Analysis Of Effective Composition In Rosa Roxburghii Tratt. And Its Bioactivity In Vitro

As a medicine edible plant,Rosa roxburghii Tratt.have a long history in our country.Modern research shows that Rosa roxburghii is effective in anti-aging,anti-oxidation,immuneregulation,anti-atherosclerosis,anti-cancer,and detoxification.However,at present,systematic research on the efficacy and material basis of Rosa roxburghii is still limited at home and abroad,and its quality control has not received attention.Exaction,purification and analysis of effective component were studies,and the biological activity of the main composition was investigated.The main research contents and results was showed as follows:（1）An extraction assay applying microwave-assisted enzymatic treatment for polysaccharides in Rosa roxburghii was developed using response surface methodology.Specific conditions（microwave power,575 W;microwave time,18min;liquid-to-material ratio,13.5:1;and enzyme dose,6.5 g/mL）generated an experimental yield of 36.29%,Purification with a DEAE-52 cellulose column generated two fractions,PR-1（from 6.2×10³ to 7.4 KDa）and PR-2（from 559.8 to106.6 KDa）.The monosaccharide composition of PR-1 was determined by HPLC using a 1-phenyl-3-methyl-5-pyrazolone precolumn derivatization method.The result showed that PR-1 contained mannose,ribose,rhamnose,glucosamine hydrochloride,glucuronic acid,galacturonic acid,glucose,galactose,arabinose and fucose with molar percentages of 2.1%,0.54%,2.1%,0.26%,1.5%,22.7%,24.0%,26.4%,19.6%and 0.89%,respectively.PR-2 contained Man,Rib,Rha,GluUA,GalUA,Glu,Gal,Ara with molar percentages of 3.2%,0.8%,3.2%,2.2%,34.8%,12.1%,24.0%and19.4%,respectively.In addition,Using cell culture system in vitro,this research was mainly focused on antitumor activity in vitro of polysaccharides extracts from Rosa roxburghii.The results showed that polysaccharides could effectively inhibit the proliferation of human hepatocellular carcinoma cells HepG-2 and that flavonoids extracts had time-dose dependent on the inhibition of cell proliferation.（2）The extraction and antioxidant activity of polyphenols was studied.With extraction yield of polyphenols as the index,the effect of different extraction methods was researched,and the ultrasonic assisted extraction method was ultimately found to be the optimal method due to the highest polyphenol content using it.The optimum extraction conditions were obtained by the response surface method（RSM）and under the optimum conditions,the polyphenol extraction rate reached 19.78%.Furthermore,the polyphenol extraction was extracted and divided into four extraction parts,namely petroleum ether part,chloroform part,ethyl acetate part and n-butanol part,respectively.With the the DPPH radical scavenging activity,OH radical scavenging ability,O₂^-anion scavenging activity,total antioxidant activity and reducing capacity as the index,the antioxidant activity of four extraction parts was investigated and the the results showed that polyphenol content and the antioxidant activity of ethyl acetate part was all the highest.It can be concluded that the antioxidant activity of Rosa roxburghii Tratt.may be related to the polyphenol content.Additionally,the polyphenol purification process was also optimized by the sifting of 16 kinds of macroporous resins.AB-8 macroporous resin was ultimately chosen to be the most suitable resin for purifying polyphenols from Rosa roxburghii.Then,with loading sample concentration,sample solution pH,loading sample volume,eluent concentration as the single factors,the optimum purification process was determined and the purified polyphenol content increased to 60.34%.（3）Study on fingerprint of Rosa roxburghii Tratt.Firstly,the UV spectra of 18batches of Rosa roxburghii were determined and compared.The result showed that there was significant difference in peak position and absorbance of Rosa roxburghii from different growing area.Moreover,with the chemometric method,the UV spectrum of different origin of Rosa roxburghii was analyzed by cluster analysis and principal component analysis.Based on the process above,the UV fingerprint chromatogram of 18 batches of Rosa roxburghii from different origin was established and results showed that this method could cluster the Rosa roxburghii collected from different origin.Additionally,the infrared（IR）spectrum of 18 batches of Rosa roxburghii in the range of 4000⁴00 cm^-1 was measured using infrared spectroscopy technology.The results showed that there is much difference in the location,feature and intensity of the characteristic absorption peaks of the IR spectrum of different batches.Thus,the quality of Rosa roxburghii from different origin can be identified and classified.Finally,the HPLC fingerprint of polysaccharides extracted from Rosa roxburghii was established,and 18 common peaks were identified.Eighteen batches of polysaccharide from Rosa roxburghii were evaluated by the HPLC fingerprint chromatograph,and the similarity was more than 0.95.In summary,the research in this paper takes Rosa roxburghii as the research object and conducts a series of research on its main active ingredient polysaccharides and polyphenols.The results provide important basis for further development and comprehensive utilization of Rosa roxburghii,and provided important theoretical reference and basis for the quality control standard of Rosa roxburghii.