【作者】 郑海杰；

【导师】 张艺； 赖先荣；

【作者基本信息】 成都中医药大学 ， 药物化学， 2010， 硕士

【摘要】 黄连始载于《神农本草经》,列为上品,具有清热燥湿、泻火解毒的功效。根据中医临床辨证施治的需要,具有酒炒、姜炒、吴茱萸制、醋炙等多种炮制方法。自《名医别录》起,历代本草均有黄连治疗消渴病的记载,酒蒸黄连长于治消渴,明代李时珍《本草纲目》谓“治消渴,用酒蒸黄连”。本文以酒蒸黄连治疗“消渴病”为切入点,采用多成分分析相结合的研究方法,从整体上对酒蒸黄连改善胰岛素抵抗(IR)的药效物质基础、炮制工艺和质量控制方法进行研究。目的：探索酒蒸黄连改善IR效应的活性成分,筛选酒蒸黄连的最佳炮制工艺,并建立针对性的酒蒸黄连质量控制方法。方法：1、采用HPLC法建立同时测定酒蒸黄连及其它炮制品中6种生物碱的含量方法,并分别建立酒蒸黄连及其它炮制品的HPLC指纹图谱,以及酒蒸黄连的体内血清指纹图谱。2、采用U*6(64)均匀设计表,以改善3T3-L1脂肪细胞IR模型的葡萄糖利用率为指标,筛选酒蒸黄连最佳炮制工艺。并采用多元回归数理统计方法,分析黄连炮制品化学指纹图谱与改善IR效应的相关性,分析酒蒸黄连血清药物化学研究中的入血成分。以酒蒸黄连改善IR效应主要生物碱含量为指标筛选其最佳炮制工艺。3、建立同时测定酒蒸黄连的6种生物碱含量的新方法,并参考《中国药典》2010年版第一部黄连项下的方法,建立其薄层鉴别方法,并完成相应检查项的内容(水分、灰分、浸出物),初步建立酒蒸黄连的质量控制方法。结果：1、建立了黄连炮制品的HPLC指纹图谱,共标定了12个色谱峰,确定7-12号色谱峰分别为：盐酸药根碱、盐酸非洲防己碱、盐酸表小檗碱、盐酸黄连碱、盐酸巴马汀、盐酸小檗碱,各炮制品相似度和夹角余弦均大于0.9,但欧氏距离存在差异,主成分投影分析可将酒蒸黄连分类在一起。建立了黄连炮制品中6种生物碱的含量测定方法,6种生物碱的线性关系良好(r>0.999),加样回收率在95%-105%之间,RSD<2%,主成分投影分析可以将酒蒸黄连分类在一起,酒蒸黄连主成分得分最高为0.9412,单因素方差分析发现盐酸非洲防己碱、盐酸表小檗碱和盐酸小檗碱是酒蒸黄连与饮片和其它炮制品存在差异性的成分。建立了酒蒸黄连的血清指纹图谱,通过与混合对照品色谱图对照,鉴定出峰4-7分别是盐酸非洲防己碱、盐酸表小檗碱、盐酸巴马汀、盐酸小檗碱。2、以改善3T3-L1脂肪细胞IR模型的葡萄糖利用率为指标,均匀设计筛选结果为：黄酒20%,闷润2小时,蒸制8小时和；多元回归分析结果表明：盐酸药根碱,盐酸非洲防己碱,盐酸表小檗碱,盐酸小檗碱与IR模型的葡萄利用率密切相关；以可能的药效物质基础的总含量为指标,均匀设计筛选最佳炮制工艺结果为：黄酒20%,闷润2小时,蒸制8小时。3、建立了酒蒸黄连的薄层鉴别方法,测定了其6种生物碱的含量以及其水分、灰分、浸出物的含量。结论：1、采用HPLC法建立同时测定黄连炮制品中6种生物碱含量方法和指纹图谱,可用于黄连炮制品的质量控制。黄连炮制品间的化学成分存在差异,其差异性成分是盐酸非洲防己碱、盐酸表小檗碱和盐酸小檗碱。2、初步筛选出了酒蒸黄连的最佳炮制工艺,建立的酒蒸黄连新的的质量控制方法,为其质量控制和临床应用提供了依据和保障。更多还原

