Pharmacodynamic Substances Studies Of Active Fractions From Panax Notoginseng (Burk.) F.H. Chen & Fragmentation Behaviors Of Quinochalone Compounds From Carthamus Tinctorius L. And HSYA's Internal Distribution Studies

The thesis included two parts of studies on pharmacodynamic substances studies of active fractions from Panax notoginseng(Burk.) F.H.Chen and studies of fragmentation behaviors of quinochalone constituents from Carthamus tinctorius L.and HSYA's internal distribution.P.notoginseng(Burk.) F.H.Chen is one of the most commonly used traditional Chinese medicinal plants.Its roots(San-Qi) have been widely used for treatment of cardiovascular diseases,inflammation,fatigue,different body pains,and internal and external bleeding due to injury.Through extensive phytochemical and pharmacological studies,triterpenoid saponins have long been recognized as the main bioactive constituents of P.notoginseng.The type of saponins is mainly of dammarane-type (ginsenoside).Numerous studies,including fragmentation hehaviours,on-line analysis, determination and metabolism,have been reported which forcused on major constituents. However,the TCM is a complicated system,except for major constituents,there are a lot of minor or even trace constituents.Therefore,development of reliable analytical method for minor triterpenoid saponins is of great significance for elucidating pharmacodynamic substances of P.notoginseng and its related preparations.In the study reported here,fragmentation behaviors of triterpenoid saponins from P. notoginseng were investigated by ESI-MS~n and high-performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry(HPLC/ESI-MS~n). Analyses revealed that fragment ions from glycosidic and cross-ring cleavages can give a wealth of structural information regarding the nature of the aglycone,sugar types,the sequence and linkage information of sugar units.According to the summarized fragmentation rules,identification of triterpenoid saponins from the roots of P. notoginseng could be fulfilled,even when reference standards were unavailable. Furthermore,minor and trace constituents were enriched and detected by eliminating the major constituents in one of the saponin fractions.As a result,a total of 151 saponins, including 56 new trace ones,were identified or tentatively characterized from saponin fractions based on their retention times,HPLC/HRMS,HPLC/ESI-MS~n fragmentation behaviors and comparison with data in the literatures.These studies can assist in the effective analyses of saponins of this type and benefit the structural identification of similar trace constituents in fractions.This research provides a model for extensively screening and structural characterization of bioactive constituents in plant extracts.It also provides an informative fingerprint of P. notoginseng.Carthamus tinctorius L.belongs to the family of Compositae,which is widely distributed in the province of Henan,Zhejing,and Sichuan,etc.in China,and its flowers are used as traditional Chinese medicine for treatment of heart and brain blood vessel diseases.Quinochalones are the major bioactive constituents of Carthamus tinctorius L. It shows higher polarity,therefore,it isolation and purification is difficult.ESI is a relatively new technique and is ideally suited to the introduction of polar,thermally labile compounds into a mass spectrometer.In this study reported here,fragmentation behaviors of quinochalones from C. tinctorius were investigated by ESI-MS~n and HPLC/ESI-MS~n.And an efficient on-line identification system was established for constituents of this type.After the characteristic fragmentation patterns in positive and negative ion mode,and UV spectra of these compounds were summarized,HPLC-HR-MS,HPLC-DAD-ESI-MS~n of active fraction from C.tinctorius were measured.This technique can be utilized for the consecutive analysis of metabolites.Hydroxysafflor yellow A(HSYA),a C-Glycosyl quinochalcones isolated from the safflower,exhibited extensive biological activities such as anti-cerebral ischemia effect. However the actual site of action of HSYA was still unclear and it was less study on whether the HSYA would travel through the BBB(blood brain barrier) or not.Therefore,based on the mass spectrum schizolysis of C-Glycosyl quinochalcones, the anti-cerebral ischemia activities study of HSYA in plasma,different part of the brain and intra-cellular was developed.As an anti-cerebral ischemia drug,we investigated the distribution characteristics of HSYA in the brain at various group,different time,not alike take medicine way.The result showed the HSYA as low polarity compound,can travel through BBB at model animal and control group.In addition,the distribution of HSYA expressed significantly variance in the different part of brain,take medicine way and time.The research should do well to deeply mechanism study of anti-cerebral ischemia of HSYA.