Xuan Hai, Hage David S
Chemistry Department, University of Nebraska, Lincoln, NE 68599-0304, USA.
J Sep Sci. 2006 Jul;29(10):1412-22. doi: 10.1002/jssc.200600051.
The binding and chiral separation of R- and S-propranolol was investigated on a new type of alpha1-acid glycoprotein (AGP) column. This column was prepared through the controlled and mild oxidation of AGP, followed by the immobilization of this protein to hydrazide-activated silica. The effects of temperature, pH, ionic strength, and organic modifiers on the retention and separation of R- and S-propranolol were investigated on this column. Both the association equilibrium constants and number of binding sites for R/S-propranolol on the AGP column were found to increase with temperature and affect the measured retention factors for these compounds. Regarding the other factors, a change in the organic modifier concentration was found to give the largest change in retention and separation. It was found through these studies that both coulombic and hydrophobic interactions played important roles in determining the retention of R- and S-propranolol on the AGP column. The efficiency and separation impedance of this system were also considered. Under the final optimum conditions identified in this study, it was possible to separate R- and S-propranolol with a resolution of greater than 1.38 in less than 5 min on a 4.1 mm I.D. x 5 cm column.
在一种新型的α1-酸性糖蛋白（AGP）柱上研究了R-和S-普萘洛尔的结合及手性分离。该柱是通过对AGP进行可控的温和氧化，然后将这种蛋白质固定到酰肼活化的硅胶上制备而成。在此柱上研究了温度、pH、离子强度和有机改性剂对R-和S-普萘洛尔保留和分离的影响。发现AGP柱上R/S-普萘洛尔的缔合平衡常数和结合位点数均随温度升高而增加，并影响这些化合物的测定保留因子。关于其他因素，发现有机改性剂浓度的变化对保留和分离的影响最大。通过这些研究发现，库仑相互作用和疏水相互作用在决定R-和S-普萘洛尔在AGP柱上的保留方面都起着重要作用。还考虑了该系统的效率和分离阻抗。在本研究确定的最终最佳条件下，在一根内径4.1 mm×5 cm的柱上，不到5分钟就能以大于1.38的分离度分离R-和S-普萘洛尔。
