HPLC法测定车前草中大车前苷和毛蕊花糖苷含量

目的: 建立测定车前草中大车前苷和毛蕊花糖苷含量的色谱方法,用于指导车前草药材的质量控制。方法: 在中国药典色谱条件的基础上,增加毛蕊花糖苷为指标性成分,改变流动相、柱温等条件,建立同时测定大车前苷和毛蕊花糖苷的含量测定方法。色谱柱:Waters Xbridge C₁₈(4.6 mm×250 mm,5 μm);流动相:0.05% 三氟乙酸(A)-乙腈(B),梯度洗脱(0~35 min,12%B→13%B;35~45 min,13%B→15%B;45~50 min,15%B→17%B);流速:1 mL·min^-1;柱温:40 ℃ ;检测波长:330 nm;进样量:10 μL。结果: 大车前苷质量浓度在0.49~250 mg·L^-1 范围内,毛蕊花糖苷质量浓度在0.50~255 mg·L^-1 范围内,线性关系良好。在稳定性、精密度试验中,大车前苷的RSD 分别为0.22% 和0.36%,毛蕊花糖苷的RSD 分别为0.15%和0.31%;在重复性试验中,车前和平车前中大车前苷的RSD 分别为0.25% 和0.45%,毛蕊花糖苷分别为0.20% 和0.11%。采用改进后的色谱条件和药典色谱条件测定车前草中大车前苷的含量,P<0.01,结果具有显著性差异;改进后的色谱条件对于大车前苷及其类似物具有更好的分离效果。结论: 改进后的色谱条件结果准确,重复性好,可以用于车前草质量控制。

Objective: To establish an HPLC method for the determination of plantamajoside and acteoside in Plantaginis Herba,so as to instruct the quality control of plantain herb. Methods: Base on the chromatography condition in Chinese Pharinacopoeia,acteoside was used as a chemical maker for plantaginis Herbs. The mobile phase and the column temperature have been optimized to simultaneously determine these two compounds. HPLC analysis was carried out with a Waters Xbridge C₁₈ column(4.6 mm×250 mm,5 μm). The HPLC mobile phase consisted of 0.05% aqueous trifluoroacetic acid (A) and acetonitrile (B) with a gradient program (0-35 min,12%B→13%B;35-45 min,13%B→15%B;45-50 min,15%B→17%B) at the flow rate of 1 mL min^-1,the column temperature was 40 ℃,the detection wavelength was 330 nm,and the injection volume was 10 μL. Results: Good linearity of the proposed method was shown in the range of 0.49-250 mg·L^-1 for plantamajoside and 0.50-255 mg·L^-1 for acteoside. In the test of stability and precision,the RSDs of plantamajoside and acteoside were respectively 0.22%,0.36% and 0.15%,0.31%. In the test of repeatability,the RSDs of plantamajoside and acteoside in P. asiatica L. and P. depressa Willd. were respectively 0.25%,0.45% and 0.20%,0.11%. It was found that the quantity of plantamajoside is significantly different from the method in the Chinese pharmacopeia (P<0.01). The developed method showned adequate separation for pantamajoside from its analogue. Conclusion: The developed method is accurate and repeatable,and therefore suitable for the quality control of Plantaginis Herba.