Abstract Salicin, a glycoside isolated from Salix Alba, has provided us with most potent weapon, Acetyl-salicylic acid for killing pain. Separation of salicin was achieved on Agilent HPLC with Aclipse XDB-C18, 5 μm and 150mm X 4.6mm, injection volume 5 μl at flow rate of 1ml/min by using a binary pump system. The ratio of mobile phase was 80 ml: 20ml (Acetonitrile: Water). The linearity was determined by six different concentrations of Salicin in triplicate and calibration curve was plotted in range of 5-30 μg/ml. The Coefficient of determination (r2) of salicin was 0.998 for HPLC with best resolution and sensitivity at the absorption maxima of 253nm. The average percentage Recovery of Salicin in Ayurvedic preparation was found to be 100±1%. Specificity of the method was determined and the method was specific for estimation of Salicin in the Ayurvedic preparation. The observations and results obtained for each validation parameter including specificity, method precision (repeatability), intermediate precision (ruggedness), linearity and accuracy lie well within the acceptance criteria.
Keywords Salicin, Salix Alba, HPLC, Validation