Study On Correlation Between Antibacterial Activity And Chemical Compositions Of Trollius Ledebouri Reichb Extractive

Trollius chinensis Bunge,perennial root herb,belongs to L.in Ranunculaceae family.Its flower is bitter in taste,cold in nature and fragrant in smell.It has many effects such as heat-clearing,detoxification,carrion prevention and callus.It is well known that it's mainly used in the clinical course of treatment of blade injury,wounds with much pus,swollen sore throat,etc.Modern pharmaceutical research and clinical application shows that Trollius chinensis Bunge mainly treat upper respiratory infection,pharyngitis,amygdalitis and bronchitis by antibacterial,antiviral,anti-tumor,antioxidant activity,etc.In 2003,Trollius chinensis Bunge was selected as the prevention of severe acute respiratory syndrome(SARS)for one of the compound prescription drugs in disease.Trollius chinensis Bunge is used widely.Most of its compounds have been separated and identified,and study on its biological activity mainly concentrate on the flavonoids,organic acids and alkaloids.But all kinds of compounds in the pharmacological effects as individual contribution has not been clarified.Trollius chinensis Bunge possessing well biological activity and widely used,results in the deficiency and destruction of its wild resources.Therefore,it is necessary to find out the sources of its alternative medical materials,in order to expand sources of medical materials for Trollius chinensis.Trollius ledebouri Reichb mainly distributes in northeast of Heilongjiang and Inner Mongolia,eastern Siberia and the far east of the Soviet Union as well.Till now,the preliminary chemical research on Trollius ledebouri Reichb has been carried on,and flavonoids,organic acids,sterols,the chemical compounds isolated from it,have been involved in preliminary activity research.It is found that Trollius ledebouri Reichb also possesses antiviral,anti-inflammatory,antioxidant biological activity,etc.This paper,on the basis of previous researches,applied modern scientific and technical methods,for the research on Trollius ledebouri Reichb as follows:1.By UV,TLC,IR and HPLC,we established comparative study on the composition of Trollius ledebouri Reichb and Trollius chinensis Bunge,and determined Trollius ledebouri Reichb and Trollius Bunge by TLC and features identification.By total flavonoid as the evaluation index,we detected total flavonoid content within different extractive solvent,showing that the total flavonoid content between Trollius chinensis Bunge and Trollius ledebouri Reichb had no significant difference.The fingerprint of Trollius ledebouri Reichb compared with Trollius chinensis Bunge within different solvents,under the same conditions,respectively,showed 22 common peaks by HPLC.Through macroscopical features of IR fingerprint,we analyzed Trollius chinensis Bunge and Trollius ledebouri Reichb extracted by different solvents,showing that the various extraction parts of Trollius chinensis Bunge and Trollius ledebouri Reichb by the same solvent have basically the same ingredients.The research results provided the reference base for Trollius ledebouri Reichb medical material identification and quality monitoring.2.By silica gel,ODS,polyamide column chromatography and HPLC,semi-preparative HPLC,the modern separation methods and techniques,we isolated the chemical compounds of Trollius ledebouri Reichb and identified their structures.Among 10 isolated monomer compounds,by 1H-NMR and 13C-NMR spectrum analysis and literature comparison,finally four flavonoids were identified in structure:Orientin(1),Vitexin(2),Cytisoside(3),2-(4-methoxy-phenyl)-5,7-hydroxy-8-[6-O-(?-D-glucopyranosyl)-?-D-glucopyranosyl]-4-1-benzopyran-4-one(4).Compound 3,4 were found in this genus for the first time.3.After identifying effective compounds of Trollius ledebouri Reichb,we used cefalexin and Trollius chinensis Bunge as positive control,to examine correlation between chemical composition and antibacterial activity of Trollius ledebouri Reichb.The inhibition test in vivo and in vitro on S.aureus,E.coli,P.aeruginosa,salmonella and Sh.dysenteriae show that ethanol and water extractive of Trollius chinensis Bunge both worked to some extent,and particularly the total extractive worked the best.The bacteriostasis of orientin and vitexin on E.coli and P.aeruginosa is significant,while on other bacterial strains is not obvious,indicating that the bacteriostasis may be a synergistic effect through various compounds.The study provides scientific reference for the feasibility of Trollius ledebouri Reichb.as medicine instead of Trollius chinensis Bunge and the possibility of further exploiting resources of Trollius ledebouri Reichb.