Summary.A simple and convenient chromatographic method of simultaneous separation, identification, and quantitative determination of thimerosal (TM) (preservative) and aluminum (adjuvant) in vaccines and pharmaceuticals by reversed phase highperformance liquid chromatography (RP-HPLC) with visible (VIS) detection was developed and validated. Due to postcolumn derivatization with dithizone, any interference from matrix was excluded. Similarly, a possibility of on-column decomposition of dithizonates was eliminated. Evaluated detection limits were 0.3 μg TM and 3.0 μg Al, which correspond to the smallest, but possible to recognize, visible peak.