摘要
目的建立复方苦参合剂的HPLC指纹图谱,并测定其有效成分苦参碱、氧化苦参碱、没食子酸、甘草苷、甘草酸的含量,为制剂的质量控制提供实验依据。方法采用Green ODS-AQ C18色谱柱(4.6mm×250mm,5m),以乙腈-0.5%磷酸溶液进行梯度洗脱,流速1mL/min,检测波长为235nm;采用“中药色谱指纹图谱相似度评价系统”对10批制剂的指纹图谱进行相似度评价,对比对照品与样品中各个峰的保留时间确定制剂中主要特征峰的化学成分,通过分析初步确定制剂中各味成分的归属,采用主成分分析(PCA)和正交偏最小二乘判别分析(OPLS-DA)对测定结果进行识别,并对制剂中的苦参碱、氧化苦参碱、没食子酸、甘草苷、甘草酸进行含量测定。结果考察了复方苦参合剂HPLC指纹图谱的精密度、重复性及稳定性,结果均良好,相似度评价中10批复方苦参合剂的指纹图谱相似度为0.822~0.983,标定出31个共有峰,并指认出5个主要特征峰,分别为苦参碱、氧化苦参碱、没食子酸、甘草苷、甘草酸。通过PCA可将10批复方苦参合剂分成3类,可看出不同产地、不同批次药材之间的质量存在差异,进一步采用OPLS-DA筛选出差异成分。结论建立复方苦参合剂的指纹图谱方法稳定可靠,相较于现有质量标准中含量测定方法,该方法更佳,可作为复方苦参合剂质量控制的参考指标,为复方苦参合剂的质量控制及临床用药方面的研究提供实验依据。
Objective To establish the HPLC fingerprint of compound matrine,oxymatrine,gallic acid,liquiritin and glycyrrhizic acid,and provide experimental basis for quality control.Methods Green ODS-AQ C18(4.6 mm×250 mm,5m)column,gradient elution with acetonitrile-0.5%phosphate acid solution.The flow rate at 1mL/min,and the detection wavelength was 235 nm.The fingerprints of 10 batches of preparations were established.The Traditional Chinese Medicine Chromatographic Fingerprint Similarity Evaluation System was used to evaluate the similarity of the fingerprint of 10 batches of preparations.The main chemical components of the main characteristic peaks in the preparations were determined by comparing the retention time of each peak in the control substance and the sample,and the attribution of each flavor component in the preparations was preliminarily determined by analysis.Principal component analysis(PCA)and orthogonal partial least squares discriminant analysis(OPLS-DA)were used to identify the determination results,and the content of matrine,oxymatrine,gallic acid,liquiritin and glycyrrhizic acid in the preparation was determined.Results The precision,repeatability and stability of the compound HPLC fingerprint map were investigated,and the results were good.The similarity of the 10 approved formula was 0.822~0.983,31 common peaks were identified,and 5 main characteristic peaks,including matrine,oxymatrine,gallic acid,liquiritin and glycyrrhizic acid were identified.The 10 batches of Compound Kushen Mixture preparations could be divided into 3 categories by PCA,and the quality differences between different regions and different batches could be seen,and the difference components were further screened by OPLS-DA.Conclusion The established fingerprint method of Compound Kushen Mixture is stable and reliable,compared with the content determination method in the existing quality standard,it can be used as a reference index in the quality control of compound bitter ginseng agent,providing an experimental basis for the quality control of Compound Kushen Mixture and clinical medication research.
作者
陈思迪
唐佩莉
杨思成
黄菊
岳凤芹
彭伟
孙婉瑾
范恒
李森
段雪云
CHEN Si-di;TANG Pei-li;YANG Si-cheng;HUANG Ju;YUE Feng-qin;PENG Wei;SUN Wan-jin;FAN Heng;LI Sen;DUAN Xue-yun(Hubei University of Chinese Medicine,Wuhan 430065;Hubei Provincial Hospital of TCM/Affiliated Hospital of Hubei University of Chinese Medicine/Hubei Academy of Traditional Chinese Medicine,Wuhan 430061;Union Hospital Affiliated to Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430022)
出处
《湖北中医药大学学报》
2024年第3期36-40,共5页
Journal of Hubei University of Chinese Medicine
基金
国家自然科学基金项目(项目编号:81774093)
湖北省重大疑难疾病中西医临床协作项目(鄂卫办通[2023]2号)
全国中药特色技术传承人才培训项目(国中医药人教函[2023]96号)。
