利胆醇在新型多肽键合环糊精硅胶手性柱上分离条件的优化

Optimized Conditions of 1-Phenylpropanol Separation on the New Chiral Stationary Phase of Peptide Bonding β-Cyclodextrin

将胰蛋白酶酶解牛血清白蛋白产生的多肽与环糊精键合,用羰基咪唑法键合到硅胶上,得到新型多肽键合环糊精硅胶手性固定相.反相液相色谱法拆分手性化合物利胆醇,考察了流动相的pH梯度变化时间、离子强度以及流速对利胆醇手性拆分的影响,探讨了该新型多肽键合环糊精硅胶手性柱对利胆醇的手性识别机理及利胆醇在该柱上的手性拆分重复性.实验表明:流动相组成为60 mol/L磷酸盐缓冲液;pH梯度为1 min 5.6,10 min 6.4;流速为0.5 mL/min时分离效果最佳.

A new derivate of β-cyclodextrin with peptides was prepared, where the peptides were obtained from the enzymolysis of bovine serum albumin （BSA） in the presence of trypsin. The derivate was bonded with silica gel to give a novel chiral stationary phase for high performance liquid chromatography （HPLC）. The chiral separations of （ ＋ ）1-phenylpropanol was investigated using an re-phase RP-HPLC system with this column. Several important HPLC conditions were studied including variability of gradient time of pH value of phosphate buffer, ionic intensity, and mobile phase velocity. The chiral recogniztion mechanism and the repetition on chiral separate of 1-phenyl-propanol on the new stationary phase were probed. Experiments shown that the best condition of 1- phenylpropanol on the new chiral stationary phase （CSP） is mobile phase, 60 mmol/L phosphate, gradient pH buffer, 1 min pH 5.6, 10 m in pH 6.4, flow rate, 0.5 mL/m in.