Studies On Eucommia Folium Extract Granule And Pharmacodynamic Evaluation

Eucommia ulmoides Oliv is a plant of Eucommiaceae, Eucommia genus. Its bark was used as traditional herbal medicine, with sweet-warm character and various pharmacological benefits, such as nourishing liver and kidney, intensifying bones and muscles, tocolysis.Eucommia ulmoides Oliv has good curative effect on kidney deficiency, waist weakness,rheumatism and quickening uneasiness. However, peeling the bark of Eucommia ulmoides is harmful for its growth. Modern research indicates that Eucommia folium mainly contains phenylpropanoid, iridoid glycosides, flavonoids, phenols, and has significant effects like anti-bacteria, anti-inflammation, enhance immunity and anti-hypertension and so on. Because the main active ingredients and pharmacological actions of Eucommia leaves are almost the same with that of bark, and Eucommia folium is cheaper and easier to get, it has important potential value for application. This study optimized the technology for Eucommia folium extraction and preparation, Eucommia folium extract granule preparation, quality control and pharmacodynamic evaluation, with the purpose of providing scientific basis for further new veterinary drug declaration and clinical application.(1) Eucommia folium extraction technology optimization : On the basis of single factor experiment, using Eucommia folium extract yield and chlorogenic acid transfer rate as the indexes for evaluation, Box-Behnken central composite design as the method for Eucommia folium extraction technology optimization. According to the results, the optimal technology for Eucommia folium extract preparation was determined as follows: ethanol concentration 50%, liquid-solid ratio 10:1, extraction temperature 97?, extract for 4 times,30 minutes once. According to confirmatory experiments, under the condition, the mean composite score of Eucommia folium extract yield and chlorogenic acid transfer rate was99.08, well matched with the predictive value.(2) Eucommia folium extraction quality control: Thin layer chromatograph(TLC)and high performance liquid chromatography(HPLC) were used for determining the category and content of chlorogenic acid in Eucommia folium extract. It showed that the Eucommia folium extraction and chlorogenic acid standard displayed the same fluorescent spots in the corresponding chromatographic position of TLC, and the negative control has no interference.The results showed that the standard curve of chlorogenic acid determination was y=2887462x–148005(r=0.9998), with a good linear relationship when the injection volumeranged 1.05-6.30?g. These results indicated that the method was accurate, stable and repeatable.(3) Eucommia folium extract granule preparation: Our preliminary test screened the types and the proportion of excipient. The comprehensive score of granule forming rate, pile concentration, solubility, dormant angle and humidity-absorption was used as index. L9(34)orthogonal test was taken to optimize the preparation process of Eucommia folium extract granules. Results indicated that the best preparation process for granule were: extract : actose :extrin 2:2:1, ethanol concentration 85%, dosage of ethanol 24%. Confirmatory experiments indicated, the comprehensive score of Eucommia folium extract granule was 95.23 in this condition, similar with the predictive value.(4) Eucommia folium extract granule quality control: Identifying and determinating chlorogenic acid from granule with thin layer chromatography(TLC) and high performance liquid chromatography(HPLC). The moisture content of Eucommia folium extract granule was 2.5%, the molding rate was 91.83%, and the critical relative humidity(CRH)was 74%,conforming to the requirements of the Veterinary Pharmacopoeia, and should be kept in the environment(relative humidity < 74%). It displayed the same fluorescent spots in the corresponding chromatographic position of TLC with chlorogenic acid standard, negative control without interference. The detecting methods of Eucommia folium extract granule were highly accurate, repeatable, and stable with HPLC.(5) anti-bacterial test in vitro of Eucommia folium extract granules: Using agar punch method to determine the granule antibacterial circle diameter on Escherichia coli,Staphylococcus aureus and Salmonella, tube double dilution method to detect the minimal inhibitory concentration(MIC) and minimal bactericidal concentration(MBC). Results showed that granule solution was highly sensitive to Staphylococcus aureus, moderately sensitive to Salmonella performance for, insensitive to Escherichia coli. Among them, the MIC for Staphylococcus aureus and Salmonella were 15.6mg/mL and 31.25mg/mL, and MBC for them were 31.25mg/mL and 62.5mg/mL, respectively.