The analytical chemistry team routinely purifies samples with material in the range of milligrams to grams. Major and minor components (< 0.1 percent) have been isolated for additional testing and characterization. The teams automated sample-processing protocol allows for dispensing into 1D barcoded vials, Matrix 2D barcoded tubes and/or 96-well plates for efficient tracking, storing and testing. The entire process from receiving the sample to final plating is completed in less than one week. While the group uses a variety of liquid chromatographs to determine identity and purity, single quadruple liquid chromatography/mass spectrometry instrumentation is utilized for high-throughput automated analysis. Due to the wide variety of analytes tested, the teams range of analytical detectors includes ultraviolet (UV), mass spectrometry (MS; positive and negative mode), evaporative light-scattering detector and fluorescence (ELSD). Time-of-flight mass spectrometry (TOF/MS) is employed to achieve formula confirmation and identity determination of unknowns. Methods development with the chiral chromatography screening protocol includes analysis on seven different chiral stationary phases. Use of several mobile phase conditions enables development of the best chiral separation. The group routinely determines chemical purity and enantiomeric purity. The use of an inline chiral detector facilitates the determination of relative optical rotation. Matching the stationary phase of analytical columns with the corresponding stationary phase in both semi-preparative and preparative columns enables sample purification on a scale of up to hundreds of milligrams. The isolated enantiomers are returned as powders after chiral chromatography analysis to ensure purity of the material. As a means of storing and organizing all analytical data, Analytical Chemistry researchers developed Sample Management and Resource Tracking (SMART), which is a proprietary laboratory information management system (LIMS). SMART is a Web-based management software utilizing a central server to store data from integrated instrumentation and automation equipment accessed through a unified Web browser. While the initial focus was analytical data tracking and sample management, SMART has evolved into an all-encompassing system for sample submission, compound purification, compound processing, sample registration, compound inventory, sample tracking, data retrieval and data management. The Analytical Chemistry group has initiated two new programs as an expansion of their available services: fragment-based screening by NMR and sample analysis by high-throughput MS. Fragment-based screening is a complementary approach to high-throughput screening in the discovery of chemical entities for development into therapeutics. We have obtained fragment libraries that will be screened against protein targets associated with various disease states using established NMR techniques. For high-throughput MS analysis, we are utilizing a RapidFire 365 MS system, which will allow for efficient and expedient analysis of chemical targets within various biochemical and ADME assays.
| Liu, Li; Tang, Manshu; Walsh, Martin J et al. (2015) Structure activity relationships of human galactokinase inhibitors. Bioorg Med Chem Lett 25:721-7 |
