反相离子对高效液相色谱法测定猪肉中呈味核苷酸

赵艳; 汪泓吉; 张凤枰; 杨发树; 刘耀敏; 范秀丽

doi:10.13386/j.issn1002-0306.2016.02.004

反相离子对高效液相色谱法测定猪肉中呈味核苷酸

Determination of five flavor nucleotides in pork by ion-pair reversed-phase high performance liquid chromatography

摘要

摘要: 建立反相离子对高效液相色谱法测定猪肉中呈味核苷酸5’-尿苷酸二钠（5’-UMP）、5’-胞苷酸二钠（5’-CMP）、5’-鸟苷酸二钠（5’-GMP）、5’-肌苷酸二钠（5’-IMP）和5’-腺苷酸二钠（5’-AMP）含量的方法。应用ZORBAX SB-C₁₈色谱柱,以0.01 mol/L KH₂PO₄溶液（含1.45 mmol/L四丁基硫酸氢铵,p H4.0）-乙腈（97∶3,V/V）为流动相进行洗脱,紫外检测器在254 nm处进行检测,外标法定量。结果表明,5种呈味核苷酸线性关系良好,相关系数均为0.9999,回收率在92.5%¹06.0%范围内,相对标准偏差在1.75%⁴.26%之间,5’-UMP、5’-CMP、5’-GMP、5’-IMP和5’-AMP检出限分别为2.0、2.0、2.0、3.0和3.0 mg/kg。该方法操作简便、灵敏度高、重现性好,适用于猪肉中呈味核苷酸含量的测定。

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.