Molnár-Institute for Applied Chromatography, R&D, Schneegloeckchenstr 47, Berlin 10407, Germany.
J Chromatogr A. 2010 May 7;1217(19):3193-200. doi: 10.1016/j.chroma.2010.02.001. Epub 2010 Feb 6.
The present paper describes a multifactorial optimization of 4 critical HPLC method parameters, i.e. gradient time (t(G)), temperature (T), pH and ternary composition (B(1):B(2)) based on 36 experiments. The effect of these experimental variables on critical resolution and selectivity was carried out in such a way as to systematically vary all four factors simultaneously. The basic element is a gradient time-temperature (t(G)-T) plane, which is repeated at three different pH's of the eluent A and at three different ternary compositions of eluent B between methanol and acetonitrile. The so-defined volume enables the investigation of the critical resolution for a part of the Design Space of a given sample. Further improvement of the analysis time, with conservation of the previously optimized selectivity, was possible by reducing the gradient time and increasing the flow rate. Multidimensional robust regions were successfully defined and graphically depicted.
本文描述了一种基于 36 次实验对 4 个关键 HPLC 方法参数（梯度时间（t(G)）、温度（T）、pH 值和三元组成（B(1):B(2)））的多因素优化。通过系统地同时改变这四个因素，研究了这些实验变量对关键分辨率和选择性的影响。基本要素是梯度时间-温度（t(G)-T）平面，在洗脱液 A 的三个不同 pH 值和洗脱液 B 中甲醇和乙腈的三个不同三元组成之间重复该平面。这样定义的体积可以用于研究给定样品设计空间的一部分的关键分辨率。通过减少梯度时间和增加流速，可以在保持先前优化的选择性的同时，进一步缩短分析时间。成功地定义并图形化表示了多维鲁棒区域。
