Studies On Extraction And Biological Activities Of Total Alkaloids Of Seeds Of Ammopiptanthus Mongolica

In the present study, there are alkaloids, organic acids, flavone, isoflavone,stilbenes and other components in seeds of Ammopiptanthus mongolica. Alkaloids arethe main bioactive and toxiferous chemicals. But there was no report about theextraction process and pharmacology of total alkaloids of seeds of Ammopiptanthusmongolica until now. On these grounds, this article not only make a study on theextraction process of total alkaloids of seeds of Ammopiptanthus mongolica, but alsodo the analysis of acute toxicity, even more determine the vitro antiviral activity of thetotal alkaloids, hoping that the results could establish foundation for developing andutilizing the total alkaloids of seeds of Ammopiptanthus mongolica. The obtainedmain contents are as follows:1. Extraction technology of total alkaloids of seeds of Ammopiptanthus mongolicaTotal alkaloid is extracted by the hot dipping process and utrasonic-assistedextraction method from seeds of Ammopiptanthus mongolica with acid water. Usingthe orthogonal experimental designed method, taken the evaluating target by theweight of total alkaloids of seeds of Ammopiptanthus mongolica, determined theoptimal conditions of the hot dipping process and utrasonic-assisted extractionmethod. And compared the extraction efficiency of total alkaloids of seeds ofAmmopiptanthus mongolica that was extracted by the two methods, to pick out thebest extraction line. The optimal condition of hot dipping process is the pH value ofhydrochloric acid solution1.5、the ratio of material to solvent1:30、the extractingtemperature80℃、the extracting time3h. The optimal condition of utrasonic-assistedextraction method is the pH value of hydrochloric acid solution2.5、the ratio ofmaterial to solvent1:30、the extracting temperature30℃、the extracting time1.5h、the extracting frequency60Hz.Under the optimal condition mentioned-above, theextraction rate of total alkaloids of hot dipping process was1.926%, and theextraction rate of total alkaloids of utrasonic-assisted extraction method was2.868%.2. Acute toxicity experiment of total alkaloids of seeds of Ammopiptanthus mongolicaThe acute toxicity experiment was carried out by total alkaloid being poured to the throat of mice by oral cavity. LD50of total alkaloids of seeds of Ammopiptanthusmongolica in mice was determined by modified Karber’s method. The result showsthat the LD50of total alkaloids of seeds of Ammopiptanthus mongolica to the mice byoral administration was588.71mg/kg, its95%confidence limit was513.93～674.37mg/kg.3. Study on the vitro antiviral activity of total alkaloids of seeds of AmmopiptanthusmongolicaThe virulence of BVDV was measured by observing cytopathic effect (CPE).The cytotoxicity of total alkaloids of seeds of Ammopiptanthus mongolica to MDBKwas assayed by observing cytopathic effect and MTT colorimetry. RBV was used aspositive reference drug, by using cytopathic effect and MTT colorimetry, theinfluences of total alkaloids on reproduction and adsorption and synthesis of BVDVwere investigated. Results: The TCID50of BVDV was10-5.29; When the concentrationof total alkaloid≦0.39mg/mL, total alkaloids of seeds of Ammopiptanthusmongolica to MDBK was innocuity, and cell survival rate was115.34%at theconcentration of total alkaloid being equal to0.39mg/mL. Compared with the viruscontrol group, total alkaloids of seeds of Ammopiptanthus mongolica (0.16mg/mL、0.32mg/mL)could directly kill BVDV, inhibit the reproduction of BVDV, protectMDBK infected by BVDV, increase the cells proliferated activity, and inhibit thecytopathic effect. But the blocking effect of total alkaloid on BVDV was notobserved.In conclusion, the extraction efficiency of utrasonic-assisted extraction methodobviously surpasses hot dipping process, so utrasonic-assisted extraction method ischoosed as the extraction technique of total alkaloids of seeds of Ammopiptanthusmongolica. Total alkaloids of seeds of Ammopiptanthus mongolica is the low toxicmatter and has remarkable antiviral effect.