Niclosamide is a member of the class of salicylanilides. The chemical formula is C13H8Cl2N2O4. The HPLC analysis was performed using the Thermo Ultimate 3000 HPLC system. In a 60 ml round bottom flask, add 5 g of niclosamide to 15 ml ethanol and 20 ml H2O The solubility was checked at room temperature using the HPLC analysis procedure described below. To this solution add with continuous stirring 12 g KOH pellets. Stirr until dissolved (about 10 min) then heat the reaction mixture to 50 ° C while continuing to stir for 3 hours.

After 3 h at 50 ° C, let the reaction mixture cool down to rt over night. Filter the precipitate material by vacuum filtration system through sintered filter funnel into a 100 mL sep funnel collecting the solid residue in a beaker. Rinse back funnel and filter Weigh residues if performing assay quantitatively or collect for immediate use if assaying qualitatively by HPLC as described below: If necessary, add a few drops of water to stop formation of salt crystals to the evacuated flask.

HPLC analysis was performed using an Agilent 1100 series system equipped with a Zorbax SB-C18, 4.6 mm x 250 mm analytical column thermostatted 25 ° C., and UV detector set at 254 nm). The integration times were 1 sec (linear gradient 0). ).). -4 min 30% A / B, 5-20 min 20% A / B, 20-25 min 30% A / B, 25-34 min 40% A / B and 34-39 min 50% A / B) using the following mobile phase: 2.5 mM ammonium acetate in water (Solvent A): formula: 0.2% trifluoroacetic acid (Solvent B). The injection volume was 10 μL and the flow rate was 1.4 ml / min.