高效液相色谱法同时测定徐长卿提取物中6种酚酸类成分含量

Simultaneous Determination of Six Phenolic Acids in the Extract of Cynanchi Paniculati Radix et Rhizoma by HPLC

目的建立同时测定徐长卿提取物中绿原酸、香草酸、橙皮苷、丹皮酚原苷、白前苷B及丹皮酚含量的高效液相色谱法。方法色谱柱为AgilentEclipse XDB C_(18)柱(250 mm×4.6 mm,5μm),流动相为0.2%甲酸水溶液-乙腈(梯度洗脱),流速为1.0 mL/min,检测波长为275 nm,柱温为40℃。结果绿原酸、香草酸、橙皮苷、丹皮酚原苷、白前苷B及丹皮酚的质量浓度分别在3.9575~158.3μg/mL、2.4625~98.5μg/mL、2.56~102.4μg/mL、2.8625~114.5μg/mL、2.64~105.6μg/mL、3.1175~124.7μg/mL范围内与峰面积线性关系良好(r≥0.9996,n=6);精密度试验结果的RSD分别为1.15%,0.83%,1.79%,2.05%,1.72%,1.31%(n=6);加样回收率分别为98.47%,98.74%,98.83%,98.45%,98.51%,98.69%,RSD分别为1.30%,0.95%,1.15%,0.83%,1.26%,1.51%(n=6);3批样品中的平均含量分别为1.151,0.434,0.715,1.175,0.925,1.542 mg/g(n=3)。结论所建立的方法简便快捷、准确可靠,可用于徐长卿相关产品的质量评价。

Objective To establish a high-performance liquid chromatography(HPLC)method for the simultaneous determination of chlorogenic acid,vanillic acid,hesperidin,paeonolide,vincetoxicosid B and paeonol in the extract of Cynanchi Paniculati Radix et Rhizoma.Methods The chromatographic column was Agilent Eclipse XDB C_(18) column(250 mm×4.6 mm,5μm),the mobile phase was 0.2%formic acid aqueous solution-acetonitrile(gradient elution),the flow rate was 1.0 mL/min,the detection wavelength was 275 nm,and the column temperature was 40℃.Results The linear ranges of chlorogenic acid,vanillic acid,hesperidin,paeonolide,vincetoxicosid B and paeonol were 3.9575-158.3μg/mL,2.4625-98.5μg/mL,2.56-102.4μg/mL,2.8625-114.5μg/mL,2.64-105.6μg/mL and 3.1175-124.7μg/mL(r≥0.9996,n=6),respectively.The RSDs of precision tests were 1.15%,0.83%,1.79%,2.05%,1.72%and 1.31%(n=6),respectively.The recovery rates of the above six phenolic acids were 98.47%,98.74%,98.83%,98.45%,98.51%and 98.69%with RSDs of 1.30%,0.95%,1.15%,0.83%,1.26%and 1.51%(n=6),respectively.The average contents of the above six phenolic acids in the three batches of samples were 1.151,0.434,0.715,1.175,0.925 and 1.542 mg/g(n=3),respectively.Conclusion The established method is simple,rapid,accurate and reliable,which can be used for the quality evaluation of related products of Cynanchi Paniculati Radix et Rhizoma.