Small Peptides, And Carbohydrates By Capillary Electrophoresis Separation

Capillary electrophoresis(CE) is powerful separation technology,which has been rapidly developed on the basis of traditional separation and chromatography.It has many advantages including high performance,high speed and small sample consumption,and can be used to separate many compounds such as amino acids,peptides,proteins,and even whole cells and virus particles.In addition,CE has many varieties of flexible separation modes,versatile separation buffers and low consumption of buffers.As a branch of CE,nonaqueous capillary electrophoresis(NACE) not noly 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 polypeptide,the polysaccharid,the medicine and its metabolite and so on.At present,however,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 reseacher with reference to the choice rule of the solvent system and the optimized method research in future.Carbohydrate is a kind of substance which widely exists in nature and in organism.Over a long period of time,it has been considered that the main function of carbohydrate is to provide energy.But the newly emerging research on carbohydrate biochemistry shows that it has many other biological activities and functions besides energy-providing.Polypeptide and carbohydrates are important components of life organism,So that their have great value in biology,medicine,food,health and other fields to do research on them by Capillary Eleetrophoresis.Chemical derivation has been developed in the separation of polypeptide and carbohydrates, as these biological molecules have no or weak ultraviolet absorption and the low contents cannot be detected directly by UV detection.In this thesis,new labeling reagents have been synthesized and applied to the separation of oligopeptides and carbohydrates by capillary electrophoresis, the results are satisfactory.The thesis consists of three chapters.Chapter OneThe developments of capillary electrophoresis(CE) and nonaqueous capillary electrophoresis(NACE) and the applications in life science,medical drugs,environmental science and many other fields have been simply introduced.At the same time,the separation strategy of analyte by capillary electrophoresis and the related basic knowledge are reviewed,too.Chapter TwoThe separations of oligopeptides by NACE were investigated.The principal contents of this chapter consist of two parts.2.1 A simple and rapid method for the separation of derivatized oligopeptides using CEOC as derivatization reagent by NACE has been developed.With the mixed methanol and acetonitrile as the elution solvent,a satisfactory separation for the 4 oligopeptides derivatives could be obtained when ammonium acetate and acetic acid were used as the electrolyte.The effects of several key factors such as the proportion between MeOH and ACN,the concentrations of ammonium acetate and acetic acid on separation were also evaluated in this experimentation.2.2 A simple and rapid method for the separation of derivatized oligopeptides using 2-(11H-benzo[a]carbazol-11-yl)ethyl carbonochloridate(BCEC-C1) as derivatization reagent by non-aqueous capillary electrophoresis has been proposed.The separation conditions have been studied and optimized,the optimum conditions are as follows: 0.7mol/L acetic acid,60 mmol/L ammonium acetate buffer,voltage 30kV,column temperature at 20℃and DAD detection at 279 nm.The samples are introduced atmospherically with an injection time of 8 s.The results indicate that 5 derivatized oligopeptides achieve baseline resolution under the proposed conditions.Chapter ThreeA simple and rapid method for the separation of derivatized carbohydrates using 1-Aminoanthracene as derivatization reagent by capillary zone electrophoresis(CZE) has been developed.Under the CZE mode,the separation conditions have been studied and optimized.In optimal conditions,9 monosaccharide derivatives are separated within 23 min. The proposed method has been applied to the determination of monosaccharides in Nitraria samples with satisfactory results.