title: A high-speed liquid chromatography/tandem mass spectrometry platform using multiplexed multiple-injection chromatography controlled by single software and its application in discovery ADME screening.
abstract: BACKGROUND
  Multiplexed liquid chromatography (LC) coupled with multiple-injection-chromatogram acquisition has emerged as the method of choice for high-speed discovery bioanalysis, because it significantly reduces injection-to-injection cycle time while maintaining the chromatography quality. Historically, systems utilizing this approach had been custom built, and therefore relied on custom software tools to communicate with multiple vendor software for system control, which lacked transferability, flexibility and robustness.
  
  
  METHODS
  In this study, we refined a multiplexed bioanalytical system previously reported, by implementing open-deck auto-sampler manifold and multiple-injection-chromatogram acquisition, all on a commercially available system with single software control.
  
  
  RESULTS
  As a result of these improvements, the developed LC/tandem mass spectrometry (MS/MS) method on the system was nearly three times faster than the previous method, while demonstrating comparable analytical accuracy, precision and robustness. This system has been evaluated for in vitro ADME screening assays including metabolic stability, CYP inhibition and Caco-2. The biological data generated on the developed system displayed good correlation with those from the previous LC/MS/MS approaches.
  
  
  CONCLUSIONS
  The developed platform demonstrated applicability to the in vitro screening assays evaluated and has been successfully implemented to support the high-throughput metabolic stability assay, with a significantly improved bioanalytical throughput, capacity and data turnaround.
authors.affiliation: Bristol-Myers Squibb, Research and Development, 5 Research Parkway, Wallingford, CT 06492, USA. jun.zhang1@bms.com
authors.forename: Jun
authors.initials: J
authors.lastname: Zhang
authors.forename: Marianne
authors.initials: M
authors.lastname: Vath
authors.forename: Cheryl
authors.initials: C
authors.lastname: Ferraro
authors.forename: Ying
authors.initials: Y
authors.lastname: Li
authors.forename: Keeley
authors.initials: K
authors.lastname: Murphy
authors.forename: Tatyana
authors.initials: T
authors.lastname: Zvyaga
authors.forename: Harold
authors.initials: H
authors.lastname: Weller
authors.forename: Wilson
authors.initials: W
authors.lastname: Shou
mesh_terms: D000818:Animals; D018938:Caco-2 Cells; D002853:Chromatography, Liquid / Q000295:instrumentation / Q000379:methods*; D051544:Cytochrome P-450 CYP3A / Q000032:analysis / Q000378:metabolism; D055808:Drug Discovery / Q000379:methods*; D006801:Humans; D016014:Linear Models; D051379:Mice; D008956:Models, Chemical; D011433:Propranolol / Q000032:analysis / Q000737:chemistry; D051381:Rats; D015203:Reproducibility of Results; D012984:Software; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D011433:Propranolol; D051544:Cytochrome P-450 CYP3A