A novel miniature cold vapor generation atomic fluorescence spectrometry was developed using amorphous Teflon AF 2400 tube as the gas-liquid separator and detection cell for the determination of Hg in tobacco,based on the Teflon AF 2400 tube′s excellent gas transport behavior and high UV transparence.The effects of length of Teflon AF 2400,type and concentration of reducing agent,type and concentration of acid medium,flow rate of sample and Ar on the system were investigated in detail.Under the optimal conditions,the calibration curve was linear in the range of 007-10 μg/L with detection limit of 002 μg/L.The relative standard deviation（RSD） was 33% for eleven consecutive measurements of 050 μg/L Hg standard solution.The novel system showed the advantages of micro scale,low sample consumption and high sensitivity.The method was successfully used in the determination of Hg in tobacco sample with recoveries of 97%–102%.