华佗豆的红外指纹图谱和薄层色谱鉴别及酚酸类成分含量测定

Infrared Fingerprint,TLC Identification and Content Determination of Phenolic AcidComponents of Calonyction muricatum(Linn)G.

目的建立瑶药华佗豆药材的红外指纹图谱、薄层色谱鉴别和酚酸类含量测定的方法。方法采用红外光谱法构建10批华佗豆药材的指纹图谱,采用相似度分析、聚类分析、主成分分析及偏最小二乘判别分析对光谱数据进行分析;采用薄层色谱法对华佗豆中的绿原酸、异绿原酸A、咖啡酸进行定性鉴别;采用高效液相色谱(HPLC)法同时测定新绿原酸、绿原酸、咖啡酸、隐绿原酸、异绿原酸A、异绿原酸C的含量。结果红外光谱法可推测出华佗豆含有有机酸、黄酮类等成分,红外指纹图谱共标定9个共有峰,相似度评价>0.999。聚类分析、主成分分析结果显示分为两大类,产地S1、S2、S3聚为一类,其余产地为一类;偏最小二乘判别分析筛选出5种差异性成分(VIP>1);建立的薄层色谱的供试品和对照品显示位置一致,且斑点清晰,分离度好;华佗豆中6种酚酸类成分线性关系均良好(r≥0.9992),平均加样回收率97.77%~102.59%,相对标准偏差(RSD)均≤2.90%。结论建立的华佗豆药材薄层色谱和红外指纹图谱方法简便可行、结果稳定;建立的华佗豆中6种酚酸类成分的含量测定方法结果可靠,为该药材的质量控制研究奠定科学基础。

Objective To establish the method of infrared fingerprint,TLC identification and content determination of phenolic acid components of Yao medicine Calonyction muricatum(Linn)G.Methods The infrared fingerprint of 10 batches of Calonyction muricatum(Linn)G were established by infrared spectroscopy.The spectral datas were analyzed by similarity analysis,infrared spectroscopy(HCA),principal component analysis(PCA)and Partial-least-squares discriminant analysis(PLS-DA).Chlorogenic acid,heterochlorogenic acid A and caffeic acid of Calonyction muricatum(Linn)G were identified by TLC.The contents of neochlorogenic acid,chlorogenic acid,caffeic acid,cryptochlorogenic acid,isochlorogenic acid A and C were determined simultaneously by HPLC method.Results It could be suggested that organic acids,flavonoids and other compounds of Calonyction muricatum(Linn)G by infrared spectroscopy and nine common peaks were calibrated by infrared fingerprint;the similarity evaluation was above 0.999;the results of cluster analysis(CA)and principal component analysis(PCA)showed that it could be clustered into 2 categories,including S1,S2 and S3 were clustered into one categoriy and the rest were one.5 differential components(VIP>1)were selected by Partial-least-squares discriminant analysis;the test and control samples of TLC showed consistent locations the spots were clear,with good separation degree;the six components of Calonyction muricatum(Linn)G showed good linear relationship(r≥0.9992),average sample recovery rate 97.77%-102.59%,and RSD less than 2.90%.Conclusion The TLC and infrared fingerprint were simple and stable,and the results of the six components were reliable,which can lay a scientific foundation for the quality control of the materials.