祖卡木颗粒中8个成分的含量测定及聚类分析

Content Determination of 8 Constituents in Zukamu Granules and Cluster Analysis

目的:建立同时测定祖卡木颗粒中8种成分含量的方法。方法:采用HPLC法,色谱柱:Agilent Eclipse Plus C18柱(250 mm×4.6 mm,5μm),流动相:乙腈(A)-0.05%磷酸水溶液(B),梯度洗脱;流速:1.0 ml·min-1;检测波长:245 nm(大黄素、甘草酸铵)、276 nm(没食子酸、甘草苷)、309 nm(对甲氧基肉桂酸乙酯)、360 nm(烟花苷、槲皮素、山奈酚);柱温:35℃;进样量:10μl。采用SPSS 22统计软件对含量测定结果进行聚类分析。结果:没食子酸、甘草苷、烟花苷、槲皮素、山奈酚、对甲氧基肉桂酸乙酯、甘草酸铵和大黄素检测质量浓度线性范围分别为3.566~71.328,5.475~109.492,0.466~9.328,7.367~147.344,0.349~6.976,1.528~30.568,7.340~146.796,3.959~79.188μg·ml-1(r>0.9990);平均加样回收率为95.7%~99.4%,RSD为0.65%~1.44%(n=6);12批样品聚类分析为2类。结论:本法简便、准确、重复性好,可用于祖卡木颗粒的鉴定与评价,作为祖卡木颗粒质量控制的重要科学依据之一。

Objective:To establish a method for the simultaneous determination of 8 constituents in Zukamu granules.Methods:An HPLC analysis was performed on an Agilent Eclipse Plus C18(250 mm×4.6 mm,5μm)column with the column temperature at 35℃and the sample size was 10μl.The mobile phase was acetonitrile(A)-0.05%acetic acid solution(B)with gradient elution at the flow rate of 1.0 ml·min-1 and the detection wavelength was 245 nm for ammonium-glycyrrhizinate and emodin,276 nm for gallic acid and liquiritin,309 nm for ethyl-4-methoxycinnamate,and 360 nm for kaempferol-3-O-rutinoside,quercetin and kaempferol.The cluster analysis was conducted for the results of content determination by SPSS 22 statistical software.Results:The linear range was 3.566-71.328μg·ml-1 for gallic acid,5.475-109.492μg·ml-1 for liquiritin,0.466-9.328μg·ml-1 for kaempferol-3-O-rutinoside,7.367-147.344μg·ml-1 for kaempferol,0.349-6.976μg·ml-1 for quercetin,1.528-30.568μg·ml-1 for ethyl-4-methoxycinnamate,7.340-146.796μg·ml-1 for ammonium-glycyrrhizinate and 3.959-79.188μg·ml-1 for emodin r>0.9990.The average recoveries were 95.7%-99.4%and the RSDs were 0.65%-1.44%(n=6).The samples of 12 batches of Zukamu granules could be divided into 2 categories.Conclusion:The method is simple and accurate with good repeatability,which can be used for the identification and evaluation of Zukamu granules,and provides important scientific basis for the quality control of Zukamu granules.