变波长高效液相色谱法同时检测葡萄酒中4种合成色素的研究

Simultaneous determination of four synthetic colorants in wine by a HPLC method with variable wavelength detector

【目的】建立一种可同时测定葡萄酒中不允许添加的4种人工合成着色剂（柠檬黄、日落黄、胭脂红和苋菜红）的变波长高效液相色谱检验方法,为葡萄酒中不允许添加人工合成着色剂的检测提供参考。【方法】将葡萄酒样品分别用液-液萃取法、聚酰胺吸附法、直接进样法进行前处理,对变波长高效液相色谱法的检测条件进行优化,并采用该方法进行精密度和加标回收率试验。【结果】葡萄酒样品采用聚酰胺吸附法进行前处理,优化的色谱条件为：Agilent ZORBAX SB-C18（4.6 mm×250 mm,5μm）反相色谱柱,流动相A液为甲醇,流动相B液为醋酸铵溶液（0.02 mol/L）,流速为1.0 mL/min,梯度洗脱,变波长检测葡萄酒样品中柠檬黄、苋菜红、胭脂红、日落黄的检测波长分别为430 nm（0-2.2 min）,230 nm（2.50-3.95 min）,480 nm（3.95-5.00 min）和480 nm（3.95-5.00 min）,柱温35℃,检测时间为5 min,进样量为20μL。对葡萄酒样进行的精密度及加标试验表明,以上4种人工色素的相对标准偏差（RSD）为0.73%-1.51%,回收率为87.83%-107.42%。【结论】建立了一种能同时检测葡萄酒中4种人工合成色素的变波长高效液相色谱法,该方法简单快速,精确可靠,重复性好。

【Objective】 A HPLC method with variable wavelength detector for simultaneous determination of four synthetic colorants not permitted to add into wine by national standards,namely tartrazine,sunset yellow,carmine and amaranth,was developed to give reference for detection of synthetic colorants in wine.【Method】 Wine samples were pretreated by liquid-liquid extraction,polyamide absorption,and direct injection;the detection conditions of a HPLC method with variable wavelength detector were then optimized,and the recovery of standard addition and the precision were also tested by the developed method.【Result】 Wine samples were pretreated by polyamide absorption.The optical chromatographic conditions：5 μm particle size reversed phase column Agilent ZORBAX SB-C18（4.6 mm×250 mm） was used and the mobile phase consisted of solution A（methanol） and solution B（ammonium acetate 0.02 mol/L） with gradient elution and variable wavelength detection,and the detection wavelengths for tartrazine,sunset yellow,carmine and amaranth were 430 nm（0-2.2 min）,230 nm（2.50-3.95 min）,480 nm（3.95-5.00 min）,480 nm（3.95-5.00 min）,respectively.The flow rate was 1.0 mL/min,column temperature 35 ℃,detection time 5 min,and the injection volume 20 μL.When the four standards of the colorants were added into the wine respectively,the average relative standard deviation（RSD） was 0.73%-1.51%,and the average recovery 87.83%-107.42%.【Conclusion】 A HPLC method,with variable wavelength detector developed to simultaneously determine four synthetic colorants not permitted in wine,is a simple,fast,reliable and reproducible one.