【Abstract】 Coptis chinensis Franch, first recorded in Shen Nong’s Herbal, has the function of clearing hear, depriving dampness, and detoxicating. According to the needs of TCM diagnosis and treatment, it has processed with rice wine, ginger juice, evodia rutaecarpa decoction, vinegar and other processing methods. Since the the book of Ming yi bie lu,the Rhizoma Coptidis steamed with rice wine have superiority advantages in the treatment of diabete in many medicine books. Such as Ben cao gang mu record"Treat diabete use Rhizoma Coptidis steamed with rice wine" etc.. It can be seen, Rhizoma Coptidis steamed with rice wine treat diabete, has sufficient practice basis.In preliminary studies on Rhizoma Coptidis steamed with rice wine, we find that it has preventive and therapeutic effect on diabetes B. In this paper we use Rhizoma Coptidis steamed with rice wine with the therapeutic action on diabete as breakthrough point, We determine multicomponent contents in Rhizoma Coptidis steamed with rice wine, establish fingerprint of Rhizoma Coptidis steamed with rice wine, determine the biological activity in Rhizoma Coptidis steamed with rice wine, Analysis correlation between them. Aim to do macro research on Active ingredient which improve Insulin resistance, processing technology and quality evaluation of Rhizoma Coptidis steamed with rice wine.Objective:Explore active ingredients which improve Insulin resistance of Rhizoma Coptidis steamed with rice wine, select its best processing technology, establish its quality control methods.Method:1.Establish fingerprint of processed products of Rhizoma Coptidis, determination method for six alkaloids in processed products of Rhizoma Coptidise, serum fingerprint of Rhizoma Coptidis steamed with rice wine by HPLC.2.Use IR model glucose utilization as indicator, select its processing technology. Analyze relativity between the fingerprint and bioeffect by statistical method of multiple regression, refer components into the serum of serum fingerprint, be sure the active ingredient of Rhizoma Coptidis steamed with rice wine. Use total content of three alkaloids which related IR as indicato, select the best processing technology of Rhizoma Coptidis steamed with rice wine.3.Based on Determination of Rhizoma Coptidis in Chinese Pharmacopoeia edited in 2010, determine six alkaloids in Rhizoma Coptidis steamed with rice wine, establish its TLC method, complete its Check items (moisture content, ash content, extractive content. Establish its quality control method.Result:1.Fingerprint of processed products of Rhizoma Coptidis was established. 12.chromatographic peaks was demarcated:7. jatrorrhizine,8. columbamine,9. epiberberine,10. coptisine,11. palmatine,12. berberine. they have a good similarity by Similarity analysis and angle cosine analysis, but, they are different by euclidean distance analysis, Rhizoma Coptidis steamed with rice wine was classed together by principal component projection points analysis. The determination method for six alkaloids was established, The six alkaloids have a good linear relationship (r>0.999) and recoveries(95%-105,RSD<2%). Rhizoma Coptidis steamed with rice wine was classed together by Principal component projection points analysis, it has the highest Principal component score 0.9412, Columbamine, epiberberine and berberine are the different components in processed products of Rhizoma Coptidis by variance analysis. Serum fingerprint of Rhizoma Coptidis steamed with rice wine was established, Columbamine, epiberberine, palmatine, and, berberine were the into blood components.2.Use IR model glucose utilization as indicator, processing technologies are rice wine 20%, moistened 2h, steamed 8 h and rice wine 60%, moistened 2h, steamed 8 h. Jatrorrhizine, columbamine, and, berberine related to IR model glucose utilization. Use total content of three alkaloids which related IR as indicator, processing technology is rice wine 20%, moistened 2h, steamed 8 h.3.The quality Standard of Rhizoma Coptidis steamed with rice wine was Initially established.Conclusion:1.The fingerprint and determination method for six alkaloids established by HPLC can be used to control the quality of processed products of Rhizoma Coptidis, the contentsof Chemical composition in processed products of Rhizoma Coptidis are different, the different composition may be columbamine, epiberberine and berberine. Serum fingerprint established by HPLC can be used to study Rhizoma Coptidis steamed with rice wine, Columbamine, epiberberine, palmatine, and, berberine were the into blood components.2.The processing technology of Rhizoma Coptidis steamed with rice wine was selected, The quality Standard Initially established can be used for the quality Control of Rhizoma Coptidis steamed with rice wine, provide reference and basis for quality Control and clinical application of Rhizoma Coptidis steamed with rice wine.更多还原