HPLC双波长切换法同时测定加味益母草膏中5个成分的含量

Determination of five components in modified Yimucao ointment by HPLC dual-wavelength method

目的:建立HPLC双波长切换法同时测定加味益母草膏中益母草碱、芍药苷、芦丁、阿魏酸和毛蕊花糖苷5个成分含量。方法:采用Phenomenex Luna C_(18)色谱柱(250 mm×4.6 mm,5μm),以甲醇-0.1%磷酸水溶液为流动相,梯度洗脱(0~8 min,12%A→23%A;8~25 min,23%A→38%A;25~50 min,38%A→50%A),流速为1.0 mL·min^(-1);检测波长为230 nm(益母草碱、芍药苷、芦丁)和322 nm(阿魏酸、毛蕊花糖苷),进样量为20μL,柱温为30℃。结果:益母草碱、芍药苷、芦丁、阿魏酸和毛蕊花糖苷的平均含量分别为1.586、0.348、0.186、0.036、0.034 mg·g^(-1),5个成分线性关系良好(r>0.9995),平均加样回收率为98.8%~101.3%,RSD为1.1%~2.0%。结论:此方法高效、简便,易操作,可同时测定加味益母草膏中5个成分的含量,检测结果准确度高,精密度好,可用于加味益母草膏的质量控制,也可为加味益母草膏的进一步研究及临床应用提供科学依据。

Objective:To develop an HPLC dual-wavelength method for thesimultaneously determination of five components of leonurus,paeoniflorin,rutin,ferulic acid and verbascoside in modified Yimucao Ointment.Methods:Using methanol-0.1%phosphoric acid solution as mobile phase and gradient elution,flow rate was 1.0 mL·min^(-1).Detection wavelengths were 230 nm for leonurus,paeoniflorin,rutin,and 322 nm for ferulic acid andverbascoside.Column temperature was 30℃.Injection volume was 20μL.Results:The five components of leonurus,paeoniflorin,rutin,ferulic acid and verbascoside had a good linear relationship(r>0.9995),the average recovery was 98.8%-101.4%,and the RSD was 1.1%-2.0%.The average contents of five components were 1.586,0.348,0.186,0.036,0.034 mg·g^(-1),respectively.Conclusion:This method is efficient,simple and easy to operate,and can be used to determine the content of 5 components in modifiedYimucao Ointment at the same time.The results are accurate and precise.It can also provide scientific basis for further research and clinical application of modified Yimucao Ointment.