A high performance liquid chromatography(HPLC) method was developed for the determination of polyoxin residue in paddy soil and water. The paddy water was adjusted to pH 8.0 with ammonia water，and extracted with acetic ether to remove organic impurities. The water phase was concentrated to constant. The paddy soil was extracted using alkaline mixed with solution of carbinol-water(70∶30)，and concentrated to constant. The above extracts were separated on a C18 hydrophilic stainless steel chromatographic column(AQ-C18,4.6 mm×250 mm,5 μm) and detected with ultraviolet detector(UVD).The quantification analysis of polyoxin was performed by the external standard.The other optimal conditions were as follows:mobile phase:carbinol- water(13∶87);flow rate:1.0 mL/min for first 0-8 min，and 0.3 mL/min for the next 8-16 min，column temperature:30 ℃，detection wavelength:272 nm. Under the optimal conditions，the calibration curve was linear between peak area and polyoxin concentration in the range of 0.05-2.00 mg/L with correlation coefficient of 0.998 8.The limit for detection for polyoxin was 1.0×10-9 g，and the detection limits of polyoxin in paddy soil and water were 0.05 mg/kg and 0.05 mg/L，respectively.The spiked recoveries of polyoxin in paddy water and soil at spiked concentrations of 0.06,0.60,1.00 mg/kg were in the range of 97%-99% and 95%-97%,with RSDs of 0.71%-2.4% and 1.7%-4.5%，respectively.This method showed the advantages of easy operation，good separation effect and high accuracy，and could meet the requirements for pesticide residue detection.