The Non-aqueous Capillary Electrophoresis And Liquid Chromatography For The Separation Of Amines And Peptide Compounds

Capillary electrophoresis (CE) and high performance liquid chromatography (HPLC) are modern analytical technologies, which all have some advantages of high efficiency, fast analysis, high automaticity and little sample consumption. Being given great concerns by separation and analysis scientists, they have been the most-watched and most advanced technologies in biochemistry and analytical chemistry in last ten years, especially in recent years.As a branch of CE, nonaqueous capillary electrophoresis (NACE) not only has the advantage of CE, but also has its own unique characteristic and the superiority comparing with the aqueous phase capillary. The research scope is also continuously expanding, and has covered the basic research and the analysis application fields related to the separation of inorganic ion, neutral material, the organic acid, the medicine and its metabolite and so on. At present, the physico-chemical properties of very many nonaqueous solutions are not very clear. Moreover, the organic reagent types are various. All of these problems can bring certain difficulty to the choice of reagent. But, they can provide many subjects for the researcher with reference to the choice rule of the solvent system and the optimized method research in future.Amino acids and oligopeptides are important components of life organisms, so the research of their separation by NACE is of great significance on the research in protein, polypeptide, medicine, food, health, and other fields. However, amino acids and oligopeptides have no or weak ultraviolet absorption and the low contents, so they can not be detected directly by UV detection. At present, the widely used derivatization reagents are o-phthalaldehyde (OPA), 9-fluorenylmethyl chloroformate (FMOC), dansyl chloride (DNS), but these reagents have also been reported some various shortcomings in their application, such as short detection wavelengths, poor stability of reagents and serious interference in the determination of biological samples. Aromatic amines have brought great risks in our life because of their toxicity and potential carcinogenicity. So it is very necessary to establish the fast, simple and effective determinations of aromatic amines.The novel labeling reagent has been synthesized and applied to the separation of amino acids and oligopeptides by nonaqueous capillary electrophoresis. The separations of derivatized aromatic amines by high performance liquid chromatography are investigated. The thesis consists of four chapters.Chapter one:The developments of capillary electrophoresis (CE) and its applications in pharmaceutical analysis, biomolecular separation, genetic engineering, clinical medicine and many other fields have been simply introduced. At the same time, the separation strategy of analyte by nonaqueous capillary electrophoresis and the related basic knowledge are reviewed.Chapter two:A simple and rapid method for the separation of amino acids by nonaqueous capillary electrophoresis using 10-ethyl-9-oxo-9,10-dihydroacridine-2-sulfonyl chloride (EASC) as derivatization reagent has been developed. The effect of several key factors on the separation is studied. Under the proposed conditions, a good baseline resolution for 18 derivatized amino acids is achieved.Chapter three:A simple and mild method for the separation of oligopeptides based on a condensation reaction with 10-ethyl-9-oxo-9,10-dihydroacridine-2-sulfonyl chloride (EASC) as labeling reagent with nonaqueous capillary electrophoresis has been developed. The results indicate that five derivatized oligopeptides can be achieved baseline resolution under the proposed conditions.Chapter four:An overview of the development and basic theory about high performance liquid chromatography and HPLC coupled with mass spectrometry technique. Pre-column derivatization methods for the sensitive determination of aromatic amines with 2-[2-(7H-dibenzo [a,g]-carbazol-13-yl)] isopropyl chloroformate (DBCPC-C1) as labeling reagent followed by HPLC-MS analysis have been developed. The optimization of derivatization and chromatographic separation conditions using DBCPC-C1 as labeling reagent are evaluated. Then, the established method has applied to analyze the real samples, the results show that the method has great feasibility.