潜阳育阴颗粒UPLC指纹图谱和多成分定量方法的建立

Establishment of UPLC fingerprint and quantitative method of multi-components for Qianyang yuyin granules

目的建立潜阳育阴颗粒的指纹图谱和多成分定量方法。方法采用超高液相色谱(UPLC)法,以马钱苷峰为参照,建立15批潜阳育阴颗粒的UPLC指纹图谱,确定共有峰并采用《中药色谱指纹图谱相似度评价系统(2004A版)》评价相似度;进行聚类分析(CA)、主成分分析(PCA)、偏最小二乘法-判别分析(PLS-DA),并以变量重要性投影(VIP)值大于1.0为标准筛选影响潜阳育阴颗粒质量的差异性成分;采用UPLC法测定差异性成分和哈巴苷、阿魏酸、杯苋甾酮的含量。结果15批潜阳育阴颗粒样品有18个共有峰,相似度均大于0.96;指认峰6为莫诺苷,峰7为马钱苷,峰11为金丝桃苷,峰12为二苯乙烯苷,峰13为阿魏酸,峰14为杯苋甾酮,峰16为哈巴俄苷,峰17为肉桂酸,峰18为大黄素。CA和PCA结果均显示,15批样品可分为6类,S1、S2、S4~S6为一类;S3为一类,S7、S8为一类,S9、S10为一类,S11~S14为一类,S15为一类;PLS-DA结果显示,峰3、9、6(莫诺苷)、7(马钱苷)、15、12(二苯乙烯苷)、16(哈巴俄苷)的VIP值均大于1.0。15批样品中,哈巴苷、哈巴俄苷、莫诺苷、马钱苷、二苯乙烯苷、阿魏酸、杯苋甾酮的含量分别为0.336~0.563、0.205~0.377、0.692~0.837、0.433~0.562、1.294~2.030、0.020~0.027、0.167~0.235 mg/g。结论成功建立了潜阳育阴颗粒的UPLC指纹图谱和多成分定量方法,可用于提高和完善该制剂的质量控制标准;莫诺苷、马钱苷、二苯乙烯苷、哈巴俄苷等可能是影响该制剂质量的差异性成分。

OBJECTIVE To establish a fingerprint and quantitative method of multi-components for Qianyang yuyin granules.METHODS Using the peak of loganin as reference,fingerprints of 15 batches of Qianyang yuyin granules were established by ultra-high liquid chromatography(UPLC).The common peaks were identified and the similarity analysis was performed by Similarity Evaluation System of Chromatographic Fingerprint of TCM(2004A edition).Cluster analysis(CA),principal component analysis(PCA)and partial least square-discriminant analysis(PLS-DA)were performed,and the differential components affecting the quality of Qianyang yuyin granules were screened by the criterion of the value of variable importance in the projection(VIP)>1.0.UPLC was used to determine the contents of the differential components and harpagide,ferulic acid,cyasterone.RESULTS A total of 18 common peaks were identified from the fingerprints of 15 batches of samples,and the similarities were all higher than 0.96.It was identified that peak 6 was morroniside,peak 7 was loganin,peak 11 was hyperoside,peak 12 was stilbene glycoside,peak 13 was ferulic acid,peak 14 was cyasterone,peak 16 was harpagoside,peak 17 was cinnamic acid,and peak 18 was emodin.The results of CA and PCA showed that the 15 batches of samples could be divided into 6categories.S1,S2,S4-S6 were grouped into one category,S3 was grouped into one category,S7-S8 were grouped into one category,S9-S10 were grouped into one category,S11-S14 were grouped into one category,S15 was grouped into one category.VIP values of peak 3,9,6(morroniside),7(loganin),15,12(stilbene glycoside)and 16(harpagoside)were greater than 1.0.Among 15 batches of samples,the contents of harpagide,harpagoside,morroniside,loganin,stilbene glycoside,ferulic acid and cyasterone were 0.336-0.563,0.205-0.377,0.692-0.837,0.433-0.562,1.294-2.030,0.020-0.027,0.167-0.235 mg/g,respectively.CONCLUSIONS The UPLC fingerprint and multi-components quantitative analysis of Qianyang yuyin granules are successfully established,which can be used to improve the quality control standard of the preparation;morroniside,loganin,stilbene glycoside,harpagoside and other components could be the differential components affecting the quality of this preparation.