超高效液相色谱法检测西红花中非法添加的染料金橙Ⅱ

目的建立检测中药西红花中非法添加的金橙Ⅱ的液质联用鉴别法和超高效液相色谱含量测定法。方法用超高效液相色谱法分离中药西红花中非法添加的金橙Ⅱ,用离子阱进行定性鉴别,流动相为乙腈-0.01 mol/L醋酸铵溶液(30∶70),离子化模式为ESI(+),碰撞能量为30 V;用超高效液相色谱法测定金橙Ⅱ含量,流动相为乙腈-0.01 mol/L醋酸铵溶液(30∶70),检测波长484 nm。结果在拟订的质谱条件下,金橙Ⅱ产生M+H-Na]+分子离子峰,二级质谱的特征碎片离子明显;定量检测方面,进样量与峰面积的线性范围为5.435～745.0 ng,r=0.999 0,平均加样回收率为98.29%,RSD为1.25%(n=5)。结论液质联用法定性准确、可靠,液相色谱含量测定法简便、快速、准确。

QualitatiVe and Quantitative Determinaion of Illegally Added Acid Orange Ⅱ in Croci Stigma by UPLC

Objective To establish an identification method of LC-MS/MS and a quantification method of ultra performance liquid chromatography-mass spectrometry（UPLC-MS/MS） for determining illegally added acid orange Ⅱin Croci Stigma.Methods Illegally added acid orange Ⅱin Croci Stigma was separated by UPLC and the qualitative identification was performed by ion trap.The mobile phase was consisted of acetonitrile-0.01 mol/L ammonium acetate（30 ∶70）.The ionization mode was ESI（＋） and MS/MS collision energy was 30 V.Then,the content of acid orange Ⅱwas determined by UPLC with the mobile phase of acetonitrile-0.01mol/L ammonium acetate（30 ∶70） and the detection wavelength was 484 nm.Results Under the experimental condition of mass spectrum,acid orange Ⅱproduced the M＋H-Na]＋molecular ion peaks and apparently revealed the characteristic fragment ion of second order mass spectrum.In the quantitative determination,the linearity between the sample size and the peak area was over the range of 5.435-745.0 ng,r=0.999 0.The average recovery rate was 98.29%,RSD was 1.25%（n=5）.Conclusion The LC-MS/MS method is accurate and fast for the qualitative identification.The UPLC method is simple,rapid and accurate for the content determination