Discovery Chemistry, Merck & Co., Inc., 213. E. Grand Avenue, South San Francisco, California 94080, United States.
Discovery Chemistry, Merck & Co., Inc., 770 Sumneytown Pike, West Point, Pennsylvania 19486, United States.
Anal Chem. 2022 Jun 14;94(23):8309-8316. doi: 10.1021/acs.analchem.2c00750. Epub 2022 Jun 3.
The pharmaceutical industry is increasingly faced with challenging separations of complex crude reaction mixtures at the microscale that require the adoption of new platforms for rapid target isolation, impurity determination, and quantitation. In this study, we describe an online microscale one- or two-dimensional liquid chromatography (1D/2D-LC) system with heart-cutting and multi (triple) detector triggering in either dimension to address this need. The advantages of charged aerosol detection (CAD) are discussed for the direct quantitation of limited quantity samples, without utilizing a second analytical instrument or gradient compensation pump. In addition to the significant time and cost savings, there is no minimum recovery requirement that exists when compared to gravimetric methods for accurate microscale quantitation. This platform has been successfully used to purify 0.5-5.0 mg scale reactions in 96- or 384-well reaction plates with a gradient time of 4 min per sample. Separations performed in both dimensions are complete in less than 12 min, including trapping and column equilibration time. The isolated arrays of small-quantity investigational compounds at a high purity enable rapid exploration of chemical reaction parameters and synthetic route scouting for biological target validation.
制药行业越来越多地面临着在微尺度下分离复杂的粗反应混合物的挑战，这需要采用新的平台来快速实现目标分离、杂质检测和定量分析。在本研究中，我们描述了一种在线微尺度一维/二维液相色谱（1D/2D-LC）系统，该系统具有切分和多维（三重）检测器触发功能，可满足这一需求。讨论了带电气溶胶检测（CAD）的优势，用于直接定量有限量的样品，而无需使用第二台分析仪器或梯度补偿泵。与重量法相比，该方法不仅显著节省了时间和成本，而且不存在准确微尺度定量所需的最小回收要求。该平台已成功用于在 96 孔或 384 孔反应板中以 4 分钟/样品的梯度时间纯化 0.5-5.0 毫克规模的反应。在不到 12 分钟的时间内完成两个维度的分离，包括捕获和柱平衡时间。高纯度的小批量研究化合物的分离阵列能够快速探索化学反应参数和合成路线，以验证生物靶标。
