In HPLC method, Methanol and Water (75:25 % v/v) was used as mobile phase, at a flow rate of 1.0 ml/min, on HPLC system containing UV- detector with Openlab EZchrome software and Kro-masil C18, 250 mm X 4.6 mm, 5 µm. The detection was carried out at 258 nm. The method gave suitable retention time i.e. 3.60 min for chlorohexidine. The results of analysis in the method were validated in terms of Filter study, Solution stability, specificity, Linearity, accuracy, precision (Repeatability and intermediate precision), limit of detection, limit of quantification and robust-ness. A simple and precise method was developed for the assay of chlorohexidine in bulk drug and in tablet formulation. The method needs regular reagents for doing analysis and also less time consuming, it can be per-formed routinely in industry for routine analysis of bulk drug and marketed product of chlorohexidine.