title: Integrated quantitative and qualitative workflow for in vivo bioanalytical support in drug discovery using hybrid Q-TOF-MS.
abstract: BACKGROUND
  UHPLC coupled with orthogonal acceleration hybrid quadrupole-TOF (Q-TOF)-MS is an emerging technique offering new strategies for the efficient screening of new chemical entities and related molecules at the early discovery stage within the pharmaceutical industry. In the first part of this article, we examine the main instrumental parameters that are critical for the integration of UHPLC-Q-TOF technology to existing bioanalytical workflows, in order to provide simultaneous quantitative and qualitative bioanalysis of samples generated following in vivo studies.
  
  
  METHODS
  Three modern Q-TOF mass spectrometers, including Bruker maXis™, Agilent 6540 and Sciex TripleTOF™ 5600, all interfaced with UHPLC systems, are evaluated in the second part of the article. The scope of this work is to demonstrate the potential of Q-TOF for the analysis of typical small molecules, therapeutic peptides (molecular weight <6000 Da), and enzymatically (i.e., trypsin, chymotrypsin and pepsin) cleaved peptides from larger proteins.
  
  
  CONCLUSIONS
  This work focuses mainly on full-scan TOF data obtained under ESI conditions, the major mode of TOF operation in discovery bioanalytical research, where the compounds are selected based on their pharmacokinetic/pharmacodynamic behaviors using animal models prior to selecting a few desirable candidates for further development. Finally, important emerging TOF technologies that could potentially benefit bioanalytical research in the semi-quantification of metabolites without synthesized standards are discussed. Particularly, the utility of captive spray ionization coupled with TripleTOF 5600 was evaluated for improving sensitivity and providing normalized MS response for drugs and their metabolites. The workflow proposed compromises neither the efficiency, nor the quality of pharmacokinetic data in support of early drug discovery programs.
authors.affiliation: Bristol-Myers Squibb, Research & Development, Bioanalytical & Discovery Analytical Sciences, Princeton, NJ 08543, USA. asoka.ranasinghe@bms.com
authors.forename: Asoka
authors.initials: A
authors.lastname: Ranasinghe
authors.forename: Ragu
authors.initials: R
authors.lastname: Ramanathan
authors.forename: Mohammed
authors.initials: M
authors.lastname: Jemal
authors.forename: Celia J
authors.initials: CJ
authors.lastname: D'Arienzo
authors.forename: W Griffith
authors.initials: WG
authors.lastname: Humphreys
authors.forename: Timothy V
authors.initials: TV
authors.lastname: Olah
mesh_terms: D002110:Caffeine / Q000032:analysis / Q000737:chemistry; D055808:Drug Discovery / Q000295:instrumentation* / Q000379:methods*; D013058:Mass Spectrometry; D010455:Peptides / Q000032:analysis / Q000737:chemistry; D054852:Small Molecule Libraries / Q000032:analysis / Q000737:chemistry; D012984:Software
chemical_list: D010455:Peptides; D054852:Small Molecule Libraries; D002110:Caffeine