59.html Selleck NVP-BGJ398 incubated for 30 min, as per the procedure described by Amit Bansal et al. Finally the nanosuspensions

was centrifuged and lyophilized with 2% mannitol. Compatibility of ddi and BSA, were analyzed using FT-IR (Fourier transform infrared) spectroscopy, Shimadzu Corporation, Japan by the potassium bromide disc method (1:100). The DSC (differential scanning calorimetry, MettlereToledo star 822 systems, Switzerland) thermogram of drug and lyophilized nanoparticles gives information regarding the physical properties and melting point of the drug. Scanning electron microscopy was performed to characterize the Sclareol surface morphology of the prepared nanoparticles to detect their morphological character of nanoparticles. This was done by placing freeze dried nanoparticles on brass stub then were gold-coated to render them electrically conductive and examined under the Scanning Electron Microscope at 20 kV (JSM 6100 JEOL, Tokyo, Japan). The particle size and zeta potential of didanosine albumin nanoparticles was determined by dynamic light scattering, using a Malvern system, with vertically polarized supplied by Helium/Neon laser (red laser) operated at 4 mM, 633 nm. The samples were dispersed in distilled water and taken in clear disposable zeta cell. The experiments were

performed with non-invasive backscatter technology at a temperature of 25.0 ± 0.1 °C at a detection angle of 173° to the incident beam. Freshly prepared nanosuspensions were centrifuged at 20,000 rpm for 30 min and the amount of unincorporated didanosine in supernatant liquid was measured. The % entrapment efficiency (EE) and % drug loading were calculated according to the following formula. %EE=[Amountofdrugactuallypresentinnanoparticles/amountofdrugactuallyadded]×100 %Drugloading=[(Totalamountofdrugadded−amountofunbounddruginsupernatantliquid)/totalamountofdrugadded]×100 The dug release studies were carried out by dialysis method. A known quantity of nanoparticles equivalent to 10 mg of the drug was taken in a cellulose dialysis bag (molecular weight cut off 5 kDa, Himedia, India) and added 5 ml of pH 7.4 phosphate buffer.