Study On Extraction,Purification And Antimicrobial Activity Of Aklaloid From Veratrum Nigrum

Veratrum nigrum is Liliaceous plants of the genus hellebore, our country as early as in ancient times have been study about the Veratrum nigrum. Compendium of Materia Medica, Sichuan Materia Medica records Veratrum nigrum medicine medicine. The foreign There are reports in the mid-twentieth century, the state of idaho, United States of America local herders found abnormal status quo of their range lamb born deformity is higher than in other regions, and the performance of monocular head. Later American poisonous plants research scientist Keeler of this deformity status quo long-term research and recognized with the1964Veratrum of steroidal alkaloids is the main ingredient to cause the lanb monocular deformity.and later its alkaloids conducted in-depth research, and found to have lower blood pressure, lowering blood pressure, anti-thrombotic, antic-cancer activity, at home and abroad have Veratrum alkaloids activity in-depth research, especially in the field of anti-cancer research is the most popular. And the extraction and purification of various Veratrum alkaloids,our country containing abundant Veratrum species, so research scenaria is quite broad.This article have more systematic study of Veratrum alkaloids of Veratrum nigrum, aims to fully develop and utilize our rich medicinal resources, to provide a scientific basis for the development of our country have their own intellectual property rights of chinese herbal medicines. This article extracted from the Veratrum nigrum purified purity the higher veratramine and jervine, and the use of high performance liquid chromatography find in the Veratrum nigrum containing the anti-cancer constituents cyclopamine, the use of the response surface methodology optimize vertramine and jervine extraction conditions, HPLC analysis of these two components of the determination of the content, and final the content of the veratrum alkaloids anti-bacterial activity of the two substances purified. The main contents of the study and experimental results are as follows:(1) The first chapter extraction,purification and analysis of the chemical composition of the Veratrum nigrum. Under alkaline condition,extracted with ethanol,and n-hexane/ethyl acetate(v/v=7/3) the extract wasconcentrated liquid, the use of high performance liquid chromatography analysis of the extraction liquid. HPLC condition:column waters Symmetry C18column (250mm×4.6mm,5um); mobile phase of acetonitrile:0.05%formic acid=20/80-35/6512minutes flow rate:1.0ml/min; detection wavelength:215nm; column temperature:30℃. Veratrum plants in the presence of cyclopamine, the extract after purified by silica gel chromatography, eluted with hexane:acetone (0.5%triethylamine)(v/v=7:3) and hexane:acetone (0.5%triethylamine)(v/v=1:1) as eluent, two white solid precipitated-UV and mass spectrometry, nuclear magnetic resonance methods to detect and identify the precipitated white solid are veratramine and jervine.(2) Separation the purified from a higher purity veratramine and jervine dubbed in five different concentration gradient are4mg/ml,2mg/ml,1mg/ml,0.5mg/ml,0.25mg/ml,use high performance liquid spectrum for two standard curve. Then extracted three times veratramine and jervine peak area to calculate the veratramine and jervine content. The results obtained0.429%and0.2856%the veratramine and jervine content.(3) In order to optimize the extraction process of the veratramine and jervine, the response surface methodology to optimize the extraction time, extraction temperature and solid-liquid ratio, and the establishment of the mathematical model. In this paper, quinoa the veratramine and jervine extraction optimization, the single factor expriment based on combination of response surface methodology to analize the main factors affect the amount of amine extraction of veratramine and jervine, resulting in optimum extraction condition. Veratramine optimum extraction condition2.0h,extraction temperature75℃,solid-liquid ratio1:9.0(g/ml), under optimum conditions,20g dry quinoa reed rhizome extract veratramine amount98.23mg worth with the predicted error is2.66%, jervine optimum extraction conditions for the extraction time3.5h, extraction temperature of45.0℃,the solid to liquid ratio of1:9.0(g/ml), the actual amount of extract the20g vreatrum dried root58.23mg predicted by the model is worth the error is3.05%.(4) In this paper, filter paper method of veratramine and jervine antimicrobial activity of the two compounds studied. The selected strains are Bacillus subtilis, Escherichia coli, White aspergillus and Staphylococcus aureus. By comparing five different concentration of20mg/ml and10mg/ml,5mg/ml,2.5mg/ml,of1.25mg/ml veratramine and jervine ring size four bacteriostatic anti-bacterial extent. The result show that a certain concentration of veratramine four kinds of bacteria have different levels of anti-bacterial activity, the most significant dialogue Aspergillus anti-bacterial2.5mg/ml anti-bacterial effect. Below this concentration a few experimental concentration no inhibitory effect on E.coli, Staphylococcus aureus and Bacillus subtilis has anti-bacterial effect in the concentration of10mg/ml. The jervine dialogue inhibition of Aspergillus most significantly,2.5mg/ml inhibitory effect, the best inhibition zone. Referral-Finnish amine no inhibitory effect on E.coli, the concentration of5mg/ml anti-bacterial activity against Staphylococcus aureus, Bacillus subtilis inhibitory concentration of1.25mg/ml.