Study On The Chemical Constituents Of Gui-Zhi-Tang By LC-MS And Evaluation Of Its Anti-inflammatory Activity

Traditional Chinese medicine (TCM) formulae are the main forms of TCMs in clinic, combining several TCMs together to improve therapeutic efficacy and reduce side-effects. Study on their effective constituents has always been a hot topic in TCM research field. Gui-Zhi-Tang originating from Shang-Han-Lun, has been considered as the most important formula. It is comprised of five crude drugs, i.e., Cinnamomi ramulus, Glycyrrhizae radix et rhizoma preparata cum melle, Paeoniae radix alba, Jujubae fructus and Zingiberis rhizoma recens. In this thesis, the chemical constituents in Gui-Zhi-Tang was identified by LC-MS, and the constituents with anti-inflammatory was studied. They were presented as follows:A rapid resolution liquid chromatography-quadrupole time-of-flight mass spectrometry (RRLC-Q-TOF-MS) combined with rapid resolution liquid chromatography-diode array-ion trap mass spectrometry (RRLC-DAD-IT-MS) method was applied to the qualitative analysis of Gui-Zhi-Tang. A total of 187 constituents were detected. By contrast with retention time, exact mass, UV spectra, matching diagnostic ions, and fragmentation patterns,144 compounds were confirmed or assumed from Gui-Zhi-Tang decoction tentatively, among which 31 compounds were confirmed unambiguously by comparing with corresponding authentic standards.Macroporous resin column chromatography method and preparative liquid chromatography method were utilized for dissecting Gui-Zhi-Tang into 64 components. Anti-inflammatory activity of these components was evaluated by determining the production of NO in LPS induced RAW 264.7 cells. As a result,34 components presented anti-inflammatory activity to some extent.34 active components of Gui-Zhi-Tang were analyzed by RRLC-DAD-IT-MS, and the anti-inflammatory activity of 13 pure compounds was verified experimentally. As a result, the NO inhibition rate IC50 of licochalcone A in LPS induced RAW 264.7 cells was (3.9 ± 0.6) μM. At the concentration of 50.0 μM, liquiritin and ononin presented anti-inflammatory activity to some extent, while paeoniflorin and rutin presented anti-inflammatory activity at the concentration of 200.0 μM. This method could reduce the blindness of active consitutuent found to some extent.