Rapid, precise, accurate, and reproducible methodology using ultra-high-pressure liquid chromatography (UHPLC) for the analysis of heroin and basic impurities is described. The determination of heroin, morphine, O3-monoacetylmorphine, O6-monoacetylmorphine, codeine, acetylcodeine, noscapine, and papaverine is accomplished using reversed-phase chromatography (RPC), employing a 1.7 μm Acquity UPLC BEH C18 column (2.1 mm × 150 mm) with a phosphate buffer (pH 1.6)-acetonitrile gradient and PDA detection. The target analytes are well resolved from each other and most adulterants in less than 20 min. For the few instances when adulterants interfere with target analytes, a 1.7 μm Acquity CSH Fluoro-Phenyl (2.1 mm × 150 mm) column is utilized with the same gradient conditions. The reported methodology can detect impurities as low as 0.02% relative to heroin, and is well suited for heroin profiling.