Study On The Analytical Methods Of Active Components In Herbal Medicine And Biological Samples And Their Applications

The modern analytical instruments, especially the development of gas chromatograph-mass spectrometer (GC-MS), high performance liquid chromatograph-diode array detection (HPLC-DAD), high performance liquid chromatograph-mass spectrometer (HPLC-MS), provide us much useful information. With the help of new analytical theory and technique, the combination of chromatographic separation and spectral or mass spectrum characterization provides us powerful tool for the resolution of analytical problems on biology, biochemistry, pharmacology and environmental science. The recent developments of high-dimensional chemometrics resolution methods provide a powerful tool for us to deal with the difficult "black system". Therefore, the aim of the thesis is to develop new analytical methods for the analysis of active components in the complex systems, such as herbal medicine and biological samples, with GC-MS and LC-MS hyphenated chromatography in mass spectra data mining and hyphenated chromatography.1. Application of hyphenated chromatographic resolution methods to herbal medicine systems (chapter 2 to chapter 3). With chemometrics resolution methods and relative chemometrics methods, the active components in herbal medicine can be identified and quantified. By use of chromatographic and spectral or mass spectrum information, fingerprint of herbal medicine can be obtained. In chapter 2, Constituents of essential oil in main root of Angelica sinensis were identified and quantified by gaschromatography-mass spectrometry (GC-MS) combined with subwindow factor analysis (SFA) and the related chemometric methods. Constituents of essential oil in two different parts of Angelica sinensis, main root and root fibre, were also compared with spectrum-oriented correlative chromatography (SCC) and related chemometric methods. In chapter 3, HPLC flavonoid fingerprints of methanol extract of Astragali Radix from four different resources were studied with HPLC-DAD and LC/ESI-MS.2. Qualitative analysis and quantitative analysis of active components in herbal medicine with hyphenated HPLC-DAD and LC/ESI-MS methods (chapter 4). The research, witch looks for guiding compounds for the possible new medicines, is very important. Because the separation process for active components from pharmaceutical plants and sea is very time-consuming with the traditional methods, it is very important to develop high effective, fast, sensitive and selective methods for the analysis of active components in natural plants. In chapter 4, a method for identification and determination of nucleosides in Cordyceps sinensis and its substitutes by high performance liquid chromatography with photodiode array detection and mass spectrometric detection was developed. Adenine, hypoxanthine, adenosine and cordycepin are the main nucleoside components in Cordyceps sinensis and its substitutes, so it is necessary to develop a useful method for analysis of above components. A simple and rapid isocratic LC/MS coupled with electrospray ionization (ESI) method for simultaneous separation and determination ofadenine, hypoxanthine, adenosine and cordycepin in Cordyceps sinensis (Cs) and its substitutes was also developed.3. Qualitative analysis and quantitative analysis of active components in biological samples with hyphenated HPLC-DAD and LC-MS methods (chapter 5 to chapter 7). The development of biotic science can't be achieved without the help of modern analytical technique. Thus, new problems are aroused for biotic scientist and analyst. Because there are no proper methods, much work on foundational and clinic research can't be done. It is imperative that the advanced, reliable, sensitive, accurate, fast analytical methods for the determination of active components in biological samples be developed. These methods can be used for qualitative analysis and quantitative analysis of low concentration of drug and its study on pharmacokinetics and bioequivalent character. In chapter 5, a simple, sensitive and fast isocratic high-performance liquid chromatography -mass spectrometry method coupling with an atmospheric pressure chemical ionization (APCI) was developed for simultaneous separation and determination of L-arginine (ARG), N°, N°-dimethylarginine (ADMA) and N°, N"0- dimethylarginine (SDMA) in biological samples. The method was successfully used for the determination of ARG, ADMA and SDMA in human urine and plasma with satisfactory result. In chapter 6, a sensitive, fast, accurate and selective liquid chromatographic-electrospray ionization mass spectrometric method was developed and validated for the determination of finasteride in human plasma.The method was used to estimate the pharmacokinetics of finasteride after oral administration of a 5mg tablet of finasteride to 24 healthy volunteers. In chapter 7, a simple, sensitive and fast reversed-phase high-performance liquid chromatography-mass spectrometry coupling with an electrospray ionization (ESI) interface method was described for the quantitative determination of adenosine in human synovial fluid. The method was applied to determination of adenosine in some synovial fluids of patients affected by rheumatoid arthritis and for study on correlation between adenosine and patients affected by rheumatoid arthritis.