固相萃取-高效液相色谱法测定自制饮料中10种合成色素的含量

Determination of 10 synthetic pigments in homemade beverage by solid phase extraction and high performance liquid phase method

建立一种固相萃取-高效液相色谱法测定自制饮料中10种常见合成色素含量的方法。样品经25%氨化甲醇提取,冷冻沉淀法除蛋白后,采用固相萃取柱净化富集,以Agilent5HC-C18(2)(4.6mm×250mm,5μm)为色谱柱进行分离,甲醇和0.02mol/L乙酸铵为流动相进行梯度洗脱,二极管阵列检测器[扫描范围(190～700)nm],选择254 nm作为检测波长,外标法定量分析。结果表明,10种常见合成色素的线性关系良好,线性范围为(0.1～5.0)mg/L,相关系数为0.9996～0.9999;检出限为(0.012～0.024)mg/L,平均回收率为91.2%～101.4%(n=3),相对标准偏差为0.6%～2.4%。该方法操作简单、快速、结果准确性高,可广泛用于自制饮料中10种常见合成色素的含量测定。

A method of determination of 10 common synthetic pigments in domestic beverage by solid phase extraction and HPLC was established. The sample was extracted by 25% ammonia methanol.After freezing precipitation, the protein was removed and enriched by solid phase extraction column and separated by Agilent 5 HC-C18(2)(4.6 mm×250 mm, 5 μm). Methanol and 0.02 mol/L ammonium acetate undergo gradient elution for the flow phase, diode array detector [scan range(190~700)nm], 254 nm was selected as the detection wavelength, and the external standard method was quantitatively analyzed.The results showed that the linear relationship of 10 common synthetic pigments was good, the linear range was 0.1 mg/L to 5.0 mg/L, and the correlation coefficient was 0.9996 to 0.9999;The detection limit was 0.012 mg/L to 0.024 mg/L, the average recovery rate was 91.2% to 101.4%(N=3), and the relative standard deviation was 0.6% to 2.4%. The method was simple, fast and accurate, and could be widely used in the determination of 10 common synthetic pigments in homemade beverage.