高效液相色谱法同时测定独活中8种香豆素类化合物的含量

Simultaneous Determination of 8 Coumarins in Angelicae Pubescentis Radix by HPLC

目的建立高效液相色谱(HPLC)法同时测定独活中东莨菪内酯、二氢欧山芹素、补骨脂素、花椒毒素、香柑内酯、蛇床子素、异欧前胡素和二氢欧山芹醇当归酸酯8种香豆素类成分含量的方法,比较5批独活中8种香豆素类化合物的含量差异,为独活的质量控制提供参考。方法采用Intertsil ODS-SP C18色谱柱(250 mm×4.6 mm,5μm),以乙腈(A)-0.1%磷酸水溶液(B)为流动相,梯度洗脱,流速1.0 mL·min^(-1),检测波长320 nm,柱温30℃,进样量10μL。结果独活中8种香豆素类成分均可实现较好的分离,东莨菪内酯、二氢欧山芹素、补骨脂素、花椒毒素、香柑内酯、蛇床子素、异欧前胡素和二氢欧山芹醇当归酸酯的线性范围分别为3.375~108.00、3.375~108.00、14.75~472.00、127.50~4080.00、2.484~79.50、39.25~1256.00、4.453~142.50和36.625~1172.00μg·mL^(-1),线性关系良好(r≥0.9996);平均加样回收率在98.20%~101.74%之间,RSD均小于3%。5批独活样品中8种香豆素类成分含量差异明显。结论该方法简便准确、可靠稳定、重复性较好,可用于独活中多种香豆素类成分的含量测定,为独活的质量标准的提升提供实验依据。

ObjectiveA method for simultaneous determination of 8 coumarins(scopoletin, columbianetin,psoralen,methoxsalen,bergapten,osthole,isoimperatorin,columbianadin)in root of Angelica pubescens by high performance liquid chromatography(HPLC)was established. The content differences of 8 coumarins in 5 batches of Angelica pubescens were compared to provide reference for the quality control of Angelicae pubescentis radix.MethodsHPLC analysis was performed on Intertsil ODS-SP C18 column(250 mm×4.6 mm,5 μm). The mobile phase was acetonitrile(A)-0.1% phosphoric acid solution(B) with gradient elution. Detection wavelength was320 nm,and flow rate of mobile phase was 1.0 mL·min;. The column temperature was 30 ℃,and injection volume was 10 μL.ResultsThe 8 chemical constituents of roots of Angelica pubescens could be well separated. The linear ranges of scopoletin, columbianetin, psoralen, methoxsalen, bergapten, osthole, isoimperatorin, and coumbianadin were 3.375-108.00, 3.375-108.00, 14.75-472.00, 127.50-4 080.00, 2.484-79.50, 39.25-1 256.00,4.453-142.50,and 36.625-1 172.00 μg·mL^(-1),respectively,with good linear relationships(r ≥ 0.999 6).The average recoveries ranged from 98.20% to 101.74%,RSDs<3%. The contents of 8 coumarins in 5 batches of Angelica pubescens roots were significantly different.ConclusionThe method is convenient and accurate,reliable,and reproducible. It can be used for the determination of coumarins in Angelica pubescens root, and provide experimental basis for improving the quality standard of Angelicae pubescentis radix.