Huang Hai-Qiang, Zhang Xi, Xu Zhi-Xiu, Su Juan, Yan Shi-Kai, Zhang Wei-Dong
Department of Natural Medicinal Chemistry, School of Pharmacy, Second Military Medical University, Shanghai, China.
J Pharm Biomed Anal. 2009 May 1;49(4):1048-55. doi: 10.1016/j.jpba.2009.01.011. Epub 2009 Jan 20.
A rapid resolution liquid chromatography coupled with evaporative light scattering detection method was established for simultaneous determination of six saikosaponins, namely saikosaponin a, saikosaponin c, saikosaponin d, 6''-O-acetylsaikosaponin a, 3''-O-acetylsaikosaponin d and 6''-O-acetylsaikosaponin d in Bupleurum. The analysis was performed by using an Agilent Zorbax SB-C18 column (1.8 microm, 3.0 mm x 50 mm i.d.) at gradient elution of water and acetonitrile, and the saikosaponins were well separated within 12 min, which provided about a fourfold reduction in analysis time by comparing a conventional high performance liquid chromatography method. Owing to their low ultraviolet absorption, the saikosaponins were detected by evaporative light scattering. The standard curves to quantify the saikosaponins were constructed by the log-log plot, which showed good linearity with the correlation coefficients exceeding 0.9954. The detection limits and quantification limits ranged in 8.38-25.00 microg/mL and 25.13-45.00 microg/mL, respectively. Satisfactory intra-day and inter-day precisions were achieved with the relative standard deviation (R.S.D.) less than 6.58%, and the average recoveries obtained were in the range of 96.9-100.4%. In addition, MeOH-1.0% (v/v) pyridine was found to be the best the extraction solvent when compared to MeOH and MeOH-1.0% (v/v) ammonia water. A total of 23 samples of roots of Bupleurum from different species or locations were examined with this analytical method, and their chemical profiles provided information for the chemotaxonomic investigation. The results demonstrated that the analytical method is highly effective for the quality evaluation of Bupleurum species.
建立了一种快速液相色谱-蒸发光散射检测法，用于同时测定柴胡中6种柴胡皂苷，即柴胡皂苷a、柴胡皂苷c、柴胡皂苷d、6''-O-乙酰基柴胡皂苷a、3''-O-乙酰基柴胡皂苷d和6''-O-乙酰基柴胡皂苷d。采用Agilent Zorbax SB-C18柱（1.8μm，3.0mm×50mm内径），以水和乙腈为流动相进行梯度洗脱，12分钟内柴胡皂苷得到良好分离，与传统高效液相色谱法相比，分析时间缩短了约四倍。由于柴胡皂苷紫外吸收低，采用蒸发光散射检测器进行检测。通过对数-对数图构建柴胡皂苷定量标准曲线，线性关系良好，相关系数均大于0.9954。检测限和定量限分别为8.38-25.00μg/mL和25.13-45.00μg/mL。日内和日间精密度良好，相对标准偏差（R.S.D.）小于6.58%，平均回收率为96.9-100.4%。此外，与甲醇和甲醇-1.0%（v/v）氨水相比，甲醇-1.0%（v/v）吡啶是最佳提取溶剂。采用该分析方法对23个不同品种或产地的柴胡根样品进行了检测，其化学图谱为化学分类学研究提供了信息。结果表明，该分析方法对柴胡属植物的质量评价具有高效性。
