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目的:建立复方黄黛片中青黛的薄层色谱(TLC)鉴别方法及高效液相(HPLC)法同时测定靛蓝、靛玉红含量。方法:TLC法对复方黄黛片中青黛进行定性鉴别。HPLC法对复方黄黛片中靛蓝及靛玉红的含量进行测定,色谱柱Agilent TC-18色谱柱(5μm,4.6mm×250mm);流动相甲醇-水(75:25);流速1mL·min~(-1);柱温25℃;检测波长292nm。结果:TLC结果表明青黛标准药材、靛蓝、靛玉红及供试品斑点清晰,阴性空白无干扰。HPLC结果表明,靛蓝在0.208~1.04μg、靛玉红在0.088~0.44μg范围内呈良好的线性关系,平均回收率分别为99.95%(RSD=1.59%)和99.30(RSD=1.86)。结论:TLC和HPLC试验结果表明,试验方法简便、准确,可用于复方黄黛片中青黛的鉴别及靛蓝和靛玉红含量的测定。
OBJECTIVE: To establish a thin layer chromatography (TLC) identification method and high performance liquid chromatography (HPLC) method for the simultaneous determination of indigo and indirubin in Fufang Huangdai tablets. Methods: The TLC method was used for the qualitative identification of Compound Indigo Naturalis. HPLC method was used to determine the contents of indigo and indirubin in the compound Huangdai tablets. The mobile phase was methanol-water (75:25) with a mobile phase of 1 mL · min and the mobile phase was Agilent TC-18 (5 μm, 4.6 mm × 250 mm) ~ (-1); column temperature 25 ℃; detection wavelength 292nm. Results: The TLC results showed that indigo, indirubin, indigo and indigo natural samples were clear spots, negative blank without interference. The results of HPLC showed that there was a good linear relationship between indigo and indirubin in the range of 0.088-0.44μg, the average recoveries were 99.95% (RSD = 1.59%) and 99.30 (RSD = 1.86), respectively. Conclusion: TLC and HPLC test results show that the test method is simple and accurate, and can be used for identification of Indigo Indigo and indirubin content of compound Huangdai tablets.