Abstract: A method was established for the determination of five flavor nucleotides in pork by ion-pair reversedphase high performance liquid chromatography. Separation and quantification was achieved by using a ZORBAX SB-C_(18) column,and a mobile phase of 0.01 mol/L KH_2PO_4 solution(contain 1.45 mmol/L tetrabutylammonium hydrogen sulfate, p H4. 0)- acetonitrile( 97 ∶ 3, V / V) were used. The detection wavelength was 254 nm. The external standard calibration curves were used for quantification. There were good linear correlations for five flavor nucleotides(R~2>0.9999). Average spike recovery of five flavor nucleotides was 92.5% ~106.0%, with relative standard deviations 1.75%~4.26%,the limit of detection for 5'-UMP,5'-CMP,5'-GMP,5'-IMP and5'-AMP being 2.0,2.0,2.0,3.0 and 3.0 mg/kg,respectively. This method was simple,sensitive,reproducible and suitable for the determination of flavor nucleotides in pork.