方案摘要
方案下载应用领域 | 制药/生物制药 |
检测样本 | 其他 |
检测项目 | |
参考标准 | 暂无 |
Purifying a large number of compounds by liquid chromatography usually requires screening experiments and manual interaction during upscaling to preparative conditions. The Agilent Automated Purification software facilitates the workflow of analytical scouting to preparative purification by smart algorithms and calculated scale-up of separation gradients, all integrated in an intuitive interface. This Technical Overview demonstrates the purification of a compound library using the Agilent 1260 Infinity Automated LC/MSD Purification System controlled by the Automated Purification software.
Abstract
Purifying a large number of compounds by liquid chromatography usually requires screening experiments and manual interaction during upscaling to preparative conditions. The Agilent Automated Purification software facilitates the workflow of analytical scouting to preparative purification by smart algorithms and calculated scale-up of separation gradients, all integrated in an intuitive interface. This Technical Overview demonstrates the purification of a compound library using the Agilent 1260 Infinity Automated LC/MSD Purification System controlled by the Automated Purification software.
Introduction
Preparative liquid chromatography (LC) is a technique that is frequently applied in pharmaceutical chemistry or early stages of drug discovery. Depending on the compound of interest, optimum chromatographic conditions usually need to be determined by an analytical scouting run before large amounts of sample are purified. This process can be time-consuming, especially with the high number of samples typical in pharmaceutical chemistry.
The Agilent Automated Purification software facilitates this scale-up process, and supports analysts in speeding up their workflows, providing true automatic scale-up from analytical to preparative gradients. An intuitive interface guides the user along the complete workflow including analytical scouting, scale-up, focused gradient generation, fraction collection and review, and export of results. Smart algorithms check the validity of user inputs on-the-fly, and prevent the user from proceeding if conflicting parameters are entered. Two different views offer either the complete settings for method developers or a restricted set of parameters for operators or walk-up users (Easy Prep mode). Once all details of the task are entered, the workflow can be started either in fully automatic mode or with the option to manually change parameters and review analytical results before proceeding with the preparative runs.
The workflow in the Automated Purification software consists of an analytical scouting run, an (automatic or manual) evaluation step, and a preparative run with fraction collection. During the scouting run, diluted samples are analyzed by a generic gradient on the analytical flow path of the Agilent 1260 Infinity Automated LC/MSD Purification System. In the evaluation phase, the compounds of interest are identified by target masses specified in the analytical sequence, and aligned with the signal of the UV detector. These data can be imported from any analytical Agilent LC/MSD System. Based on the matching signals, the virtual elution point of each target compound is calculated. Software algorithms generate a focused gradient that is optimized for the calculated elution point of the target compound. The purification is then run on the preparative flow path of the system, applying the calculated focused gradient for each sample, enabling optimized separation and collection of the target compound.
This Technical Overview demonstrates the analytical-to-preparative workflow with the Automated Purification software using a sample library of eight compounds. Impure samples were separated and purified by peak-based fraction collection triggered by a combination of ultraviolet (UV) and mass selective detector (MSD) signals. Fraction purity and recovery of the injected sample amounts were measured as performance markers.
Conclusion
The Agilent Automated Purification software is a versatile add-on to the Agilent OpenLAB CDS ChemStation that facilitates the workflows of purification laboratories. All crucial steps from analytical scouting to fraction collection and data review are combined in an intuitive user interface. With truly automated scale-up, on-the-fly calculation of focused gradients, and import and export functionalities, the Automated Purification software is a valuable tool to speed up the process of LC purification for experienced and novice users. This Technical Overview demonstrates the purification of a compound library on an Agilent 1260 Infinity Automated LC/MSD Purification System controlled by the Automated Purification software. Even in fully automated operation, fraction purity and recovery are typically above 90 %.
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