高效液相色谱法测定四角蛤蜊、菲律宾蛤仔中呈味核苷酸

摘要

建立高效液相色谱法(high performance liquid chromatography,HPLC)同时测定四角蛤蜊、菲律宾蛤仔中呈味核苷酸5’-尿苷酸(5’-UMP)、5’-鸟苷酸(5’-GMP)、5’-肌苷酸(5’-IMP)和5’-腺苷酸(5’-AMP)含量.采用Waters Atlantis T3(4.6 mm×250 mm,3μm)色谱柱,以0.01 mol/L KH2PO4溶液-甲醇为流动相梯度洗脱,紫外检测器在260 nm 处进行检测,外标法进行定量.结果表明:各呈味核苷酸线性关系良好,相关系数R2均为0.9999,回收率在96.19％ ～103.25％内,相对标准偏差在1.08％ ～ 1.28％内.四角蛤蜊与菲律宾蛤仔中均含有4种呈味核苷酸,其中5’-AMP含量最多,其次是5’-IMP和5’-UMP,而5’-GMP含量最低.该方法简单,重现性好、灵敏度和准确度高,适用于四角蛤蜊、菲律宾蛤仔中呈味核苷酸含量测定,为研究贝类风味及贝类食品的开发提供一定的依据.%A method was established for determination of flavor nucleotides in Mactra veneriformis and Ruditapes philippinarum by high performance liquid chromatography.Methods:Separation and quantization were perfored by using a Waters Atlantis T3 column,with 0.01 mol/L KH2PO4-methanol as the mobile phase in gradient elution.The detection wavelength was 260 nm.Results:The external standard calibration curves were used for quantization.A good linear correlations for flavor nucleotide was shown with R2 ＞ 0.999 9.The average spike recovery of flavor nucleotides was 96.19％-103.25％,with relative standard deviations of 96.19％-103.25％.Conclusion:The method is simple,with good reproducibility,sensitivity and high accuracy.It is suitable for determining flavor nucleotides in Mactra veneriformis and Ruditapes philippinarum.The method provideds a basis for the research of shellfish flavor and the development of shellfish food.