Study On Extraction And Separation Of Bioactive Compounds In Leaves Of Chaenomeles Speciosa S. Nakai

Chaenomeles speciosa S.Nakai(called Xuancheng pawpaw)is a kind of precious Traditional Chinese Medicine,but nowadays only fruit-a small part of it,can be roughly utilized and thus resources use percentage is extremely low with much waste.In a word,the phenomenon mentioned above lead to negative effects on the protection of germplasm resources.To make good use of Xuancheng Pawpaw,extraction and separation-purification technology were studied on main bioactive compounds--Oleanolic Acid(OA)in leaves of Xuancheng pawpaw in this paper,and a quick method to separate and to test OA-High Performance Capillary electrophoresis(HPCE)was stated.Based on the qualitative experiment of Thin Layer Chromatography(TLC), Oleanolic Acid(OA)in leaves and fruits of Xuancheng pawpaw were found and to the contrary in branches.And then rate and the purity were taken as the target and the Comparisons of four solvents were demonstrated(methanol,95%ethanol,ethyle acetate,acetone),as result,95%ethanol as extraction solvent had a highest yield of 10.63%,acetone as extraction solvent had a best purity of 12.64%.Based on effects above combined with the toxicity and the experimental cost,95%ethanol was chosen as the optimized solvent.Then extracted through such approaches as refluxing extraction,microwave assisted extraction,ultrasonic wave assisted extraction,Infusion extraction and Soxhlet extraction.Such approaches mentioned above were optimized,as a result,the microwave assisted extraction was the best extraction technique,the parameter as follows:80%ethanol as extraction solvent,extraction time for 3min,ratio of material to liquor 1/16 g/ml,microwave power 100W,the yield is 16.06%,the purity reaches 12.60%.OA was separated and purified by recrystallization and macroreticular resin chromatography separation,and chose the concentration of desorbing solvent.It shows that 70%ethanol as desorbing solvent was best,and the two separation and purification methods were employed to get OA and the purity were 89.22%and 87.7%respectively.A quick method to separte and to test OA-High Performance Capillary electrophoresis(HPCE)was stated with its conditions:20mMKH₂PO₄-40mM sodium borate buffer system,containing 15%(v/v)methanol,pH7.0,injection pressure 0.5p.s.i, The separating voltage was 25kV,column temperature 25℃,and detection at 195 nm. The result shows that OA was well detected at 8.97min.And.the content of OA both in fallen leaves and fresh leaves were determinated,1.39%and 1.58%respectively by HPCE.