波长切换HPLC同时测定车前草中5个活性成分的含量

Simultaneous Determination of 5 Active Components in Plantaginis Herba by HPLC with Wavelength Switching

目的:建立多波长切换HPLC同时测定车前草中大车前苷、毛蕊花糖苷、异毛蕊花糖苷、木犀草素和芹菜素5个活性成分的含量测定方法。方法:采用Waters Symmetry C18色谱柱(250 mm×4. 6 mm,5μm),以乙腈(A)-0. 1%甲酸水(B)溶液为流动相,梯度洗脱,检测波长为330 nm(0～29 min检测大车前苷、毛蕊花糖苷和异毛蕊花糖苷)、350 nm(29. 01～31 min检测木犀草素)和338 nm(31. 01～35 min检测芹菜素),流速为0. 8 m L·min^(-1),柱温为25℃。结果:在所建立的色谱条件下各成分的专属性良好,无干扰峰;大车前苷、毛蕊花糖苷、异毛蕊花糖苷、木犀草素和芹菜素的线性范围分别为42. 84～428. 40μg·m L^(-1)(r=0. 999 7)、35. 36～353. 60μg·m L^(-1)(r=0. 9995)、18. 40～184. 00μg·m L^(-1)(r=0. 999 5)、0. 856～8. 56μg·m L^(-1)(r=0. 999 6)和0. 108～1. 08μg·m L^(-1)(r=0. 999 6);平均回收率(n=6)分别为98. 62%(RSD=0. 87%)、98. 09%(RSD=1. 10%)、98. 19%(RSD=1. 16%)、95. 52%(RSD=1. 03%)、92. 34%(2. 00%); 6批车前草样品中5种成分的含量范围分别为1. 51～6. 78、2. 53～18. 77、0. 60～2. 92、0. 03～0. 18、5. 34～12. 36μg·g^(-1)。结论:该方法操作简单、准确、灵敏度高、重复性好,为车前草的质量控制提供参考依据。

Objective： To establish a method for simultaneous determination of plantamajoside,acteoside,isoacteoside,luteolin and apigenin in Plantaginis herba by HPLC with wavelength switching. Methods： The samples were separated on a Waters Symmetry C18 column（ 250 mm×4. 6 mm,5 μm） by a gradient elution using acetonitrile（ A） and 0. 1% formic acid（ B） as mobile phase. The detection wavelength was set at 330 nm（ 0-29 min,for plantamajoside,acteoside,isoacteoside）,at350 nm（ 29. 01-31 min,for luteolin）,at 338 nm（ 31. 01-35 min,for apigenin）. The flow rate was 0. 8 m L·min-1 and the column temperature was 25 ℃. Results： The specificity of the established method was good,and there was no interference peaks. The linear ranges of plantamajoside,acteoside,isoacteoside,luteolin and apigenin were 42. 84-428. 40 μg·m L-1（ r =0. 999 7）,35. 36-353. 60 μg·m L-1（ r= 0. 999 5）,18. 40-184. 00 μg·m L-1（ r= 0. 999 5）,0. 856-8. 56 μg·m L-1（ r= 0. 9996） and 0. 108-1. 08 μg·m L-1（ r = 0. 999 6）,respectively. The average recoveries（ n = 6） were 98. 62%（ RSD = 0. 87%）,98. 09%（ RSD = 1. 10%）, 98. 19%（ RSD = 1. 16%）, 95. 52%（ RSD = 1. 03%） and 92. 34%（ RSD = 2. 00%）,respectively. The contents of five components in 6 batches for samples were 1. 51-6. 78,2. 53-18. 77,0. 60-2. 92,0. 03-0. 18 mg·g-1 and 5. 34-12. 36 μg ·g-1, respectively. Conclusion： The established method is simple, accurate, high sensitive and well reproducible,which could provide a theoretical basis for the quality control of Plantaginis herba.