Acetylamino Sugars And Sialic Acid Prior To Column Derivatization HPLC-FLD (UV) -MS Analysis

The important roles of saccharides in a variety of organisms have become more recognized, so there are considerable interests in research of carbohydrate over the past recent years. The popular HPLC technique has become an important technology in methodology research of sugar. HPLC and HPLC-MS method has become the most important method for carbohydrate analysis in analytical chemistry. Although there are many HPLC methods for the detection of carbohydrate, most of them were used for common monosaccharide analysis. Sugar derivatives are important compounds with a lot of physiological activities and functional properties, and fascinating insight has recently been focused on them. There are two problems in carbohydrate analysis: lack of sensitive detectors and difficulties of separation. Derivatization was often employed to change structure of sugars, so ultraviolet or the fluorescence examination could be used for HPLC separation. This article has put emphasis on extraction method and detection method for sugar derivatives, and novel HPLC methods for carbohydrate analysis have been established, and also have been applied to analyze biological samples successfully.Chapter one: The present situation of carbohydrate research were summarized, including biological functions of sugar, difficulties in analysis work, the application of HPLC technique in carbohydrate research, the derivatization development in carbohydrate analysis.Chapter two: An efficient and sensitive method of HPLC using 1-(2-naphthyl)-3-methyl-5-pyrazolone as derivatization reagent coupled with UV detection and online MS identification was established for the analysis of common N-acetylated amino sugars. The pivotal parameters affecting the liberation of N-acetylated amino sugars from real samples were investigated with response surface methodology(RSM). The developed method was successfully applied to analyze carbohydrates in Potentilla anserina L. and milk-based products, and exhibited satisfactory applicability.Chapter three: A novel, sensitive and convenient method, utilizing an excellent probe 2-[2-(7H-dibenzo[a,g] carbazol-7-yl)-ethoxy] ethyl carbonylhydrazine(DBCEEC) as pre-column derivatization reagent prior to HPLC coupled with fluorescence detection and online mass spectrometry identification, has been developed for determination of sialic acids. The mass spectrum was used to explain the derivatization principle, and the pivotal parameters affecting the release of sialic acids from serum and the derivatization were investigated with RSM, and changes in the sialic acid content of serum were measured.Chapter four: An archetypical Zirconium-based Metal-organic framework material, UiO-66-NH2, has been synthetized and characterized in a dispersive micro-solid-phase extraction procedure(DSPE)in combination with HPLC with fluorescence detection for the preconcentration and detection of sialic acids in serum samples. Extraction parameters were investigated and properly optimized. The DSPE method has better extraction results than traditional sample preparation methods.