on 10-17-201208:45 AM - edited on 10-15-202109:29 AM by AnalyteGuru
Cook K, Dreyer M, Murphy K. Scientific Poster Purpose: Plasma protein binding determination.
Methods: Rapid equilibrium dialysis, HRAM, and online extraction.
Results:Protein binding determination was conducted in both human and rat plasma for a wide spectrum of low to high binding compounds such as Gabapentin (literature value: <3%), Levofloxacin(24-38%) and Warfarin(98-100%).In an effort to create a generic workflow, incubations were performed at a single concentration for the compound library analyzed. Utilizing a Transcend TLX-2 system with a single generic online Turbo Flow method, the analytical time per sample was 1.35 minutes. The throughput was further enhanced through fullscan HRAM analys is made possible using an Exactive non-hybrid Orbitrap MS. The total time to analyze the entire 16 compound sample set(n=4), for both human and rat, was approximately 5 hours. The calculated peak area ratios were completed using the quantitation software package, LCquan, from Thermo Scientific. The summarized data was compared to literature values provided in Goodman & Gilman’s, The Pharmacological Basis of Therapeutics (10thed.). Overall, the data acquired agreed with literature values with acceptable reproducibility using either a simple mean or well to well calculation. The time savings provided with this workflow is useful in both a discovery and development environment due to the elimination of off-line sample preparation, chromatographic multiplexing, and removing compound optimization required for triple-stage quadrupole instruments.