Studies On Separation And Purification Of Chemical Constituents From Traditional Chinese Medicine Salvia Miltiorrhizae Bunge And So On

Traditional Chinese medicine is the precious wealth of the Chinese nation,which has a unique advantage and role in disease prevention and health care.However,its composition is more complex and the content of effective component is relatively low,which make the separation and purification difficult.As a result,the clinical application and pharmacological research has been greatly restricted.Thus,establishing a more simple and efficient method of the separation and purification of the active ingredients from Chinese herbal medicine is particularly important.In this paper,the novel SFC methods and PHPLC methods were established about separating the main active ingredients of traditional Chinese medicine Danshen,Caodoukou and Pericarpium Citri Reticulatae,radix peucedani.The purities of the compounds was determined by high performance liquid chromatography(HPLC),and the structures of the compounds were identified by ultraviolet spectrum(UV)and nuclear magnetic resonance(NMR)analysis.So as to provide reference for further development and application of this kind of medicine.1.Separation and purification of five tanshinones from Salvia miltiorrhiza Bunge by1.Separation and purification of five tanshinones from Salvia miltiorrhiza Bunge by preparative supercritical fluid chromatography In the experiment,90% ethanol was used to extract the tanshinones from Salvia miltiorrhiza.Then using SFC to isolate and purify the crude extract.Experiment parameters were optimized on an analytical scale through a series of single-factor experiments including the type of the stationary phase,the type and content of organic modifiers,the flow rate of the mobile phase,the column pressure,and the column temperature.A preparative SFC method was then established on the basis of analytical SFC conditions by chromatographic amplification formula.Finally,five components were obtained with high purities as determined by HPLC.Their structure were identified by NMR and MS analysis.The thermodynamics of chromatographic process was also studied and the result revealed that the SFC separation process was enthalpically driven.2.Separation and purification of four components from Alpinia Katsumadai Hayata by preparative supercritical fluid chromatographyIn the experiment,70% ethanol was used to extract the active ingredients from Caodoukou.Then using SFC to isolate and purify the crude extract.Experiment parameters were optimized,such as the effects of column,modifier,flow rate and so on.Compared of the retention behavior of compositions on the column,we got the optimum operation conditions to separate four components from Caodoukou.Then a preparative SFC method was established on the basis of analytical SFC conditions.Finally,four components were obtained with high purities as determined by HPLC.They were identified as alpinetin,pinocembrin,cardamonin and alnustone according to NMR analysis.The thermodynamics of chromatographic process was also studied and the result revealed that the SFC separation process was enthalpically driven.3.Isolation and purification of five flavonoids from Pericarpium Citri Reticulatae by preparative high performance liquid chromatographyIn the experiment,Pericarpium Citri Reticulatae was first extracted by 85% ethanol to obtain the crude extract.And then the crude extract was extracted by ethyl acetate to obtain the water-soluble and lipid-soluble fractions.To establish a PHPLC method for separation and purification of flavonoids from the two fractions,experiment parameters were optimized preliminarily until the suitable conditions were obtained.Under the suitable conditions,five compounds obtained purification through separation.They were identified as naringin,hesperidin,nobiletin,3,5,6,7,8,3',4'-heptamethoxyflavone,and tangeretin by NMR and MS analysis.4.Isolation and purification of coumarins from Radix Peucedani by PHPLCPHPLC was applied to the preparative separation of coumarins with function from Radix Peucedani.The experiment was performed on a PHPLC with a C18(250 mm×25.4mm I.D.,10 ?m)column,After various parameters of PHPLC were optimized preliminarily,the suitable conditions were obtained.Under the above conditions,three compounds obtained purification through one-step separation.They were identified as praeruptorin A,praeruptorin B and praeruptorin E by UV,NMR and MS analysis.