Chloupek R C, Hancock W S, Snyder L R
Genentech, Inc., South San Francisco, CA 94080.
J Chromatogr. 1992 Mar 6;594(1-2):65-73. doi: 10.1016/0021-9673(92)80313-j.
Computer simulation was used to optimize the separation of a tryptic digest of recombinant human growth hormone using reversed-phase high-performance liquid chromatography in a gradient mode. DryLab G/plus software modelled the retention behavior of the complex tryptic digest mixture as a function of gradient conditions, based on data from two experimental gradient runs. The theoretical optimum separation conditions were rapidly obtained and reproduced experimentally. Resolution did not simply increase as gradient steepness was decreased, rather, an intermediate gradient time provided maximum sample resolution. The simulation results also indicate that the method is reasonably rugged, with little change in the separation expected for different high-performance liquid chromatography systems, and changes in the separation can be compensated by a change in the gradient steepness. Computer simulation can also be useful to quickly reoptimize conditions for a new column, if it fails to provide the same separation.
采用计算机模拟方法，以梯度模式通过反相高效液相色谱对重组人生长激素的胰蛋白酶消化产物进行分离优化。DryLab G/plus软件基于两次实验梯度运行的数据，将复杂的胰蛋白酶消化混合物的保留行为模拟为梯度条件的函数。快速获得了理论上的最佳分离条件并通过实验进行了重现。分辨率并非简单地随着梯度陡度的降低而增加，相反，中等梯度时间可提供最大的样品分辨率。模拟结果还表明该方法具有较好的耐用性，不同的高效液相色谱系统预期分离变化很小，并且分离变化可通过梯度陡度的改变来补偿。如果新色谱柱无法提供相同的分离效果，计算机模拟对于快速重新优化其条件也很有用。
