Studies On The Extraction Process Of Extract From Semen Hoveniae And Extracts’ Effect On Exercising Ability Of The Mice Under The Condition Of Low Pressure And Hypoxia

The research subject of this thesis is the study of high altitude anoxia-resistantHovenia Dulcis Thunb tablets. Previous research has proved the activity of the waterextracts from semen hoveniae in resisting fatigue, anoxia, coldness and heat. Furthermore,its water extracts can be made into tablets. The author’s task for this thesis is to optimizethe extracting and refining process and explore the anti-fatigue activity and possiblefunctions mechanism of semen hoveniae extracts in high altitude environment.This experiment has adopted the method of High Performance LiquidChromatography (HPLC) for qualitative and quantitative analysis. Our study makes acomparison of Relative Retention Time (RRT) among comparative samples to identifymain flavonoid compounds. with regard to another compound, it had been enriched,purified, and analyzed through spectrum as well. Meanwhile, This study adopted thefactorial design of two factors and three levels to optimize the material liquid ratio andextraction time of semen hoveniae in water by taking the contents of three main flovonoidsfrom the extracts as indicators, followed by conducting one-factor parallel experiment tooptimally select extraction times, alcohol concentration during the alcohol depositingprocess and the time for that process as well. In the ethyl acetate extract process, the ratioof ethyl acetate to EEE water solution by volume is1:1, we extracted for5times, thecontents of flovonoids from every single time were compared. On the other way, Male ICRmice are randomly divided into many groups, namely control group, model group，Nuodikang capsule positive group, six groups of Hovenia dulcis Thunb extracts1and2in high, middle and low dosage. Some comparisons were made among different effectsbrought by those dosages on the mice’s swimming time to provide direct evidence of theextracts’activity in resisting anoxia and fatigue at high altitude and to observe the extracts’capability in interfering with the mice’s UN,MDAconcentration, the activity ofCAT,GSH-PX, and T-AOC after the swimming test in a simulated high-altitudeenvironment, realizing the preliminary exploration of pharmacological activity mechanismfor the semen hoveniae extracts.Main Chromatographical peaks for the extract solution in High Performance LiquidChromatography (HPLC) were correspondent with chromatographical peaks fordihydromyricetin, dihydroquercetin, myricetin and quercetin with consistent RelativeRetain Time.(RRT). The spectrum data for the enriched compound and nuclear magnetic resonance hydrogen spectrum data were in accordance with the spectrum data of(2S,3S)-3,5,7-Trihydroxy-2-(3,4,5-trihydroxyphenyl)chroman-4-one. The results of theProcess experiment showed that the material liquid ratio and the extraction time are twofactors statistically significant for the result of water extraction with the combination of1:12for the material liquid ratio and0.5hours for the extraction time as the best optimizedcombination.(p<0.05). The two factors interacted with each other. After3times ofextraction with extraction volume for the third time taking up8.28%of the volume forthree times of extraction and the total transfer ratio of flavones reaching50.7%, it can besafely concluded that three times of extraction is the most appropriate way. After theprecipitation of ethanol in2times,3times,4times of volume with95%in concentration,there is no statistical significance for the differences between total flavone transfer ratio.However, there is statistical significance by comparing the ethanol two times of volumewith highest extraction rate with the other two groups. The differences of flavones transferratio and extraction ratio brought by precipitation for8hours,10hours,12hoursrespectively have no statistical significance. The contents of flovonoids from every singletime were less than5%of the total after extraction for4times in the ethyl acetate extractprocess. The results of Swimming Test for mice under simulated high-altitude environmentdemonstrated that middle dosage group and high dosage group taking semen hoveniaeextracts1and high dosage group taking semen hoveniae extracts2had longer swimmingperformance than model group and the difference had statistical significance. Meanwhile,the UN concentration in serum in high dosage group taking semen hoveniae extracts1andmiddle, low dosage groups taking semen hoveniae extracts2were lower than theconcentration for the model group, which had statistical significance. Meanwhile, theMDAconcentration in hepatic tissue in high and middle dosage group taking semenhoveniae extracts1and in high and middle dosage group taking semen hoveniae extracts2were lower than the concentration for the Model group, which had statistical significance.The activity of CAT in hepatic tissue in high,middle and low dosage group taking semenhoveniae extracts1and in high dosage group taking semen hoveniae extracts2were higherthan the concentration for the Model group, which had statistical significance. The activityof GSH-PX in hepatic tissue in high,middle and low dosage group taking semen hoveniaeextracts1and in high,middle dosage group taking semen hoveniae extracts2were higherthan the concentration for the Model group, which had statistical significance. The T-AOCin serum in high dosage group taking semen hoveniae extracts1and hoveniae extracts2 were lower than the concentration for the model group, which had statistical significance,while the T-AOC of the model was higher compared with the control group.Test results showed that: it can be confirmed that setting material liquid ratio at1:12and three times of extraction with0.5hours for each time can yield the highest extractionratio for the flavones in the water extraction process. Choosing ethanol two times ofvolume for precipitation process would satisfy the principle of thrift. Theethonol-depositing time was about12hours. Extraction for4times was enough the ethylacetate extract process. Under the high-altitude environment, both semen hoveniae extracts1and2have the activity to enhance the athletic ability of mice. Moreover, both of themcan reduce serum UN concentration and serum T-AOC of mice after swimming. At thesame time, Hoveniae extracts1and2showed the ability of bringing down theconcentration of MDAand raising the activity of CAT and GSH-PX in hepatic tissue ofmice after swimming.