A sensitive and specific high-performance liquid chromatographic-tandem mass spectrometric (HPLC-MS-MS) method was developed for the determination of 3-hydroxypropylmercapturic acid (3-HPMA) in human urine. Samples were extracted using ENV+ cartridges and then injected onto a C8 Superspher Select B column with acetonitrile and formic acid as eluent (5:95, v/v). N-Acetylcysteine was used as internal standard for HPLC-MS-MS. Linearity was given in the tested range of 50-5000 ng/ml urine. The limit of quantification was 50 ng/ml. Precision, as C.V., in the tested range of 50-5000 ng/ml was 1.47-6.04%. Accuracy ranged from 87 to 114%. 3-HPMA was stable in human urine at 37 degrees C for 24 h. The method was able to quantify 3-HPMA in urine of non-smokers and smokers.