Studies On The Chemical Material Basis Of Evodia Rutaecarpa (Juss.) Benth. For Antagonising Helicobacter Pylori

Evodia rutaecarpa benth is the dry and nearly mature fruit of one of the followingrutaceae plants: Evodia rutaecarpa (Juss.) Benth, rutaecarpa (Juss.) benth. Var.bodinier（iDode）Huang or E.rutaecarpa(Juss.) benth.Var.offcinalis (Dode) Huang. Inclinic, it is used in the treatment of vomiting, acid regurgitation, stomach duck painand other diseases. As the main pathogenic microbial of chronic gastritis, peptic ulcerand gastric cancer, helicobacter pylori(HP) has already been listed as one of the ClassOne carcinogenic factors by World Health Organization(WHO). In this paper, theauthor studied the active anti-helicobacter pylori components containing in evodiarutaecarpa benth and got the following results:1. Found that evodia rutaecarpa benth has the best anti-helicobacter pyloriactivityWith agar diffusion method, the author tested the anti-helicobacter pylori activityof26traditional Chinese medicines, such as coptis, golden cypress and magnoliaofficinalis, etc. It turned out that all the26traditional Chinese medicines haveanti-helicobacter pylori activity Among which, evodia rutaecarpa benth has the bestanti-helicobacter pylori activity and hericiμm erinaceus is next to it.2.Optimized the preparation method of the active anti-helicobacter pyloricomponents containing in evodia rutaecarpa benthAfter studied the effect of solvent to the extraction of the active anti-helicobacterpylori components containing in evodia rutaecarpa benth, the author optimized theextraction temperature, solvent dosage and extraction times by the use of orthogonaldesign. It turned out that the best extraction condition as follows: ethyl acetate is usedas the extraction solvent under the temperature of70℃;the volμme of ethyl acetate isthree times of that of the object to be extracted; the extraction is conducted three timeswith the duration of each extraction as one hour.The author chose to use silica gel Gand Sephadex LH-20colμmn chromatography as well as HPLC to separate andpurify the active anti-helicobacter pylori components containing in evodia rutaecarpabenth and got6monomeric compounds.3. Confirmed the structures of the anti-helicobacter pylori compoundscontaining in evodia rutaecarpa benthVia the data analysis of ultraviolet spectrμm, infrared spectrμm, electrosprayionization mass spectrμm, nuclear magnetic resonance spectrμm as well as physico -chemical properties, it is confirmed that the structures of the anti-helicobacter pyloricompounds containing in evodia rutaecarpa benth are respectively as follows: limonin,rutaecarpine,1-methyl-2-nonyl-4(1H)-quinnolone,1-methyl-2-undecyl-4(1H)-quinnol-one, dihydroevocarpine and evocarpine.4. Established the HPLC method for the determination of the content of theanti-helicobacter pylori compounds containing in evodia rutaecarpa benthThe chromatographic condition for simultaneously determining the3anti-helicobacter pylori compounds containing in evodia rutaecarpa benth, namely,limonin, rutaecarpine and evocarpine, is confirmed via laboratory screening asfollows: chromatographic colμmn (Boston Crest ODS Colμmn（4.6mm×250mm，5μm）); mobile phase: acetonitrile-water (gradient elution with45%-95%acetonitrile);velocity:1.0mL/min; examination wavelength:240nm; colμmn temperature: roomtemperature; the chromatographic condition of the other compounds, namely,1-methyl-2-nonyl-4(1H)-quinnolone,1-methyl-2-undecyl-4(1H)-quinnolone,anddihydroevocarpine is determined as follows: chromatographic colμmn (Boston CrestODS Colμmn（4.6mm×250mm，5μm）); mobile phase: acetonitrile-water (gradientelution with60%-95%acetonitrile); velocity:1.0mL/min; examination wavelength:240nm; colμmn temperature: room temperature.5. Conducted the test for the anti-helicobacter pylori activity of the compoundscontaining in evodia rutaecarpa benthAfter tested the anti-helicobacter pylori activity of the compounds containing inevodia rutaecarpa benth with agar diffusion method, it is found that all the6sampleshave excellent in vitro anti-helicobacter pylori activity and the minimμmbacteriostatic concentrations of limonin, rutaecarpine,1-methyl-2-nonyl-4(1H)-quinn-olone,1-methyl-2-undecyl-4(1H)-quinnolone,dihydroevocarpine and evocarpine arerespectively as0.03mg/mL,0.05mg/mL,0.02mg/mL,0.01mg/mL,0.01mg/mL and0.01mg/mL while their minimμm bactericidal concentrations are respectively as0.05mg.ML,0.05mg/mL,0.03mg/ML,0.02mg/mL,0.01mg/mL，and0.01mg/mL.