Cox G B
Prochrom Inc., Indianapolis, IN 46278.
J Chromatogr. 1992 May 22;599(1-2):195-203. doi: 10.1016/0021-9673(92)85473-7.
The mass-overloaded separation of bovine and porcine insulins has been studied in the reversed-phase gradient elution mode. Strong solute-solute displacement effects have been found, which are related to the efficiency of the column used. Low flow-rates and small particle diameters maximise the displacements, as well as improve the resolution between the parent insulin and its desamido contaminant. The gradient slope did not substantially affect the separation between the parent insulins, but, due to the relative "S" values of the solutes, an increase in gradient slope improved the separation of the parent insulin from its desamido compound. An optimum pore size of 150 A was found for the insulins. Experiments to optimise the loadability were not carried out, but a recovery of 90% at a purity of 99.5% was obtained with a loading of 12 mg/g of porcine insulin.
已在反相梯度洗脱模式下研究了牛胰岛素和猪胰岛素的质量过载分离。发现了强烈的溶质-溶质置换效应，这与所用色谱柱的效率有关。低流速和小粒径可使置换最大化，并提高母体胰岛素与其脱酰胺杂质之间的分离度。梯度斜率对母体胰岛素之间的分离影响不大，但由于溶质的相对“S”值，梯度斜率的增加改善了母体胰岛素与其脱酰胺化合物的分离。发现胰岛素的最佳孔径为150埃。未进行优化负载量的实验，但在猪胰岛素负载量为12 mg/g时，获得了纯度为99.5%时90%的回收率。
