A simple, precise and rapid high-performance liquid chromatography (HPLC) method has been developed and validated for the simultaneous determination of hexamidine di-isetionate (HMI), chlorhexidine digluconate (CGH) and chlorocresol (CLC) in pharmaceutical preparations. The chromatographic conditions comprised of a reversed-phase C18 column (250 x 4.6 mm), 5 μ with a mobile phase consisting of a mixture of acetonitrile : phosphate buffer solution in the ratio (35 : 65). Flow rate was 1.3 mL min-1. Detection was carried out at 220 nm. Results have shown good separation of the three components: HMI (tR= 2.11 min), CGH (tR= 3.25 min) and CLC (tR= 7.15 min). The linear regression analysis data for the calibration plots showed good linear relationship with limit detection of hexamidine di-isetionate, chlorhexidine digluconate and chlorocresol is 0.18, 0.15 and 0.28 µg, respectively. Limit of quantitation, correlation coefficient, slope and intercept were studied. The method was validated for precision, recovery, ruggedness and robustness. As the method could effectively separate the determination of each in composite form, it can be employed as a stability-indicating one. Egypt. J. Appl. Sci., 25 (2010) 7-11.