富含蛋白食品中的8种合成色素的 高效液相色谱测定方法

目的 建立高效液相色谱法测定富含蛋白食品中合成色素的方法。方法 样品匀浆后采用乙醇:氨水:水(7:2:1)溶液提取, 经低温冷冻去除蛋白后, 水浴加热浓缩除氨, 采用聚酰胺固相小柱萃取净化, 以甲醇和20 mmol/L乙酸铵溶液为流动相, 通过Agilent extend-C18色谱柱梯度洗脱分离, 可见光区段多波长信号检测柠檬黄、日落黄、苋菜红、胭脂红、赤藓红、诱惑红、亮蓝、靛蓝8种合成色素, 保留时间定性、外标法定量。结果 建立的测定8种合成色素的方法在0.6~10 mg/kg内曲线线性关系良好, 相关系数为0.995以上, 检测限介于0.1~0.4 mg/kg, 不同加标浓度下回收率介于75%~95%之间, 变异系数在3.6%~9.3%之间。结论 该方法用于高蛋白食品中合成色素的测定, 具有操作简便、灵敏高、准确性好、无杂质干扰等优点。

Determination of 8 synthetic food colorants in protein-rich samples with high performance liquid chromatography

Objective To establish a method for the detection of synthetic colorants in protein-rich processed food with high performance liquid chromatography. Methods Sample homogenates were extracted by ethanol:ammonium hydroxide:water (7:2:1), and frozen for deproteinization, further incubated in water to remove ammonium, purified by solid phase extraction with polyamine column. Extract containing 8 synthetic colorants (lemon yellow, sunset yellow, amaranth, ponceau 4R, erythrosine, allura red, brilliant blue, and indigo carmine) was separated by Agilent extend-C18 with methanol and 20 mmol/L of ammonium acetate as the mobile phase. Compounds were detected under multi-wavelength measurement. Results Measurement on 8 synthetic colorants with the developed method showed an excellent linear relationship between 0.6~10 mg/kg, with the correlation coefficient more than 0.995, and the limit of detection between 0.1~0.4 mg/kg. Under different spiked concentration, the recoveries were determined between 75%~95%, and relative standard derivations were between 3.6%~9.3%. Conclusion The developed method can be applied in the detection of synthetic colorants in protein-rich processed food, with easy operation, high sensitivity, excellent accuracy and less disturbance.