蒲黄中1种新染色掺假色素的确认及9种橙黄色色素的检测方法研究

Identification of a New Adulterated Dye and Study on Detection Method for 9 Orange-yellow Dyes in Typhae Pollen

目的:确认蒲黄中1种新染色掺假色素,并建立同时检测9种橙黄色色素的方法。方法:采用薄层色谱法(TLC)和液相色谱-质谱联用法对新染色掺假色素进行确认;采用TLC和高效液相色谱法(HPLC)检测9种橙黄色色素:色谱柱为Welch AQ-C18,流动相为乙腈-0.05 mol/L乙酸铵溶液(梯度洗脱),流速为1.0 m L/min,检测波长为432 nm(柠檬黄、皂黄、金橙O、碱性橙Ⅱ)和484nm(日落黄、酸性橙Ⅰ、金橙Ⅱ、橙G、碱性橙21),柱温为35℃,进样量为10μL。结果:金胺O、柠檬黄和新染色掺假色素TLC图斑点清晰,分离度好,阴性对照无干扰;新染色掺假色素为皂黄。柠檬黄、日落黄、橙G、酸性橙Ⅰ、金橙Ⅱ、金胺O、皂黄、碱性橙21和碱性橙Ⅱ的TLC图斑点清晰,分离度好,阴性对照无干扰;检测限分别为0.30、0.20、0.33、0.16、0.19、0.19、0.31、0.26、0.30mg/kg。结论:本研究所建方法可快速准确地对蒲黄中染色掺假色素和橙黄色色素进行定性检测。

OBJECTIVE：To identify a new adulterated dye,and to establish simultaneous detection method for 9 orange-yellow dyes in Typhae Pollen. METHODS：TLC and LC-MS were used for the identification of the new adulterated dye;TLC and HPLC method were used for determining 9 orange-yellow dyes. The determination was performed on Welch AQ-C18 column with mobile phase consisting of acetonitrile-0.05 mol/L ammonium acetate solution（gradient elution） at the flow rate of 1.0 m L/min.The detection wavelength were set at 432 nm（lemon yellow,metanil yellow,gold orange O,basic orange Ⅱ） and 484 nm（sun-set yellow,acid orange Ⅰ,gold orangeⅡ,orange G,basic orange 21）. The column temperature was 35 ℃.The sample size was 10 μL. RESULTS：TLC spots of auramine O,lemon yellow and new adulterated dye were clear and well separated without interference from negative control;new adulterated dye was identified as metanil yellow. TLC spots of lemon yellow,sunset yellow,orange G,acid orangeⅠ,gold orangeⅡ,auramine O,metanil yellow,basic orange 21 and basic orangeⅡwere clear and well separated without interference from negative control. The limits of detection were 0.30,0.20,0.33,0.16,0.19,0.19,0.31,0.26,0.30 mg/kg. CONCLUSIONS：The established method is able to detect adulterated dye and orange-yellow dye in Typhae Pollen rapidly and accurately.