Nikki Dare, Richard Lee, Karl Demmans, Anne Marie Smith
Introduction
High throughput experiments (HTE) are complex studies used for product screenings and reaction optimizations. Although, reaction success can be assessed simply by the presence of the product synthesised, chemists will want to know the extent of success and the quantitative amount of the product generated. Specialized Windows based applications are often use for the design of these experiments, however, these applications do not incorporate analytical data analysis or quantitation workflows, and require separate data analysis solutions. Moreover, these applications are chained to the specific workstations, and require the user to be present in the same laboratory. Here, we present an all-in-one, browser-based application that supports quantitation for HTE, including automation for data retrieval and data processing.
Methods
To support HTE, data processing and quantitation modules were developed within the HTE design application to support all major vendor LC/UV/MS systems. Quantitation experiments can be designed within the application, including the selection of instrument type (vendor LC, LC/DAD, LC/UV/MS), sample definition (calibration, samples, blanks, system suitability), and target lists. The application automatically processes the data based on the target list and processing parameters, and the user can visualize the results including, chromatograms, spectra, tabulated results, and calibration plot. Data can also be reprocessed individually within the browser-based application, or users can change processing parameters and initiate batch reprocessing. Results are stored in a database, and any user-initiated changes are stored as part of the audit trail.
Preliminary Data
High throughput experiments, along with the subsequent quantitation studies are designed within the browser-based application, bypassing the CDS software for design and data management. This allows the user to create these experiments from any location, independent of where the data lies. Users can designate the appropriate samples including, calibration samples, blanks and system suitability. Compound target lists are supported through several methods, 1) manual text entry, 2) compound look up if integrated into an internal chemical database, and 3) SD file (drag and drop). It also supports a chemical structure drawing program where users can draw structures, load mol files, or enter InChI notation. Retention times can also be entered for each target if desired to ensure component identity.
As the study layout is defined, a sequence file is generated for the selected CDS. The system provides a unique name for each sample, thus, each data file has a unique identifier that the system uses to reconcile sample information to data file. The application incorporates automation services to retrieve and automatically process data based on user-set parameters as the data files are acquired. Regression analysis is performed based on user preferences, plots are generated, and can be viewed within the application for review.
Users can review the processed chromatograms and spectra within the browser application, where they have the ability to alter data processing parameters and initiate batch reprocessing on the entire set or subset of data. Moreover, for more granular data processing needs, the application incorporates a manual data processing interface where the user has full control to adjust baseline, peak detection, and integration.
The browser-based design of the application allows it to be platform independent and the underlying technology enables it to be compatible with all major LC/UV/MS vendors.
Novel Aspect
Vendor-neutral, browser-based application supporting quantitation of LC, LC/UV, and LC/UV/MS datasets for high throughput experiments.