Preparation And Evaluation Of Poly (L-lysine) Stationary Phase In HPLC

High performance liquid chromatography analysis has a wide range of applications. More than seventy percent of organic compounds can be analysized by high performance liquid chromatography. Especially for high boiling point molecules, polar or poor thermal stability of compounds it shows a great advantage. High performance liquid chromatography played a huge role in food and drug inspection, industrial wastewater and agricultural production, environmental pollutants. In order to expand the detection scope of chromatography analysis, people do the research in many ways:firstly, develop new sample pretreatment methods, such as solid phase extraction, solid phase micro-extraction method; secondly, develop new chromatographic separation modes. Recently hydrophilic interaction Chromatography has been concerned; thirdly, prepare new stationary phase.90% of the stationary phase in HPLC is bonded phase silica gel. People separate different compounds through bonding different functional groups to the silica gel, such as chiral stationary phases for the analysis of chiral compounds, hydrophilic stationary phase for separation of polar compounds.This work focus on resolving the problem that hydrophilic compound is difficult to be separated on RP-C18 column. A novel poly lysine chemically bonded silica stationary phase was sunthesized. It is found that the new phase can be used under both reversed phase chromatography and hydrophilic interaction chromatography mode. Both polar and weakly polar substances can be separated in mixed-mode chromatography. The following aspects included:1. Lysine monomers was changed to lysine anhydride monomer, which bonded to amino modified silica surface and the poly-lysine bonded silica stationary phase is obtained. The effect of pH, buffer concentration and the volume of acetonitrile in the mobile phase on the retention of the test compounds on the new stationary phase were studied. We came to a conclusion:the reaction mechanism between the test compounds and stationary phase is a mixed mechanism, including hydrophobic interaction mechanism, ion exchange, hydrogen bonding interaction and so on. Aniline compounds, phenol compounds, salbutamol sulfate and clenbuterol were separated well.2. Gynostemma bonded-, Polydextrose bonded-, poly (1-lysine) bonded-and pure silica stationary phase were evaluated for per aqueous liquid chromatography.