液相色谱-质谱联用法测定罂粟壳中5种生物碱的含量

Determination of 5 Alkaloids in Papaver Hull by HPLC-MS

目的:建立一种液相色谱-质谱联用法,测定罂粟壳中吗啡、可待因、罂粟碱、蒂巴因、那可丁5种生物碱的含量。方法:采用液相色谱/质谱联用仪(三重串联四级杆质谱),色谱柱以十八烷基硅烷键合硅胶为填充剂(规格为2.1×100mm,5μm),以乙腈-0.1%甲酸水(7:3)为流动相,流速0.4mL/min,柱温40℃。结果:5种生物碱在此条件下分离效果良好,吗啡、可待因在25~250ng/mL范围内,罂粟碱、蒂巴因、那可汀在5~50ng/mL范围内与其峰面积呈良好线性关系,相关系数r分别为0.99597、0.99686、0.99998、0.99905、0.99762。定量限(Limit of Quantitation, LOQ)分别为2.4×10-2mg/kg、2.4×10-2mg/kg、6×10-4mg/kg、6×10-4mg/kg、6×10-4mg/kg。回收率、重复性、稳定性均良好。结论:建立了一种液相色谱-质谱联用法,测定罂粟壳中吗啡、可待因、罂粟碱、蒂巴因、那可丁5种生物碱的含量,该方法高效快捷,分离效果好,定量准确稳定,为加强罂粟壳药材的质量监管提供技术支撑。

Objective: A liquid chromatography-mass spectrometry(LC-MS) method was established for the determination of morphine, codeine, papaverine, Thebaine and Noscapine in Papaver Hull. Methods: Liquid chromatography/mass spectrometry(triple tandem quadrupole mass spectrometry) was used. The column was filled with Octadecyl silane bonded silica gel(specifications were 2.1×100 mm, 5μm), the mobile phase was acetonitrile-0.1%formic acid water(7: 3). flow rate was 0.4 mL/min, column temperature was 40℃. Results: The separation effect of five alkaloids is good under these conditions, Morphine, Codeine in the range of 25~250 ng/mL, Papaverine, Thebaine and Noscapine in the range of 5~50 ng/mL showed a good linear relationship with the peak area. The correlation coefficients was 0.99597、0.99686、0.99998、0.99905、0.99762. The LOQ was 2.4×10-2 mg/kg、2.4×10-2 mg/kg、6×10-4 mg/kg、6×10-4 mg/kg、6×10-4 mg/kg. The recovery, repeatability and stability were all good. Conclusion: A liquid chromatographic mass spectrometry was established to determine the content of five alkaloids in Papaver Hull. The method is efficient and fast, the separation effect is good, the quantity is accurate and stable, and can provide technical support for strengthening the quality control of Papaver Hull.