UPLC-MS/MS法测定鄂西北地区不同产地重楼中9种皂苷类成分的含量

Simultaneous Determination of Nine Components in Rhizoma Paridis From Different Producing Areas in Northwest of Hubei by UPLC-MS/MS

目的：建立超高效液相色谱-串联质谱法（UPLC-MS/MS法）测定重楼中9种皂苷类成分的含量,并对鄂西北地区不同产地的重楼药材中9种成分含量进行比较。方法：采用Ultra Bi Ph色谱柱（2.1 mm×100 mm,5μm）,流动相A（0.1%甲酸）-B（0.1%甲酸乙腈溶液）,洗脱梯度：0-1 min,5%-52%B,1-6 min,52%-56%B,6-7min,56%-95%B,7-9 min,95%-5%B,9-10 min,5%-5%B;流速为0.3 m L/min。采用电喷雾离子源（ESI）,多反应监测（MRM）,结合正负离子扫描切换模式检测;利用所建立的方法对鄂西北地区18个重楼样品中皂苷类成分进行含量测定。结果：重楼皂苷Ⅰ、重楼皂苷Ⅱ、重楼皂苷Ⅲ、重楼皂苷Ⅴ、重楼皂苷Ⅵ、重楼皂苷Ⅶ、重楼皂苷H、纤细薯蓣皂苷和17-羟基纤细薯蓣皂苷的线性范围分别为：2.393-2450、5.312-2720、5.078-1300、1.174-2045、3.701-3790、2.524-2585、2.476-1267、1.189-2435、2.524-1292 ng/m L,各成分在线性范围内呈良好的线性关系（r〉0.99）;定量限分别为：2.393、5.312、5.078、1.174、3.701、2.524、2.476、1.189、2.524 ng/m L;日内和日间精密度RSD均小于5%,平均加样回收率均在93.1%-108.5%范围内,RSD均小于5%。样品含量测定结果显示,9种成分在18个样品中均能检测到,各成分在不同样品中含量差异较大。结论：该方法简单、快速、灵敏,重复性好,可以用于重楼药材中9种皂苷类成分的含量测定;通过对皂苷类成分含量比较,发现鄂西北地区重楼药材中皂苷类成分的含量差异较大,适宜作为种植基地的为竹溪县。

Objective To establish an UPLC-MS/MS method for simultaneous determination of 9 components in rhizoma paridis and compare the contents of 9 components in rhizoma paridis from different producing areas in northwest of Hubei province.Methods The chromatographic separation was performed on Ultra BiPh-C18 column （2.0 mmx 100 mm, 5 txm） with gradient elution using 0.1% formic acid in water as mobile phase A and 0.1% formic acid methyl cyanide as mobile phase B（0-1 min,5%-52%B, 1-6 min,52%-56%B,6-7 min,56%-95%B,7-9 min,95%-5%B,9-10 min,5%-5%B） ; the flow rate was 0.3 mL/min.Electrospray ionization source（ESI） and multiple reaction monitoring（ MRM）were used combined with positive and negative scanning switch. Results The liner range of pariphyllin I , pariphyllin 11 , pariphyllin m , pariphyllin V, pariphyllin VI, pariphyllin VII, pariphyllin H, gracillin and 17- hydroxy gracillin were 2.393 - 2450,5.312 - 2720,5.078 - 1300,1.174- 2045,3.701 - 3790,2.524 - 2585,2.476 - 1267,1.189 - 2435 and 2.524 - 1292 ng/mL, respectively.The liner correlations of 9 components were good during their liner range, （ all r〉0.99） .The limits of quantitation were 2.393,5.312,5.078,1.174,3.701,2.524,2.476,1.189 and 2.524 ng/mL,respectively.The RSDs of Intra-day and inter-day precision were less than 5.0%.The average recovery rates of 9 componetits was between 93.1% and 108.5% with RSD of less than 5.0%.All the 9 active components could be detected in the 18 samples, but the contents of the components were obviously differences.Conclusion This method was simple ,rapid,sensitive and accurate ,which can be used to determine the contents of 9 active components in rhizoma paridis.The results showed that the contents of saponins in rhizoma paridis were varied from different producing areas of northwest of Hubei province, in which Zhuxi county would be the best area for planting rhizoma paridis .