| ObjectiveThere are about 155 genera,1500 species in Rutceae,mainly distribute-d in tropical and subtropical regions,including Melicope ptelefolia?Champ.ex Benth.?Hartley.Melicope ptelefolia?Champ.ex Benth.?Hartley,also named "San-ya-ku","San-cha-hu","San-zhi-qiang","San-cha-ku","San-ya-hu".In China,located in Zhejiang province,Fujian province,Jiangxi province,Guangdong province,Guangxi province,Hainan province.Melicope ptelefolia is mainly used for treatment of injuries,trauma,abscess and eczema.At present Melicope ptelefolia has been widely used in"Jinmeiganmaopian","shuanglongfengshidiedagao",rupianxiaoj iaonang,sanjiuweitai,sanjiuganmaoling and other traditional Chinese medicine patent prescription.Meanwhile Melicope ptelefolia is a kind of herbal tea ingredient in Lingnan areaabout 100 years already,also is a kind of traditional food material in Lingnan region and southeast Asia ingredients,so it can be considered as a health care product for the development and utilization.Modern study has found that Melicope ptelefolia not only have the anti-inflammatory effect,analgesic effect,antioxidant effect and protect liver effect,antibacterial effect,etc.The literature have been reported that alkaloids,benzopyrans,flavonoids and other chemicals have been isolated from the Melicope ptelefolia.However,the main effective component of Melicope ptelefolia is not clear,quality standard has not been set,in order to further develop it's medicinal value,expounds active ingredients.We systemic researched the chemical constituents of Melicope ptelefolia petroleum ether part,chloroform part and ethyl acetate part,and to establish the detected method of main compound.MethodsThe stems of Melicope ptelefolia been in room temperature immersion 6 times with 90L of 95%ethanol,and then vacuum solvent,and then get extract?675g?.To suspended in the 1L of water,and then used petroleum ether,chloroform,ethyl acetate,n-butanol to extractied and concentrated,get petroleum ether extract?45g?and chloroform respectively extract?300g?,ethyl acetate extract?75g?,n-butanol extract?110g?.Used silica gel,ODS,SephadexLH-20,silica gel separation technology to separated and purified the petroleum ether,chloroform and ethyl acetate parts to obtain monomeric compound.Integrated use of mass spectrometry?MS?and nuclear magnetic resonance spectroscopy(1H-NMR and 13C-NMR,DEPT,1H-1H-COSY,HSQC,HMBC)and other modern spectrum and spectroscopy techniques to identified the chemical structure of the monomer compounds.Use MTT method to test the cytotoxicity of new compound.Used HPLC method to determined the content of the new compo-und,chromatographic conditions as follows:chromatographic column:Kromasil C18 chromatographic column?4.6mmx250mm,5microns?;Mobile phase:m ethanol:water?42:68?;The flow rate:1.0 mL,min;Sample quantity:20?L;detection wave length of 210 nm;Column temperature ?;ResultsThere was 22 compounds isolated from the 95%ethanol extracts of petroleum ether,chloroform and ethyl acetate parts,and 22 compounds were identified,including 5 steroids,10 coumarins,2 anthraquinones,1 flavone,two Hydrocarbons-aliphatics and two alkaloids compounds.The compounds were ordinal named numbers for 1-22,the compound 15 is new compound,named pteleifolosin D,and another 12 compounds?1,2,3,6,7,8,9,12,13,15,17,19?were isolated from Melicope ptelefolia for the first time.The new compound didn't show cytotoxic activity of MCF-7,SW-480,HeLa.Analyzed the content of pteleifolosin D?3-?2',3'-dihydroxy?isopentyl-7-hydroxycoumar?by HPLC.The results showed that the linear range of pteleifolosin D was 0.24-2.64?g-ml-1 and the average recovery was 99.96%?RSD=1.61%,n=6?with an average content of 0.085mg·g-1?Qingyua n?.But the average level of the another 3 batches is 0.055 mg-g-1,it is lowe r than 0.085mg g-1?Guangdong Qingyuan?.The content of Pteleifolosin D in Melicope ptelefolia is different from the origin,picking time etc. |
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