基于多成分含量测定结合化学计量学考察甘草不同提取方法对药效成分的影响

Study on effects of different extraction methods of licorice based on multi-component content determination and chemometrics

以9个产地甘草药材为原料,通过水浴回流法、超声波辅助法和微波辅助法获得甘草药材提取物,采用超高效液相色谱(UPLC)法测定甘草药材中芹糖甘草苷、甘草苷、芹糖异甘草苷、甘草查尔酮B、甘草素、刺甘草查尔酮以及甘草酸的含量,并建立指纹谱图。通过主成分分析(PCA)和正交偏最小二乘法—判别分析(OPLS-DA)研究不同提取方法对甘草药材溶出成分的影响。结果表明:不同提取方法建立的甘草指纹谱图中有30个共有峰,通过对照品指认出芹糖甘草苷、甘草苷、芹糖异甘草苷、甘草查尔酮B、甘草素、刺甘草查尔酮和甘草酸7个峰;不同提取方法甘草药材的相似度为0.832~0.985;利用PCA和OPLS-DA分析将不同提取方法甘草药材划分为3类,分别对应3种提取方法,共确定9个差异标志物,影响显著性排序分别为峰25>峰12(芹糖异甘草苷)>峰20>峰9>峰14>峰18>峰30>峰13>峰29。9批甘草药材中7种药效成分的含量采用微波辅助法提取时最高,分别是超声波辅助法的113%~135%,水浴回流法的115%~150%。微波辅助法对甘草药材溶出成分最好。

The experiment was carried out by using licorice as raw material and extraction of 9 batches of licorice by water bath reflux, ultrasonic-assisted and microwave-assisted method.The content of celery glycyrrhizin, glycyrrhizin, celery isoliquiritin, licorice chalcone B,glycyrrhizin, licorice chalcone and glycyrrhizic acid in licorice was determined by ultra performance liquid chromatography(UPLC),and the fingerprint was established.And then, the principal component analysis(PCA) and orthogonal partial least squares-discriminant analysis(OPLS-DA) were used to find the effects of different extraction methods on the dissolution components of licorice.The result shows there are 30 common peaks in the fingerprints established by different extraction methods of licorice,and 7 peaks of celery glycyrrhizin, glycyrrhizin, celery isoliquiritin, licorice chalcone B,glycyrrhizin, licorice chalcone and glycyrrhizic acid are identified by the reference substance.The similarity of licorice medicinal materials with different extraction methods was 0.832~0.985.The licorice medicinal materials with different extraction methods were divided into three categories by PCA and OPLS-DA analysis.A total of 9 differential markers were identified: peak 25>peak 12(apioside isoliquiritigenin)>peak 20>peak 9>peak 14>peak 18>peak 30>peak 13>peak 29.The contents of seven active ingredients in licorice extracted by microwave-assisted method were the highest, which was 113%~135% by ultrasonic method and 115%~150% by water bath method.