Preparation And Application Of Saccharide Functionalized Polymer Monolithic Column

Saccharide,a natural compound with unique chemical structure and biological function,is widely distributed in nature.Because of its multi-hydroxyl network structure,saccharide was widely applied in the separation of polar compound,chiral compound,glycoprotein,glycopeptide and so on.Till to now,most saccharide functionalized columns are base on silica packed column but with many shortcomings?such as complicated preparation process,narrow range of pH?,which cannot meet the need of separation of biomolecular.Monolith column is novel porous material formed by in-situ polymerization of mixture cosist of monomer,cross-linking,pogrogen and initiator.Thanks to complicated pore crosslinking network,monolith column shows outstanding chromatographic performance and good biocompatibility,making it suitable for separation of biomolecule.In this study,a novel monosaccharide and disaccharide functionalized monolith column were prepared,and used for the rapid separation of samples including biosample and drug.Chapter one,the definition,structure characteristic and chromatographic application of saccharide was firstly introduced.Till to now,saccharide functionalized silica packed column was widely used in each field.Secondly,monolith column was detailedly described involving classification and advantages/disadvantages.At the end,a short review about immobilization methods for saccharide was presented.Chapter two,combined with click reaction,a novel synthesis route for saccharide functional monomer was designed,which was suitable for the modification of linear saccharide.And then,PMA-Glucose and PMA-Maltose was successfully synthesized by this route and identificated by IR,MS,¹H-NMR,¹³C-NMR.Chapter three,PMA-Glucose was used to prepare the poly?PMA-Glucose-co-MBA?monolith.In order to obtain better permeability and column efficiency,the dosage of monomer,cross-linker,porogens were optimized.Then,retention mechanisms,physical-chemical properties of poly?PMA-Glucose-co-MBA?monolith were investigated.The results showed that poly?PMA-Glucose-co-MBA?monolith exhibited good retention of polar compound meaning a stronge hydrophile interaction between stationary phase and analytes.And some polar compounds?such as nucleosides,phennols,phenylformic acids?were successfully separated by this column.Chapter four,PMA-Maltose was used to preparation of the poly?PMA-Maltose-co-MBA?monolith by mixing with MBA?cross-linker?,methanol and water?porogens?,AIBA?initiator?.Then the dosage of monomer,cross-linker,porogens was optimized to obtain good permeability and column efficiency.Retention mechanism,physical-chemical properties of poly?PMA-Maltose-co-MBA?monolith were also investigated.The same as poly?PMA-Glucose-co-MBA?,poly?PMA-Maltose-co-MBA?monolith exhibited better separation of polar compound.Chapter five,the ability of glycopeptide enrichment of poly?PMA-Glucose-co-MBA?monolith and poly?PMA-Maltose-co-MBA?monolith was inverstigated.A total of 31 and 18 glycopeptides were detected from IgG after enriched by poly?PMA-Glucose-co-MBA?monolith and poly?PMA-Maltose-co-MBA?monolith,respectively.Chapter six,summarized this study and briefly described the future application prospects of the saccharide functionalized polymer monolith...