加压溶剂提取-HPLC法测定北沙参中6种香豆素类化合物

Simultaneous determination of six coumarins in Radix Glehniae using pressurized liquid extraction and HPLC

目的建立高效液相色谱法测定不同产地北沙参中6种香豆素类化合物。方法用加压溶剂提取技术和高效液相色谱法对北沙参药材进行分析。色谱柱:Kinetex core-shell-C_(18)column(50 mm×4.6 mm,2.6μm),流动相:乙腈-水梯度洗脱,流速:0.4 mL·min^(-1),柱温:30℃,检测波长:310 nm,进样量:20μL。考察该方法的专属性、标准曲线和定量下限、精密度与回收率、稳定性以及重现性。结果北沙参中6种主要香豆素类成分,骨脂素、花椒毒素、佛手柑内酯、欧前胡素、cnidilin和异欧前胡素分离良好,分别在0.50~10.00,1.00~20.00,0.32~6.40,0.25~5.00,0.05~1.00,0.17~3.50μg·mL^(-1)质量浓度内线性关系良好(r≥0.999 2),且回收率较高(均在92.8%~99.2%内)。不同产地北沙参药材中香豆素类成分含量存在明显差异。结论用加压溶剂提取和高效液相色谱法能快速、有效地分析北沙参中6种香豆素类成分,方法准确可靠,重复性好,结果稳定,可作为该药材质量控制的方法。

Objective To develop a HPLC method for simultaneous determination of six coumarins in Radix Glehniae from different sources.Methods The samples were analyzed by HPLC after pressurized liquid extraction（PLE）.The chromatographic conditions were as following：The column was Kinetex core- shell- C_（18） column（50 mm × 4.6 mm,2.6 μm）,the mobile phase consisted of acetonitrile- water using gradient elution,the flow rate was 0.4 mL · min^-1 and the column temperature was 30 ℃.The detection wavelength was set at 310 nm and the sample injection volume was 20 μL.The method was validated by investigating its specificity,linearities and limit of detection,precision,extraction recovery,stability and repeatability.Results Six components were determined by HPLC,including psoralen,xanthotoxin,bergapten,impera—torin,cnidilin and isoimperatorin.The linearities of them were good（r≥0.999 2） in the concentration ranges of 0.50- 10.00,1.00-20.00,0.32- 6.40,0.25- 5.00,0.05- 1.00,0.17- 3.50μg · mL^-1,respectively.The coumarins showed the overall recoveries ranging from 91.7%-107.6%.The contents of six compounds of Radix Glehniae from different areas had an obvious difference.Conclusion Six coumarins in Radix Glehniae were analyzed by PLE and HPLC.The method has good specificity,reproducibility and high sensitivity,which can be applied for the quality control of Radix Glehniae.