HPLC-DAD法测定血竭饮片中添加苏丹红Ⅰ、Ⅱ、Ⅲ、Ⅳ及808猩红

目的：建立HPLC-DAD对血竭饮片中非法添加苏丹红Ⅰ、Ⅱ、Ⅲ、Ⅳ及808猩红的定性和同时定量方法。方法：采用Kromasil100-5 C₁₈色谱柱（250 mm×4.6 mm，5 μm），以乙腈-水（90：10）为流动相，流速1.0 mL·min^-1，检测波长478 nm（苏丹红Ⅰ）和520 nm（苏丹红Ⅱ、Ⅲ、Ⅳ、808猩红）。结果：5个色素分离度良好；苏丹红Ⅰ、Ⅱ、Ⅲ、Ⅳ、808猩红质量浓度分别在0.512 8～20.512 8 μg·mL^-1（r=1.000）、0.496 5～19.859 4 μg·mL^-1（r=1.000）、0.495 1～19.805 8 μg·mL^-1（r=1.000）、0.477 1～19.082 0 μg·mL^-1（r=1.000）和0.506 4～20.255 4 μg·mL^-1（r=1.000）内线性关系良好；精密度RSD小于0.30%；3个加样浓度的平均回收率（n=6）为98.9%～106.6%；重复性RSD为0.51%～1.8%；5个成分在48 h内保持稳定，RSD小于3.0%。20批样品中，3批为阳性样品，其中1批掺入苏丹红Ⅳ，2批掺入808猩红，含量分别为3.0、3.8、2.0 mg·g^-1。结论：该方法简便准确，可用于血竭饮片中非法添加苏丹红Ⅰ、Ⅱ、Ⅲ、Ⅳ、808猩红5个合成色素的定性鉴别和含量测定。

Objective: To establish an HPLC-DAD method for simultaneous qualitative and quantitative analysis of sudan Ⅰ,sudan Ⅱ,sudan Ⅲ,sudan Ⅳ and 808 scarlet illegally added in Draconis Sanguis. Methods: The separation was performed on a Kromasil 100-5 C₁₈(250 mm×4.6 mm,5 μm)column and the mobile phase consisted of acetonitrile-water(90:10)at a flow rate of 1.0 mL·min^-1. The detection wavelength was 478 nm(sudan Ⅰ)and 520 nm(sudan Ⅱ,sudan Ⅲ,sudan Ⅳ and 808 scarlet). Results: The five synthetic pigments were well separated. The linear ranges of sudanⅠ,sudanⅡ,sudanⅢ,sudanⅣ,808 scarlet were 0.512 8-20.512 8 μg·mL^-1(r=1.000),0.496 5-19.859 4 μg·mL^-1(r=1.000),0.495 1-19.805 8 μg·mL^-1(r=1.000),0.477 1-19.082 0 μg·mL^-1(r=1.000)and 0.506 4-20.255 4 μg·mL^-1(r=1.000). The results of the precision tests were satisfactory with relative standard deviations(RSDs)all below 0.30%. The average spike recoveries(n=6)at three concentration levels were 98.9%-106.6% and RSDs were 0.51%-1.8%. The results of the stability tests were satisfactory and the five synthetic pigments remained stable in 48 hours with all RSDs below 3.0%. Among 20 batches of samples,3 batches were positive samples,including 1 batch mixed with Sudan Ⅳ and 2 batches mixed with 808 scarlet,with the content of 3.0,3.8 and 2.0 mg·g^-1,respectively. Conclusion: The method is simple and accurate,which can be used for qualitative and quantitative determination of sudan Ⅰ,sudan Ⅱ,sudan Ⅲ,sudan Ⅳ and 808 scarle illegally added in Draconis Sanguis.

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