染色蒲黄中柠檬黄、酸性黄36检查方法的建立

目的：建立蒲黄用柠檬黄、酸性黄36染色的定性检查方法。方法：采用TLC法，对染料柠檬黄、酸性黄36进行初步检查；采用HPLC-DAD法进行验证，采用C₁₈（150 mm×4.6 mm，5 μm）色谱柱，以甲醇（A）-0.2 mol·L^-1乙酸铵溶液（B），梯度洗脱（0~10 min，12% A，88% B； 10~15 min，12% A → 90% A，88% B → 10% B； 15~20 min，90% A → 12% A，10% B → 88% B； 20~30 min，12% A，88% B），在428 nm波长检测柠檬黄，在422 nm波长检测酸性黄36；采用HPLC-MS法对2种色素的检出进行确证，采用乙腈-0.02mol·L^-1乙酸铵（35∶65）流动相系统，ESI+扫描模式。结果：薄层色谱斑点清晰，分离度好，专属性较强。HPLC-DAD法供试样品中被检成分与对照品保留时间一致，且UV图谱一致。HPLC-MS法供试样品中相关分子离子峰及碎片特征与2种对照品一致。结论：本文所建立的方法可用于蒲黄用柠檬黄、酸性黄36染色的检查。关键词:蒲黄；柠檬黄；酸性黄36；薄层色谱；高效液相色谱-二极管阵列检测器；高效液相色谱-质谱联用中图分类号：R 917文献标识码：A文章编号：0254-1793（2017）04-0670-07doi：10.16155/j.0254-1793.2017.04.18

Objective: To establish inspection method of Typhae Pollen illegally dyed with tartrazine and metanil yellow. Methods: TLC was used to preliminary test of tartrazine and metanil yellow. Then the results were verified by HPLC-DAD method. Separation was performed on a C₁₈ column(150 mm×4.6 mm,5 μm) with methyl alcohol as mobile phase A and 0. 2 mol·L^-1 ammonium acetate as mobile phase B using a gradient elution program (0-10 min, 12%A, 88%B; 10-15 min, 12%A → 90%A, 88%B → 10%B; 15-20 min, 90%A → 12%A, 10%B → 88%B;20-30 min,12%A,88%B). The detection wavelengths were 428 nm for tartrazine and 422 nm for metanil yellow. HPLC-MS was applied to confirmation with acetonitrile-0.2 mol·L^-1 ammonium acetate(35:65) as the mobile phase and positive electrospray ionization as the detection mode. Results: In TLC test,tartrazine and metanil yellow could be identified specifically by clear and well separated spots. In HPLC-DAD test,the retention times and UV spectra of the detected peaks of the test solution corresponded to those of the reference solution. In HPLC-MS test,the molecular ions and fragments of the detected peaks of the test solution corresponded to those of the reference solution. Conclusion: The method proposed in this paper could be used to test Typhae Pollen dyed with tartrazine and metanil yellow.

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