高效液相色谱法测定四角蛤蜊、菲律宾蛤仔中呈味核苷酸

Determination of flavor nucleotides in Mactra veneriformis and Ruditapes philippinarum by high performance liquid chromatography

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 KH_2PO_4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R^2均为0.999 9,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 KH_2PO_4-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 R^2 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.