Preparation And Pharmacodynamic Studyof Compound Pulastilla Chinensis Granule

Pulastilla chinensis decoction is preferred by the traditional Chinese Medicine prescription for curing dysentery, which is widely used in clinical and veterinary clinic. Therefore, in the present study, the Pulastilla chinensis decoction was prepared compound Pulastilla chinensis granular (CPCG). We have studied preparation process, quality standard, stability, toxicology, target animal safety, pharmacodynamics and clinical effect of CPCG, results showed that:To establish the preparation technology of CPCG,the first extraction time, time ratio and the amount of water is optimized, and different CPCG prescription were studied. The results indicated that the optimum extraction technology of CPCG is:decocting3times with8times amount of water, the duration of each time is120min,80min and80min, respectively. Mixed the dry ointment powder and dextrin by the ratio of1:1, with90%ethanol as solvent, the granulars are easily formed. Pilot studies show CPCG process is stable, easy to operate, adapted to large-scale production.The content of berberine hydrochloride in CPCG was tested, and the analysis method is verified by HPLC. The linear relationship of berberine hydrochloride from0.025-0.250mg/mL is y=46972x+28.184, R2=0.9999.The average recovery rate was96.63%, RSD was1.02%. The results showed that the method is simple, accurate, repeatable, thus the method can be used as the quality control standard.Accelerated test, illumination test and long-term test were used to study the stability of CPCG The test results showed that the mainindexes of CPCG including the character, identification, granularity, moisture content, solubility, the content of berberine hydrochloride had no obvious change after accelerating test, illumination test and long-term test, the stability properties in all experimental conditions meet the quality standard requirements.Acute toxicity test and sub chronic toxicity test were used to study the toxicological safety of CPCG. Results showed that the LD50of CPCG on Kunming mice is5.0g/kg·bw, and belongs to actually non-toxic drug. Sub chronic toxicity test showed that the body weight, hematological indexes, biochemical indexes of blood and organ morphology of the rats had no significant change after fed with CPCG in dose of6.0,12.0,24.0g/L for30days. These results showed that CPCG has higher security in the dose of clinical range.To evaluate the clinical safety of CPCG the chicken were administrated in0,1times (2.0g/L),3times (6.0g/L),5times (10.0g/L) recommended dose of CPCG. Chicken signs, weight gain, feed efficiency, hematological indexes, biochemical indexes, histology pathology were observed. The results showed that the CPCG administration to the target animal chicken with5times water administration is safe.Micro-dilution method was adopted to determine minimum inhibitory concentration and minimum bactericidal concentration of CPCG for drug-resistant Escherichia coli, Staphylococcus aureus, Salmonella. Mice infected by drug-resistant E. coli were used to observe preventive effect of CPCG. The MIC and MBC of CPCG to drug-resistant Escherichia coli, Staphylococcus aureus, and Salmonella are6.25mg/ml,3.12mg/ml,12.50mg/ml and12.50mg/ml,6.25mg/ml,12.50mg/ml. CPCG experimental group of mice infected with drug-resistant E. coli have a protective effect in varying degrees and a dose-response relationship. The results showed that CPCG has good antibacterial effect on drug-resistant Escherichia coli, Staphylococcus aureus and Salmonella, and has a good protective effect on mice infected with drug-resistant E. coli. Its effect is the same with2%of ciprofloxacin hydrochloride soluble powder.Effect of CPCG on chicken Escherichia coli disease was tested using artificial induced pathological model of avian Escherichiacolibacillosis. Results showed that there are significant prevention effect on artificial infected chicken Escherichia coli disease in high (4.0g/L) and medium dose group (2.0g/L) of CPCG, the effective rates were83.33%and76.67%, respectively, the effect was equivalent to that of compound sulfamonomethoxine sodium soluble powder.In short, CPCG is stable and reliable quality, good safety performance, and has a significant effect on chicken infected E.coli disease. It is a safe, effective and controllable quality of veterinary drugs in new formulations.