A stability-indicating HPLC analytical method for the anticancer agent dibromodulcitol (DBD, Mitolactol, NSC-104800) has been developed that will completely resolve the compound from its degradation products. A 5 μm octadecylsilane analytical column was used in conjunction with a refractive index detector, with a mobile phase of 98:2 water/methanol. The limit of DBD detection was determined to be 250 ng (5 μg/ml) with a signal to noise ratio of 2:1. Intraday variation of the method, as percent relative standard deviation, was 4.37% (12.5 μg/ml) and 0.973% (250 μg/ml), and interday variation was 3.93% (250 μg/ml). Comparison with potentiometric titration of bromide after digestion of DBD with NaOH indicated that the method was more sensitive and specific than titration. The method has been used in tablet content analysis, as well as degradation studies of DBD in solution.