Studies On Quality Of Folium Mori And The Purification Of Active Components

Folium Mori(leaves of Morus alba L),the traditional medicine,contain varieties of functional compounds,such as minerals,vitamins,dietary fiber,amino acids, steroids,flavonoids,alkaloids,polysaccharide and so on,with activities of exhibiting a wide range of biological effects,including reducing blood glucose,hypertensive, antihypedipidemia,antioxidant,enhanceing immunity,anti-inflammatory,antivirus and anticancer activity.Folium Mori have been the hot spot in medicine and food area.In order to provide basis for the development of Folium Mori resource,the present study was to determine the contents of alkaloids and polysaccharide,as well as the separation and purification of the main active components from Folium Mori. Furthermore,the inhibitory effect of Folium Mori alkaloids extract on alpha-glucosidase activity and the inhibitory type were also investigated.1.Determination of alkaloids and polysaccharide of Folium MoriObjective:To determine and compare the contents of total alkaloids, 1-deoxynojirimycin(DNJ)and polysaccharide in Folium Mori from different collected time,plant sites,varieties and medicinal part.Methods:The content of total alkaloids was measured by Spectrophotometry based on the reaction of alkaloid with Remecke salt and the contents of DNJ was determined by HPLC-ELSD.The HPLC analysis was performed on Agilent ZORBAX NH2 column(4.6mm×250mm,5μm),the mobile phase was CH₃CN:H₂O(V/V=80:20),the flow rate was 1.0 ml·min^-1,the temperature of column was 30℃and the injection volumn was 20μl.The ELSD condition were as follows:the temperature of drift tube was 55℃,the gas pressure was 1.5 Bar,the value of gain was 7.0.The polysaccharide was deternined by phenol-sulfuric acid method.Results:The contents of polysaccharide in Folium Mori samples harvested in winter were higher than that of other seasons,while the alkaloids varied respectively. Meanwhile the contents of alkaloids and polysaccharide varied obviously among Folium Mori from different plant sites,different varieties and different medicinal part.Conclusion:The contents tested components are influenced by collection season, plant site,varieties and medicinal part.2.Studies on the separation and purification of the active compands from Folium Mori.by resinObjective:To investigate the technological parameters of the purification process of the active components from Folium Mori.by macroreticular and cation positive ion resin.Methods:The Chitosan was used for the clarification of the extraction,and the flocculation conditon was studied.Continuous separation by column chromatography (macroreticular resin and cation positive ion resin),the flavonides,alkaloids and polysaccharide was isolated from the extraction simultaneously.The static absorption capacity,static elution ratio,static elution capacity of 2 types cation positive ion resins were studied respectively and compared in order to find the better cation positive ion resin.The technical process for purification of the active components with the macroreticular resin and cation positive ion resin,was screened by purity and yield of respective product.Results:Compared with the traditional alcohol-precipitation,Chitosan can efficiently reserve active components at the higher rate.Q2 cation positive ion resin owned better absorption and eluation parameters for alkaloids.The absorption ratio was 35.81mg·g^-1.After the optimization,the yield of total flavonois was 65.15% and product purity was 71.86%;the yield of total alkaloids was 74.59%and product purity was 51.09%;the yield of polysaccharide was 69.94%and product purity was 60.11%。Conclusion:The R1 type macroreticular resin and Q2 cation positive ion resin showed better comprehensive absorption property.It can be used to purify the extract of Folium Mori.3.Study the inhibitory effect of Folium Mori alkaloids extract on alpha-glucosidase activity in vitroObjective:To investigate the inhibitory effect of Folium Mori alkaloids extract on alpha-glucosidase activity and the inhibitory type.Methods:Alpha-glucosidase activities were examined with or without Folium Mori alkaloids extract,Ramulus Mori extract and acarbose(positive control)in vitro, and the IC₅₀were calculated.Results:The Folium Mori extract,Ramulus Mori extract and acarbose inhibited the activity of alpha-glucosidase with IC₅₀0.075mg/ml,0.142mg/ml,0.709mg/ml respectively.The enzyme inhibiton style was reversible-competitive.Conclusion:The Folium Mori extract inhibited the activity of alpha-glucosidase remarkably,and the effect was much higher than that of Ramulus Mori extract and acarbose.