克咳片的HPLC指纹图谱研究及聚类分析

Study on HPLC Fingerprint and Cluster Analysis of Keke Tablets

目的:建立克咳片的高效液相色谱(HPLC)指纹图谱,并进行聚类分析,考察克咳片高效液相图谱的一致性。方法:采用HPLC法,色谱柱为Waters Sunfire C18(250 mm×4.6 mm,5μm),流动相为乙腈-0.1%磷酸溶液(梯度洗脱),流速为0.8ml·min^-1,检测波长为210 nm、250 nm,柱温为30℃,进样量为10μl。以甘草酸铵为参照,测定10批克咳片样品的HPLC图谱,采用《中药色谱指纹图谱相似度评价系统》(2012 A版)对10批样品进行相似度评价,确定共有峰。采用SPSS 22.0软件进行聚类分析。结果:10批样品的HPLC图谱有24个共有峰,相似度均>0.90;经验证,10批样品HPLC图谱与对照指纹图谱具有较好的一致性。10批样品可聚为两大类。结论:所建指纹图谱可为克咳片的质量评价提供参考。

Objective:To establish an HPLC fingerprint of Keke tablets,and study the consistency of HPLC chromatograms of Keke tablets by cluster analysis(CA)method.Methods:An HPLC method was adopted.The determination was performed on a Waters Sunfire C18(250 mm×4.6 mm,5μm)column with the mobile phase consisting of acetonitrile-0.1%formic acid solution(gradient elution)at the flow rate of 0.8 ml·min^-1.The detection wavelengths were 210 nm and 250 nm.The column temperature was 30℃,and the sample size was 10μl.Using glycyrrhizic acid ammonium salt as the reference,the HPLC fingerprints of the 10 batches of Keke tablets were determined.The similarity of the 10 batches of samples was evaluated by using TCM Chromatographic Fingerprint Similarity Evaluation System(2012 edition),and the common peak was identified.CA was conducted by using SPSS22.0 software.Results:There were 24 common peaks in the HPLC chromatograms of the 10 batches of samples.The similarity of HPLC fingerprints of the 10 batches of samples was higher than 0.90 and the HPLC fingerprint was in good agreement with the control fingerprint.The 10 batches of samples could be clustered into 2 categories.Conclusion:The established fingerprint can provide reference for the quality evaluation of Keke tablets.