Drug Quality and Registration Group, Faculty of Pharmaceutical Sciences, Ghent University, Ghent, Belgium.
J Sep Sci. 2010 Mar;33(6-7):728-39. doi: 10.1002/jssc.200900476.
A wide variety of hydrophilic interaction chromatography (HILIC) stationary phase surface chemistries are currently available. Although their selectivity can be considerably different, column comparison or clustering using peptides is limited. In this study, ten pharmaceutically relevant model peptides are analyzed on seven different HILIC columns (bare silica, amide, poly-hydroxyethyl aspartamide, diol and zwitterionic) for the evaluation of their performance and classification. The responses examined include single and multiple responses: plate number, asymmetry factor, LOD, geometric mean resolution, resolution product, time corrected resolution product, peak capacity and chromatographic response function. Column classification was performed using hierarchical clustering and principal component analysis. Moreover, the overall performance quality of the HILIC columns was compared using a linear desirability function. Hierarchical clustering and principal component analysis showed consistent clusters. The zwitterionic phase was clustered apart from the other HILIC columns and both poly-aspartamide columns were clustered together. In addition, the two bare silica phases represent two different clusters, and thus different selectivities. Overall, the responses showed the best performance for one of the bare silica columns (Alltima-Alltech), followed by the zwitterionic phase (ZIC)-HILIC. Thus, these columns, belonging to different clusters, were found to be the best performing systems in pharmaceutical peptide analysis for the selected peptide set.
目前有多种亲水相互作用色谱（HILIC）固定相表面化学可供选择。尽管它们的选择性可能有很大的不同，但使用肽进行柱比较或聚类受到限制。在这项研究中，对十种与药物相关的模型肽在七种不同的 HILIC 柱（裸硅胶、酰胺、聚羟乙基天冬酰胺、二醇和两性离子）上进行分析，以评估它们的性能和分类。考察的响应包括单一和多个响应：塔板数、不对称因子、LOD、几何平均分辨率、分辨率产物、时间校正分辨率产物、峰容量和色谱响应函数。使用层次聚类和主成分分析对柱进行分类。此外，还使用线性适宜度函数比较了 HILIC 柱的整体性能质量。层次聚类和主成分分析显示出一致的聚类。两性离子相与其他 HILIC 柱分离，两个聚天冬酰胺柱聚集在一起。此外，两个裸硅胶相代表两个不同的聚类，因此具有不同的选择性。总的来说，这些响应表明，在所选择的肽组中，其中一个裸硅胶柱（Alltima-Alltech）和两性离子相（ZIC）-HILIC 的性能最佳。因此，这些属于不同聚类的柱被发现是药物肽分析中表现最好的系统。
