| Chaenomeles speciosa S. Nakai(called Xuancheng pawpaw)is a kind of fruit with a medicinal function,it is rich in nutrients and abundant of active substances.Especially,it is rich in organic acids(OA) and ursolic acid(UA),these two belongs to triterpenes(saponins),which has conspicuous pharmacological effects and biological activities. To use Chaenomeles speciosa S. Nakai resources effectively, papers on the Anhui region of the basic components of Chaenomeles speciosa S. Nakai measured to study the extraction of triterpenes from Chaenomeles speciosa S. Nakai, include isolation and purification process, utilize Pre-HPLC to get the high purity monomer of OA and UA In addition, also discussed the antimicrobial effect of the triterpenes to conduct research.This study is to provide a theoretical foundation for the processing of Chaenomeles speciosa S. Nakai and further application of the active substances.1.The nutrient component in fruits of Chaenomeles speciosa S. Nakai was determined, it contained of water,2.582%of total sugar,2.924%of organic acid,1.752%of crude fiber,1.264%of protein,0.086%of OA,0.040%of UA,1.321%of pectin.2. A new method for quality and content determination of OA and UA in Chaenomeles speciosa S. Nakai at the same time-analysis high performance liquid chromatography(HPLC) was developed.The mobile phase composition, the flow rate and detection wavelength were optimized. The optimum operation parameters were selected as follows:a Phenomenex C18Column(250mm×4.6mm i.d.，5μm) was used with the mobile phase being Methanol-water(85:15,v/v),flow rate being0.5mL/min,determinating wavelength being210nm,the column temperature being25℃,and the injection volume being5μL.3.The paper gives extraction optimization of triterpenes from Chaenomeles speciosa S. Nakai.Selected ethanol concentration, extraction time,temperature and solid-liquid ratio as test factors to design the orthogonal test.The result showed that the best condition of extraction is as follows:80%ethanol, temprature55℃,solid-liquid ratio1:20, supersonic wave treatment for20min.The extraction was purified more than once, the content and purity were determined with analysis HPLC. The result showed that the purity which extracted by N-butanol water saturated solution was41.41%the purity which purified by macroporous resin was71.98%then,the reversed-phase silica column method was used,and the purity of the triterpenes samples was reach up to93.56%.The content of OA and UA in Chaenomeles speciosa S. Nakai,the proportion of OA and UA was2.14～ 2.22:1,RSD value was1.39%.4. A preparative high performance liquid chromatography(Pre-HPLC) method was successfully used to separate OA and UA from Chaenomeles speciosa S. Nakai, the result was validated by analysis HPLC. It showed that97.50%OAmonomer and97.27%UA monomer were separated respectively. The mobile phase composition,the mobile phase,the flow rate,the detection wavelength and injection volume were optimized. The optimum operation parameters were selected as follows: a pursuit XRs C18Column(250mm×212mm,5μm) was used with the mobile phase being Methanol-water(80:20,v/v),flow rate being20mL/min,determinating wavelength being200nm,the column temperature being25℃,and the injection volume being20mL.5.The antimicrobial effect of the triterpenes in Chaenomeles speciosa S. Nakai showed that it has broad-spectrum antimicrobial function，it had anti-activity for G+and G-. The triterpenes of different purity had anti-activity for Bacillus subtilis,and when its purity was improved,its anti-activity was also improved the minimal antibacterial concentration of the triterpenes which was purified by reversed-phase silica column was1.56mg/mL Compared the antibacterial effect of41.41%triterpenes in different pH value,the result showed that the anti-activity of triterpenes for Bacillus subtilis was best in pH value is3Compared the antibacterial effect of41.41%triterpenes for Bacillus subtilis in different temperature, the result showed that the thermal stability for triterpenes was stable relatively. |
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