| Camellia oleifera Abel,a perennial woody oil crop,mainly distributed in Guangxi,Anhui,Jiangxi and other southern regions,the seeds are mainly used for the production of Camellia oleifera oil.Camellia Cake is a kind of dreg,which is rich in flavonoids and other active ingredients.Through extraction,isolation and identification for oil,we find that flavonoid glycosides to aglycones kaempferol is rich in oil tea Camellia,and the raw material drug of kaempferol can be obtained by hydrolyzing flavonoid glycoside.In addition,as camellia oleifera is rich in saponin,and we can only extract and separate saponin from camellia oleifera by the existing technology,we can try to grope for a workable process to separate tea saponin and kaempferol from camellia oleifera.In this paper,we make the camellia oleifera as raw material,and have finished studies on extraction and isolation identification of camellia oleifera,and explored two feasible process to make the coproduction of tea saponin and kaempferol.The main research contents and conclusions are as follows:1 The extraction process of camellia oleifera.We use alcohol extraction,and focus on the effects of solid-liquid ratio,ethanol concentration and extraction time on extraction rate of flavonoids.The optimum extraction process is obtained by using the design and analysis experiments of Design Expert8.0 software,which is:the ethanol concentration is 70%,the extraction time is 2.08h,and the charge to liquid ratio is 1:25;2 Chemical compounds are obtained by the technology of the combination of plant chemistry and spectroscopy,which are:kaemferol-3-O-[2-O-β-D3-galactopyran-osyl-6-O-α-L-rhamnopyra-nosyl]-β-D-glucopyranoside,kaemferol-3-O-[2-O-β-D-glu copyranoside-6-O-α-L-rhamnopyranosyl]-β-D-glucopyranoside,kaemferol-3-O-α-L-r hamnopyranosyl-(1→6)-p-D-glucopyranoside and kaemferol.3 Hydrolysis process of camellia oleifera flavonoids.We use the method of water extraction to extract the polysaccharides in Camellia oleifera,and focus on the effects of solid-liquid ratio,extraction temperature and time on the extraction rate of polysaccharides.The optimum hydrolysis condition is obtained by using the design and analysis experiments of Design Expert 8.0 software,which is:the hydrolysis temperature is 80℃,the hydrolysis time is 1.5 h,and the concentration of hydrochlor--ic acid is 8 mol/mL.4 Comprehensive extraction process of Camellia saponin and kaempferol.Technology one:Firstly,obtaining the crude extract by using alcohol to extract the defatted Camellia dreg,and then,hydrolyzing the liquid by hydrochloric acid.We can get kaempferol by concentrating the ethyl acetate extraction,and get tea saponin by purifying the water phase by AB-8.The experimental results are as follow:the tea saponin yield is 3.53%and the purity is 97.4%,the kaempferol yield is 0.52%and the purity is about 90.7%.Technology two:Firstly,the alcohol extraction of Camellia oleifera should be condensed,and be extracted by isopropyl alcohol,and then,we will get camellia oleifera flavonoids after the extract is concentrated.We can get tea saponin by purifying the water phase dissolution,and get kaempferol by concentrating the extraction which is obtained by hydrochloric acid hydrolysis and ethyl acetate extraction.The experimental results are as follow:the tea flavonoids yield is 0.68%,the tea saponin yield is 6.15%,and the purity is 95.2%;the kaempferol yield is 0.43%,and the purity is 91.3%.5 We mainly study the toxicity and anti-inflammatory effects of tea,tea saponin,flavonoids kaempferol on cells,and results shows that tea saponin and flavone of Camellia oleifera have a inhibitory effect on the expression of NO,and show a good dose dependence.The inhibitory effect of kaempferol on PGE2 is the best when the concentration of it is 20 g/mL which is induced by LPS,and the content of PGE2 in cell supernatant is 41.6 pg/mL at the same time.Tea saponin and flavone of Camellia oleifera have a inhibitory effect on the expression of PGE2,but the dose dependence is not obvious.We test the cell toxicity of tea saponin,camellia flavone and kaempferol by MTT method,and find a less toxic which shows a great potential in drug research and development.We have finished studies on extraction and isolation identification of camellia oleifera flavonoids,and explored two feasible process to make the coproduction of tea saponin and kaempferol,and study the toxicity and anti-inflammatory effects of tea,tea saponin,flavonoids kaempferol on cells.Results in this paper will provided the foundation of experiment and theory for their further development and application. |
