We report an approach for assessing human exposure to bisphenol A (BPA), which involves measuring the glucuronide in urine sample that were subjected to a novel size-exclusion flow extraction method. The present approach includes the addition of 13C(12)-BPA, enzymatic deconjugation, and the proposed sample preparation method. The sample solution is separated and detected by liquid chromatography-mass spectrometry (LC-MS). The following are used for the LC-MS: a reversed-phase separation column, electrospray ionization (ESI), negative mode, and single ion monitoring (SIM) with m/z 227 for BPA and m/z 239 for 13C(12)-BPA. The detection limit was 0.1 ng ml(-1) and the calibration curves (0.45-90 ng ml(-1)) had correlation coefficients exceeding 0.999. To urine samples requiring deglucuronidation, beta-glucuronidase was added followed by incubation at 37 degrees C for 3 h. After the enzymatic treatment, the samples were subjected to the extraction in the reversed-phase (ODS) and size-exclusion (GPC) modes. It was possible to extract, clean up and concentrate BPA in a single run of 20 min by means of the novel extraction method. The method enables the determination of standards and may be applied to the detection of trace amounts of BPA in human urine samples.