A new method for the adsorption and preconcentration of bismuth in urine was described. The methodology combines determined using a hydride generation-atomic absorption spectrometry (HG-AAS) with pre-concentration and separation of the bismuth on the nano-barium-strontium titanate powder coated by dithizone (BST-dithizone). The experimental conditions for the pre-concentration and separation of the bismuth, including pH value of the medium, shaking time, eluent condition and co-existing ions have been investigated. The result showed that the bismuth in urine could be adsorbed on the BST-dithizone. The adsorption percentage was affected by the pH value of medium and shaking time. In the medium of pH 6.0, the adsorption capacity of BST-dithizone to bismuth (III) was 13.4 mg·g-1 when the shaking time was 10 min. The bismuth adsorbed on the BST-dithizone could be completely eluated by 2 mol·L-1 HNO3. The enrichment factor was 100. The detection limit of bismuth (III) was 5.1 ng·L−1.The method has been applied to the pre-concentration and separation of bismuth in the human urine and tap water with satisfied results.