Synthesis And Application Of Amino Acid Chiral Ligand-exchange Chromatography Stationary Phase

Through the study of life sciences, people found the enantiomers have different physiological and pharmacological activity in a living body. The significance of chiral separation is to explore pharmacological and toxicological activity of drugs, and to prepare drugs of low toxicity and low side-effect. In the current the High Performance Liquid Chromatography(HPLC) has become the most effective way to separate chiral Drugs. The chiral ligand exchange chromatography stationary phase is the most effective means of separating underivatized amino acids, and is used widely for its environmental protection and better economy.In this article, two new bonded chiral ligand-exchange chromatography stationary phases of L-phenylalanine chiral ligand-exchange chromatography stationary phase(CSP-1) and L-proline chiral ligand-exchange chromatography stationary phase(CSP-2) had been synthesized by using 3-aminopropyl triethoxysilane reagent and trigonal cyanuric chloride as the link arm, and using silica gel as the carrier. We also got specific conditions of synthesizing the two chiral stationary phase. Then the chiral stationary phases were characterized by elemental analysis, thermal analysis and infrared analysis, and the experimental data showed the new stationary phase of CSP-1 and CSP-2 is success to synthesis.The CSP-1 has separated five pairs of enantiomers: DL-Malic acid, DL-Isoleucine, DL-Tryptophan, DL-Valine and DL-Chlorpheniramine. The effect of resolution on the type and concentration of the metal ions and the buffer solution, flow rate and p H of the mobile phase were systematically investigated in the two stationary phase that have already synthesized. Through the experiments we obtained the central metal ion of copper ions and phosphate buffer solution is the best choice, and the preferable concentration of phosphate buffer solution is 0.02mol/L. But when separating different enantiomers, the flow rates of mobile phase are different. We obtained the p H has effects on enantioseparation by studying the amino acids of isoleucine. Based on the experience of separation of CSP-1, the effect of separating chlorpheniramine was systematically investigated by using CSP-2. The better conditions of separetion of CSP-2: the concentration of copper ion is 0.7mmol/L, the concentration of buffer concentration is 0.01mol/L, the p H of mobile phase is 7.0 and the temperature of column is 35 ℃. Under these conditions, the separation of chlorpheniramine is R = 2.85, and is higher than the standard resolution.In the two stationary phases, CSP-2 is better than CSP-1 on enantioseparation of chlorpheniramine, but in the number of separeting enantiomers the CSP-1 is superior the CSP-2.