目的以β-环糊精为流动相添加剂,拆分积雪草药材及其总苷中羟基积雪草苷和积雪草苷,建立羟基积雪草苷和积雪草苷的定量测定方法,同时建立积雪草总苷的高效液相色谱特征图谱。方法采用Dikma Diamon-silTMC18(250 mm×4.6 mm,5μm)色谱柱;流动相为乙腈-2 mmol/Lβ-环糊精溶液(24:76);柱温30℃;体积流量1.0 mL/min;检测波长205 nm;理论塔板数按积雪草苷峰计算应不低于4 000。结果 β-环糊精的加入很好地分离积雪草药材及其总苷中的羟基积雪草苷和积雪草苷,满足了定量分析的要求,其中有效成分羟基积雪草苷和积雪草苷的线性范围分别为0.181~18.1μg(r=0.999 98)和0.195~19.5μg(r=0.999 98),积雪草药材的平均加样回收率(n=9)分别为101.2%和101.6%,积雪草总苷的平均加样回收率(n=9)分别为100.3%和99.3%,11批次的积雪草总苷样品的高效液相色谱特征图谱中确定6个共有峰作为定性的指标峰。结论 本法简便快速、结果准确、重现性好,可用于积雪草药材及其总苷的质量控制。

Objective To separate the isomers of madecassoside in Centellae Herba and its total glucosides,determine madecassoside and asiaticoside in them,and establish an HPLC specific chromatogram for Centellae Herba total glucosides by using β-cyclodextrin as mobile phase additives.Methods The Dikma DiamonsilTM C18 column(250 mm×4.6 mm,5 μm) was served as analytical column and acetonitrile-2 mmol/L β-cyclodextrin(24:76) as mobile phase at a flow rate of 1.0 mL/min,the detection wavelength was 205 nm and column temperature was 30 ℃.Theoretical plate number of asiaticoside was over 4 000.Results The isomers in Centellae Herba and its total glucosides were separated with the additives of β-cyclodextrin,meeting the requirements of quantitative analysis.In which,madecassoside and asiaticoside showed a good linearity in the range of 0.181—18.1 μg(r=0.999 98) and 0.195—19.5 μg(r=0.999 98) each.The mean recovery of madecassoside and asiaticoside in Centellae Herba was 101.2% and 101.6%(n=9),and that of its 11 batches of total glucosides was 100.3% and 99.3%,respectively.Under the qualitative conditions,the six common peaks in HPLC specific chromatogram of its 11 batches of total glucosides could be used as marker peaks for qualitative identification.Conclusion The analytical method is proved to be convenient and accurate,which can be used for the quality control of Centellae Herba and its total glucosides.