| Liquid chromatography-mass spectrometry(LC-MS)features both the high-performance separation ability of liquid chromatography and the qualitative superiority of mass spectrometry.In particular,the LC-MS technology with API interface has been widely applied in many fields,such as medicines,biochemistry and environmental chemistry.In this thesis,ultrahigh performance liquid chromatography,electrospray ionization mass spectrometry,high resolution time-of-flight mass spectrometry and tandem mass spectrometry were applied to study the methods for the components analysis of traditional Chinese medicines and the recognition of the characteristic components of indole biotransformation.Firstly,high performance liquid chromatography-time-of-flight mass spectrometry(HPLC-TOF/MS)was used to detect the ethanol-water extracts from the roots of two stellera chamaejasme(Euphorbia Fisheriana and Euphorbia ebracteolata Hayata).Different HPLC separation conditions were optimized.The components were then qualitatively analyzed by mass spectrometry correctly.Specifically,fifty-five components in Euphorbia Fisheriana and eighty-three components in Euphorbia ebracteolata Hayata were analyzed.The main components in both Euphorbia Fisheriana and Euphorbia ebracteolata Hayata were identified as diterpene lactones which were the primary components causing stellera chamaejasme's anti-tumor efficacy.Secondly,a rapid qualitative method based on HPLC-TOF/MS and a quantitative method based on HPLC-MS/MS were established respectively to analyse the complex components of traditional Chinese medicines.Using these two mothods,the main components in Yangxinshi tablets and Chanfukang gragules were succefully analyzed qualitatively and quantitatively.Specifically,a total of thirty compounds in Yangxinshi tablets and twenty-four compounds in Chanfukang gragules were obtained,and sixteen chemical constituents in Yangxinshi tablets and thirteen chemical constituents in Chanfukang gragules were identified and quantified without any deviation from using the existing standard reference substances.These two novel analytical methods can be used for simultaneous qualitative and quantitative analysis of multi-components,providing technical support for quality control and pharmacological research of traditional Chinese medicines.Thirdly,a rapid quantitative method based on HPLC-MS/MS was established to study the pharmacokietics of Chanfukang granules in rats.The pharmacokinetic differences were compared between stachydrine and leondrine in single administration and in compounds of Chanfukang granules,respectively.The experimental results indicate that the well-known compatibility law of compound prescription of traditional Chinese medicines,i.e."reducing toxin and increasing efficiency",can be successfully explailed through the viewpoint of pharmacokinetics.The bioavailability of stachydrine and leondrine in the form of Chanfukang granules was higher than that of single component,which was more beneficial to the exertion of pharmacological activity.Finally,a method basd on HPLC-TOF/MS was established to analyze the metabolites during the biotransformation of indole induced to produce indigoids by the phenol-degrading strains PI1 and PI2.The results indicated that both strains could catalyze the formation of four indigoids with the same prominent molecular ion peak at m/z 261.0670 and a molecular formula of C16H10N2O2,including indigo and a purple product,2-(7-oxo-lH-indol-6(7H)-ylidene)indolin-3-one.Isatin and 7-hydroxyindole were also detected as the intermediates.Thus,the results indicated possible pathways for the production of indigoids from indole.Moreover,the optimal conditions for the production of indigo from indole were determined using response surface methodology.Maximum of 11.82 ± 0.30 mg/L and 17.19 ± 0.49 mg/L indigo were achieved by strain PI1 and PI2,respectively. |
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