乌拉尔甘草的UPLC指纹图谱研究

Study on UPLC Fingerprint of Glycyrrhiza uralensis

目的:建立乌拉尔甘草的超高效液相色谱(UPLC)指纹图谱。方法:采用UPLC法。色谱柱为Waters CORTECS UPLC C18,流动相为乙腈-0.1%甲酸溶液(梯度洗脱),流速为0.3 m L/min,检测波长为254 nm,柱温为30℃,进样量为1μL。以甘草酸为参照,测定27批药材样品的UPLC图谱,采用《中药色谱指纹图谱相似度评价系统》(2004 A版)进行相似度评价,确定共有峰,并对27批药材样品进行聚类分析。结果:27批药材样品的UPLC图谱有20个共有峰;除S2、S4、S19、S21、S22、S24药材样品的相似度小于0.90外,其余21批药材样品相似度均大于0.90;经验证,该21批药材样品UPLC图谱与对照图谱具有较好的一致性。27批药材样品可聚为3类,S24为Ⅰ类,S2、S4、S12、S19、S21、S22为Ⅱ类,其余为Ⅲ类。结论:该研究所建指纹图谱可为甘草的质量评价提供参考。

OBJECTIVE：To establish UPLC fingerprint of Glycyrrhiza uralensis. METHODS：UPLC method was adopted.The determination was performed on Waters CORTECS UPLC C18 column with mobile phase consisted of acetonitrile-0.1% formic acid（gradient elution）at the flow rate of 0.3 m L/min. The detection wavelength was set at 254 nm,and column temperature was 30 ℃. The sample size was 1 μL. Using glycyrrhizic acid as control,UPLC chromatograms of 27 batches of sample were determined. Similarity evaluation was conducted by using TCM Chromatogram Fingerprint Similarity Evaluation System（2004 A edition） to determine common peak and conduct cluster analysis of 27 batches of samples. RESULTS：There were 20 common peaks in UPLC chromatograms of 27 batches of samples. The similarity degree of S2,S4,S19,S21,S22,S24 were less than 0.90,the others samples were more than 0.90. After validation,UPLC chromatograms of 21 batches of batches of samples were in good agreement with control fingerprint. 27 batches of samples were clustered into 3 categories,in which S24 was categoryⅠ;S2,S4,S12,S19,S21,S22 were categoryⅡ;other were categoryⅢ. CONCLUSIONS：Established fingerprint can provide reference for quality evaluation of G. uralensis.