The purpose of this study was to use various atomic absorption spectroscopic techniques to investigate the amount of biological trace metals, including selenium, manganese, cobalt, chromium and zinc, in the urine of Blackfoot disease (BFD) patients. Urine specimens from BFD patients were collected, acidified with a mixed acid solution, purified and preconcentrated with a solid phase extraction cartridge before being digested in a microwave oven at controlled conditions prior to analysis. The optimum composition of the mixed acid was established according to the analysis of standard known sample. The level of urinary selenium and zinc was determined by hydride atomic absorption spectrophotometry (HAAS) and flame atomic absorption spectrophotometry (FAAS), respectively, whereas manganese, cobalt and chromium level were determined using graphite furnace atomic absorption spectrophotometry (GFAAS) with stabilized temperature platform furnace conditions. The validity of the analytical methods was confirmed by the analysis of standard known samples. The result showed that total urinary Mn, Co, Cr and Zn levels in BFD patients were significantly different than those in normal subjects (Mann-Whitney test, p<0.05). But the total urinary Se level in BFD patients did not differ from those in normal control subjects (p>0.05). The possible connection of these elements with the etiology of the disease is discussed.