Preparation Of Chromatographic Stationary Phases With Special Morphology And Function

With special physical and chemical characterization, nanoparticles have showed great potential in preparation of chromatographic stationary phase. To accomplish the separation of complex or special samples such as proteomics sample and chiral drugs, the development of special surface functionalized stationary phases has become focus of research in analytical chemistry and material science recently.Nanoparticles with different shapes were utilized in preparation of special functionalized nano chromatographic stationary phase. CEC column packed by SBA-15rod was successfully prepared, because the special shape of the rod SBA-15can form certain framework during packing procedure. To take both advantages of monolith and nanoparticles, different kinds of nanoparticles were embedded into the matrix of monolith. The prepared nanoparticles incorporated hybrid monolithic columns have high column efficiency up to290000plates/m, because of the large surface area of nanoparticles. Ion exchange (IE) and reverse phase (RP) separation mechanisms offered by aminopropyl-functionalized nanoparticles and butyl methacrylate (BMA) monomers were combined by incorporation. So the hybrid monolithic column has RP/IE mixing separation mechanism and showed great potential in separation of natural complex products, such as Chinese medicine.Kinds of macrocyclic antibiotics bonded chiral stationary phases (CSP) have been prepared by surface modification of macrocyclic glycopeptide antibiotics. Macrocyclic antibiotics can be directly immobilized by reaction of amino group and epoxy group on the surface of stationary phase or indirectly immobilized to the stationary phase with amino-group using glutaraldehyde as spacer. The prepared macrocyclic antibiotics boned CSPs showed satisfactory ability in chiral separation. Based on the high activity of epoxy group of glycidyl mehtacrylate monolith, the novel eremomycin boned CSP was prepared by a one-step derivatization process. Five chiral amino acids and a chiral drug were baseline separated. The eremomycin CSP showed great ability for separating the amino acid containing phenyl group. Meanwhile, vancomycin was bonded to silica particles by indirectly modification. Five racemic compounds have been separated by the prepared vancomycin CSP in HPLC mode. Investigation of separation conditions showed that the retention behavior and chiral selectivity of racemic compounds were significantly affected by organic modifer. concentration and pH of buffers and temperature.To develop the stationary phase with high selectivity in protein recognition, molecular imprinting technology was used in preparation of protein imprinted particles. First, surface-imprinted silica beads were prepared based on metal coordination for the separation of protein. Because the imprinted cavities were located on the surface, the recognition sites were accessible for the template proteins, so the adsorption of target protein reached equilibrium within30min. Second, based on complex structure of protein,"soft" and flexible recognition sites that can allow "induced fit" of the target proteins were created by a novel surface imprinting technique. Compared to classic protein imprinting, the surface imprinted silica with "soft" recognition sites has better selectivity in recognition of target protein. Finally, exploratory research was carried out in preparation of protein molecular imprinted colloid array (MICA).