Study On Content Determination By HPLC-ELSD And Preparation Method Of T19 In Different Parts Of Ginseng

Ginseng is a genus of the family of Araliaceae,Originated from the "east Asian-North America" of the third ancient tropical geohistory tertiary mountains,such as,southwest area of China,East area of Asia and North America.Among them,Genuine producing area of the ginseng mainly distributs in northeast of China,north of Korea,and the the part of north American covered by the quaternary glacier,which result in that there are only American ginseng and dwarf ginseng now.However,distribution range of notoginseng which compares to American ginseng and dwarf ginseng is extremely small,notoginseng mainly distributes in Yunnan province,Guangxi province,Jiangxi province,Sichuan province of China,etc.Ginseng is one of the important plants in Panax ginseng,and is a good tonic.Composition of ginseng is very complicated,which is rich in ginseng saponin,ginseng polysaccharide,ginseng protein,amino acid and flavonoid,etc.Modern pharmacological studies show that ginsenoside is the most important effective components of Panax ginseng.Now there are more than 40 kinds of ginsenoside monomers which have a variety of modern pharmacological activities.At present,ginseng has been widely used in the world,which is mainly used in pharmaceutical,food,health care and skin care.Because of the extensive pharmacological effects of ginseng,it has been a series of in-depth study after our group found ginsenoside T19 in 1990.It is found that ginseng has a very good hypoglycemic activity.Therefore,Our group wants to study the hydrolysis process to improve its content in the original plant,so as to preparation large quantity of T19,which has great social and economic significance for the utilization and development of new medicinal resources of Panax ginseng.On the basis of the research results of the chemical constituents and pharmacological activities of Panax ginseng in the early stage of the research group,the content of the Dammaranes protopanaxtriolsaponinswas established for the first time by the shudy.Using chromatographic column model Agilent C18(250 mmx4.6 mm,5?m)to peRform HPLC-ELSD analysis,mobile phase was Methanol-water(70:30,v/v)to carry on the elutio,the flow rate was 1.0 mL/min,ELSD gasification chamber temperature was 87.0? carrier gas velocity was 2.20 L/min,sample volume was 10 ?L for testing.The experimental result shows that the established method has a good linear relationship between the protopanaxtriolsaponin in Panax ginseng.It has strong specificity,precision,repeatability,stability and recovery rate that fulfill the requirements.It can be used for the determination of the content of Dammaranes protopanaxtriolsaponins in Panax.In the present study,the content of ginsenoside 20(R)-T19 in theoriginal plant was improved by hydrolyzing the total saponins and optimizing the acid hydrolysis process from the stems and leaves of Panax ginseng.Single factor experiments and orthogonal experiments were used to select the best experimental scheme for the preparation of 20(R)-T19 by acid hydrolysis.The experimental results show that the main factors affecting the hydrolysis arereaction temperature>reaction time>the concentration of Hydrochloric acid>the concentration of methanol>hydrochloric acid concentration.The optimum process is Methanol addition 3ml,addition of hydrochloric acid 15ml,hydrochloric acid concentration is 60%,reaction temperature is 35? and the reaction time was 2h.In this experiment,the optimum process was used to enlarge the experiment,and then the T19 was purified by silica gel column elution.Column chromatography elution conditions is Dichloromethane:Ethanol=12:1,which makes the value of Rf around 0.3.The thin layer detection is Dichloromethane:Ethanol=10:1,which makes the value of Rf around 0.5.After many column chromatography separation and purification,refinement of T19 which was got greater than 95%purity,yield of T19 was about 4.96%.