Chemical Constituents And Pharmacological Effects Of Three Frogs In Heilongjiang Province

Heparin is an animal mucopolysaccharide and has many physiological activity.This studyapplies liver in rana amurensis boulenger,rana chensinensis,rana nigromaculata as raw materialand extracts heparin, mainly adopts ultrasonic water extraction and ultrasonic alkali extraction of thecrude polysaccharides in the livers of rana, and then purifies and refines to get more pure heparin,determines and compares the heparin anticoagulant activity by the two methods. The results andconclusions of our research are as follows:Detect the basic components in the liver:Determine the content of the moisture, crude fat,crude protein and ash in the livers by national standard test methods.The results are respectively:rana amurensis boulenger:7.33%,3.59%,12.12%,2.27%; rana chensinensis:6.02%,9.50%,6.02%,9.50%; rana nigromaculata:6.04%,10.79%,6.04%,4.96%. Detecting the content ofbasic components is to have a basic understanding of the components in the liver and to providebasic data for subsequent experimentsEstablish the extraction method of heparin: The content of polysaccharide is determined byphenol-sulfuric acid method with glucose as reference substance,The best technological conditionsof ultrasonic water extraction and ultrasonic alkal extraction are determine by the single factor testand orthogonal test,the best technological parameters of water extraction shows that:the formerultrasonic heating time is10minutes, ultrasonic time is15minutes,the ratio of material to liquid is1:25, alcohol consumption is1.5times;the best technological parameters of ultrasonic alkali showsthat:the former ultrasonic heating time is40minutes, ultrasonic time is10minutes,the ratio ofmaterial to liquid is1:20, the concentration of lye is1.5mol/L..The purpose is to extract extremelypolysaccharide from the rana liver and improve the yield of heparinPurifine and refine:Two times hydrogen peroxide hydrocarbonylation purifies crudepolysaccharides getting by water extraction and alkali extraction,hydrogen peroxide solution isadded into the sample solution by two times,which the purpose is oxidation decoloration,removingimpurities and obtaining purer heparin.Determine the content of the previous and posteriorpurification heparin by sulfuric acid hydrocarbonylation,The content of heparin in rana amurensisboulenger,rana chensinensis and rana nigromaculata are respectively:water extraction crudeheparin1.99%,1.45%,1.99%, water extraction refined heparin0.225%,0.107%,0.140%,alkali extraction crude heparin2.02%,1.95%,2.02%,alkali extraction refined heparin0.215%,0.144%,0.137%.Heparin potency of rana amurensis boulenger,rana chensinensis and rana nigromaculata withAzure A dyeing method are as follows: water is63.12,57.2,93.48, U; alkali is72.76,123.84,144.6,UInfrared spectrum analysis:Analysing extracts in livers of three ranas by infrared spectrum,candetermine the existence of the heparin, and the heparins have same types, similar structure.Theheparin getting by ultrasonic alkal extraction than by ultrasonic water extraction has higher purityand relatively less impurities.Purified heparin has higher purity than raw heparin,less impurity peaksand more obvious characteristic peaks.The heparins of rana chensinensis and rana nigromaculataare easier purification and higher purityAnticoagulant activity analysis:Applicating prothrombin (PT) analyses anticoagulant activityin the samples of heparin, heparins in the livers of three ranas have better anticoagulant activity, theultrasound water and ultrasound alkali extraction have less effect on the heparin anticoagulantactivityThis topic studies the conten of heparin in liver of three ranas and anticoagulant activity,andfinally confirms the existence of the heparin, and the content is higher.The study not only laies thefoundation for the further study of heparin in the liver, but also provides a new way to the researchof antithrombotic drugs...