Research On Effective Sleep Improvement Parts Of Tilia Amurensis Rupr. And Its Chemical Consitituents

Linden is a traditional medical plant in civil.Its flower and bark have relieving superficies by cooling effect,while it is used commonly to as sedative,hynponosis and antidepressive drugs.Tilia amurensis Rupr.isparticular species of Tiliaceae in Asia,but its pharmacal reports are seldom seen.Some aspects of pharmacognosy and chemical constituents of sedative and hypnic part were studied on the base of anti-inflammatory,analgesia and antibiosis experiments.Active components of improving sleep in T.amurensis Rupr.inflorescences were studied by bioassay-guided separation,and mechanism of action was elucidated.Firstly,extracts of T.amurensis Rupr.inflorescenceswere divided into several parts according to the polarity and sedative and hypnic part was screened by experiments which was coordinated with barbiturates drugs.There was sedative and hypnic effects in the acetic ether part,whereas,no finding in other parts.Effective part of T.amurensis Rupr.inflorescences causes best effect at 0.54g/kg during seven days' medication by consideration of dose-effect and time-effect relationship.Secondly,chemical composition of effective part of T.amurensis Rupr.inflorescences were studied.(1)Major components of acetic ether part of T.amurensis Rupr.inflorescences were exosyndromed by HPLC-MS/MS and MSE,and thirty-nine ion peaks were detected under positive,negative mode,twenty-four coponents were identified.Eighty pecent of compounds were flavone with mother nucleus,such as campherol,quercetin,isorhamnetin,acacetin,and considerable procyanidin and its polymers were found.(2)Acetic ether part was isolated by repeatedly column chromatography,ODS column chromatography,Sephadex LH-20 column chromatography and preparative HPLC,seventeen compounds were isolated.Moreover,structures of isolated compounds were identified by 1H-NMR,13C-NMR,HSQC,HMBC and Q-TOF-MS.Forteen compounds,including seven flavone compounds and seven other compounds were isolated from acetic ether part.In fourteen compounds,four compounds were firstly found in Tiliaceae,additionally,fourteen compounds were firstly found in T.amurensis Rupr.(3)Assay method of three components including astragalin,tiliroside and isoquercitrin in acetic ether part of T.amurensis Rupr.flower were established,ZORBAX SB-C18 chromatographic column(4.6mm×150mm,5?m)was adopted,gradient elution,mobile phase:methanol-water-0.1%phosphoric acid;rate:1mL/min;Linear relation of three components was well,extent of average recovery was 95%?100,97%and RSD 2.7%-4.6%.Finally,sedative and hypnic effects of three compounds(astragalin,tiliroside,isoquercitrin)were evaluated by experiments which was coordinated with barbiturates drugs.Compared with control group and effective part group,there were distinguished sedative and hypnic effects in the tiliroside and isoquercitrin groups for extend mice sleep time and shorten sleep incubation period.So tiliroside and isoquercitrin are active components for improving sleep effect of T.amurensis Rupr.inflorescences.It provides experimental advice for active components on improving sleep of T.amurensis Rupr.inflorescences,at the same time,lays the foundation for the medical development and resource exploitation of T.amurensis Rupr.