HPLC测定决明子中决明子苷A，B含量

摘要

OBJECTIVE:To establish a HPLC assay for determining cassiaside A and B in the seeds of Cassia obtusifolia L.METHODS:The dried powder of the seeds of the plant was defatted with CHCl3 in a Sothles apparatus followed by extraction with MeOH.The MeOH extra was injected for analysis.A μ-Bondapak C18 column (3.9nm×300mm,10μm) was used with a mobile phase of acetonitrile-water-THF-acetic acid (20∶76.5∶3.0∶0.5).The flow rate was 1.0ml.min-1.A UV detector at 278nm was used.The peaks of cassiaside A and B were identified by co-chromatography and spectrophoto metry with their standards.RESULTS:The peas of cassiaside A and B in the chromatogram from seeds C. obtusifolia are pure.The retention time of A and B were 12.13 min and 13.27 min,respectively.The standard curves of A and B were linear in the range from 0.25 to 1.25 μg (r=0.9999) and 0.125 to 0.625 μg (r=0.9999),respectively.The average recoveries were (96.83±3.53)% for A and (97.78±2.16)% for B.CONCLUSION:The method is sensitive and specific.It is suitable for analysis of cassiaside A and B in the medicinal materials.%目的：建立决明子中降血脂有效成分决明子苷A，B(以下简称苷A，B)的HPLC定量测定方法。方法：决明子药材经氯仿脱脂后再由甲醇提取，甲醇提取液经释释后直接进样分析；在μ-Bondapak C18柱上，以乙腈--水-四氢呋喃-冰醋酸(20∶76.5∶3.0∶0.5)为流动相，流速为1ml.min-1，检测波长为278nm，测定了样品中苷A、B含量，并采用光谱法和色谱法对A、B峰的色谱纯度进行了鉴定。结果：苷A、B的保留时间分别为12.13min和13.27min，经鉴定苷A、B峰均为单一物质色谱峰：苷A、B的标准曲线分别在0.25～1.25μg(r=0.9999)和0.125μg～0.625μg(r=0.9999)之间有较好线性关系，苷A、B的加样回收率分别为(96.83±3.53)%和(97.78±2.16)%。结论：本法专属性强，可用于决明子中决明子苷A、B的含量测定。