四神丸中3种非法添加染色成分检测方法研究

Detection methods of three illegal dyeing components in Sishen pills

建立四神丸中胭脂红、酸性红73、赤藓红等3种非法添加染色成分的检测方法。TLC法采用乙酸乙酯-正丁醇-乙醇-氨水-水(2∶2∶3∶1∶1)为展开剂,日光下检视;HPLC法采用Agilent 5TC-C_(18)(2)色谱柱(4.6 mm×250.0 mm,5μm),以乙腈-0.02 mol/L乙酸铵溶液为流动相,梯度洗脱,检测波长为510 nm,流速1.0 mL/min,柱温30℃。结果显示,14批四神丸样品中均未检出阳性样品。模拟阳性样品薄层色谱斑点清晰,阴性对照无干扰;胭脂红、酸性红73、赤藓红分别在12.92~77.57μg/mL、8.32~49.92μg/mL、5.28~31.68μg/mL范围内线性关系良好(r≥0.9996),平均回收率分别为93.24%、97.20%、94.47%,RSD分别为2.60%、2.62%、2.05%(n=6)。该方法操作简单、准确,可用于四神丸中3种非法添加染色成分的检测。

To establish a method for the detection of three illegal dyeing components in Sishen pills,including carmine,acid red 73 and erythrosine,ethyl acetate-n-butanol-ethanol-ammonia-water(2∶2∶3∶1∶1)was used as the developing solvent in TLC,and checked under sunlight;Agilent 5 TC-C_(18)(2)chromatographic column(4.6 mm×250.0 mm,5μm)was used in HPLC method,acetonitrile-0.02 mol·L^(-1) ammonium acetate solution was used as the mobile phase,gradient elution,the detection wavelength was 510 nm,the flow rate was 1.0 mL·min-1,and the column temperature was 30℃.The results showed that no positive detection was found in the 14 batches of Sishen pill samples.The TLC spots of the simulated positive sample were clear,and the negative control had no interference;the linear relationship of carmine,acid red 73 and erythrosine was good in the range of 12.92-77.57μg·mL^(-1),8.32-49.92μg·mL^(-1) and 5.28-31.68μg·mL-1,respectively(r≥0.9996),the average recovery was 93.24%,97.20%and 94.47%with the RSD of 2.60%,2.62%and 2.05%,respectively(n=6).The method is simple and accurate,and can be used for the detection of three illegal dyeing components in Sishen pills.