Research On The Application Of LC-MS And NMR In Qualitation Of Chemical Constituents In Zhi-Zi-Chi-Tang Analogous Formulae

The modernization study of formula is an important part of TCM’s modernization.Moreover,the studies on the chemical constituents of TCM formula is the basis of formulae’s modernization research.Zhong-Jing-Fang is regarded as a main object for the modernization of formula by its rigorous compatibility and excellent therapeutic effect.Zhishi-Zhizi-Chi-Tang,Zhizi-Gancao-Chi-Tang,and Zhizi-Baipi-Tang are three TCM formulae from Shang-Han-Lun.However,the constituents in the three formulae have been rarely reported.In this work,these three formulae were selected as the research object,and the main topics of this thesis are as follow:1.The LC-IT-MS and LC-Q-TOF-MS methods were applied to the qualitative analysis of ZZGCCT and ZZBPT,a strategy based on semi-preparative LC guided by LC-MS was used to prepare the target isomers in microscale for the following off-line NMR analysis,including 1D-NMR and 2D-NMR.A total number of 148 constituents were detected,and 119 ones were identified tentatively or unambiguously.Among them,20 constituents were characterized unambiguously by comparing with the reference substances.In addition,21 constituents without reference substances were prepared and their structures were unambiguously identified by MS,1D-NMR,and 2D-NMR(including three novel compounds:two triterpenoid glycosides and one flavonoid glycoside).Among them,compounds 122,125,and 129,as well as 131,136,and 140,134 and 145,48 and 50,62a,62b,93,and 97,were several groups of isomers.The fragmentation patterns of alkaloids and phenolic acid derivatives of genipin gentiobioside detected in the two formulae were summarized.2.The qualitation of chemical constituents in ZSZZCT was first carried out by LC-IT-MS and LC-TOF-MS.A total of 79 chromatographic peaks were detected,and 39 compounds were confirmed or assumed,including 8 iridoid glycosides,5 pyronane monoterpenoid glycosides,17 flavonoid glycosides,2 organic acids,2 cyclic peptides,1 alkaloid and four other categories of compounds.The source from which crude drug of all the constituents were assigned according to the LC-IT-MS data.The strategy proposed in the second chapter was used in prepareing three polymethoxy flavonoids.