Background: Toluenediamines are harmful substances. Toluene-2,4-diamine has been assigned to Carcinogen 1B hazard class, pursuant to Regulation (European Community – EC) No. 1272/2008 of the European Parliament and of the Council, and toluene- 2,6-diamine to Mutagen 2 hazard class. The main routes of exposure to toluene-2,4-diamine and toluene-2,6-diamine are via the respiratory tract and the skin. Toluene-2,4-diamine and toluene-2,6-diamine occur in the work environment in Poland. The aim of this study was to develop and validate a method for the determination of toluene-2,4-diamine and toluene-2,6-diamine that allows the simultaneous determination of their concentrations in the workplace air by personal sampling. Material and Methods: Determination of toluene-2,4-diamine and toluene-2,6-diamine derivatives in acetonitrile were carried out by means of liquid chromatography with a diode assay detector. The method involves passing amine-containing air through sulfuric acidtreated glass fiber filter, washing out the substance settled on the filter, using water and solution of sodium hydroxide, followed by the extraction with toluene, reaction with 3,5-dinitobenzoyl chloride, replacement of dissolvent with acetonitrile and analysis of obtained solution. Results: The method developed in this study enables the researcher to determine the content of toluene-2,4-diamine and toluene-2,6-diamine in the presence of other hazardous substances. In the specified measuring range (2.88–57.6 μg/ml) calibration curves are linear. Under the optimized conditions of determination, the limit of detection (LOD) values achieved: 51.36 ng/ml for toluene-2,4-diamine and 52.93 ng/ml for toluene-2,6-diamine. Conclusions: This method makes it possible to determine the concentration of toluene-2,4-diamine and toluene-2,6-diamine in the workplace air within the specified measuring range of 0.004–0.08 mg/m³ (for air sample volume of 720 l). Med Pr 2017;68(4):497–505