高效液相色谱法测定面条中柠檬黄、日落黄含量的不确定度分析

Uncertainty analysis determination of citrine and sunset yellow in noodle by HPLC

目的建立一种高效液相色谱法测定面条中柠檬黄和日落黄含量的不确定度分析方法。方法选用无水乙醇-氨水-水(7:2:1)为样品的提取溶剂,色谱柱为Waters SymmetryC18柱(250 mm×4.6 mm,5μm),流速1.0mL·min-1,柱温:35℃,流动相以甲醇为A相,以含0.02 moL·L-1乙酸铵溶液、1%冰醋酸和0.2%三乙胺的水溶液(pH=4)为B相,进行梯度洗脱,考查实验过程中对含量的测定有影响的所有因素,确定不确定度来源,建立结果数学模型,计算各个不确定度分量,从而合成不确定度。结果拟合标准曲线过程中产生的不确定度最大,其次是样品处理过程中产生的不确定度,仪器产生的不确定度最小。结论本方法客观、准确,适用于高效液相色谱法测定面条中柠檬黄和日落黄含量的不确定度计算,也可用于计算含量实验方法的不确定度,在实际工作中有较强的实用性。

Objective To establish a method for the determination and uncertainty analysis of citrine and sunset yellow in noodles by HPLC. Methods The samples was extracted by the solution of absolute ethyl alcohol, ammonia water and clear water (with the ratio of 7∶2∶1), and separated by liquid chromatographic column of Waters Symmetry?C18 column (250 mm × 4.6 mm, 5 μm) flow rate 1.0mL/min, column temperature 35℃, using methanol-an aqueous solution of 0.02 mol/L ammonium acetate, 1%acetic acid and 0.2% triethylamine(pH=4)as mobile phase. The factors of affecting the determination of citrine and sunset yellow were investigated, the sources of uncertainty were identified, the mathematic models were established, the components of uncertainty were calculated and the uncertainty was combined. Results The greatest influence on the combined uncertainty was the standard curve, the second influence on the combined uncertainty was the sample processing and the minimum influence on the combined uncertainty was the instruments. Conclusion The HPLC method is objective and accurate for determination and uncertainty analysis of citrine and sunset yellow in noodles and calculation of uncertainty in the experimental method of content. It has strong practicability in the practical work.