Takeda San Diego, Inc., San Diego, CA 92121, USA.
J Chromatogr A. 2011 May 20;1218(20):3080-8. doi: 10.1016/j.chroma.2011.03.041. Epub 2011 Apr 12.
A new analytical two-dimensional supercritical fluid chromatography/mass spectrometry system (2D SFC/SFC/MS) has been designed and implemented to enhance the efficiency and quality of analytical support in drug discovery. The system consists of a Berger analytical SFC pump and a modifier pump, a Waters ZQ 2000 mass spectrometer, a set of switching valves, and a custom software program. The system integrates achiral and chiral separations into a single run to perform enantiomeric analysis and separation of a racemic compound from a complex mixture without prior clean up. The achiral chromatography in the first dimension separates the racemate from all other impurities, such as un-reacted starting materials and by-products. Mass-triggered fractionation is used to selectively fractionate the targeted racemic compound based on its molecular weight. The purified racemate from the achiral chromatography in the first dimension is then transferred to the chiral column in the second dimension to conduct the enantiomeric separation and analysis. A control software program, we coined SFC2D, was developed and integrated with MassLynx to retrieve acquisition status, current sample information, and real time mass spectrometric data as they are acquired. The SFC2D program also monitors the target ion signal to carry out mass-triggered fractionation by switching the valve to fractionate the desired peak. The 2D SFC/SFC/MS system uses one CO(2) pump and one modifier pump for both first and second dimension chromatographic separations using either gradient or isocratic elution. Similarly, a preparative 2D SFC/SFC/MS system has been constructed by modifying an existing Waters preparative LC/MS system. All components except the back pressure regulator are from the original LC/MS system. Applications of the 2D SFC/SFC/MS methods to the separation and the analysis of racemic pharmaceutical samples in complex mixtures demonstrated that an achiral separation (in first dimension) and a chiral separation (in second dimension) can be successfully combined into a single, streamlined process both in analytical and preparative scale.
一个新的分析二维超临界流体色谱/质谱系统（2D SFC/SFC/MS）已经被设计和实现，以提高药物发现中的分析支持的效率和质量。该系统由 Berger 分析型 SFC 泵和改性剂泵、Waters ZQ 2000 质谱仪、一组切换阀和一个定制软件程序组成。该系统将非手性和手性分离集成到一个单一的运行中，以进行对映异构体分析和从复杂混合物中分离外消旋化合物，而无需预先进行净化。第一维的非手性色谱将外消旋体与所有其他杂质（如未反应的起始原料和副产物）分离。质量触发的馏分收集用于根据分子量选择性地分离目标外消旋化合物。从第一维非手性色谱中纯化的外消旋体然后转移到第二维手性柱中进行对映异构体分离和分析。开发并集成了一个控制软件程序 SFC2D，与 MassLynx 一起检索采集状态、当前样品信息和实时质谱数据，以便在采集时获取。SFC2D 程序还监测目标离子信号，通过切换阀来实现质量触发的馏分收集，以分离所需的峰。2D SFC/SFC/MS 系统使用一个 CO2 泵和一个改性剂泵用于第一维和第二维色谱分离，采用梯度或等度洗脱。同样，通过修改现有的 Waters 制备型 LC/MS 系统构建了一个制备型 2D SFC/SFC/MS 系统。除了背压调节器之外，所有组件都来自原始的 LC/MS 系统。将 2D SFC/SFC/MS 方法应用于复杂混合物中外消旋药物样品的分离和分析表明，非手性分离（在第一维）和手性分离（在第二维）可以成功地组合成一个单一的、流线型的过程，无论是在分析还是制备规模上。
