高效液相色谱法测定酱料、酱油中的诱惑红含量的研究

摘要

目的：鉴于现有测定食品中诱惑红含量的国标方法操作繁琐，准确度难以保证，实验旨在建立快速高效并准确地使用高效液相色谱（HPLC）测定酱料、酱油中诱惑红含量的方法。方法：实验采用诱惑红在可见光区最强吸收波长510 nm作为测试波长，实验证明一定浓度的氨水-乙醇溶液作为提取液的提取效果最好。实验以“直接萃取法”提取样品中的诱惑红，操作简单有效。再以乙酸氨水溶液与乙腈-甲醇-水溶液组成流动相，梯度洗脱，C18色谱柱分离。实验探讨了过滤滤膜的选择。结果：诱惑红在0．05～10 mg／L浓度范围内，线性回归系数达0．9995以上，样品添加回收率在94％～108％之间，相对标准偏差RSD＜2．5％（n＝6）。该方法诱惑红的检出限为0．15 mg／kg，远优于国标方法的检出限25 mg／kg的水平。实验证明选用PTFE滤膜，不会对诱惑红色素造成损失。讨论：该方法操作简单高效，准确度高，所选用的色谱条件良好，谱图中的目标物几乎没有干扰，灵敏度高，回收率良好且稳定，优于国标方法，值得推广。%As the GB method about the determination of Allura red is so complicated and the accuracy of the GB method is unsure,a method for the determination of Allura red in sauce and soy sauce based on high performance liquid chromatography (HPLC)is wanted to be established.The method will be easy to operate and be more accurate.The test shows that the strongest wavelength of visible light of Allura red is 510 nm.And the result shows that ethanol-ammonia water solution is the best solution that extracts Allura red in sauce or soy sauce rather than pure water or ammonia water solution.The determination of Allura red in sauce or soy sauce can be extracted by ethanol-ammonia water solution, with removing the oil by n-hexane.The Allura red can be determined by HPLC using the acetonitrile-methanol-water mobile phase,gradient elution,C18 column separation,and 510 nm as wavelength. The standard curve shows a good linearity with concentration of 0.05~10 mg/L.The linearly dependent coefficient can be above 0.9995.And the recovery is 94% ~ 108%.The RSD is<2.5%(n= 6).The method detection limit is 0.15 mg/kg.The method has the advantages of simple opera, high sensitivity,good accuracy and good reproducibility.It is very suitable for the determination of Allura red in sauce.This method is superior to the GB method about determination of Allura red,and worth promoting.