【作者】 徐倩倩；

【导师】 李继昌；

【作者基本信息】 东北农业大学 ， 基础兽医学， 2008， 硕士

【摘要】 细菌性犊牛腹泻对畜牧业生产存在很大的威胁,抗菌药物的泛滥使其治疗更加困难,中药制剂以其独特的优点越来越显现出巨大的作用。本课题从常用抗细菌性犊牛腹泻中药中选出黄连、黄芩、黄柏、乌梅、石榴皮、双花、苦参、沙参、丹参、甘草、地榆、诃子、连翘十三味中药,应用其水提或醇提药液采用纸片法对犊牛腹泻主要致病菌大肠杆菌O₈、魏氏梭菌B型、魏氏梭菌C型、鼠伤寒沙门氏菌等进行了体外抑菌试验。结果显示,醇提黄连与乌梅所得药液对四种主要致病菌均有较好的抑制作用。根据中兽医辨证施治,标本兼治原则,选用白术、木香、干姜、甘草与之配伍应用,以达到相辅相成,提高药效的目的。根据临床使用情况及制剂稳定性,选用较为常见的颗粒剂。提取路线为:黄连、乌梅以盐酸小檗碱的含量及干膏量为指标,采用正交试验法优选了黄连、乌梅的最佳乙醇回流提取工艺;对于白术、木香、干姜、甘草含有挥发油成分的药物,采用共水回流法提取挥发油,以挥发油溶出率为指标,对加水量、浸泡时间、提取时间、粉碎程度进行了单因素考察,优选挥发油最佳提取工艺;利用β-CD包合挥发油,以包合物收得率和挥发油利用率为指标,通过正交设计优选出最佳包合工艺。白术、干姜、木香、甘草提取挥发油后的药液还含有水溶性有效成分,以甘草中的甘草酸铵含量及干膏量为指标,通过正交设计优选出最佳处理工艺。通过辅料的选择、用量和制备环境湿度指标的考察确定制粒成型工艺条件为:黄连、乌梅用8倍量60%乙醇提取3次,每次1h:白术、干姜、木香、甘草,粉碎成粗粉,不浸泡,加8倍量水,共水回流提取6h:挥发油包合工艺为β-CD:挥发油为4:1;β-CD:水为1:8;包合温度40℃;包合时间30min;剩余药液浓缩至0.85g·mL^-1,3000r·min^-1离心15min,12倍60%乙醇进行醇沉,静置16h。黄连、乌梅提取药液及白术等提取挥发油后药液合并减压浓缩制成干膏,与包合物混合,按1:1添加糊精,95%乙醇作为润湿剂,环境湿度在70%以下制粒。紫外分光光度法测定颗粒中盐酸小檗碱的含量,三批样品均在58mg·g^-1以上。对颗粒的粒度、含水量、溶化性等指标进行测定,均符合《中国兽药典》2005年版颗粒剂项下规定。加速稳定性试验结果表明,按以上工艺所制颗粒,性状、定性鉴别、粒度检查、含量测定等项在六个月内未发生改变。总之,通过以上研究,确定了治疗细菌性犊牛腹泻的中药颗粒剂的组方、制备工艺,为该制剂的进一步研究（如药效学、药代动力学、毒理学和作用机制等）提供基础。更多还原

【Abstract】 Calf diarrhea caused by bacteria threaten husbandry greatly,and the abuse of antibacterials makes its cure more difficult,traditional Chinese medicine plays an important role for its special superiority.13 traditional Chinese medicines were chosen in this topic,they were:Fructus mume,Fructus forsythiae,Radix scutellariae,Flos lonicerae,Radix adenophorae,Pericarpium granati,Rhizome picrorhizae,Radix sanguisorbae,Radix et rhizome glycyrrhizae,Fructus chebulae,Radix et rhizoma salviae miltiorrhizae,Cortex phellodendri amurensis and Radix sophorae flavescentis, which were usually used to resist bacteria.Bacteriostasis experiments in vitro were done to test the effects of those traditional Chinese medicines to resist the main pathogenic bacteria,which were Escherichia coli O₈,Clostridieum welchii type B,Clostridieum welchii type C and Salmonella typhimurium,by Scrip method.Physic liquors were extracted by water or alcohol.The results showed that alcohol extracted Rhizome picrorhizae and Fructus mume both had significant inhibitory effects to the four main pathogenic bacteria.Chose Atractylodes macrocephala, Zingiberis rhizoma,Costustroot and Radix et rhizome glycyrrhizae to match with them,based on the Chinese medical science theory:cure both appearance and substance and determine the treatment based on differentiation of symptoms and signs,so as to complement each other and enhance pharmacodynamic actions.According to the clinical use and stability of preparation,drug granules were choosen as their form of medication.Extraction flow-sheet was:Rhizome picrorhizae and Fructus mume were extracted by alcohol,the optimum extraction condition was investigated with orthogonal design method by studying the content of berberine and extracta sicca;as to volatile oil contained in Atractylodes macrocephala,Zingiberis rhizoma and Costustroot,used distillation with water method to extract,and wrapped it up withβ-cyclodextrin（β-CD） to stable the preparation and ensure the therapeutic effect.Radix et rhizome glycyrrhizae could promote the dissolution of volatile oil,so it was extracted with those three medicines.The optimum extraction condition was determined by observing the imbitition amoun,soaking time,extracting time and crashing degree, based on the dissolution ratio of volatile oil.The optimum extraction condition was investigated with orthogonai design method by determing the yiels of clathratio and utilization ratio of volatile oil;there were water-solubility effective ingredient in the physic liquor of Atractylodes macrocephala,Zingiberis rhizoma,Costustroot and Radix et rhizome glycyrrhizae after extracting volatile oil,the orthogonal design method was used again by determining the content of glycyrrhizic acid and extracta sicca.Decided granulate forming technology by observing the choosen and usage amount of adjuvant and ambient humidity.The result was:Rhizome picrorhizae and Fructus mume were extracted three times by 8 times 60%alcohol,1 hour per time; Atractylodes macrocephala,Zingiberis rhizoma,Costustroot and Radix et rhizome glycyrrhizae were crushed into coarse powder,no soak,extracted 6 hours with 8 times water;clathration technology of volatile oil was:proportion ofβ-CD and volatile oil was 4:1,and that ofβ-CD and water was 1:8 clathratio for 30min under 40℃;remaining physic liquor was condensed into 0.85g·mL^-1,centrifugated 15min by 3000r·min^-1 and precipitated by adding 12 times 80%alcohol, and standing for 16h.Alcohol extracted physic liquor of Rhizome picrorhizae and Fructus mume and remaining physic liquor of Atractylodes macrocephala et aL after extracting volatile oil affiliated and decompression condensed into dry mastic,then mixed with clathrate,added partes aequales dextrine,took 95%alcohol as wetting agent,granulate on the ambient humidity below 70%.Ultraviolet spectrophotometry was used to measure the content of berberine hydrochloride in the granula.Indexes such as granularity,moisture capacity and dissolve ability were evaluated, which were consistented with drug granules regulations of <Chinese Veterinary Pharmacopeia> edited in 2005.Accelerated stability test results showed that charactors,qualitation,granularity and contents of main effective ingredient of granula according to technology above had no change in six months.Studies above had determined the prescription and prepare technology of traditional Chinese medicine to cure calf diarrhea caused by bacteria,set groundwork for the further studies,such as pharmacodynamics,pharmacokinetics,toxicology and mechanism of action and so on.更多还原