【作者】 袁波；

【导师】 李发美；

【作者基本信息】 沈阳药科大学 ， 药物分析学， 2006， 博士

【摘要】 复方大青叶注射剂由大青叶、金银花、羌活、拳参和大黄5味药材加工制成。具有清瘟解毒的功效，用于急、慢性肝炎，流行性感冒，腮腺炎的治疗。本论文以复方大青叶注射剂及大青叶、金银花、羌活、拳参、大黄5味药材为研究对象，对其质量控制方法进行研究，为全面控制和评价复方大青叶注射剂的质量，提供了方法学依据。首先，对大青叶药材进行了质量控制方法研究。采用LC／MS方法，对大青叶药材水溶性部分的化学成分进行了研究，通过与胞苷、鸟苷、尿苷和次黄嘌呤标准品的色谱和质谱行为比较，确证了大青叶水提液含有胞苷、鸟苷、尿苷和次黄嘌呤，在此基础上，建立了四种化学成分的含量测定方法，并对不同产地大青叶药材中4种成分含量进行了测定；以10批安徽白云山药厂板蓝根基地产大青叶药材为样品，尿苷为参照物，建立了标准色谱指纹图谱，标定了大青叶药材指纹图谱中的19个共有峰。采用国家药典委员会的“中药色谱指纹图谱相似度评价系统”进行相似度计算，结果表明：10批药材的相似度在0.95以上，不同产地大青叶指纹图谱间存在差异。因此，规定凡符合药典标准的大青叶药材按给定色谱条件下所得指纹图谱的相似度达到0.90以上者，可作为合格药材入药。其次，对金银花、羌活、拳参和大黄药材进行了指纹图谱研究，为与注射剂和中间体的相关性研究奠定了基础。采用HPLC方法，建立了四种药材的标准色谱指纹图谱。标定了四种药材药材指纹图谱中的共有峰，其中金银花有16个，羌活中有17个，拳参中有6个，大黄中有12个。第三，对复方大青叶注射剂进行了质量控制方法研究。建立了复方大青叶注射剂中尿苷、绿原酸、咖啡酸、大黄素和大黄素甲醚的定量分析方法；采用高效液相色谱法，建立了复方大青叶注射剂的标准指纹图谱，以尿苷为内参比峰，确认了20个共有指纹峰，用国家药典委员会的“中药色谱指纹图谱相似度评价系统”对不同企业不同批次的注射液与共有模式进行相似度计算，确定了本品的相似度阈值为0.85。通过对比各单味药材水煎液、阴性对照液、中间体和注射液指纹图谱，确定了药材和制剂、中间体和制剂之间的相关性。使通过药材、制剂工艺和中间体来控制复方大青叶注射剂的质量成为可能。最后，对复方大青叶注射剂进行了药动学和药效学研究。采用HPLC-UV法测定了大鼠腹腔注射复方大青叶注射剂后血浆中绿原酸的浓度，初步探讨了绿原酸在大鼠体内的药物动力学过程；进行了注射剂的体外抑菌、抗内毒素、抗炎及解热作用的试验。结果表明：复方大青叶注射液不仅对革兰细菌有较强的抑菌作用，而且对某些真菌也有较强的抑菌作用；抗炎和解热作用非常明显。这对大青叶注射剂在临床上的广泛应用具有一定的指导意义。总之，本论文将中药成分化学、分析化学、化学统计学、中药药理学和计算机技术相结合，采用HPLC方法及LC／MS技术，建立了大青叶、金银花、羌活、拳参、大黄及其复方制剂质量的控制方法，对大青叶、金银花、羌活、拳参、大黄及其复方制剂质量控制进行了深入、系统的研究，为依据指纹图谱整体控制复方中药制剂的质量作了有意义的探索。更多还原

【Abstract】 The injectio folii isatidis composita was prepared by Isatis indigotica Fort, Lonicera japonica Thunb, Polygonum bistorta L, Notopterygium incisum Ting ex H. T. Chang, Rheum palmatum L.. It had the effects of bacteriostasis, anti-virus etc. and was used clinically to treat influenza, acute and chronic hepatitis, epidemic parotitis. In this thesis, the quality control method of five herbs and injectio folii isatidis composita were studied for the evaluation of the injection.Firstly, the quality control method of Folium Isatidis was investigated. The water-soluble components of Folium Isatidis were studied and four compounds named cytidine, hypoxanthine, uridine, guanosine was characterized by LC/MS method by comparing the LC/MS behaviors with the standard substances. The method for determination of the four compounds was developed and used to determine the contents in different Folium Isatidis samples. The 10 batches of Folium Isatidis from Anhui Baiyunshan Pharmaceutical Corporation were evaluated. Uridine was chosen as the reference substance. 19 "common peaks" were achieved after analyzing the samples. 10 batches of samples were evaluated by "Similarity Evaluation System" for chromatographic fingerprint of TCM of the Chinese Pharmacopoeia Committee and the results showed that similarities is more than 0.95. The similarity of the FoliumIsatidis above 0.9 was considered as superior herbs.Secondly, the fingerprint of Isatis indigotica Fort, Lonicera japonica Thunb, Rheum palmatum L., Notopterygium incisum Ting ex H. T. Chang, Polygonum bistorta L were also studied. The standard fingerprints of the four herbs were developed by HPLC method. The 16, 17, 6 and 12 "common peaks" in the fingerprints for Isatis indigotica Fort, Lonicera japonica Thunb, Rheum palmatum L., Notopterygium incisum Ting ex H. T. Chang, Polygonum bistorta L, respectively. The samples were evaluated by "Similarity Evaluation System" for chromatographic fingerprint of TCM of the Chinese Pharmacopoeia Committee. The results showed that the similarity varies with the sources of the herbs and the fingerprint can be applied to the evaluation of the herbs.Thirdly, the quality control method of injectio folii isatidis composita was investigated. The determination method of the uridine, chlorogenic acid, caffeic aicd, emodin and physcion in the formulation was developed. With Uridine as reference substance, the HPLC-FP of injectio folii isatidis composita was established. There were 20 "common peaks" in the fingerprints. The similarities of the various batches of injection were analyzed by the "Similarity Evaluation System". The similarities threshold of 0.85 was set up for the samples. By comparing the fingerprints of each herb decoction, intermediate and injection, the correlation of peaks among their fingerprints were found.Finally, the pharmacokinetics and pharmacology of injectio folii isatidis composita were studied. The concentration of chlorogenic acid in rat plasma after peritoneal administration of the injection was determined by the HPLC method. The pharmacokinetics of chlorogenic acid in rats was elucidated. The antibiotics, bacteriostasis, and anti-inflammatory activities of injectio folii isatidis composita were also studied. The results indicated that injectio folii isatidis composita have antibiotics effects not only to bacterial spectrum but also to some kinds of epiphyte and strong anti-inflammatory effects. The study of the activities of folii isatidis composite provide a scientific basis for the clinical usage of the formulation.In a word, by the combination of chemical TCM, analytical chemistry, statistics and computer technology, and the application of HPLC and HPLC/MS method to the quality control of raw herbal materials of Folium Isatidis, Flos Lonicerae Radix et Rhizoma, Rhizoma et Radix notopterygii and Rhizoma Bistortae and the injectio folii isatidis composita were developed and provided a significant exploration for quality assessment of TCMs based on chromatographic fingerprint.更多还原