Study On Extraction And Biological Activities Of Total Flavonoids From The Seeds Of Ammopiptanthus Mongolicus

The genus Ammopiptanthus, a survivor of the evergreen broadleaf shrub in thecentral Asia desert from theTertiary period. It belongs to the tribe Thermopsideae of the Leguminosae. Ammopiptanthus comprises twospecies, these are Ammopiptanthus mongolicus (Maxim.) Cheng f. and Ammopiptanthus nanus (Popov.) Cheng f.which is widely distributed in Mongolia, Ningxia, Gansu Kizilsu Autonomous Prefecture in the southwest ofXinjiang province and neighboring central Asia. Alkaloids and Flavonoids are the characteristic chemicalconstituents of Ammopiptanthus, According to the literature. Numerous studies show that most flavonoids havegood antiviral, antitumor, anti-cancer, anti-inflammatory, antimicrobial physiological and antioxidant activity.In this thesis, the total Flavonoids from the seeds of Ammopiptanthus mongolicus including extraction technology;acute toxicity experiment; inhibit BVDV in vitro were studied systematically. The key technological parameters onextracting, purifying and determining were optimized. The optimal condition for ultrasonic extraction ofFlavonoids from seeds of Ammopiptanthus was investigated by Uesponse Surface Analysis. The content ofFlavonoids in the sample as research index was tested by spectrophotometry and optimal extraction process oftotal Flavonoids and its determination method was set up. This experiment was conducted to evaluate the acutetoxicity of the total Flavonoid from Seeds of Ammopiptanthus in mice, and to determine the medium lethal dose(LD50). The experiment was designed to investigate antivirus mechanism of the total flavonoids from the Seeds ofAmmopiptanthus on Bovine viral diarrhea virus (BVDV) by MTT and CPE. Drug safely and drug action weremeasured by three administration routes of virus-after-hypericin, virus before hypericin and virus-and-hypericin,respectively.1. The results showed that the operating efficiency of extracting total Flavonoids from the Seeds ofAmmopiptanthus is remarkably increased by Response surface analysis and utrasonic-assisted extraction method.The optimal parameters of the extraction process were as follows: extraction time (50min), solid-liquid ratio (1:20),extraction temperature (40℃), ethanol volume fraction (70%). The crude extract is flavonoids component wasidentified by the color reaction. The content of flavonoids in the seeds of Ammopiptanthus mongolicus was0.326%.2. The medium lethal dose of the total Flavonoids from Seeds of Ammopiptanthus was2004mg/kg, itsconfidence limit was1693.78～2372.47mg/kg. The main acute toxicity symptoms and signs of dead mice arenegligent action, then arising from unsteady gait, abdominal lying sleeping, shortness of breath, continuousconvulsions, and eventually to death. Mice were poisoned with the total Flavonoid from Seeds of Ammopiptanthus, the main organ, such as the heart, liver, spleen, lung and kidney were harmed in certain extent, but the continuedaffect on mice is small.3. Experimental data indicated that BVDV TCID50was10-7.733/0.1mL. When the concentration of totalflavonoids≦0.187mg/mL, total flavonoids of seeds of Ammopiptanthus mongolicus to MDBK was innocuity, andcell survival rate was115.34%and more than100%at the concentration of total alkaloid being equal to0.187mg/mL. It indicates not only no toxicity to cells, but also can promote cell proliferation. Low the drugconcentration0.187mg/mL, the proliferation of BVDV was effectively inhibited by hypericin soluble powderwhen drug and BVDV were added at the same time. Compared with the control group cells of virus and drug meanwhile was in good condition, integrity cell monolayer and had small amount of round cells and dead cells, whilethe other routes of administration had marked CPE when the drug concentration low0.187mg/mL.In conclusion, utrasonic-assisted extraction method is the best suitable method for extracting total Flavonoidsfrom the seeds of Ammopiptanthus mongolicus, It was Identified by color reaction and the method that themeasure of content of flavonoids of the seeds of Ammopiptanthus mongolicus was build up. The result Showedthat The method is convenient and reliable. It can be used as a quality control method of total flavonoids of theseeds of Ammopiptanthus mongolicus.Total Flavonoids of seeds of Ammopiptanthus mongolicus is the low toxicmatter and has remarkable antiviral effect. To establish a foundation for researching and developing drugs wereantiviral and provide important experimental data and scientific basis.