超高效液相-串联四极杆质谱联用法同时测定并比较甘草各部位中8种成分含量

Simultaneous Quantification and Comparison of 8 Components in Different Parts of Glycyrrhiza uralensis Using Ultra-High Performance Liquid Chromatography-Triple Quadrupole Mass Spectrometry

建立同时检测甘草中8种成分:22-羟基-甘草次酸、3-epi-甘草次酸、甘草次酸、甘草酸、甘草苷、芒柄花黄素、异甘草酚、甘草香豆素的超高效液相色谱-串联四极杆质谱联用法。选用胆酸作为三萜成分的内标,相思子素2'-O-β-芹菜糖苷作为酚类化合物的内标。甘草样品经粉碎用甲醇(含内标液)超声提取,采用安捷伦ZORBAX RRHD C18柱(50 mm×2.1 mm,1.8μm),以甲醇-0.1%甲酸溶液为流动相,梯度洗脱,以电喷雾电离源,采用超高效液相-串联四极杆质谱联用动态多反应监测模式测定。结果表明,8种成分检出限均小于0.048 8μg/mL,定量限均小于0.195 2μg/mL,在0.048 8~12.500 0、0.048 8~12.500 0、0.012 2~12.500 0、0.048 8~50.000 0、0.048 8~50.000 0、0.012 2~12.500 0、0.012 2~15.000 0、0.195 2~12.500 0μg/mL范围内,22-羟基-甘草次酸、3-epi-甘草次酸、甘草次酸、甘草酸、甘草苷、芒柄花黄素、异甘草酚、甘草香豆素的峰面积与质量浓度呈良好线性关系,相关系数均大于0.99;方法回收率为97.2%~113.0%间;其日内及日间精密度实验的相对标准偏差分别为2.63%~4.47%及1.75%~3.72%。该方法灵敏度高、稳定性强、操作简便、快捷、准确,可用于甘草各部位及其相关食品质量控制。

An ultra-high-performance liquid chromatography-triple quadrupole mass spectrometry(UPLC-QQQMS) with dynamic multiple reaction mode was developed to simultaneously quantify 8 constituents including 22-hydroxyglycyrrhetinic acid, 3-epi-glycyrrhetinic acid, glycyrrhetinic acid, glycyrrhizic acid, liquiritin, glycycoumarin, formononetin and isoglycyrol in Glycyrrhiza uralensis. Cholic acid and abrusin 2'-O-β-apioside were used as the internal standards for triterpenoid and phenolic components, respectively. The internal standards were added to methanol before being used to extract samples. The separation was performed on an Agilent ZORBAX RRHD Eclipse Plus C18 column(50 mm × 2.1 mm, 1.8 μm) with methanol-0.1% formic acid as the mobile phase at a flow rate of 0.4 mL/min. As a result, the linear ranges of 22-hydroxy-glycyrrhetinic acid, 3-epi-glycyrrhetinic acid, glycyrrhetinic acid, glycyrrhizic acid, liquiritin, formononetin, isoglycyrol and glycycoumarin were 0.048 8–12.500 0, 0.048 8–12.500 0, 0.012 2–12.500 0, 0.048 8–50.000 0, 0.048 8– 50.000 0, 0.012 2–12.500 0, 0.012 2–15.000 0 and 0.195 2–12.500 0 μg/mL, respectively, with correlation coefficients higher than 0.99. The average recovery rate of the developed method ranged from 97.2% to113.0% and the intra-day and inter-day relative standard deviations(RSDs) were 2.63%–4.47% and 1.75%–3.72%, respectively. This method is sensitive, simple, fast, and accurate, and can be used for the quality control of different parts of Glycyrrhiza uralensis and related food products.