高效液相色谱法同时测定甘草总黄酮口腔贴膜中甘草查尔酮A和甘草素含量

Simultaneous Determination of Licochalcone A and Liquiritigenin in Glycyrrhizic Flavone Oral Patch by HPLC

目的建立同时测定甘草总黄酮口腔贴膜中甘草查尔酮A和甘草素含量的高效液相色谱法。方法色谱柱为Aglient HC-C18柱(250 mm×4.6 mm,5μm),流动相为乙腈(A)-0.1%磷酸溶液(B),梯度洗脱(0~10 min时20%A~30%A,10~30 min时30%A~45%A,30~40 min时45%A~60%A,40~50 min时60%A~90%A),流速为1.0 m L/min,检测波长为276 nm(0~30 min)、365 nm(30~50 min),柱温为30℃,进样量为10μL。结果甘草查尔酮A、甘草素的质量浓度分别在4.16~166.40μg/m L和0.384~15.360μg/m L范围内与峰面积线性关系良好,精密度、稳定性、重复性试验的RSD均小于3%;平均加样回收率分别为99.04%和98.81%,RSD分别为1.06%和1.13%(n=9)。结论该方法操作简便、专属性强、准确度高,可用于甘草总黄酮口腔贴膜中甘草查尔酮A和甘草素的含量测定。

Objective To establish a high-performance liquid chromatography( HPLC) method for simultaneous determination of licochalcone A and liquiritigenin in Glycyrrhizic Flavone Oral Patch. Methods The chromatographic column was Aglient HC-C18 column( 250 mm ×4. 6 mm,5 μm),the mobile phase was acetonitrile( A)-0. 1% phosphoric acid solution( B) with gradient elution( 0-10 min,20% A-30% A;10-30 min,30% A-45% A;30-40 min,45% A-60% A;40-50 min,60% A-90% A),the flow rate was 1. 0 m L/min,the detection wavelengths were 276 nm( 0-30 min) and 365 nm( 30-50 min),the column temperature was 30 ℃,and the injection volume was 10 μL. Results The linear ranges of licochalcone A and liquiritigenin were 4. 16-166. 40 μg/m L,0. 384-15. 360 μg/m L,respectively. RSDs of precision,stability and repeatability tests were less than 3%. The average recoveries of licochalcone A and liquiritigenin were 99. 04%( RSD = 1. 06%,n = 9) and 98. 81%( RSD = 1. 13%,n = 9),respectively. Conclusion The method is simple,specific and accurate,which can be used for the content determination of licochalcone A and liquiritigenin in Glycyrrhizic Flavone Oral Patch.