The Synthesis Of Alanine-Deoxy-Fructose Of Red Ginseng And Study On Its Pharmacological Activity About Anti-diabetic

This research investigates ADF formed during Maillard reaction and aimed at a new anti-diabetic agent.The experiment determinates the content of ADF,optimizes its synthetical process and determinates the inhibition ratio on maltin,all of which provide a theoretical basis for research and development of a new anti-diabetic agent.It is found that the content of ADF was extremely low during the determination of the content of ADF from red ginseng.It took much labor and wasted raw material to extract ADF for further study on its bioactivity.In order to obtaining a great quantity of ADF,it was synthesized in vitro in this experiment.The materials were alanine and glucose,and glacial acetic acid as the menstruum,the ratio of alanine and glucose was1:1and the reaction was progressed in a water bath of80℃for1hour.Then the mixture was placed in evaporating dish to remove glacial acetic acid at40℃.In this research a new separation system is built on the basis of previous studies.The crude ADF was obtained by macroporous adsorptive resins,and purified by Sephadex LH-20.At last,the content was determined of90%by HPLC analysis.The product identified as the target compound by using physical and chemical properties,TLC,IR,MS and NMR.There are ELSD,pre-column derivatization and post-column derivation to detect alanine.It was stable and accurate by using Pre-column derivatization was used in this experiment because of its stablility and accuracy.Chromatographic conditions were a Venusil-AAcolumn used specially for analysis amino acid (5μm,4.6mm×250mm,column), temperature:30℃,UV detector:211nm,sample volume:20ul.Phosphate buffer solution and chromatography methanol were mobile phase A、B respectly.Mobile phase A:B=70:30,flow rate:0.5ml/min with constant gradient elution.This research investigated the conversion rate of ADF from three factors:reaction temperature,reaction time and substrate.It’s found that when at80℃and the ratio of glucose and alanine:1:1the conversion rate was at the highest.The conversion rate of ADF was the best at60min by testing reaction time from10min to70min.So choosing the optimum reaction time needs further study.Optimizing the synthetic process by exploying uniform experiment for the conversion rate of ADF. Through the statistical analysis data it was found that the conversion rate was38.55%at the best condition.The new process method worth popularizing because of its high stability and good repeatability through stability test.It provides a theoretical basis for large scale preparation of ADF to study biological and pharmacological activity further.This paper also discussed the inhibition of ADF on α-Glucosidase.DMSO(1mg/mL) and α-Glucosidase(1U/mL) were added to tribasic potassium phosphate buffer,37℃water-bath for10min,put different concentration of sample solution,added PNPG(50mmol/ml) to the reaction,and Na2CO3was added to end the reaction at last.Measured its absorbance at400nm.It found that when the concentration of sample solution was12.5mg/ml, the maximal inhibition rate reached60.79%.The liveless of α-Glucosidase was restrained,it’s hydrolysis descended,and the content of blood glucose decreased as well.So ADF is effective on anti-diabetic.