Study On The Preparation Of Traditional Chinese Medicine "Qin Baidu" Granule

Riemerella anatipestifer disease is the contagious disease caused by Riemerella anatipestifer (RA).It is also known as the infectious serositis of duck, duck septicemia etc.RA infect the ducklings in1-8weeks of age and the ducks present acute or chronic septicemia. Its clinical manifestations are variety of the eye and nasal secretion pantings, coughs, diarrheas, head and neck symptoms such as tremor or skew etc. The mortality of duck is up to40%.Clinical commonly use antibiotics, chemical drug or vaccine to treat it, but serious drug residues, drug resistance and a variety of pathogenic RA serotype vaccine generally application restrictions continue to highlight the problem. Studying on the Chinese herbs against Riemerella anatipestifer in vitro first just to find the Chinese herbs which have good results. Doing the in vitro bacteriostasis experiment toform the compound prescription of traditional Chinese medicine.So choosing these Chinese lerbs to form compound ash powder. To extract the effective ingredients in the prescription by using the modern extraction method. The index for Chinese herbal including extract yield,minimal inhibitory concentration(MIC),and minimum bactericidal concentration (MBC).Screening extraction technology of the active ingredients of compound Cortex Fraxini powder. Observing the compound Cortex Fraxini extract by the optimal extraction technology which has effect on mouse infected RA so as to determine the best treatment concentration. To process the effective ingredients of Chinese medicine compound as granules and to establish the quality standard of the granule preparation. The main results are as follows:1. Using two times the tube dilution method to determine the MIC and MBC of the extracts against Riemerella anatipestifer. Cortex Eucommiae, Radix et Rhizoma Rhei, Radix Isatidis, Fructus Citri Sarcodactylis, Cortex Fraxini and Isatidis Folium have good effects on RA in vitro. The MIC of these lerbs are all15.6mg/ml. Choosing Cortex Eucommiae, Radix Isatidis and Cortex Fraxini to combine. Through a combination of antimicrobial we can discovery Cortex Eucommiae combined with Cortex Fraxini have additive effects,Radix Pulsatilla combined with Cortex Eucommiae and Cortex Fraxini were independent of the united, Radix Pulsatilla combined with Radix et Rhizoma Rhei performance antagonism. Cortex Eucommia,Cortex Fraxini and Radix Pulsatilla can form a prescription inhibition on Riemerella anatipestifer.2.Single factor test was used to take extraction solvent as single-factor study. Extact yield of effctive parts, MIC and MBC of clinical isolates of RA. The results showed that50%,60%and75%of concentration of ethanol were selected to screen the best extraction techonogy by orthogonal design in the next step.3.In order to screen the best extraction techonogy, the orthogonal design table L9(34) was used. Soild-liquid ratio (A), extraction time(B), each time of extraction (C) and concentration of ethanol (D) were selected as the four factors and extact yield of effctive parts, MIC and MBC of clinical isolates of RA. The best extraction techonogy A3D3C1B2, which meaned that soild-liquid ratio was1:10, extraction time was2, spent half an hour each time and the concentration of ethanol was75%, was identified by analysis of orthogonal experiment.4.To observe the curative effect of compound "Qin Baidu"extract on the of mouse infected RA. Doing the preliminary tests on mouse,to determine the optimal attack germs concentration is1×109CFu/mL. The result of the formal test showed that the medium dose group of compound"Qin Bai Du" extract has obvious therapeutic effect and the dead rate of mouse is35%which is lower than the ceftiofur group.But the high group of compound "Qin Baidu" extract and the low group of compound "Qin Baidu"extract have no effect which dead rate of mouse are95%. So the compound "Qin Baidu"extract medium dose group to the drug concentration is1time of compound "Qin Baidu"extract clinical dose has obvious effect of anti infection.5. Adjuvant material and the nature of various aspects of compound "Qin Baidu"granule were evaluated. In order to screen the appropriate proportion of liquid(the concentration was2gram extracteach milliliter) and accessories, the preparation method of granule was used. The results showed that when the proportion of liquid and accessories was1:4,the quality of the soft material and the appearance of the granule were both meet the standards of Pharmacopoeia. Then, taking the appearance, clarity and water content as idexes and the amount of soluble starch, a-lactse, dextin and the different concentration of HPMC as factors, to evaluate the optimum formula. The results showed that The optimal preparation condition of granule as follow:content of soluble starch, a-lactse and dextin was56%,14%and30%respectively, when0.3%HPMC was used as adhesives. Under the optimum formula, the nature of various aspects of compound Cortex Fraxini granule as follow:the granule color was uniform, unqualified was less, clarity was good and the water content was3.2%. Indexes above were all comply with the quality requirements of ganule of chinese medcine.6. Established the method of determination contents of aesculetin in compound Cortex Fraxini granule by SH MADZU LC-2010C HT (CLASS-VP6.13multi)of reverse phase high-performance liquid chromatography. Methanol was used as solvent of the granule. Chromatographic conditions of aesculetin were these that Ture Kromasil C18column(4.6mm×250mm,5μm) with methanol-0.1%phosphoric acid (5:95, v/v) mixture as the mobile phase was adopted, flow rate was1.0mL·min-1, the detecting wavelength was set at334nm, the column temperature was25℃and the sample size was10μL. The result showed that a good linear relationship emerged between the range of21.60μg·mL-1-129.60μg·mL-1of the concentration of esculetin (r=0.9999, n=6), the average recovery rate was98.29%and RSD was0.69%(n=6). The average aontents of aesculetin in compound "Qin Baidu"granule was0.26mg·g-1.