DES-HPLC法同时测定桂枝中香豆素、桂皮酸和桂皮醛的含量

DES-HPLC to simutaneously determine the contents of coumarin,cinnamic acid and cinnamaldehyde in Cinnamomi Ramulus

目的建立绿色环保的低共熔溶剂萃取-高效液相色谱(DES-HPLC)法同时测定桂枝药材中的香豆素、桂皮酸和桂皮醛。方法氯化胆碱与1,4丁二醇在物质的量比为1∶4时组成的低共熔溶剂对桂枝中的香豆素、桂皮酸和桂皮醛有较高的提取效率,最佳提取条件为提取剂用量25 mL,超声(功率:250 W,频率:40 k Hz) 25 min,分析条件为以Phenomenex C18(250 mm×4. 60 mm,5μm)分离,用磷酸的体积分数为0. 05%的水溶液-乙腈(体积比为70∶30)洗脱,流速:1. 0 mL·min^(-1),检测波长:290 nm,柱温:25℃。结果香豆素在0. 430~17. 2 mg·L^(-1)、桂皮酸在0. 202~8. 08 mg·L^(-1)、桂皮醛在16. 4~98. 4 mg·L^(-1)内线性关系良好,三者的平均回收率分别为103. 4%(w,RSD=3. 5%)、103. 2%(w,RSD=4. 2%)和98. 4%(w,RSD=5. 5%),样品中香豆素、桂皮酸、桂皮醛的含量质量分数分别为(0. 218±0. 007)%、(0. 0616±0. 0007)%和(1. 43±0. 01)%,测定结果与《中华人民共和国药典》(2015年版)方法的测定结果无显著性差异。结论该方法精密度与重复性良好,可用于测定桂枝中的香豆素、桂皮酸和桂皮醛含量。

Objective To develop a green and efficient extraction and determination of coumarin,cinnamic acid and cinnamaldehyde in Cinnamomi Ramulus by the deep eutectic solvent extraction-high performance liquid chromatography（ DES-HPLC） method. Methods A deep eutectic solvent called CB2 that was made from a 1∶ 4 mixture of Choline chloride and 1,4-Butanediol using a heat-stiring method. The optimal extraction condition was extracted by ultrasound under 250 W for 25 min using 25 mL CB2. The analysis was performed on a Phenomenex C18 column（ 250 mm × 4. 60 mm,5 μm） with the elution of 0. 05%（ V∶ V） aqueous phosphoric acid-acetonitrile（ V∶ V = 70∶ 30） at a flowrate of 1. 0 mL·min-1. Column temperature was maintained at 25 ℃ and detection wavelength was set at 290 nm. Results Coumarin,cinnamic acid and cinnamaldehyde had good linear relationships in the ranges of 0. 430-17. 2,0. 202-8. 08,16. 4-98. 4 mg·L-1,respectively. The average recoveries were 103. 4%（ w,RSD = 3. 5%）,103. 2%（ w,RSD = 4. 2%） and 98. 4%（ w,RSD = 5. 5%）,respectively. The contents were（ 0. 218 ± 0. 007） %（ w）,（ 0. 061 6 ± 0. 000 7） %（ w）and（ 1. 43 ± 0. 01） %（ w）,respectively. There was no significant difference from the results of Chinese Pharmacopoeia（ 2015） using methanol as extraction reagent. Conclusion The method is precise and accurate,which is suitable for the extraction and determination of coumarin,cinnamic acid and cinnamaldehyde in Cinnamomi Ramulus.