Analytical Research and Development, MRL, Merck & Co., Inc., Rahway, NJ, 07065, USA.
Analytical Research and Development, MRL, Merck & Co., Inc., Rahway, NJ, 07065, USA.
J Chromatogr A. 2020 Jul 5;1622:460895. doi: 10.1016/j.chroma.2020.460895. Epub 2020 Jan 16.
Baseline separation and analysis of multicomponent mixtures of closely related pharmaceuticals using single column selectivity can often be challenging, requiring the combination of orthogonal stationary and mobile phase methods to monitor all the species and optimize reaction outcomes. In recent years, two-dimensional liquid chromatography (2D-LC) has become a valuable tool for improving peak capacity and selectivity. Though powerful, standard 2D-LC instrumentation and software can often lead to tedious method development and has a requirement for very specific expertise that is poorly suited for a fast-paced industrial environment. In this regard, the introduction of an automated online 2D-LC setup that could screen multiple columns in both dimensions without manual intervention will undeniably serve to streamline column/mobile phase selection and secure the viability of 2D-LC as a mainstay instrument for industrial applications. Herein, we introduce and investigate a multicolumn online 2D-LC approach that simplifies column screening and method development dramatically. This setup incorporates 6-position column selection valve technology whose functionality enables us to combine multiple columns in the first and second dimensions. This strategy in conjunction with diode array detection (DAD) in both dimensions and mass spectrometry (MS) acquisition in the second dimension serves to explore different columns and mobile phases as a framework for screening targeted compounds in multicomponent mixtures without having to perform chromatographic purification. Multiple online heart cutting achiral RPLC - achiral RPLC and achiral RPLC - chiral RPLC coupled to DAD and ESI-MS methods combining several stationary phase selectivity in an automated fashion are successfully applied to the separation and analysis of complex mixtures of drug substances, where in many instances, traditional 1D-ultra-high performance liquid chromatography (UHPLC) fails or delivers sub-optimal results. This automated online multicolumn 2D-LC workflow enables rapid and efficient identification of column/eluent combinations, as well as sample analysis across multiple columns in both dimensions overnight with a single click.
基线分离和分析密切相关的多组分药物混合物，使用单一柱选择性通常是具有挑战性的，需要正交固定相和流动相方法的组合来监测所有的物种和优化反应结果。近年来，二维液相色谱（2D-LC）已成为提高峰容量和选择性的有效工具。虽然功能强大，但标准的 2D-LC 仪器和软件往往会导致繁琐的方法开发，并且需要非常特定的专业知识，这对于快节奏的工业环境来说是不适合的。在这方面，引入一种自动化的在线 2D-LC 装置，可以在无需人工干预的情况下在两个维度上筛选多个柱子，这无疑将简化柱/流动相的选择，并确保 2D-LC 作为工业应用的主要仪器的可行性。在此，我们介绍并研究了一种多柱在线 2D-LC 方法，该方法大大简化了柱筛选和方法开发。该装置采用 6 位柱选择阀技术，其功能使我们能够在第一维和第二维中将多个柱子组合在一起。这种策略与二维中的二极管阵列检测（DAD）和二维中的质谱（MS）采集相结合，用于在不进行色谱纯化的情况下筛选多组分混合物中的目标化合物，作为探索不同柱子和流动相的框架。多个在线心脏切割手性 RPLC-非手性 RPLC 和非手性 RPLC-手性 RPLC 与 DAD 和 ESI-MS 方法相结合，以自动化的方式组合了几种固定相选择性，成功地应用于药物混合物的复杂混合物的分离和分析，在许多情况下，传统的一维-超高效液相色谱（UHPLC）失败或提供次优结果。这种自动化的在线多柱 2D-LC 工作流程能够快速有效地识别柱/洗脱液组合，以及在两个维度上的多个柱子上进行样品分析，只需点击一次即可在一夜之间完成。
